Biological Effects of Ultraviolet Radiation

with Emphasis on Human Responses to

Longwave Ultraviolet
Biological Effects of Ultraviolet Radiation
with Emphasis on Human Responses to
Longwave Ultraviolet

JOHN A. PARRISH, M.D.

and
R. ROX ANDERSON
Harvard Medical School

FREDERICK URBACH, M.D.

Skin and Cancer Hospital
Temple University School of Medicine

and
DONALD PITTS, 0.0., Ph.D.
College of Optometry
University ofHouston

PLENUM PRESS . NEW YORK AND LONDON

 Library of Congress Cataloging in Publication Data
Main entry under title:
UV-A: biological effects of ultraviolet radiation with emphasis on human responses to long-
wave ultraviolet.
Includes bibliographical references and index.
1. Ultra-violet rays - Physiological effect. 2. Ultra-violet rays. I. Parrish, John Albert,
1939- [DNLM: 1. Ultraviolet rays. 2. Skin-Radiation effects. 3. Eye-Radiation effects.
QT162.U4 Ul06]
QP82.2.U4Ul8 612.01448'4 78-14968
ISBN-13: 978-1-4684-2477-5 e-ISBN-13: 978-1-4684-2475-1
DOl: 10.1007/978-1-4684-2475-1

© 1978 Plenum Press, New York

Softcover reprint of the hardcover 1st edition 1978
A Division of Plenum Publishing Corporation
227 West 17th Street, New York, N.Y. 10011
All rights reserved
No part of this !look may be reproduced, stored in a retrieval system, or transmitted,
in any form or by any means, electronic, mechanical, photocopying, microfilming,
recording, or otherwise, without written permission from the Publisher
Preface

The origin of this text was a request by industry and government to summarize
the biological effects and to estimate the limits of safe exposure to longwave ul-
traviolet radiation. The specific issue was the safety of a small medium-pressure
mercury arc designed to emit UV-A (NUVA-Lite, L. D. Caulk Co., Milford,
Delaware) for photopolymerization of resinous fillings used in dentistry. How--
ever, the context grew to become a consideration of the risks and benefits to hu-
mans of electromagnetic radiation between the biologically active short UV and
the visible spectrum. We have accumulated data from our own experimental
work and from the literature and have attempted to put this information in the
perspective of known biologic effects of ultraviolet radiation as it influences hu-
mans.
Interest in the biological effects of longwave ultraviolet radiation is increas-
ing in all of the many scientific disciplines that make up the complex field of
photobiology. In order to minimize the chance for error and personal prejudice
and to maximize the use of expertise, each chapter has been reviewed by several
authorities. Some of the contributions of this group led to significant alterations
and creative additions to the chapter, and these persons deserve not only our sin-
cere gratitude but also recognition by the reader. These include Chapters 2 and 3:
Dr. Robert E. Levin, Mr. Charles P. Comeau, Mr. Donald Gonser, Dr. David
Sliney; Chapter 5: Dr. Jerry Williams, Dr. Robert Webb, Dr. Madhu A. Pathak;
Chapter 6: Dr. Farrington Daniels, Jr., Dr. Albert Kligman, Dr. Barbara Gil-
chrest, Dr. Thomas B. Fitzpatrick; Chapter 7: Dr. H. A. D. White; Chapter 8:
Dr. Donald Forbes, Dr. Reinaldo Rosario, Dr. Ernesto Gonzalez, Dr. Warwick
Morison; Chapter 9: Dr. Seymour Zigman, Dr. Mathea Allansmith, Dr. William
T. Ham, Jr.
We are also grateful to Dr. David Sliney for his expert criticism, to Patricia
Novak for her skilled editorial suggestions, and to Diane Patry, who headed the
team of typists necessary for the multiple revisions of the manuscript. Figures

v
vi Preface

were prepared by Thomas Mcinnes and Garrison Root, and Christopher Shea
provided library and technical assistance.

John A. Parrish, M.D.

R. Rox Anderson
Frederick Urbach, M.D.
Donald Pitts, D.D., Ph. D.
Contents

Chapter I THE SPECTRUM OF ELECTROMAGNETIC RADIA-

TION: UV-A IN PERSPECTIVE ............................... .

Chapter 2 SOURCES OF UV-A............................................... 7

Introduction ....................................................... 7
Solar Ultraviolet Radiation...................................... 9
Summary........................................................... 14
Artificial Sources of UV-A ............. ............. ....... .... 15
Ultraviolet Spectral Transmission and Reflection of
Common Materials.......................... .... . . . . . . .... . . . . 26
References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34

Chapter 3 RADIOMETRY OF ULTRAVIOLET RADIATION .......... 37

Introduction and General Considerations of Radiometry... 37
Detectors. . . . . ... . . . . ... .. . . . . . . . . .. . . . . . . . . . . . . . . . . .. .... . . . . . .... . . 42
Spectral Filters and Input Optics. .. . . . . . .. . . . . . . . . .. . . . . . . . ... . 48
Appendix: U.S. Manufacturers of UV-Related
Instrumentation. . .. . . . . . . . . . . ... . . . . . .... . . . . . . ... . . . . . . . .... . . 55

Chapter 4 OPTICAL PROPERTIES OF THE SKIN AND EyES....... 59

Introduction ....................................................... 59
Structure of the Skin............................................. 59
Factors Affecting Penetration and Absorption of
Ultraviolet Radiation in the Skin............................ 62
Measurements of the Penetration and Reflection
of Optical Radiation in Skin................................. 65
Ultraviolet Optics for the Eye.................................. 77
References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 81
vII
viii Contents

Chapter 5 EFFECTS OF ULTRAVIOLET RADIATION ON MICRO-

ORGANISMS AND ANIMAL CELLS......................... 85
Introduction ....................................................... 85
Effect of Ultraviolet on Cells................................... 88
DNA Repair............................................... ........ 89
Effects of UV-A ..... .......... ................................... 95
Summary........................ ................ ................... 101
References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 101

Chapter 6 IMMEDIATE AND SHORT-TERM BIOLOGIC EFFECTS

OF ULTRAVIOLET RADIATION ON NORMAL SKIN.... 107
Introduction ....................................................... 107
Erythema........ ........... ................. ................... .... 109
Histology.......................................................... 124
UV and Epidermal Macromolecular Synthesis .............. 126
Effects of UV-A on Mucous Membrane ...................... 128
Tanning ............................................................ 128
References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 134

Chapter 7 ADVERSE CUTANEOUS REACTIONS TO UV-A ......... 141

Introduction ....................................................... 141
Chemical Photosensitivity. ............................ .......... 141
Persistent Light Reactivity...................................... 149
Actinic Reticuloid . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . 149
Polymorphous Light Eruption.................................. 150
Solar Urticaria.................................................... 150
Porphyrias and Other Endogenous Photosensitization
Syndromes. .......... .......... ............................. .... 150
Melasma and Ephelides ......................................... 151
Management of UV-A-Induced Dermatoses. ............... 151
References.................................................. ....... 153

Chapter 8 SKIN AGING AND CARCINOGENESIS DUE TO

ULTRAVIOLET RADIATION ................................... 157
Introduction....................................................... 157
Incidence of Skin Cancers in Man...... . . . . . . . . . . . . . . . . . . . . . . . 158
Epidemiologic Evidence Supporting the Role of Sunlight. 159
Mechanisms of UV Carcinogenesis........................... 165
Action Spectrum of Animal Photocarcinogenesis and the
Carcinogenic Effects of UV-A .... ........ ........... ....... 166
Contents Ix

Other Factors Influencing or Associated with Development

of Skin Cancer.... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 168
Ultraviolet Radiation and Aging ............................... 171
References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . 172

Chapter 9 EFFECTS OF ULTRAVIOLET RADIATION ON THE EYE 177

Introduction ....................................................... 177
Morphology and Histology of the Cornea and the Lens.... 179
Histologic Effects of Ultraviolet Radiation. . . . . . . . . . . . . . . . . . . 185
Action Spectrum of Ocular Effects of Ultraviolet
Radiation........................................................ 188
Corneal and Lenticular Effects of Longwave
Ultraviolet Radiation.... . . ........ ..... ........ . . ............. 193
Effects of UV-A in the Retina............ .......... ............ 211
The Ocular Effects of UV-A Exposure in the Presence of
Photosensitizing Compounds (Psoralens).................. 212
Summary: UV-A Exposure of the Eye........................ 214
References......................................................... 215

Chapter 10 USES OF UV-A INVOLVING EXPOSURE OF HUMANS 221

Introduction ....................................................... 221
Therapeutic Uses of Ultraviolet Radiation.................... 221
Ultraviolet Treatment of Psoriasis. . ...... . ......... . . .... . .... 222
Photochemotherapy .............................................. 225
Oral Psoralen Photochemotherapy . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 225
UV-A-Activated Polymerization of Resinous Dental
Restorations..... ....... ....... . . ..... . ....... . . ......... ...... . 233
Diagnostic Uses of UV-A....................................... 234
References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 235

Chapter J J SAFETY MEASURES AND PROTECTION AGAINST

ULTRAVIOLET EXPOSURE .................................... 241
Introduction ....................................................... 241
Ultraviolet Exposure Safety Standards........................ 242
Sunscreens......... ...... ......... . . ...... ...... . . ......... . ....... 248
Eye Protection against Ultraviolet Radiation. ....... . . ..... .. 252
References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 254

INDEX ................................................................ 257

CHAPTER 1

The Spectrum
of Electromagnetic Radiation:
UV-A in Perspective

Continuous thermonuclear reactions in the sun's core yield a wide spectrum of

electromagnetic energy that radiates through space in all directions. This radiant
energy is called electromagnetic because it is in the form of oscillating electric
and magnetic fields. Electromagnetic radiation exhibits both wavelike (oscillat-
ing field) and particlelike (discrete packet) properties. These discrete packets, or
quanta, are called photons. The number of oscillations per second (frequency)
times the distance traveled through space per oscillation (wavelength) is the ve-
locity of light. Because all photons travel through space at this velocity,
wavelength and frequency are inversely proportional. Max Planck, in 1900, de-
termined that the energy carried by a photon is directly proportional (Planck's
constant) to its frequency. Therefore, shorter wavelength, higher frequency elec-
tromagnetic radiations consist of photons of higher energy.
The sun's radiant energy sustains all life on earth. Solar energy not only
maintains the earth's temperature but also supports the growth of photosynthetic
plants, which have the ability to convert radiant energy to chemical potential
energy. Man obtains his own body fuel by ingesting both plants and plant-eating
animals. Without sunlight, the surface of the earth would be cold, still, and com-
pletely lifeless.
Humans have evolved in sunlight and depend upon it for much more than an
indirect source of food. Our skin, eyes, blood vessels, and certain endocrine
glands respond to radiation from various portions of the electromagnetic spec-
trum of the sun. Many of our daily rhythms are dependent upon the cycles of
sunlight. The sophisticated sensory mechanism of the eye transduces absorption
of electromagnetic radiation into nerve impulses to provide an instantaneous de-
tailed concept of the environment.
2 Chapter One

Excessive amounts of any life-supporting agent can be harmful. Sunlight

can damage or kill living cells, including those of man, and gross changes may
follow this microscopic damage. Sunlight causes sunburn, skin cancer, wrinkling
and "aging" of the skin, inflammation of the eyes, and possibly cataracts.
Physiologic and anatomic alterations and instinctual responses protect the skin
and eyes, but attitudes, learned responses, customs, and lifestyle determine one's
sun exposure. Thus, humans live in a complicated ecobalance with sunlight: it is
essential to sustain life, yet excessive solar radiation may be harmful.
The effects of radiation on an organism depend on the effects of the ab-
sorbed radiation on the cells of which the organism is composed. The cellular
changes caused by exposure to electromagnetic energy are in turn caused by
chemical reactions following the absorption of photons by molecules of the cell.
Such photochemical reactions cannot take place unless radiation is absorbed
(Grotthus and Draper's law). The effect on the cells depends on the presence of
absorbing molecules within or around the cells. Absorption is a relatively precise
phenomenon. Specific molecular structures absorb radiation of a specific wave-
length or quantum energy region. Therefore, the molecular structure of
biomolecules determines their absorption of radiation of various photon energies,
or wavelengths.
Specific photochemically alerted molecules may present either benefit or
hazard to the cell or organism. For example, the production of vitamin D3 and the
production of damaged DNA may be caused simultaneously in human skin cells
by photons of the shortest wavelength region present in sunlight. Cells and or-
ganisms have therefore evolved complex mechanisms for recognition and repair
or degradation of harmful photo products and have come to depend upon useful
photo products .
It is convenient to give names to certain portions of the electromagnetic
spectrum, but it is important to remember that the biologists' practical classifica-
tions of electromagnetic energy are, in part, an example of the arbitrary and
egotistical nature of man. The names used by convention are related to a strange
mixture of a number of properties of the radiation, such as (1) wavelength, fre-
quency or photon energy; (2) spectral relation to visible light or sunlight on earth;
(3) the apparent effects of the radiation on humans; and (4) the uses man has
found for the radiation (Fig. 1-1).
Visible light (wavelengths of approximately 400-700 nm) is that radiation
that passes through the human lens to be absorbed by specialized molecules in the
retina and to stimulate central nervous system recognition of the event. Visible
light also penetrates through skin and subcutaneous tissues, but under usual cir-
cumstances such photons do not cause photochemical events that are recogniza-
ble in the form of tissue alteration or damage. Of the familiar visible light color
spectrum (Fig. 1-1), the shortest wavelengths are perceived as violet, and the
longest wavelengths are perceived as red. The invisible regions of the elec-
The Spectrum of Electromagnetic Radiation 3

COSMIC RADIO
RAYS WAVES

SHORT - MIDDLE LONG-

VACUUM VISIBLE
WAVE UV WAVE WAVE UV
UV UV LIGHT
(UVC) (UVB) (UVA)

100 200 300 400

WAVELENGTH IN NANOMETERS

Fig. I-I. Electromagnetic spectrum with expanded scale of ultraviolet radiation.

tromagnetic spectrum that neighbor the visible region are termed the ultraviolet
and infrared regions. Ultraviolet radiation consists of shorter-wavelength,
higher-energy photons than violet light, and infrared radiation consists of
longer-wavelength, lower-energy photons than red light. Both of these
wavelength regions of radiation penetrate the skin but cause different biologic
effects because of the differences between the energies of photons of ultraviolet
and infrared radiation.
When a photon is absorbed, all of the photon's energy is transferred to the
absorbing atom or molecule, and the photon no longer exists. For some period of
time, its energy is invested in the atom or molecule, which is therefore said to be
in an excited state. The mode of this excitation depends upon the amount of
energy invested (photon energy) which in turn depends upon the wavelength of
the absorbed photon. In increasing order of photon energy, the following may
occur: rotation of molecules; vibration of atoms within the molecule; changes in
the orbital shells that the molecule's or atom's electrons occupy. All three of
these alterations must occur in discrete "jumps"; hence the absorption of photons
by molecules occurs at wavelengths with photon energies equal to those of the
allowed rotational, vibrational, or electronic transitions. At very high photon
energies, electrons may be removed from the molecule, resulting in ionization of
the molecule.
Infrared photons may alter lower-energy excitationallevels of a molecule by
affecting rotational and vibrational states. Visible and ultraviolet photons may
lead to higber vibrational states or electronic excitation. Ultraviolet photons of
4 Chapter One

wavelengths shorter than about 200 nm, X-rays, gamma rays, and cosmic rays
may cause ionization.
Ultraviolet radiant energy absorbed by nucleic acids, proteins, or other
molecules within the cell may be dissipated as heat or reemitted as light. A
molecule may also be structurally altered or cleaved, or it may react with other
molecules as a result of its excitation foIlowing the absorption of ultraviolet radi-
ation. The resulting molecular alteration mayor may not lead to changes in ceIl
function, mutation, death, or other responses.
The ultraviolet radiation region of the spectrum is subdivided into several
bands in terms of phenomenologic effects. Unlike these wavelength divisions,
the effects of ultraviolet radiation do not terminate sharply at specific
wavelengths. Furthermore, the subdivisions are arbitrary and differ somewhat,
depending on the discipline involved. Photobiologists generaIly divide the ul-
traviolet spectrum into three portions, called UV-A, UV-B, and UV-C, in order
of decreasing wavelength (Figs. 1-1 and 1-2). In this text, the wavelength range
from 200 to 290 nm is caIled UV-C. Radiation of wavelengths shorter than 200
nm is mostly absorbed by air, and solar radiation of wavelengths below 290 nm
does not reach the earth's surface because of absorption by ozone formed in the
stratosphere. The band from 290 to 320 nm is called UV-B, and the band from
320 to 400 nm is called UV-A. This terminology was originally derived by Cob-
lentz in 1932, and is based on a combination of physical properties and biologic
effects of each of these ultraviolet wavelength regions. The division between
UV-C and UV-B is sometimes chosen as 280 nm, and 315 nm is sometimes cho-
sen as the division between UV-B and UV-A. Because the divisions between
UV-C, UV-B, and UV-A are neither phenomenologicaIly exact nor agreed
upon, for critical work one should define ultraviolet radiation in more rigorous
spectroradiometric terms.
Radiation in the UV -C band causes erythema of normal skin very efficiently
and can cause photokeratitis (inflammation of the cornea). UV-C is also caIled
germicidal radiation because of its effectiveness in killing one-ceIled organisms.
UV-C is often caIled shortwave UV because the wavelengths in this region are
the shortest ultraviolet radiation transmitted through air. This ultraviolet region is
the furthest from the visible spectrum and is also calledfar UV.
Solar ultraviolet radiation of wavelengths between 290 and 320 nm reaches
earth in relatively small quantities but is very efficient in causing sunburning of
human skin. For this reason, it is often referred to as the sunburn jpectrum or the
erythemal band. Because of its relative spectral position, UV-B is also called
middle UV, mid UV, or middlewave UV. Radiation of wavelengths. longer than
320 nm is relatively inefficient at causing redness of human skin. The UV -B por-
tion of the spectrum has been shown to induce skin cancer in laboratory animals
and mutations in bacteria. Epidemiologic evidence suggests strongly that solar
UV-B causes skin cancer in man. Long-term UV-B exposure is also thought to
 The Spectrum of Electromagnetic Radiation 5

solar spectrum at earth·s~!!.!~e

delayed akin erythema action spectrum
visible light
I UV-C lUV-_~1 UV-A
200 290 320 400

WAVELENGTH IN NANOMETERS

Fig. 1-2. Graphic illustration of phenomenologic definition of UV·A, UV·B, and UV-c.

be at least partly responsible for producing the changes of exposed human skin
that are commonly termed aging.
Many of the effects of UV-B and UV-C are now easily recognized and
much knowledge has accumulated regarding changes produced in molecules,
cells, tissues, and living humftn skin. Much less attention has been paid to ul-
traviolet radiation of wavelengths longer than 320 nm (UV-A, 320-400 nm).
UV -A is both melanogenic (producing skin pigment, tanning) and
erythemogenic (producing redness of skin), but the amount of energy required to
produce an effect is orders of magnitude higher than for the UV-B region. The
relative inefficiency of these longer wavelengths in producing skin erythema and
photokeratitis, the relative absence of striking germicidal properties, and, until
recently, the lack of high-intensity artificial light sources strengthened the idea
that UV-A was innocuous. UV-A is sometimes referred to as longwave UVand
long UV and is also called near UV because of its proximity to the visible spec-
trum. This spectral region has also been called the blacklight region, because its
principal use for many years was to excite fluorescent and phosphorescent sub-
stances that reradiate the absorbed energy as light in the visible spectrum.
UV-A has received more attention in the past decade for the following
reasons:
L The amount of solar UV-A reaching the earth's surface is enormously
greater than that of UV-B.
2. Photosensitivity reactions (phototoxicity and photoallergy) are mostly
mediated by UV-A.
3. High doses of UV-A can cause redness of human skin; moreover,
UV-A may potentiate or add to the biologic effects of UV-B.
4. High-intensity sources oi UV-A are now available for basic research
and clinical studies.
5. The development of sunscreens that effectively block or diminish the
highly erythemogenic UV-B permits prolonged sun exposures. Many
6 Chapter One

of these sunscreens do not significantly alter the amount of UV-A

reaching the skin. Furthermore, UV-A is transmitted by most window
glass and many plastics that do not transmit UV-B.
6. Recent studies suggest that UV -A can affect cells and microor-
ganisms.
7. UV-A-induced photopolymerization and photochemical reactions are
being used in industry to alter rubber, plastic, glass, metal, paper, and
photographs.
8. Photopolymerization of certain chemicals provides convenient ways
to apply dental and orthopedic appliances and "photocure" them in
place.
9. The use of UV-A in conjunction with photosensitizing drugs has
opened up new therapeutic possibilities in chronic skin disorders, such
as psoriasis, mycosis fungoides, and eczema.
10. There is experimental and epidemiologic evidence to suggest that
solar UV -A is one of the possible etiologic agents for certain kinds of
cataracts in humans.
This book attempts to summarize current knowledge of the biologic effects
of UV-A, especially as they may relate to humans. For perspective, biologic ef-
fects of UV-A, UV-B, and UV-C are compared throughout the book, a brief
examination of exposure safety criteria for humans is presented, and the produc-
tion and measurement of ultraviolet radiation in the laboratory is reviewed. The
need for more basic photobiologic research is emphasized throughout.
CHAPTER 2

Sources of UV-A

Introduction

The intent of this chapter is to review solar ultraviolet radiation and the major
environmental, research, and phototherapy sources of ultraviolet radiation in use
today, with particular emphasis on sources of UV-A.
Photometry is based on visible light measurements that simulate the human
eye's photopic response curve and is used extensively by light source manufac-
turers and in photography. Radiometry is not confined to the visible spectrum and
is based on direct measurements of radiant energy over a defined wavelength reg-
ion. Because the main interest is description of ultraviolet radiation, photometric
terminology will not be used here (see Chapter 3). Definitions of some funda-
mental radiometric terms used throughout this book are given in Table 2-1.1
The radiometric terms in the table may also be expressed in terms of
wavelength by adding the prefix spectral. Thus, spectral irradiance, the ir-
radiance at a particular wavelength, is given, for example, in mW/cm 2 • nm. A
plot of spectral irradiance (at a given distance and position relative to the source)
versus wavelength is extremely useful in describing sources of ultraviolet radia-
tion. The irradiance over a wavelength region of interest is simply the area under
the spectral irradiance curve in that region. "Spectral power distribution" or, es-
sentially, spectral radiant flux, is often used by lamp manufacturers to describe
the radiant flux emitted by a lamp as a function of wavelength. For a thorough
discussion of terminology and lamp data, see references 1 and 2.
The convention is often to express irradiance or radiant exposure dose in
terms of watts or joules per square meter, respectively. In most photobiologic
literature, these terms are expressed per square centimeter rather than per square
meter. Furthermore, the use ofmilliwatts (1 mW = 10- 3 W), millijoules (1 mJ =
10- 3 J), or microwatts (1 f.LW = 10 -6 W = 10- 3 mW) is also common, and
these are radiometric units most often used in this text. A less commonly used

7
8 Chapter Two

Table 2-1. Some Fundamental Radiometric Terminology

CIE
Term Units symbol a Definition Comments and synonyms
0
Wavelength A, nm,/lm A Nanometer = 10 -9 meter (also
called millimicron, mil); /lm,
micron = 10- 6 meter; A,
angstrom = 10 -1 0 meter.
Radiant energy J Qe 1 joule (J) = 1 watt (W) X 1
second (s).
Radiant flux W If>e dQe Rate of radiant energy delivery
(ff ("radiant power"). mW = 10 -3 W
/lW=lO"'W.
Radiant W/sr Ie dlf>e Describes the radiant flux emitted
intensity dw by the source into a given solid
angle (solid angle ex pressed in
steradians) .
Irradiance W/m 2 Ee ~ Radiant flux arriving over a given
dA area. In photobiology, also ex-
pressed in W/cm 2 , mW/cm 2 or
/lW/cm 2 • Note implied depen-
dence of irradiance on the angle
of the area being irradiated
relative to a beam. In a colli-
mated, uniform beam, the ir-
radiance Ee on a planar surface
varies directly with cos (), where
() = angle of incidence from a
normal to the surface ("dose-
rate," "intensity").
Radiant J/m 2 He Ee X t Usually expressed as J / cm 2 or
exposure where t = mJ/cm 2 ("exposure dose,"
(dose) exposure "dose").
time in
seconds

aThe subscript e serves to distinguish radiometric quantities from photometric quantities,

which have a v subscript. The e subscript is often dropped when only radiometric terms
are used.
Sources of UV-A 9

Table 2-2. Conversion Between Irradianee UnitsQ

W/m2 mW/em 2 p.W/em 2 erg/em 2 • s erg/m2. s

I W/m 2 I 0.1 100 1000 107

I mW/cm' 10 10" 104 10"
I p.W/cm' om 10- 3 1 10 105
I erg/cm 2 • s 10- 3 10-4 0.1 104
I erg/m2 • s 10- 7 10-" 10- 5 10-4 I

QSimilar conversions hold for radiant exposure units, if watts (W) are replaced by joules (J)
in the table, and seconds (s) are eliminated from terms expressed in ergs/unit area. s.

unit of energy is the erg (1 erg = 10- 7 J). In following any field of research
related to radiant energy, one must frequently interconvert these units of power,
irradiance, energy, and radiant exposure (see Table 2-2).

Solar Ultraviolet Radiation

Man rather curiously evolved as a furless species under tropical sunlight.

The protective mechanisms of a photoinducible pigmentation response (tanning)
and a specialized stratum corneum (outer, dead layer of the skin) presumably
reflect this evolutionary process. Despite the recent and increasing use of arti-
ficial sources of light that emit ultraviolet radiation, the sun is still the most
common source of ultraviolet exposure to humans. In certain applications, how-
ever, artificial sources may expose humans to greater ultraviolet irradiances and/
or shorter ultraviolet wavelengths than are present in nature.
Extraterrestrial "sunlight" is a relatively constant, wide spectrum of elec-
tromagnetic energy extending from wavelengths shorter than UV -C through in-
frared, microwave, and radio wavelengths. 3 - 6 The spectral irradiance curve
grossly resembles that of blackbody emission at a temperature of about 60000 K
(Fig. 2-1). Rotation of the earth and atmospheric absorption, reflection, and scat-
tering of sunlight provide conditions necessary for life and account for most of
the daily and seasonal variations in solar irradiance at the earth's surface. Ab-
sorption by stratospheric ozone and other molecules strongly attenuates
wavelengths below 300 nm, effectively preventing the biologically injurious ul-
traviolet radiation below 290 nm (UV-C) from reaching the earth. Other absorp-
tion bands of atmospheric CO 2 and H20 may also be discerned (Fig. 2-1). Solar
spectral irradiance on a cloudless day with the sun directly overhead increases
rapidly from about 295 nm to a maximum at about 500 nm and then decreases
more gradually (Figs. 2-1 and 2-2). The solar UV-A irradiance (320-400 nm)
under these conditions is about 5 m W/ cm 2.
10 Chapter Two

E
..'E
::t.

LAR IRRADIANCE CURVE OUTSIDE ATMOSPHERE

~
I SOLAR IRRADIANCE CURVE AT SEA LEVEL
-...:
w 1500 CURVE FOR BLACKBODY AT 5900 K
w
U
Z
~
o
~
CI: 1000
CI:
...J
~
CI:
ti
~ 500 -----H ' O
(I)

~........._H , O,CO ,

, CO

o~~~~~~~~WZ~~~~~~~~~~
o 0 .2 0.4 0 .6 0.8 1 .0 1.2 1.4 1 .6 1 .8 2 .0 2 .2 2 .4 2.6 2 .8 3 .0
WAVELENGTH(A)-~m

Fig. 2-1. Spectral irradiance E. of the sun at sea level. Shaded areas indicate absorption bands of the
atmospheric constituents shown. (From Gast, P.R. Section 16-1, Solar irradiance, in Handbook of
Geophysics and Space Environments, S. L. Valley, scientific editor. Air Force Cambridge Research
Laboratories, Office of Aerospace Research, United States Air Force, Hanscom Field, Bedford, MA,
1965).

Variations of the spectral distribution and irradiance of sunlight are due

mainly to variations of:

1. The thickness of the atmosphere that the sun's rays must penetrate be-
fore reaching the observer (altitude, solar zenith angle)
2. Atmospheric conditions and pollution
3. Absorption and scattering within various atmospheric layers
4. Ground reflectivity
In general, the ultraviolet region of the solar spectrum is the most variable. 7-10
Figure 2-3 is a simplified illustration of the direct radiation, neglecting scat-
tered "sky" radiation, reaching the earth at two solar zenith angles (angle of sun
measured from directly overhead). When the sun is near the horizon, its rays
Sources of UV-A 11

must traverse an atmospheric path of approximately d. sec (), where d is the thick-
ness of the atmosphere, and () is the solar zenith angle. If no atmosphere were
present, the spectral irradiance El.Ron the earth's surface relative to that of ex-
traterrestrial sunlight, )" would be simply E I.R= E), cos () (see Table 2-1, definition
for irradiance). Applying Beer's law to roughly approximate (neglecting scatter-
ing) the absorption of UV wavelengths by stratospheric ozone, the UV solar
spectral irradiance at the earth's surface is E,,= E), cos () 10- EA1[O,]dsec 9 , where E),
is the molar extinction coefficient for ozone and [0 3] is ozone concentration.
Below about 330 nm, E), increases greatly with decreasing wavelength; sec () in-
creases as () increases. Therefore, the solar spectral irradiance for wavelengths
below 330 nm decreases more rapidly as the solar zenith angle () increases than
for wavelengths above 330 nm. In other words, solar UV-B wavelengths reach-
ing the earth are more strongly dependent on the solar zenith angle than are

220

200

E 180
'",
'"
5 160
::::
~
:I. 140
w'
()
z 120
::;
0 100
~
a::
a:: 80
a::
~
...J
0 60
CJ)

40

20

o 300
WAVELENGTH, nm

Fig. 2-2. Comparison of solar spectral irradiance outside the atmosphere with that at sea level. (From
Stair, R. Measurement of natural ultraviolet radiation, historical and general introduction, in The
Biologic Effects of Ultraviolet Radiation, F. Urbach, editor. Pergamon Press, Oxford, 1969. Used
with permission.)
12 Chapter Two

SUN AT ZENITH

A MOSPHERE

J:ig. 2-3. Atmospheric path of directly transmitted solar radiation as a function of solar zenith
angle, O.

UV-A or visible wavelengths . This is one reason a primarily UV-B-caused sun-

burn develops if one is exposed around noontime. Figure 2-4 plots solar ul-
traviolet spectral irradiance at selected wavelengths during the day. It is apparent
from the figure that solar UV-A spectral irradiance is approximately 100 times
that of the 300 nm UV-B around noon. In midafternoon or midmorning, how-
ever, this ratio increases to about 500.
The dependence of the solar spectral irradiance below 330 nm upon ozone
concentration is also seen in the simple model above. Part of the seasonal fluctua-
tion of solar UV-B irradiance is due to small seasonal changes in ozone concen-
tration [O~]. The recent advent of stratospheric flight and pollutants that di-
minish the concentration of ozone has focused much attention on the formation,
degradation, and measurement of stratospheric ozone. 11 Solar ultraviolet spectral
irradiance as a function of effective stratospheric ozone thickness is given in Fig.
2-5.
Scattering of solar radiation by molecules in the atmosphere accounts for a
large percentage of the total solar ultraviolet irradiance reaching the ground. 10
The degree of molecular or Rayleigh scattering is inversely proportional to the
fourth power of the wavelength of the radiation. Shorter wavelengths are scat-
tered much more than longer wavelengths, and therefore the sky appears blue.
Ultraviolet wavelengths are scattered even more strongly than visible light. At
Sources of UV-A 13

noontime, about 30 to 50% of the total, so-called global ultraviolet irradiance is

scattered radiation, the remainder being directly transmitted through the atmos-
phere. Thus, it is possible to receive a sunburn while sitting "in the shade" if one
is still exposed to the sky. When employing the sun as a source of UV for photo-
therapy, photochemotherapy, or other relatively precise exposures, it is impor-
tant to calculate the exposures based on measurements of the global (direct plus
sky) UV irradiance.
Cloudiness and local atmospheric pollutants generally decrease the solar ul-
traviolet irradiance reaching the ground. Increased ground reflectivity may sig-
nificantly increase the ultraviolet irradiance, depending on the position of the

-4
10

-5
10

E -6
c 10
NE
~
:5:
uj
u -7
z 10
«
Q
«a:
a:
...J -8
«
a: 10
I-
U
w
a.
CJl

-9
10

-10
10 ~~~~-r-+-r~4-~~~~-+-r-r~
3 4 5 6 8 10 12 14 16 18 20

TIME OF DAY/HOURS

Fig. 2-4. Diurnal variation of global solar ultraviolet spectral irradiance at different UV wavelengths;
June at Davos, Switzerland. (Modified from Bener, P. The diurnal and annual variations of the spect-
ral intensity of ultraviolet sky and global radiation on cloudless days at Davos, 1590 m.a.s.1. Air
Force Contract AF61(052)-618, Technical Note No.2, Davos, January 1963.)
14 Chapter Two

-4
10

E
c: ~5
N 10
E
~
~
ui
t)
z
«
o
«
a:
a:
-I
«
a:
l-
t)
w
c...
(fJ

a:
«
-I
o
(fJ
-I
«co
o-I
C1
0
A C
B
-9
10
280 290 300 310 320 330 340

A (nm)

Fig. 2-5. Global solar ultraviolet spectral iQ"adiance for various ozone thicknesses. A: 0.20 cm. B:
0.24 cm. C: 0.28 cm. D: 0.40 cm. (Adapted from Environmental Impact of Stratospheric Flight-
Biologic and Climatic Effects of Aircraft Emission in the Stratosphere. National Academy of Sci-
ences, Washington, D.C., 1975.)

subject. Fresh snow cover may reflect 80% or more of solar ultraviolet radiation.
Ground reflectance is especially important because the reflected radiation may
expose parts of the human body that are generally shaded from significant solar
ultraviolet exposure, such as the eyes.

Summary

Sunlight is a highly variable but potent and common source of ultraviolet

radiation. The maximum UV-A (320-400 nm) irradiance is about 5 mW/cm 2
Sources of UV-A 15

(cloudless sky, zenith angle less than 30°) at sea level and may be greater at high
altitudes or high ground reflectivity. The spectral distribution of sunlight shifts
toward longer wavelengths at increasing solar zenith angles because of greater
subtraction of shorter wavelengths by atmospheric absorption and scattering.
Scattered (sky) ultraviolet radiation is a significant portion of the total global ul-
traviolet radiation.

Artificial Sources of UV-A

Virtually every source of light emits some UV-A radiation. Our interest
here is confined to common environmental sources of UV-A and to specialized
UV-A sources for phototherapy, industrial applications, and photobiologic re-
search. These may be divided into three general categories:
1. Gas discharge sources
a. Direct
b. Fluorescent
2. Incandescent sOurces
3. Lasers and special sources

Gas Discharges

Radiation emitted by a gas discharge is a result of the passage of an electric

current through a gas. Free electrons are driven between two electrodes within
the gas at velocities sufficient to ionize the gas atoms as the two collide. The
displaced electrons may then return to particular electronic states of the atoms,
thereby releasing the energy they have absorbed in the form of electromagnetic
radiation. Because transitions between the discrete electronic states, and the
energies associated with these states, are dependent on the nature of the gas
atoms, the emission spectra of gas discharge lamps show characteristic spectral
lines.
When the gas pressure and temperature within such a lamp are low, it emits
radiation mainly in the form of spectral lines. At high gas pressures and tempera-
tures, gas discharge lamps also emit a continuum of wavelengths between the
spectral lines because of broadening of the spectral lines by collisions between
gas atoms, and heating of the electrodes, which then become incandescent. In-
teractions during collisions between atoms broaden the lines by modifying the
energy associated with excited and ground electronic states. Gas discharge lamps
are commercially available with a variety of gases or mixtures, at different pres-
sures, and in a large variety of lamp configurations, power inputs, and envelope
materials. Arc lamps are gas discharge lamps that, in general, develop high cur-
16 Chapter Two

rent densities and temperatures near the cathode electrode. There is, however,
some debate over the precise definition of an arc discharge. The most widely
used lamps emitting significant UV-A radiation are medium-pressure (2-8 atm)
and high-pressure (over 8 atm) mercury vapor or xenon arcs or mixtures of these
two and other gases. High-pressure short-are-length lamps, also called compact
arcs, are particularly suited for collimating or focusing the radiant energy into
intense beams for use with spectral filters or monochromators. Long-arc lamps
are particularly useful for wide-area sources of radiation.
At low pressures and potential gradients, the mercury discharge lamp emits
more than 90% of its radiant output at a wavelength of 253.7 nm. These lamps
are called germicidal or cold quartz lamps. At higher pressures, the emission
spectrum shifts toward longer-wavelength spectral lines of mercury, plus con-
tinuum (Fig. 2-6). The shift toward longer-wavelength spectral lines at higher
pressure is the result of many factors, including increased self-absorption of
253.7 nm radiation by other Hg atoms within the lamp.

u
w B
I'MW
200 300 400
.
500
WAVelENGTH IN NANOMETERS

C
~.
600
I
700

z
~
0
e(
I-
a: :;)
!!: CL
-' ~
e(
I-
a: l-
I- e(
U
w ~
CL
til a:
w
w CL
>
i=
e(
-'
w
a:

300 400 500 600 700 300 400 500 600 700
WAVelENGTH IN NANOMETERS

Fig. 2-6. Emission spectra of mercury arc lamps at different gas pressures. A: Spectral power dis-
tribution of UA-2 medium-pressure lamp (250 W). Note that approximately 4% of the input power is
radiated as UV-A. (Courtesy ofR. Levin, GTE Sylvania.) B: 31 atm. C: 75 atm. D: 165 atm. E: 285
atm. (From Noel, E. B. Radiation from high pressure mercury arcs. Illuminating Engineering
36 :243-256, 1941. Copyright, Illuminating Engineering Society, 1941. Reprinted from the February
1974 issue of Illuminating Engineering with permission of the Illuminating Engineering Society of
North America.)
Sources of UV·A 17

The strong spectral line at 365 nm accounts for much of the UV -A emitted
by medium-pressure mercury lamps. Medium-pressure Hg lamps are commonly
used for lighting purposes, with a glass envelope employed to attenuate radiant
energy below 300 nm. They are also a readily available source of high-irradiance
UV-A. Attenuation of UV-A by the envelope is minimal, except in lamps that
employ a phosphor coating on the outer bulb to absorb power in the ultraviolet
and reradiate it in the red end of the visible spectrum, thus providing a more ac-
ceptable source of visible light.
Mercury lamps are also available in quartz envelopes that transmit the short-
er ultraviolet wavelengths and are then called photochemical lamps. There has
been extensive use of the large variety of these lamps in ultraviolet photobiologic
research, phototherapy, and industrial applications. At intermediate or high pres-
sures, these lamps are rich sources of 250-400 nm ultraviolet radiation. These
lamps, sometimes called hot quartz lamps, have been commonly used in der-
matologic photobiology and phototherapy.
High-pressure xenon arc lamps emit a broad continuum that extends from
about 170 nm into the infrared region,· with strong spectral lines in the near in-
frared (Fig. 2-7). If a high-pressure Xe-Hg mixture is used, the spectral lines of
mercury enhance the ultraviolet emission over that of xenon alone. With the ex-
ception of the infrared region, the spectral distribution of xenon arc emission can
somewhat resemble that of extraterrestrial sunlight (Fig. 2-7). Solar simulation
may be accomplished using a xenon arc in combination with filters that mimic the
atmospheric absorption of UV wavelengths and also attenuate the excessive in-
frared radiation. 12 Moreover, essentially any wavelength band from UV-C to the
far infrared may be obtained using a compact xenon arc, focusing optics, and
appropriate filters or monochromators.
Much of the ultraviolet emission of high-pressure xenon arcs is due to the
high plasma temperature generated, which, among other factors, corresponds to
the current density within the arc. As the current density is increased, the emis-
sion spectrum shifts toward shorter wavelengths, yielding a richer source of ul-
traviolet radiation. A practical upper limit of arc current and current density is
reached for continuous arcs because of excessive heat loading. The electrodes
that provide the arc may melt, leading to dangerous explosive failure. Most
high-pressure arc lamps may fail in this manner.
However, extremely high current densities, with correspondingly greater ul-
traviolet outputs, are produced in relatively low-pressure confined-arc xenon
flashtubes (Fig. 2_8).13,14 The confined-arc flashtube is a lamp in which the arc
fills the confines of the enclosure. In the unconfined-arc, the arc forms between
the electrodes and does not fill the confines of the enclosure. Greater current den-
sities are possible in the confined arc. The gas within a flashtube is excited by a
brief avalanche of many high-velocity electrons passing between the electrodes,
rather than by a steady current. Because heating of the lamp is largely related to
18 Chapter Two

en
t: 7
z
:::l
>-
a:
~ 6
a:
I-
OJ
a: 5
~
w'
()
z
~ 4
0
~
a:
a:
3
...J
~
a:
I-
()
w 2
Q.
en
w
>
i=
~
...J
W
a:
200 400 600 800
,
1000 1200 1400 1600
. . -..-----
1800 2000 2200
WAVELENGTH,NANOMETERS
Fig. 2-7. Spectral power distribution of 5-kW xenon compact arc lamp. Dashed curve depicts the
solar spectrum outside the earth's atmosphere. (Adapted from General Electric Co. Bulletin LD-l.)

the average power dissipated by the lamp, brief flashing creates higher current
density and UV output, without excessive heating of the lamp.
Peak UV-A irradiances over a large range (1-10 5 W/cm 2 ) may be produced
near these lamps with pulse durations ranging from a fraction of a microsecond to
several milliseconds. The biologic effects and hazards of high-intensity pulsed
UV-B and UV-A is discussed in Chapters 6 and 9. Xenon flashtubes are com-
monly used in photography and stroboscopy, visual beacons, printing, flash
photolysis, and laser excitation. Use in phototherapy has been limited and may
warrant further investigation. The flashed mode is the most efficient means of
generating UV-A from xenon arcs.
Fluorescent lamps are low-pressure mercury discharge lamps in which a
phosphor coating is applied to the inside of the envelope. Depending on the
phosphor used, some fraction of the 253.7 nm ultraviolet radiation absorbed by
the phosphor is reemitted at longer wavelengths. The efficiency may be very high
for fluorescent lamps relative to most other sources. The remainder ofthe electri-
cal input power not converted to radiant energy is lost in the form of heat by
conduction and convection. In fact, any lamp delivers to the environment all of
Sources of UV-A 19

the electrical input power as both heat and radiant energy. Most of the radiant
energy, upon absorption, creates heat. Consequently, the total heat load imposed
by a 40-W lamp of any type upon the environment is essentially the total 40 W.
Fluorescent lamps with ultraviolet-emitting phosphors are currently the
most efficient sources of UV-A. At present, there are two phosphor types com-
monly used in commercial fluorescent UV-A lamps (X and Y in Fig. 2-9A).
These lamps are also available with a visible-absorbing, UV-A-transmitting glass
envelope (BLB, lower curve of Fig. 2-9B). There is also a readily available
fluorescent "sunlamp" (FS40, Fig. 2-9C) with appreciable UV-A and UV-B
emission. Banks of fluorescent lamps are often useful when a large-area diffuse
source is desired. This-in addition to long lifetime, low heat load, and
nonexplosiveness-makes fluorescent lamps excellent for phototherapy or other

-4
30·10

28

26
r-
::J
Cl. 24
E
c ~
"-
Cfl Cfl 22
w W
...J ...J
::J ::J
0...., 0...., 20
BLACKBODY RADIATION
DISTRIBUTIONS
18 9400'K
I 7000'K
Cfl
« 16
,
...
...J

... 14
/
I

,
I
0
(
>- 12
,
l:)
a: I I
w
Z 10 I
w
...J
« 8
,I /
I

a:
u
r- I,'
I "
w 6 II
Cl. 'I
Cfl 1700amp/cm 3
4
"
'I
I,
2 1/
1/
(
0
100 300 500 700 900 1100

WAVELENGTH - nm

Fig. 2-8. Spectral emission of confined-arc xenon ftashtube at two current densities. Relative black-
body curves shown give color temperatures for both current densities. (Adapted from Goncz, J. H.
New developments in electronic ftashtubes. Instrument Society of America, Vol. 5, No. I, 1966.)
20 Chapter Two

A 0.25.,...-----------------------,

0.20

E
c 0.15
a::
w
D..
II)
....
....
« 0.10
~

0.05

0
300 400
WAVELENGTH - nm
I
500
~ /\ .
600

B 1.0

a:: BL
w
~
0
Q.
..J
«
a::
.... 0.5
u
w
Q.
II)
W
>
~
«
..J
w
a::
0
300 350 400 450
WAVELENGTH - nm

Fig. 2-9. (A) Spectral power distribution of 4-ft, Tl2, 40-W fluorescent lamps with different
ultraviolet-emitting phosphors. (B) Relative spectral power distribution of BL and BLB lamps of
equal size and power. (C) Spectral power distribution of 4-ft, Tl2, 40-W fluorescent sunlamp (FS40).
(Courtesy of R. Levin, GTE Sylvania.)
Sources of UV-A 21

,
C
E /).~,NM WATTS
c 0.15 250-290 0.3
290-320 4.0
VI
~ 320-400 2.7
~
« 400-800 1.3
~
0.10
a:
w
~
0
CL.
..... 0.05
«
a:
~
u
w
CL.
VI
0
250 350 450 550
WAVELENGTH - nm

UV exposures in vivo. A high-irradiance UV-A fluorescent lamp has recently

been developed for use in photochemotherapy of skin diseases. 15 Because of
manufacturing changes, the variety of phosphors or mixtures available, and indi-
vidual fluorescent lamp variations, one must perform spectral irradiance mea-
surements of fluorescent lamps to be used for precise research work.

Incandescent Sources

An incandescent body is defined as one that emits radiation primarily be-

cause of its temperature. Incandescent lamps operate by passing electric current
through a filament, usually tungsten. As the filament temperature rises, so does
its electrical resistance, and an equilibrium of current and temperature is reached.
Photons are emitted as a result of the increased vibrational excitation of
molecules within the filament. The total radiant flux emitted by an incandescent
source increases greatly as the temperature increases. The energy distribution of
molecular kinetic excitations, and hence the distribution of emission wave-
lengths, is a function of the temperature of the filament material. At higher temp-
eratures, increased vibrational energies and shorter emission wavelengths result
(Fig. 2-10). Wien's displacement law gives the wavelength in microns of the
emission spectrum maximum of an ideal incandescent body as

Amax =~
TCK)

Incandescent emission may be visualized qualitatively as the reverse of the ab-

sorption process at a material's surface. Thus, spectral emissivity and spectral
reflectivity of substances are inversely related. The ideal incandescent source
22 Chapter Two

UV VIS INFRARED
50~~~~--'------r----~----~r-----'

w
U
Z
~
o
«
II:

«
...J

II:
t-
U
w
Il.
IJ)

w
>
i=
«
...J
10
w
II:

o ;
200 500 1000 1500 2000 2500 3000
WAVELENGTH - nm

Fig. 2-10. Spectral radiance of blackbody emission at various temperatures. Shaded area is visible
light region; dotted curve is photopic response of the human eye. (Adapted from Elenbaas, W. Light
Sources. Crane, Russak and Co., New York, 1972.)

would be one with no reflectance over the optical spectrum (reflectivity of 0.0,
emissivity of 1.0). The rather unfortunate term blackbody is therefore used to
describe an ideal incandescent source, there being nothing "black" about a
source of light.
In general, incandescent sources are relatively weak in ultraviolet emission
compared with arc lamps, which employ both spectral emission lines and high
plasma temperatures to generate ultraviolet radiation. Most of the radiation of
common incandescent lamps is emitted in the infrared region. However, some
tungsten-halogen incandescent sources ("quartz iodide lamps") may produce
significant UV-A and some UV-B emission. These are common sources in pro-
jection systems and outdoor or stage lighting applications. Table 2-3 compares
the UV-A irradiances of sunlight and various arc, incandescent, and ultraviolet
laser sources. Common sources of UV radiation are reviewed in references 16,
17, and 18.

Lasers and Special Sources

Several lasers have recently been developed that emit intense, coherent
beams of monochromatic UV -A radiation (Table 2-4). These special characteris-
tics make UV-A lasers useful research tools.
Sources of UV-A 23

More recently, "tunable" lasers with output from 360 to 670 nm have been
developed. 19 - 22 The pulsed output from a nitrogen gas (337.1 nm) or other
laser or lamp is used to optically "pump" and induce laser action in a cell con-
taining certain dyes. Pumping energy may also be supplied by xenon flash lamps.
Wavelength selection is usually accomplished by replacing the end mirror of the
dye cell laser with a diffraction grating. The monochromaticity, high peak ir-
radiance, and nanoseconds pulse time of tunable lasers may be useful for examin-
ing biologic action spectra and the effects of high irradiances and very brief
pulsed radiation.
"Monochromatic" ultraviolet radiation is more commonly obtained by
selectively transmitting a narrow spectral region of a broadband light source such
as the compact xenon arc lamp; dispersive elements (diffraction grating or quartz
prism) or narrow bandpass filters (absorption or interference types) are com-
monly used. A complete discussion of monochromators is not presented here, as
excellent reviews are available elsewhere (see references 23-26). Mono-
chromators are essential for determinations of biologic action spectra.
For photobiologic research in the UV -A region, several light source criteria
must be met. In general, organisms and mammalian cells are several orders of
magnitude more sensitive to UV-C and UV-B wavelengths than to UV-A
wavelengths. Therefore, for meaningful results, "stray" shorter-wavelength ul-
traviolet radiation emitted by a UV -A source must generally be less than 10- 5
that of the UV -A emitted. Other experimental or practical requirements-such as
degree of monochromaticity, size of exposure sites, uniformity of irradiance over

Table 2-3. Comparison of UV-A Irradiances of Various Sources

Approximate UV-A
irradiance, mW/cm 2
Source (320-400 nm)

(Tropical) sunlight at noon (8 = 0°) 5.0-6.0

Sunlight at 3 P.M. or 9 A.M. (8 = 45°) 2.5-3.5
400-W (clear) Hg street lamp at 3 m 0_1
500-W Photoflood #2 lamp at 2_5 cm 1.0-2
Fluorescent black light (F40BL) at 2.5 cm 5.0
1000-W Xe-Hg compact arc lamp at 10 em 50-80
Nitrogen gas laser (337.1 nm) 500 average
108 peak
Sylvania fluorescent UV-A source for photochemotherapy
10.0
of psoriasis 15
24 Chapter Two

Table 2-4. UV-A Lasers

Emission wavelength (5) Typical radiant flux

Laser substance (nm) in watts

Nitrogen gas (pulsed) 337.1 0.5 average

10 5 peak
Tunable liquid dyes 360-670 0_05 average
pumped by N2 laser 5 X 104 peak
(pulsed)
With crystal doubling 200-350 0.005 average
10 3 peak
Helium-cadmium metal 325.0,441.6 0.015
vapor
Neon ion gas 332.4 0.25
(continuous)
Argon ion gas 35l.l,363.8 0.25
Krypton ion gas 350_7,356.4 0.25
Xenon fluoride 351 0.5

the exposure area, UV -A irradiance, safety aspects for exposure of humans, and
cost-generally define the type of UV-A source that is most suitable.

Spectral Filters for High-Intensity Broadband UV-A Sources

Although an ultraviolet monochromator system is necessary for determining

biologic action spectra, it is often adequate to use a broadband source ofUV-A in
experimental work if the wavelength band is well defined. In general, higher
UV-A irradiances and larger exposure areas may be achieved more easily with
broadband sources. Any number of broadband UV -A sources may be designed
for a given application, given the number and types oflamps and filters available.
In UV-A phototherapy, for example, fluorescent lamps are generally the best.
However, the most versatile broadband high-intensity UV -A experimental
source is probably the high-pressure mercury, xenon, or xenon-mercury short
arc lamp (1000-2500 W) in a housing with fast (f/1.5-f/0.7) quartz lenses that
gather and collimate the radiant energy into an intense uniform beam. Spectral
filtering of the output beam is then used to provide UV-A or other wavelength
bands of interest. Many such lamp sources are commercially available (see list of
manufacturers, end of Chapter 3).
Colored-glass or interference-type optical filters (described in Chapter 3)
may be used to selectively transmit the UV-A wavelength band of interest. Of the
Sources of UV-A 25

absorption colored-glass filters, Corning CS7-51 and CS7-60 and Schott UG-I
are commonly used for selection of UV-A. These filters may solarize quickly
under intense ultraviolet radiation and must be replaced frequently to restore their
original spectral transmission characteristics. They are not very expensive. Inter-
ference filters, in general, show better stability than colored-glass filters but are
more expensive. The main advantage of interference filters is that they may be
custom-designed for desired spectral transmission characteristics. Thus, one can
specify an interference filter to suit a particular experimental task. Regardless of
the type of filter used, the intense short-ultraviolet, visible, and infrared radiation
emitted by the source should be attenuated before reaching the final spectral
filter, otherwise the filter will be destroyed by overheating or photochemical
degradation due to absorption of these wavelengths.
The most common means of attenuating infrared radiation is to use a water
cell, which is available from most manufacturers with the source and housing. A
pump, tubing, and reservoir are required to circulate the solution and keep it from
boiling. Aqueous solutions of inorganic salts may also be used as liquid optical
filters in these systems. 27 One useful UV-A bandpass liquid filter consists of
CoS0 4 and CUS04 dissolved in 0.2% H 2 S0 4 solution. While more complicated
to use than colored-glass filters, liquid bandpass filters do not solarize and can
handle greater power levels. Another means is to use a first-surface, multilayer,
dielectric-coated (similar to interference filter coatings) mirror, often called a
dichroic mirror, maximized for UV-A reflectance at 45° incidence angle. The
broadband UV-A reflectance of these mirrors may be as high as 99%, while the
reflectance over much of the visible and infrared region is about 10%. Because
essentially no energy is absorbed by dielectric coatings (the infrared and visible
wavelengths not reflected are transmitted), overheating is not a problem. The
"hard-coating" types are vacuum-deposited at high temperatures as opposed to
the lower temperatures used for soft-coating types, and they offer the best
maintenance. Figure 2-11 illustrates, for example, a broadband UV-A source
using a UV -A dichroic mirror and collimated compact arc lamp emission.
Multiple reflections of the beam off dielectric-coated 45° mirrors may be
employed to form a very efficient and stable but expensive high-intensity broad-
band UV-A filter. An arrangement using four reflections is commercially availa-
ble with peak reflectance of the mirrors at 340 nm (Schott UV -R-340, Fig. 2-12).
The coatings in this case are deposited on black glass substrates, the first of
which may overheat at high broadband power levels. For high-intensity applica-
tions, the substrate should be a Pyrex® (Corning Glass) glass or quartz, and
shields should be placed appropriately to trap the transmitted component from
each coated mirror. When these reflective filters are used in a UV-A source, an
additional glass filter is necessary to remove stray UV-B and UV-C wavelengths.
Custom dielectric-coated mirrors, interference filters, and other thin-film-coated
optics are readily available from many optical manufacturers.
26 Chapter Two

Ultraviolet Spectral Transmission and Reflection

of Common Materials

All transmissive and reflective materials are filters in the sense that they
modify the spectral distribution of radiant power. Thus, many materials may be
used as UV-transmitting or UV-absorbing filters. For some purposes, large-area
filters as opposed to small optical filters must be specifically provided. At other
times, physically flexible filters-or filters that weigh very little, or withstand
shock, etc. -are necessary. Often some common material may be found to suit
these requirements. Finally, transmission and reflection characteristics of com-
mon materials affect levels of environmental UV -A.
Reflection from glass or plastic is a function of the index of refraction,
wavelength, and the angle of incidence of the radiation. Typically, the reflec-
tance is about 5% at an air-glass interface in the near ultraviolet when the flux is
perpendicular to the interface. The minimum UV-A reflectance for a sheet of
window glass is about 10% because of the two surfaces. The reflectance at an
air-glass interface increases rapidly at large angles of incidence as shown in Fig.
2-13. Thus, for grazing angles, considerable radiation can be reflected.
Spectral transmittance of glass is dependent on the absorbing properties of
the various constituents. Since the number of glasses and glasslike materials is
extremely large, it is difficult to generalize. Most visible transmitting glasses ab-
sorb in the UV-A or UV-B band, and nearly all absorb UV-c. Very pure quartz
(silica glass) may have virtually no attenuation at wavelengths greater than 200
nm. Many types of quartz have high transmission in the UV-C band, as illus-
trated by the example of Fig. 2-14A.
The most common glass is soda lime glass, which is sometimes used for
ultraviolet filtering. Its other uses range from "window glass" to light bulbs. The

UV-A REFLECTIVE
DICHROIC MIRROR

ABSORBER

UV-A + 10% IR-vis

" FIL TER

l i1
COMPACT ARC LAMP

UV-A

Fig. 2-11. A research source of high-irradiance broadband UV -A radiation.

Sources of UV-A 27

UV SOURCE
UV-A

COLLIMATING LENS

B 100
90
80

z 70
~ 80
III
III
iIII 50
Z 40
C
c
~ 30

...
~
Z 20
U
...
C
L
10
1.0

0.1
0.01

0.001
240 280 280 300 320 340 360 380 400 420 440
WAVElENGTH - nm

Fig. 2-12. (A) Use of the Schott UVR-340 high-efficiency broadband UV-A filter. (8) Approximate
spectral transmission of Schott UVR-340.

UV transmission cutoff tends to be near the boundary between UV-A and UV-B,
but the spectral transmittance characteristics depend on the specific glass formu-
lation, especially iron impurities, and on the thickness. Figure 2-14B shows the
transmittance of two samples of soda lime glass at 2-mm thickness, about the
thickness of single-strength window glass. Absorption is exponential with thick-
ness. Consequently, a change in thickness has the greatest effect at wavelengths
oflower transmittance. The glass of sample B in Fig. 2-14B is increased to 6-mm
thickness (dashed curve) to show the effect on spectral transmittance. Thus, one
simple method of decreasing transmittance at shorter wavelengths without ap-
 28 Chapter Two

1.0

0.75
,L// "
w
u ~AIR
z
« GLASS
I- 0.50
u
w
...
...J

W
cr 0.25

O. .-.-,---.---.-~--.---;
O· 20· 40· 60· 80·
ANGLE OF INCIDENCE - L
Fig. 2-13. Reflectance at air-glass interface as a function of angle of incidence, i, for unpolarized
light (index of refraction = 1.5).

preciably changing transmittance at longer wavelengths is to stack several layers

of the appropriate glass.
When using glass as a filter, it is necessary to obtain a spectral transmittance
curve for that type of glass and correct it to be the appropriate thickness. 28 For
example, the ultraviolet transmission of window glass may vary with the man-
ufacturer and change with time even from a particular manufacturer. The man-
ufacturing tolerance range, especially on the steep portion of the cutoff curve, is

A 1.00 ,.--_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _----,

0.75
w
(J
z
«
l-
iI- 0.50
If)
z
«a:
I- 0.25

0.00
200 250 300 400
WAVELENGTH - nm
Sources of UV-A 29

81.00

/- ........ _ /
/
015
III
(J
cl
z /
~
1= /
:i 0.50
II) I
z
~
I
.-II:
0.25
I
I
I
I
Qoo ., ./
I

250 300 360 400

WAVELENGTH - nm

C 1.00

0.75
III
(J
Z
.-.-
~

i
II)
0.50
z
~
.-
II:

0.25

O~------------------------------~------------------------------~-------------------------~
200 250 300 350
WAVELENGTH - nm

Fig. 2·14. Spectral transmittance. (A) A: 2·mm quartz (Coming 7910). B: 2-mm borosilicate (Com-
ing 7740). C: 2-mm Pyrex 774. D: 3-mm Pyrex 7331. (B) A and B: 2-mm samples of soda lime glass.
C: 6-mm sample of glass B. (C) Solarization of I-mm Pyrex 9741. A: initial. B: 230-hr exposure at
15 cm from 500-W mercury arc. (Courtesy of R. Levin, GTE Sylvania.)
30 Chapter Two

likely to be large. For critical applications, the spectral transmittance of the

specific glass samples to be used should be measured.
It should not be assumed that the transmittance is zero where the conven-
tional linear spectral transmittance curve appears to go to zero. Examining the
transmittance on a logarithmic scale (optical density) usually shows that the
cutoff with wavelength is not sharp. Appreciable power may be transmitted in a
wavelength region of low filter transmittance if the source is rich in that region.
The spectral irradiance passing through a filter is the product of filter transmit-
tance and the spectral irradiance of the source.
A photochemical reaction occurs in most glasses exposed to ultraviolet radi-
ation. This effect, which may cause the ultraviolet transmittance to decrease and
is often accompanied by a change of color, is known as solarization. Depending
on the glass composition, the temperature, the exposure time, and the
wavelengths and intensity of the irradiation, solarization may be of considerable
importance in the use of filters. Figure 2-14C illustrates the effect of solarization
on a Pyrex sample. The transmission loss due to solarization can be reversed for
many filters by proper heat treatment.
Many plastics have appreciable transmission in the near ultraviolet. Unfor-
tunately, in most cases the UV-A transmission decreases after some ultraviolet
exposure. Figure 2-15A shows, for example, the spectral transmittance of

A 1,00

0.75
w
U
Z
<{
l-
I-
:ii 0.50
(/)
z
<{
a::
I-
0,25

0.00
250 300 350 400
WAVELENGTH - nm

Fig. 2-15. Spectraltransmittance. (A) 0.025" polycarbonate sheet exposed to 8 mW/cm' UV-A. A: 0
hr. B: 200 hr. C: 500 hr. 0: 1000 hr. (B)0.020" Mylar sheet exposed to 8 mW/cm' UV-A. A:O hr. B:
250 hr. C: 900 hr. (C) 0.125" acrylic sheet (Plexiglas). A: type IIUVT. B: type G. C: type UF3. 0:
type G after 2-yr exposure to sunlight in Pennsylvania. (Courtesy of R. Levin, GTE Sylvania.)
 Sources of UV·A 31

B 1.00

0.75
UJ
U
z
«
~
~
~ 0,50
(fJ
z
«
a:
~
0,25

0,00

250 300 350 400

WAVELENGTH - nm

C 1.00

w 0,75
u
z
-4:
l-
I-
0.50
::E
IJl
Z
-4:
a:
I- 0,25

0.00
250 300 ~ 400 450
WAVELENGTH - nm

polycarbonate after various UV-A exposures. The radical changes in UV-A

transmittance caused by photochemical changes within the plastic indicate the
unsuitability of this material for filtering purposes with UV-A lamps. When it is
necessary to remove UV-B from a large-area source of UV-A, a Mylar® (Dup-
ont) sheet may be used because of its very sharp cutoff at about 315 nm. The
UV-A transmission of Mylar also decreases with increasing exposure of the My-
lar, as shown in Fig. 2-15B. Consequently, frequent changes of the plastic are
necessary to maintain both the level and the spectral distribution of the irradia-
tion.
Acrylic plastic often referred to by the terms Plexiglas® (Rohm and Haas) or
32 Chapter Two

Lucite® (Dupont), is frequently used for rigid enclosures, large-area lenses or dif-
fusers, etc. This material is available in many formulations with differing ul-
traviolet transmission characteristics. Figure 2-15C illustrates the most common
form (curve B), which cuts off in the middle of the UV -A band, as well as formu-
lations especially developed to transmit or to block ultraviolet. Similar to other
plastics, the ultraviolet-transmitting types change in spectral transmission under
exposure to ultraviolet (curve D).
It is sometimes necessary to use a protective plastic sheet that transmits ul-
traviolet radiation. Teflon® (Dupont) is one plastic that absorbs very little UV
radiation and can be used in this protective application. Since it has low UV ab-
sorbance, it also exhibits excellent stability under UV exposure. The material
commonly used is identified as Teflon FEP by most plastics suppliers. It is avail-
able in a variety of tubing and sheet sizes. For thicknesses up to at least 1 mm,
the absorption is small. Transmission is about 70% at 250 nm and 80% or greater
at 275 nm or greater.
Plastics exposed to ultraviolet radiation tend to "yellow" because the
transmission decreases in the violet and blue part of the spectrum as well as in the
ultraviolet. Manufacturers have therefore added ultraviolet-absorbing agents to
some plastics to minimize this effect. Such plastics are said to be UV -stabilized
and are useful when it is necessary to remove virtually all ultraviolet radiation
from a source of visible light.
With a few exceptions, most common nonmetallic surfaces are medium to
strong absorbers in all parts of the ultraviolet, including the UV -A, irrespective
of the visible color. White-coat plaster does have a high reflectance extending
into the UV-C, with typical values of 0.75 at 360 nm, 0.65 at 300 nm, and 0.45
at 250 nm. White Teflon is a good reflector in the UV -A band with a typical
reflectance in this region of 0.85. Fresh snow also can have a reflectance of 0.85
in this. band. Some bleached cotton and linen cloths may reach as high as 0.5
reflectance over most of the UV -A band. Figure 2-16A illustrates the reflectance
of a few typical nonmetallic surfaces.
Most metallic surfaces reflect UV-A and visible equally. The best-known
exception is silver, with a minimum of reflectance at about 315 nm. Figure 2-16B
illustrates the spectral reflectance of a few typical metal surfaces. Reflectors for
controlling and redirecting UV radiation should have a high reflectance in the
appropriate UV region. Aluminum is the most suitable reflector material for gen-
eral UV applications. Construction aluminums have low reflectance (e.g., curve
D, Fig. 2-16B). However, aluminum especially prepared for reflector applica-
tions can have a UV-A reflectance near 90% (curve A, Fig. 2-16B). Reflector
aluminum suitable for the UV is available in a variety of forms, thicknesses, and
surfaces.
The most common potent source of UV -A (320-400 nm) is sunlight, which
may produce approximately 5 mW/cm 2 of UV-A irradiance but is highly vari-
Sources of UV-A 33

A 1.00

0.75
III
c.J
Z
C J
~
c.J 0.50
...
III
....J
III
a:
0.25

C
0
300 400 500 600 700
WAVELENGTH - nm

B 100.,....._ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _.,

",,,,, -------------------------------------
A
,
075 ,,
.' B
III
c.J
Z
C
t; 050 o
....... -------_"'!"
III

III
a:
0.25 F
I
I
I

o+----------,.----------.-----------.----------~
300 400 500 600 700

WAVELENGTH - nrn

Fig. 2-16. Spectral reflectance on common materials. (A) Nonmetallic materials. A: white acrylic
paint electrostatically deposited on metal. B: white latex paint. C: white asphalt tile. D: yellow oil
paint. E: white dacron polyester. F: white polished marble. G: yellow brick. H: beige nylon. J: asbes-
tos cement siding. (B) Metals. A: aluminum reflector sheet. B: chromium. C: galvanized steel. D:
aluminum sheet 3S. E: stainless steel. F: brass. G: silver. (Courtesy of R. Levin, GTE Sylvania.)

able_ Numerous arc lamp sources, fluorescent lamps, and incandescent lamps
expose humans to various, generally lower, UV-A irradiances_ Available to the
researcher are specialized sources producing UV-A, such as solar simulators,
continuous- and pulsed-arc sources, monochromators, and lasers_ The variety of
sources and of filtering materials is constantly expanding_
34 Chapter Two

References

I. Commission Internationale d'Eclairage (ClE). Internatioiwl Lighting Vocabulary. Bureau Cen-

tral de la ClE, Paris, 1970.
2. Illuminating Engineers Society (IES). Lighting Handbook, fifth edition. Illuminating Engineer-
ing Society of North America, New York, 1972.
3. Henderson, S. T. Daylight and Its Spectrum. American Elsevier, New York, 1970.
4. Stair, R. Measurement of natural ultraviolet radiation: Historical and general introduction. In
The Biologic Effects of Ultraviolet Radiation (with Emphasis on the Skin) (F. Urbach, Ed.). Per-
gamon Press, Oxford, 1969, pp. 377-391.
5. Buettner, K. The effects of natural sunlight on human skin. In The Biologic Effects of Ultraviolet
Radiation (with Emphasis on the Skin) (F. Urbach, Ed). Pergamon Press, Oxford, 1969, pp.
237-251.
6. Robertson, N. Solar Radiation. Elsevier, New York, 1966.
7. Gates, D. M. Spectral distribution of solar radiation at the earth's surface. Science 151 :523,
1966:
8. Bener, P. The diurnal and annual variations of the spectral intensity of ultraviolet sky and global
radiation on cloudless days at Davos, 1590 m.a.s.1. Air Force Contract AF61(052)-618, Techni-
cal Note No.2, Davos, January 1963.
9. Bener, P. Investigation on the influence of clouds on ultraviolet sky radiation. Air Force Contract
AF61(052)-618. Technical Note No.3. Davos-Platz, March 1964.
10. Green, A. E. S., Sawada, T., and Shettie, E. P. The middle ultraviolet reaching the ground.
Photochem. Photobiol. 19:251-259, 1974.
11. Environmental Impact of Stratospheric Flight-Biologic and Climatic Effects of Aircraft Emis-
sions in the Stratosphere. National Academy of Sciences, Washington, D.C., 1975.
12. Berger, D. S. Specification and design of solar ultraviolet solar simulators. J. Invest. Dermdtol.
53:192-199, 1969.
13. Goncz, J. H. New developments in electronic flashtubes. Instrument Society of America, Vol. 5,
No. I, 1966.
14. Goncz, J. H., and Newell, P. B. Spectra of pulsed and continuous xenon discharges. J. Opt.
Soc. Am. 56:18-92, 1966.
15. Levin, R., and Parrish, J. A. Phototherapy of vitiligo. Lighting Design and Application 5:35-
43, March 1975.
16. Task Force on Photobiology of the National Program for Dermatology. Report on ultraviolet
light sources. Arch. Dermatol. 109:833-839, 1974.
17. Elenbaas, W. Light Sources. Crane, Russak, New York, 1972.
18. RCA Electro-Optics Handbook. RCA Corp., Lancaster, Pennsylvania, 1974.
19. Von Gutfeld, R. J. Picosecond dye laser pulses using nitrogen laser pumping. App. Physics Lett.
/8(11 ):481-482, 1971.
20. Myer, J. A., Kierstead, E., and Itzkan, l. Dye lasers in the ultraviolet. Nature 225:544-545,
1970.
21. Goldman, L. Applications of the Laser. CRC Press, Cleveland, 1973.
22. Fine, S., and Klein, E. Ultraviolet lasers. In The Biologic Effects of Ultraviolet Radiation (with
Emphasis on the Skin) (F. Urbach, Ed.). Pergamon Press, 1969 pp. 115-123.
23. Johns, H. E., and Rauth, A. M. Theory and design of high intensity UV monochromators for
photobiology and photochemistry. Photochem. Photobiol. 4:673-692, 1965.
24. Johns, H. E., and Rauth, A. M. Comparison of spectral purity and intensity of different UV
monochromators. Photochem. Photobiol. 4:693-707, 1965.
Sources of UV-A 35

25. MacKenzie, L. A., and Frain-Bell, W. The construction and development of a grating mono-
chromator and its application to the study of the reaction of the skin to light. Br. J. Dermatol.
89:251-264, 1975.
26. Satoh, Y., Irimajiri, T., Okawara, S., Shimao, K., and Seiji, J. A newly designed monoc-
hromator and action spectra of various photodermatoses. In Sunlight and Man: Normal and Ab-
normal Photobiologic Responses (M. A. Pathak, L. Harber, M. Seiji, and A. Kutika, Eds.; T.
B. Fitzpatrick, Consulting Ed.). University of Tokyo Press, Tokyo, 1974, pp. 575-591.
27. Muel, B., and Malpiece, C. Chemical filters for narrow band U. V. irradiation between 235 and
300 nm. Photochem. Photobiol. 10:283-291, 1969.
28. Koller, L. Ultraviolet Radiation. Wiley, New York, 1965.
CHAPTER 3

Radiometry
of Ultraviolet Radiation

Introduction and General Considerations of Radiometry

It is of utmost importance in any field of research to control, quantify, and objec-

tively describe the conditions of an experiment. Much of the literature in photo-
biology lacks adequate, and occasionally any, measurement of the radiation.
This is no longer acceptable. Recent advancements in electro-optics technology
have produced readily available, versatile, and relatively precise ultraviolet
radiometry equipment. This chapter reviews radiometric techniques and devices,
with emphasis on the ultraviolet region.
The basic scheme in radiometry is to employ a device that transduces its
absorption of radiant energy into a directly measurable form of energy, such as
heat, or an electric current or potential, in a definable, calibrated, and repeatable
manner. A large variety of detectors are available in many forms, covering a
large range of wavelengths, sensitivities, fields of view, and other characteris-
tics. Those most applicable for ultraviolet radiometry are discussed here in some
detail.
In practice, some type of optical filter or a monochromator is often used
between the detector and the source of radiation, which allows direct measure-
ment of irradiance over a spectral region of interest, or wavelength-scanned
spectral irradiance measurements in the case of monochromator/detector combi-
nations. Detectors or filter/detector combinations that respond equally to a wide
range of wavelengths (" flat" spectral response) are particularly useful because
total irradiance may be easily measured by the unfiltered detector, and because
the spectral response of these detectors when used with filters is easily calculated
from the filter's spectral transmission. The term spectral response refers to the

37
38 Chapter Three

signal produced by a given detector or filter/detector combination as a function of

wavelength. Calibration of radiometers or spectroradiometers consists of deter-
mining the spectral response by means of a standard source.
The National Bureau of Standards maintains ultraviolet calibration ac-
curacies of about 1.5% (using a stabilized deuterium arc source). In practice, the
calibration accuracy of most ultraviolet radiometers is somewhat less than this,
and the accuracy of actual irradiance measurements is again somewhat less, de-
pending on the circumstances of the measurements. Most manufacturers of
radiometers have optical calibration facilities traceable to the NBS standard;
however, radiometers are generally not supplied with calibrations, which should
therefore be specified when purchasing an instrument. Periodic recalibration and
instrument stability checks using a stable source of ultraviolet radiation should be
performed.
The goal of radiometry in photobiology is to define the spectral irradiance,
the exposure dose, and the geometries of the radiation striking the subject. In the
case of essentially monochromatic radiation, a single measurement of irradiance
using a calibrated radiometer sensitive to that wavelength is often adequate. In
the more common case of polychromatic (broadband) radiation, three general
methods are employed to arrive at spectral irradiance: (1) direct absolute mea-
surement with a spectroradiometer; (2) calculation of spectral irradiance from
photometric (visible light weighted to an average human eye spectral response
curve) measurements, whem the spectral radiance relative to the visible luminous
flux is known and applicable; and (3) calculation of spectral irradiance from mea-
surements of the total irradiance, when relative spectral radiance data are known
and applicable.
Direct measurement of spectral irradiance at the same position and orienta-
tion as the sample to be irradiated is the best of these three methods and is dis-
cussed here in some detail. The methods involving calculations are important be-
cause lamp manufacturers often give spectral power data for common lamps rela-
tive to luminous flux (lumens) or simply as a relative curve without calibration. If
one makes a single measurement of illumination (lumens/ft 2 = footcandles) at
the distance of interest from the bare lamps, the manufacturer's spectral data
(typically given in ILW/nm·lm) are readily converted to spectral irradiance units
by multiplying the data given by the illumination in footcandles.
The total irradiance caused by a source is the sum of spectral irradiance over
all wavelengths emitted by the source. Relative uncalibrated spectral radiance
curves, often called spectral power distribution, may therefore be reduced to
spectral irradiance data by normalizing the area under the curve and setting this
area equal to the total irradiance measured by a detector that responds equally to
all wavelengths present, that is, a "flat" spectral response detector. These two
indirect methods of arriving at spectral irradiance are prone to considerable error
and apply only to bare lamps without reflectors behind them, but they may be
Radiometry of Ultraviolet Radiation 39

useful in estimating the ultraviolet spectral irradiance that would be expected

from bare commercially available light sources.
Geometric factors, such as the solid angle subtended by the source relative
to the position of the subject, the divergence of the radiation striking the subject,
and the orientation and shape of the subject, are important considerations when
performing irradiance measurements. Since the irradiance caused by a beam of
light falling on a flat surface varies directly with the cosine of the angle of inci-
dence (Fig. 3-1A), the ideal detector for measuring irradiance on essentially flat
objects such as small areas of skin has an angular response weighted as the cosine
of the angle of incidence at the detector's face (Fig. 3-1B). It is also necessary
that the detector's field of view encompass the full solid angle presented by the
source of radiation. This is particularly important for wide-area ultraviolet
sources, such as sunlight or nearby fluorescent lamps. The detector must also be
held in the same position and orientation as the subject to be irradiated. Irradia-
tion of non flat subjects presents a special problem in radiometry.
Furthermore, the uniformity of irradiance over the exposure area should be
measured, unless source characteristics define a highly uniform irradiance, such
as with most artificial sources if used at a distance of at least 10 times the diame-
ter of the source, with no focusing optics (lenses or concave reflectors). In the
case of a small exposure area, irradiance uniformity may often be measured using
an optical pinhole placed over the detector. Relative irradiance readings are then
taken over the exposure field, and uniformity is expressed as: measured ir-
radl·ance +xm
- 70,
where X = Ii'12 X greatest differenceofpinholereadings X 100 m
average of pmhole readmgs -10.
The exposure dose (typically in J/ cm 2) delivered by a stable source of radia-
tion to a given area is simply the irradiance (in W/ cm 2) times the duration of the
exposure in seconds. In this case, one need only specify the appropriately mea-
sured spectral irradiance, the various source and subject geometries associated
with the exposure, and the exposure duration to adequately describe the delivered
exposure dose. In cases of pulsed, chopped, or unstable sources such as sunlight
over a long exposure period or on a partly cloudy day, the radiometer signal,
corresponding to irradiance, may be electronically time-integrated during the ex-
posure period. These instruments are usually referred to as photodosimeters.
It is often the case that relatively weak sources of ultraviolet radiation emit
intense visible or infrared radiation as well. The high sensitivity of living or-
ganisms to radiation of UV -B and UV -C wavelengths makes it necessary to mea-
sure accurately the low ultraviolet irradiances present. If one is using a UV-
transmissive filter/detector or monochromator/detector combination with some
seemingly small response to the longer wavelengths present, considerable error
may result. One method of ascertaining whether this type of error due to stray
light is present is to interpose an ultraviolet-absorbing, visible, and infrared-
transmitting glass filter ("long-pass" filter) between the detector and the source.
The irradiance reading due to longer wavelengths may then be noted and sub-
40 Chapter Three

Ea =E n COS a

RELATIVE RESPONSE
B
OF DETECTOR

e ANGLE OF INCIDENCE FROM

------....
NORMAL AT DETECTOR FACE
COSINE RESPONSE

20°

NARROW FIELD
OF VIEW

RELATIVE RESPONSE
OF
DETECTOR

Fig. 3-1. (A) Illustration of the Lambertian cosine law for irradiance. (B) Polar plot of detector angu-
lar response characteristics (field of view).
Radiometry of Ultraviolet Radiation 41

tracted from the former reading after compensation for the glass filter's transmis-
sion. The Schott WG series of optical filters is especially useful for this purpose.
The source employed, the subject being irradiated, and other specific factors
generally define the methods of radiometry best suited for a particular experi-
ment.
In general, the important factors and considerations that may affect
radiometry are:
Detector/Filter or Detector/Monochromator Combinations
1. Sensitivity and dynamic range (the lower and upper limits of irradiance
that may be accurately measured).
2. Signal-to-noise ratio and stray light response (unwanted signal due to
inherent noise of the detector and to detection of wavelengths other than
those for which the measurement is intended).
3. Precision (detector/ filter and electronics stability, and therefore the
short-term repeatability of irradiance measurements).
4. Accuracy (the confidence limits of irradiance measurements on an abso-
lute scale, i.e., the confidence of absolute calibration of the instrument
spectral response and angular response characteristic).
5. Useful spectral range.
6. Field of view and angular response characteristics.
7. Speed ofresponse.
8. Degree of interference from nearby sources of electromagnetic noise.
9. Cost, ruggedness, and portability.
Sources
1. Spectral irradiance at the site of exposure.
2. Size of exposure field.
3. Uniformity of exposure field.
4. Geometries of the source and its radiant output.
5. Stability of irradiance over the period of exposure.
Subjects
1. Shape and size.
2. Physical state.
3. Optical properties of absorption, transmission, reflection, and scatter of
both subject and environment. (For instance, if a UV-A-transmitting
petri dish containing tissue cultures is placed on a metal tabletop for
UV -A irradiation from above, the cells are exposed to both incident
radiation and some reflected radiation from the tabletop. If unaccounted
for, this reflected radiation may cause a large error in the accuracy of
delivered exposure doses.)
42 Chapter Three

Accurate, versatile, and practical ultraviolet radiometer systems are cur-

rently available (see list of manufacturers at end of chapter); some of the detec-
tors, filters, radiometers, and spectroradiometers applicable to the ultraviolet reg-
ion will now be discussed in some detail.

Detectors

In general, five types of detectors on the market today are most suitable for
the measurement of ultraviolet radiant energy. These are photomultipliers,
solid-state photodiodes, vacuum photodiodes, thermopiles, and the recently in-
troduced pyroelectrics. Recent advances in materials technology have caused the
pyroelectric detector to gain an increasing role in the measurement of optical
radiation. Other detectors of radiant energy such as photochemical actinometry,
thermocouples, thermometers, and ballistic thermopiles are generally less suit-
able for UV-A measurement because of various technical and practical difficul-
ties associated with these devices and methods. These methods have largely been
replaced by the more convenient, more accurate detectors listed above.

Photomultipliers

A photomultiplier is a highly sensitive vacuum tube photodetector contain-

ing a built-in current-amplifying system. Figure 3-2 is a schematic representation
of the internal structure of a typical vacuum tube photomultiplier. The incoming
radiation impinges on the semitransparent photocathode releasing electrons via
the photoelectric effect (the release of low-energy electrons from a metal caused
by absorption of photons). These electrons are accelerated through space by the
electric field between the photocathode and the first dynode. They then strike the
first dynode with sufficient energy to release additional electrons from the dynode
by the process known as secondary emission. This process is repeated at each
dynode. The net effect is an avalanche of electrons reaching the anode of the
tube, caused by a relatively small number of photons, which originally began the
avalanche. The current collected at the anode is further amplified and displayed.
The minimum photon energy (and therefore maximum wavelength) required
to release electrons at the photocathode is related to the electronic bonding
strength of the metal atoms. The minimum energy required to produce the photo-
electric effect in a given material is called the work function of the material.
Photomultiplier photocathodes are generally Cs-Sb or Cs-Sn compounds that
have a low photoelectric work function. Other photocathode materials include
Ag-O-Cs, Ag-Bi-O-Cs, Na-K-Sb-Cs, Cs-Te, and several compounds con-
taining gallium arsenide. The spectral response of the detector is related mainly
to the photocathode material.
Radiometry of Ultraviolet Radiation 43

LOW LEVEL
INCOMING IRRADIATION

PHOTOCATHODE COATING(INTERNAL)

DYNODES

SECONDARY
ELECTRON
COLLECTOR. /
ANODE

DYNODE VOLTAGE
DIVIDER RESISTORS

Fig. 3-2. Typical photomultiplier structure showing internal assembly.

The spectral range of commercially available photomultipliers extends from

180 nm through about 1100 nm (Fig. 3-3). Many of the photocathode materials
produce a wide spectral response and are therefore useful in spectroradiometers
over a wide range of wavelengths. Cs-Te photocathode detectors exhibit essen-
tially no response at wavelengths longer than about 360 nm (often called solar
blind detectors). These are extremely useful for measuring low-level UV-B or
UV-C irradiances in the presence of intense UV-A, visible, and infrared radia-
tion.
Most photomultipliers are capable of amplifying the primary photocurrent
from the photocathode by a factor of more than 10 6 • Because of this high internal
amplification and relatively low noise, photomultipliers are routinely capable of
detecting irradiances on the order of 10- 15 W/ cm 2. lithe tube is cooled to reduce
the thermally excited (dark noise) current, irradiance levels of 10- 18 W/cm 2 may
be measured, and single-photon counting is possible. This high sensitivity is not
achieved in any other electro-optical detector. Photomultipliers are ideal for use
in spectroradiometers, absorption spectrophotometers, and other instruments
where a monochromator is used to isolate a narrow wavelength region and hence,
in general, a low irradiance reaches the detector.
The disadvantages of photomultipliers are that they are fragile and require
an extremely stable high-voltage dynode power supply (less than 0.001 % voltage
deviation); both characteristics are drawbacks for a portable instrument. Photo-
multipliers also suffer (as do many detectors) from a gradual decrease in sensitiv-
 44 Chapter Three

ity due to photochemical alterations of the photocathode material. This error may
be minimized by preaging the tube with an irradiance sufficient to generate one-
third of the maximum rated photocurrent, for at least 100 hours. The tube then
exhibits a slow but more predictable decay in sensitivity.

Vacuum Photodiodes

Vacuum photodiodes are identical to photomultiplier tubes without the cas-

cade of dynodes that amplify the photoelectric current produced at the photo-
cathode. The tube simply consists of a photocathode and a single anode. Elec-
trons liberated from the photocathode via the photoelectric effect are accelerated
toward, and collected by, the anode. At a given wavelength, and over a wide
range of irradiance and current, the current across the diode is directly propor-
tional to the number of photons striking the photocathode (excellent linearity of
response). The lack of dynodes and hence an internal current amplification
mechanism necessitates use of a high-gain current amplifier in order to display
the signal; however, vacuum photodiode voltage requirements are less stringent,
and the device is more rugged than a photomultiplier because of its simpler inter-

UV-C
• UV-B UVA ____VISIBLE
-J.'-____ INFRARED
__________ ________

"'-- ,
,"
.....
,,
\
\
2 \
Cs- Te \
\
i
......
10.0
8.0
PHOTOCATHODE \ Sb-Cs
\PHOTOCATHODE
GaAs(Cs)
c
,
\ PHOTOCATHODE
E 8.0
\
> 4.0
~ Ag-O-Cs \
>
~ 2. PHOTOCATHODE'
in
z
1&1
I/)
1.0
.8
.8
.4

.2

0.1+--L-r--~-L-.r---~---r--~---.r---~--'---~--~
100 200 300 400 500 600 700 800 900 1000 1100 1200
WAVELENGTH (NANOMETERS)

Fig. 3-3. Typical spectral sensitivities of some commercially available photomultipliers.

Radiometry of Ultraviolet Radiation 45

nal construction. It is generally preferable to use a vacuum photodiode rather

than a photomultiplier except in cases of extremely low irradiance levels.
The same photocathode materials as are used in photomultipliers are
employed in vacuum photodiodes. Therefore, the spectral responses of available
vacuum photodiodes are similar to those in Fig. 3-3. Vacuum photodiodes (in
combination with calibrated filters and input optics) are often the detectors of
choice for a portable, versatile, and reasonably rugged ultraviolet radiometer sys-
tem. Several such systems are commercially available (see list of manufacturers
at the end of this chapter). The photocathodes of vacuum photodiodes age simi-
larly to those of photomultipliers, but if calibrated neutral-density filters are used
to limit the irradiance falling on the photocathode, excellent long-term stability
may be achieved.

Solid-State Photodiodes

When a conventional solid-state semiconductor junction (silicon or

selenium P-N diode) is exposed to radiant energy, a small voltage is developed
across the junction because of the generation of excess electron-hole pairs be-
tween the positive and negative junction materials. This photon-to-voltage con-
version is the basis for the solar-cell power supplies used on spacecraft. If a resis-
tor is placed across the terminals of the junction, a current is generated through
the resistor in proportion to the number of incident photons producing the volt-
age. This mode of operation of solid-state photodiodes is called the photo voltaic
mode (Fig. 3-4A).
If the junction is reverse-biased by applying a negative voltage on the anode
(P material) relative to the cathode (N material), only a small leakage current
passes in the dark. If radiant energy strikes the junction in this mode, a current
associated with the generation of excess electron-hole pairs across the junction is
produced. This mode of operation is called the photoconductive mode (Fig.
3-4B).
The spectral sensitivity of solid-state photodiodes depends on the material's
band-gap energy (the difference in energy between a material's conduction and
valence bands), similar in concept to the photoelectric work function of metals.
Both the photoconductive and the photovoltaic modes are utilized in
radiometry. Each mode has advantages and disadvantages. In the photoconduc-
tive mode, the device is both photosensitive and temperature-sensitive. As the
temperature increases, leakage currents become more of a problem. Temperature
stability in the photo voltaic mode is much better. However, the photoconductive
mode is more sensitive than the photo voltaic mode; photoconductive-mode sili-
con photodiodes are sometimes used in high-sensitivity radiometers for this
reason.
46 Chapter Three

Solid-state detectors, in ,general, are more rugged and exhibit much better
long-term stability than vacuum tube detectors. If UV irradiance levels on the
order of 5-10 mW/cm 2 impinge upon such a detector, there is a gradual deterio-
ration in sensitivity, especially for selenium detectors. Silicon photodiodes are
not as prone to changes in sensitivity because of the greater purity with which the
silicon can be manufactured. In general, the spectral sensitivity of UV -enhanced
silicon photodiodes extends from 200 nm through 1200 nm, with maximum sen-
sitivity in the near-infrared region, where the quantum efficiency (electrons of
current produced per photon absorbed) may reach 0.8 or greater (Fig. 3-5) when
used in the photoconductive mode. Use of silicon photodiodes for ultraviolet
radiometry demands excellent rejection of longer wavelengths by filters or
monochromators because of the weak ultraviolet spectral response of the detector
compared with that at longer wavelengths. Silicon photodiodes are useful for
UV-A irradiance measurements down to about 1O- 8W/cm 2 •
The stability, ruggedness, simple power supply requirements, and small
size and weight of solid-state photodiodes make them ideal detectors for portable
or very small instruments. UV-enhanced silicon photodiodes with a suitable
integrated-circuit amplifier are currently available at low cost in a single, small,
integrated package.

- . I (PHOTOCURRENTI
A
VOLTAGE OUTPUT, V V=JlEI=IR

R (RESISTORI PHOTOVOLTAIC MODE

PHOTOCONDUCTIVE MODE

lOUT =AI

Fig. 3-4. Operating modes for solid-state photodiodes. (A) Photovoltaic mode. (B) Photoconductive
mode.
Radiometry of Ultraviolet Radiation 47

0.70,....----------------------..,

~ 0.601--------------
--
~
~ 0.501-------------
>
~ 0.401-----------
z
w
en
w 0.301---------
I-
:::>
...J
o 0.201------
~ UV ENHANCED
« ~
/"
0.10 ---... _"

I I 1..1 I I I I
500 .. 600 '*'~~:-:t-""""'::-!:-:t-~~~'f7';;;!,

WAVELENGTH (nm)

Fig. 3-5. Sensitivity and spectral response of silicon photodiodes. (Modified from EG & G Applica-
tion Notes D3000B-l, 1976. EG & G, Salem, Massachusetts.)

Pyroelectric Detectors

Pyroelectric detectors are solid crystals that produce a minute current pro-
portional to the rate of change of the crystal's surface temperature. No current is
produced in the steady state. If modulated radiant energy strikes the crystal, ab-
sorption causes an increase in the surface temperature, and the current produced
is therefore proportional to the rate of change of the irradiance. This characteris-
tic is useful in rejecting ambient light and other noise when measuring pulsed
sources of radiation. However, pyroelectric detectors have the disadvantage of
requiring an optical chopper in order to detect sources of constant irradiance.
The surface temperature variations are related to changes in the absorbed
radiant energy. Coatings with strong, uniform spectral absorption are applied to
the surface of the detector, yielding a flat and wide spectral response characteris-
tic. Typical flatness of spectral response for pyroelectric detectors is about ±2%
from 300 nm to 20,000 nm. This wide and flat spectral response makes calibra-
tion accurate and simple and is the main desirable feature of pyroelectric detec-
tors.
High-gain, low-noise-current amplifiers are necessary to display the
nanoampere currents generated by pyroelectric detectors. This requirement, in
addition to the need for an optical chopper, makes pyroelectric radiometer sys-
48 Chapter Three

terns somewhat costly compared with systems using photodiodes or conventional

thermopiles. In general, pyroelectric detectors are most applicable to precision
infrared radiometry, but they can be used in the UV region if equipped with
quartz optical windows and calibrated filters for rejection of longer wavelengths.
Electrically-calibrated pyroelectric radiometers are gaining wide use as
radiometry standards.

Thermopiles

Conventional thermopiles use bimetallic thermocouple junctions wired in

series to produce a voltage corresponding to the temperature at the measuring
junctions. The thermopile is coated with a uniformly absorbing layer similar to
that used in pyroelectric detectors. The spectral response of thermopiles is simi-
larly wide and flat; however, sensitivity of thermopiles to changes in ambient
temperature induces some error. Several excellent thermopile radiometer systems
with ambient temperature-compensating circuitry and quartz windows are com-
mercially available. When using thermopiles or pyroelectric detectors to measure
the ultraviolet radiation present in broadband sources, one must completely
eliminate longer wavelengths by the filter or the monochromator. Unlike
pyroelectric detectors, thermopiles do not require an optical chopper, but they
may require frequent recalibration and must be allowed to reach thermal equilib-
rium with the environment for accurate irradiance measurements. Thermopiles
are generally not as sensitive as photodiode detectors, but they are ultimately
used to calibrate all other detectors because they are readily calibrated in absolute
terms.

Spectral Filters and Input Optics

In practice, detectors are placed behind calibrated ultraviolet-transmitting

filters (in radiometers) or monochromators (in spectroradiometers) to measure ul-
traviolet irradiance in a given spectral region. Other input optics may also be
necessary to give the filter/detector the desired angular (cosine-weighted) re-
sponse and field of view.

Filters

Solid-state ultraviolet filters can be classified into three basic types, each
with advantages and disadvantages. See Fig. 3-6 and Table 3-1 for filter ter-
minology.
Glass absorption filters are low-cost, easily obtainable filters made from
glass-melt formulations. They absorb the wavelengths not transmitted. Several
useful filters for UV-A isolation are available (e.g., Coming CS7-51, CS7-60,
Radiometry of Ultraviolet Radiation 49

and CS7-39 and Schott VG-I; Fig. 3-7A). Other useful glass filters are the so-
called sharp-cutoff Schott WG glass filter series (see Fig. 3-7B). These may be
used to essentiallY,eliminate ultraviolet radiation less than a given wavelength,
while passing longer wavelengths. Glass filters exposed to ultraviolet radiation
often show a gradual decrease in transmission and changes in spectral transmis-
sion. This phenomenon is known as solarization and may be minimized by limit-
ing the ultraviolet irradiance falling on the filter. Glass filters are commercially
available in sizes from 1 inch in diameter up to 2 ft square and from 1 to 10 mm
in thickness.
The interference filter passes a well-defined wavelength band. Typical
bandwidths of ultraviolet interference filters are from I to 20 nm. They are ex-
tremely helpful in blocking unwanted wavelengths. An interference filter uses
constructive and destructive interference to selectively transmit or reflect given
wavelengths. The filter consists of a substrate material (usually quartz or a
selected colored-glass filter for the VV) that is coated by vacuum evaporation and
condensation with various layers and thicknesses of dielectric materials or metal
films or both. The thickness of the individual layers is on the order of the
wavelength of light. Considerable control over central wavelength, bandwidth,
peak transmission, and transmission outside the pass band is gained by varying
the composition, thickness, and number of layers. This versatility is a major ad-
vantage because one can often produce an interference filter for some very
specific purpose, such as isolating an ultraviolet spectral line of mercury arcs.
Interference filters have been produced with VV bandwidths as low as 1 nm.
They typically exhibit a transmission outside the pass band of less than 10- 5 just
on either side of the central wavelength. This blocking can be further enhanced
by increasing the number of deposited layers on the substrate; blocking of better
than 10- 7 has been achieved in this manner. The multilayer coatings are usually

peak wavelength

c: I
o
'iii
.!!
I
!c: ~~----------~*-------------------~
...•..
~
half bandwidth

transmission outside of
lpsssband
.J I "_ _ "~
Wavelength. nm

Fig. 3-6. Filter tenninology for a band-pass filter.

50 Chapter Three

Table 3-1. Filter Terminology

Band-pass filter A filter that transmits a single discrete wavelength

Peak wavelength
Peak transmission
Half-power band width or full
width at half maximum
Narrow band-pass filter
Very narrow band-pass filter
Ultranarrow band-pass filter
Cutoff and cut-on wavelengths
Cutoff filter
Optical density
Spectral flatness
band.
The wavelength of maximum transmission of a
band-pass filter.
Maximum percentage of transmission in the pass
band.
The difference between those wavelengths for which
transmission equals half the peak transmission.
An interference filter having a half-power band-
width between 1% and 5% of the peak wavelength.
An interference filter having a half-power band-
width between 0.1 % and 1.0% of the peak wave-
length.
An interference filter having a half-power band-
width between 0.01 % and 0.1 % of the peak wave-
length.
The wavelengths at which transmittance equals
5% of peak.
A filter that transmits wavelengths longer than a
given spectral region with little or no transmission
of shorter wavelengths.
The logarithm of the ratio of incident intensity to
transmitted intensity; e.g., if incident intensity is
100 times the transmitted intensity,
100 100
O.D. = log - = 2.0; O.D. = log ( - )
1 %T
The transmission variation over a spectral wave-
length range expressed as a percentage.

deposited on some standard absorption-glass filter that provides blocking at

wavelengths far away from the central wavelength of transmission.
One of the major disadvantages of interference filters is high cost (typically
10-100 times that of conventional absorption-glass filters). They are also sensi-
tive to optical orientation and are most commonly used with the radiation inci-
dent at normal ± 10° to the surface of the filter. Long-term exposure to heat,
humidity, and intense UV irradiance causes a gradual deterioration of the filter
with a possible delamination of the dielectric coating, but, in general, interfer-
ence filters are rugged and stable.
Another useful filter in UV radiation experiments is the neutral-density
Radiometry of Ultraviolet Radiation 51

A 100

80 ,/ - ............
7-54 / 7-51/ \'
r 1\ ,\
~
z
w
u
II: 60 I
W
Q. II J \1\
Z
~ 40 ~ I r'\\
III
III I 7-60 I
~
III 20 I 1/
.\
z
< IT J '7 7-39 1\
II:
~ /) \~
0.10
I I,'", \ \~
I \ ' .... "
0.01 " I
200 300 400 500
WAVELENGTH - nm

WAVELENGTH.nm

Fig. 3-7. Spectral transmission of colored-glass filters useful for isolating UV-A. (A) Corning UV-
transmitting glass filters. (B) Schott "cutoff" glass filter series.
52 Chapter Three

filter. This is a filter that can be placed in the optical path to accurately attenuate
the radiation reaching the target material. Neutral-density filters attenuate all
wavelengths equally, thus, providing attenuation without affecting the relative
spectral irradiance of the incident radiation. Ideally, use of neutral-density filters
in radiometry allows a single factor of attenuation at all wavelengths. In practice,
it is advisable to calibrate neutral-density filters over a range of wavelengths and
with the specific detector or filter/detector combinations to be used. Neutral-
density filters are made in both thin-film-coated and absorption-glass construc-
tion. Good-quality thin-film types on quartz can attenuate from 260 to 1100 nm
with excellent neutrality. Neutral-density filters of stepwise optical densities
from 0.1 to 3.0 are available. Neutral-density interference filters also decay with
humidity, UV irradiance, and heat. They should be checked periodically for
transmission lind change of spectral attenuation. Fine wire screens are sometimes
used as inexpensive neutral density filters.

Input Optics

One may control the field of view of a detector/ filter combination by placing
an aperture some distance from the detector. In general, however, a 1800 field of
view and cosine-weighted response are desirable. These are generally achieved
using one or more ground quartz diffusers in front of the detector/filter, which
transmits a diffuse, approximately cosine-weighted irradiance. Figure 3-8A illus-
trates an arrangement of this type employed in International Light, Inc., radiome-
ter systems. Other types of cosine-weighted ultraviolet diffusers are also com-
mercially available.
Another common approach to achieving a cosine response is to employ an
integrating sphere, in which the radiation enters through a small entrance port
and the detector views some area of the interior of the sphere (Fig. 3-8B). The
inside of the integrating sphere is coated with a diffuse, highly ultraviolet-
reflective coating such as MgO or Eastman 6080 BaS04 powder paint. Multiple
reflections of the radiation within the sphere cause the detector to view an ir-
radiance proportional to the total radiance entering the sphere. A cosine-weighted
response is thus produced, because the radiance through a hole (the entrance
port) varies as the cosine of the angle of incidence. Integrating spheres are
somewhat bulky and the interior coating is sensitive to contamination; in portable
radiometers. a quartz or Teflon diffuser is more practical.
Spectroradiometers (detector/monochromator combinations) are indispens-
able for determining spectral irradiance. A curve of spectral irradiance versus
wavelength is determined by varying the wavelength transmitted by the mono-
chromator and recording irradiance as a function of wavelength. Portable spec-
Radiometry of Ultraviolet Radiation 53

A ROUGHLY GROUND HEMISPHERE

.........
............ ......
...... ----- ----
~--- --------- 0::::::.------ ----
~.",
.......:.,--------
ULTRAVIOLET
DETECTOR
, ............ ----------
........
" ,/'/ / L-~~~ ______________
............
............
~==~ ____________________________ ~

'" ,/
~/ QUARTZ DIFFUSER FILTER

B ULTRAVIOLET DETECTOR
INTEGRATING SPHERE
WITH INTERNAL DIFFUSE

... ...

"', ... ...

... ...
", "' ....... ~,
...

--- ---- ----

-----~--
"
-~----- -~------------
,',
'"
",
"I "
"- ... ... ... ...
... ...
...
,
""
"
\

" "\' /
,-
~---~,\' /

Fig. 3-8. UV radiometer input optics for cosine-weighted response. (A) With quartz diffuser. (B)
With integrating sphere.
54 Chapter Three

troradiometers are extremely versatile optical instruments. If a proper spectral

irradiance curve of a source is known, the irradiance over any spectral band is
easily calculated as the area under the curve in that wavelength region. Further-
more, reflection spectra or transmission spectra of materials may be obtained
with a spectroradiometer, a stable source, and known standards of transmittance
and reflectance.
The stray light characteristic of a monochromator is defined as the percen-
tage of energy transmitted through the monochromator at wavelengths other than
the wavelength for which the monochromator is set. The stray light response of a
spectroradiometer depends upon both the monochromator stray light characteris-
tic and the spectral response of the detector. The stray light response of ul-
traviolet spectroradiometers becomes extremely important when low UV spectral
irradiances must be measured in the presence of high irradiances at longer
wavelengths. For example, a 0.1 % stray light response produces gross errors in
the measurement of solar UV-B spectral irradiance.
Spectroradiometers employing ruled grating monochromators generally
have better stray light characteristics than those employing prism monochroma-
tors. Two monochromators are often employed to greatly reduce stray light, at
some expense in sensitivity. The recent development of laser-holographically
produced concave diffraction gratings has allowed for very low stray light levels
from a single monochromator. These monochromators are in general more stable
than ruled grating types because of the simplified optical design possible with a
concave grating, and they are becoming widely used in UV spectroradiometers
and other optical systems.
Spectroradiometers are somewhat more fragile than filter/detector instru-
ments because a photomultiplier is generally used and because optical alignment
of the monochromator is more critical than for a simple filter. Quartz diffusers or
integrating spheres may be used with spectroradiometers to produce a cosine-
weighted response.
Probably the most useful radiometer systems are those in which a single
high-gain, portable current amplifier with a large dynamic range may be used
with a variety of detector/ filter combinations or a spectroradiometer detector.
Special input optics, such as quartz fiber optic bundles or quartz micro-
scopic focusing optics, may be used to measUre ultraviolet spectral irradiances in
remote, extremely small, and/or hard to reach areas. It is also ppssible to use
double quartz fiber optic bundles, a stable source, and a spectroradiometer to
measure spectral transmission, reflection, or fluorescence of liquids, solids, or
gases in remote locations or of tissues in vivo and in situ. Microspec-
troradiometry, employing microscopic focusing optics in combination with spec-
troradiometers, may be used to examine optical properties of extremely small
samples, such as single cells or organelles.
Radiometry of Ultraviolet Radiation 55

Appendix: U.S. Manufacturers of UV -Related Instrumentation

Key: R = radiometer systems

D = detectors
S = ultraviolet sources
F = filters

D Clairex Electronics
Division of Clairex Corporation
560 South Third Avenue
Mount Vernon, New York 10550

F Corning Glass Works

Corning, New York 14830

S Christie Electric Corporation

3410 West 67th Street
P.O. Box 60020
Los Angeles, California 90060

D,S Ealing Corporation

Optics Division
2225 Massachusetts Avenue
Cambridge, Massachusetts 02140

R,D,S EGG, Inc.

Electro-optics Division
Salem, Massachusetts 01670

D EMI Photoelectric
P.O.Box 44
Princeton, New Jersey 08540

R,D Eppley Laboratory, Inc.

12 Sheffield A venue
Newport, Rhode Island 02841

R Gamma Scientific, Inc.

3777 Ruffin Road
San Diego, California 92123
56 Chapter Three

D Hamamatsu Corporation
120 Wood Avenue
Middlesex, New Jersey 08846

D Harshaw Chemical Co.

Crystal and Electronic Products Department
6801 Cochran Road
Solon, Ohio 44139

D H.E.!., Inc.
Jonathan Industrial Center
Chaska, Minnesota 55318

F Corion Corporation
73 Jeffrey Avenue
Holliston, Massachusetts 01746

F Heliotek, Inc.
12500 Gladstone Avenue
Sylmar, California 91342

R,D International Light, Inc.

Dexter Industrial Grf;en
Newburyport, Massachusetts 01950

D,S Laser Precision Corporation

5 West Whitesboro Street
Yorkville, New York 13495

D Northeast Associates
P.O. Box 31
Marblehead, Massachusetts 01945

F,D Optical Coating Laboratory

15251 East Don Julian Road
City of Industry, California 91746

S,R,F Oriel Corporation

1 Market Street
Stamford, Connecticut 06902
Radiometry of Ultraviolet Radiation 57

D Pacific Photometric Instruments

5745 Peladeau Street
Emeryville, California 94608

R Photo Research Corp.

3000 North Hollywood Way
Burbank, California 91502

D RCA
Electronics Components
Harrison, New Jersey 07029

S,R,F Schoeffel Instrument Corp.

24 Booker Street
/

Westwood, New Jersey 07675

F Schott Optical Glass, Inc.

York Avenue
Dury(~a, Pennsylvania 18642

D Sensor Technology, Inc.

210 12 Lassen Street
Chatsworth, California 91311

S GTE Sylvania
Danvers, Massachusetts 01923

R,D Tektronix, Inc.

P.O. Box 500 A
Beaverton, Oregon 97005

D United Detector Technology

1723 21 st Street
Santa Monica, California 90404

R Yellow Springs Instrument Co.

P.O. Box 279
Yellow Springs, Ohio 45387

A more complete directory of the optical industry is available from The Op-
tical Industry and Systems Directory, The Optical Publishing Company, Lenox
Road, Pittsfield, Massachusetts 01201.
CHAPTER 4

Optical Properties of the Skin and Eyes

Introduction

The first law of photochemistry (Grotthus-Draper law) states that no photochem-

ical (or subsequent photobiologic) reactions can occur unless radiation is ab-
sorbed. The biologic effects and microscopic alterations observed after absorp-
tion of UV radiation result from a complex sequence of biochemical reactions
and cellular responses, some of which may be distant from the site of absorption.
These chains of events must begin, however, with the excitation of important
molecules to reactive states by the absorption of photons of certain wavelengths.
The wavelengths producing an observable biologic effect (so-called action
spectrum) are theoretically those that are absorbed by a molecule or portion of a
molecule (chromophore) to initiate the photochemical events that lead to the ob-
served effect. The action spectrum may therefore yield information about the
chromophore in question, but account must be taken of the optical properties of
the tissue itself, especially the extent to which the wavelengths of interest are
absorbed by different layers or structures. The biologic action spectrum may then
be appropriately corrected f<;>r tissue optics. Unfortunately, the only case in
which optics may be rather simply modeled in human tissues is the transmission
of visible light through the eye to the retina. In practice, modeling the optical
properties of other tissues is an arduous task because of structural and optical
complexities.

Structure of the Skin

A thorough review of the anatomy of the skin and its structures is not within
the scope of this chapter (see references 1-4); however, an overview is presented
here. Figure 4-1 is a diagrammatic cross-section of the skin showing three major
tissue layers: the epidermis, the dermis, and the subcutaneous tissue. The thin,

59
60 Chapter Four

Opening of eccrine
swea t gland duct

Hair

Sebaceous glarlo "4ll.:~1!!.

Arrector pili
muscle

Blood vessel

Apocri ne
swea I gl. nd

Eccrine swea t gland

Fig. 4-1. Diagrammatic cross section of normal skin. (From Parrish, J. A. Dermatology and Skin
Care. Copyright McGraw-Hill, Inc ., 1975. Used with permission.)

outermost epidermis is composed of tightly packed cells called keratinocytes

which continually migrate outward from a germinative layer toward the surface.
In the process, they flatten , die, and cement together to form a very thin but
tough outer layer called the stratum corneum. The stratum corneum provides pro-
tection against water loss and surface abrasion and attenuates ultraviolet radiation
before it reaches living cells. The dermis is much thicker than the epidermis, has
fewer cells, and is mostly connective tissue, which provides the substance,
strength, and elasticity of the skin. Blood vessels, lymphatics, and nerves course
through the dermis. The deepest layer, the subcutaneous tissue, is mainly fatty
tissue and acts as an insulator and a shock absorber.
The thickness of skin_varies from one body region to another. Most of this
variation is accounted for by differences in the thickness of the dermis. The
Optical Properties of the Skin and Eyes 61

epidermis is relatively uniform in thickness over the body except on the palms
and the soles, where it is much thicker.

Epidermis

The epidermal keratinocytes are so named because they produce keratin, the
fibrous protective proteins ofthe skin. The exact structure of keratin is unknown.
Keratinocytes are derived from a single germinative layer of basal cells (Fig.
4-2). These stem cells divide, pushing some daughter cells outward; these daugh-
ter cells differentiate to form the prickle cells of the stratum Malpighii. In stan-
dard histologic preparations, these cells shrink during fixation and processing but
remain attached to each other by intercellular bridges called desmosomes. The
resulting appearance is a cell with "prickles," hence the name, although the cells
are not of this shape in vivo. As differentiation and outward migration continue,
the keratinocytes flatten, and granules appear within the cytoplasm, forming the
stratum granulosum. The exact nature of the functions of these granules is un-
known. The cells finally lose their nuclei, dehydrate, and flatten out into dead,
polygonal cells with a surface area about 25 times that of the basal cells. The
close packing and cementing of these flat, dead cells laden with keratin form the
tough stratum corneum. In normal skin, it may take up to 14 days for a daughter
cell of the basal layer to reach the stratum corneum and another 1-2 weeks before
it is sloughed off from the skin surface.
Specialized cells called melanocytes reside within the basal layer of the
epidermis and produce pigment granules containing melanin. Melanin, a com-
plex macromolecular protein derived from tyrosine, strongly absorbs light and

Epidermis ~~~illllb~~i~~~~i~iii~=~~Stratum
granulosum
Keratohyalin
granule
:;:.{(~---- Prickle
~~~~¥§.~~ cell

Basal cell
Dermis membrane
Blood vessel Fibroblast
fibers
Collagen

Fig. 4-2. Diagrammatic cross section of epidermis. This drawing is a magnification of part of Fig.
4-1. (From Parrish, J.A. Dermatology and Skin Care. Copyright McGraw-Hill, Inc., 1975. Used
with permission.)
62 Chapter Four

ultraviolet radiation. Dendritic processes of the melanocytes interdigitate bet-

ween keratinocytes and facilitate the transfer of pigment granules, called melano-
somes, into the keratinocytes. These melanosomes are carried outward within the
keratinocytes, and ultimately some melanin is deposited in the stratum corneum.
Racial differences in skin color are due to varying degrees of activity of melano-
cytes in producing melanin, not the number of melanocytes. Absorption of UV
radiation by melanin in the stratum corneum provides some protection against
actinic damage to the skin; the tendency to sunburn or to have certain skin can-
cers is inversely related to how much melanin is present. Increased production of
melanin is induced following sufficient sun exposure (tanning).

Dermis

The dermis is mostly a semisolid mixture of fibers, water, and a viscous gel
called ground substance. Ground substance consists largely of water and
mucopolysaccharides. There are three types of fibers present: collagen, re-
ticulum, and elastin. Collagen is a long molecule woven into fibrils that allow
stretching and contraction while maintaining tensile strength. Collagen makes up
about 70% of the dry weight of the dermis. The presence of finely branched
reticulum fibers probably serves to link bundles of collagen together. Elastin
fibers scattered through the dermis are highly elastic and presumably lend this
quality to the dermis as a whole. Scattered cells called fibroblasts produce the
fibers, proteins, and viscous materials of the dermis.
The uppermost dermal layer, the papillary dermis, contains an extensive
plexus of capillaries, lymphatics, and nerves. Scattered lymphocytes leave the
blood vessels of this layer, giving it a more cellular appearance than the underly-
ing reticular dermis. The reticular dermis is more fibrous, with fewer cells and
less ground substance, than the papillary dermis. The structure of skin is sum-
marized in Table 4-1.

Factors Affecting Penetration and Absorption of Ultraviolet

Radiation in the Skin

The complex structure of skin layers-and the presence of structures such

as hair follicles, sweat glands, and sebaceous glands-makes precise modeling
of the path of optical radiation within the tissue difficult. A schematic representa-
tion of the optical pathways of radiation in skin is shown in Fig. 4-3. Within any
given layer, four basic optical processes may occur:
1. Direct reflection at the boundar.ies of the layer due to change in index of
refraction (greatest at the air/stratum corneum interface)
Optical Properties of the Skin and Eyes 63

Table 4-1. Skin Layers

Typical
Layer thickness Basic structure

Stratum corneum 8-15 Mm Ten to twenty single-cell layers of densely packed,

flattened, dead keratinocytes, in remarkably
regular arrangement. Rich in the lipoprotein
keratin; contains melanin granules, carotenes.
Stratum granulosum 3 J.lm Granular cells between viable epidermis and stratum
corneum; two to four cell layers.
Stratum Malpighii 50-150 J.lm Ten to twenty cell layers of keratinocytes, which
produce the materials of the stratum corneum
as they differentiate and become flattened,
moving outward.
Germinative layer 5-10 J.lm Single-cell layer of columnar basal cells, which
divide to produce a continuing supply of
keratinocytes. Melanocytes also present, which
produce melanin pigment granules and transfer
these melanosomes into keratinocytes.
Dermis 1000-4000 J.lm Connective tissue composed of collagen, reticulum.
and elastin fibers and ground substance (gel).
Few cells compared to epidermis. Outermost
dermis (papillary dermis) contains many
capillaries, lymphatics, and nerves.

2. Scattering by molecules, particles, fibers, organelles, and cells within

the layer
3. Absorption (which may lead to photochemistry or dissipation of the ab-
sorbed energy via heat, fluorescence, or phosphorescence)
4. Direct transmission through the layer
In addition, it must be pointed out that because of the uneven surface
boundaries of layers in skin, the directly reflected and directly transmitted com-
ponents are diffuse. An analogy of this effect is the diffuse reflectance and trans-
mittance of a piece of ground glass as opposed to the specular reflectance and
transmittance of polished glass. When liquids of about the same index of refrac-
tion as the stratum corneum (nD-I.55) are present on the surface of the skin in
sufficient quantity to fill the many creases present, a more regular (planar) optical
interface is developed, and the directly reflected component is less diffuse. This
is why the skin may "shine" with profuse sweating. The directly transmitted
component is also less diffuse, 5 which may have some effect on the average path
length of the radiation that penetrates through the layer, and hence the biologic
response.
64 Chapter Four

INCIDENT RADIATION

DIRECT REFLECTANCE

EPIDERMAL REFLECTANCE
STRATUM CORNEUM
DERMAL REFLECTANCE
10-20)'

EPIDERMIS
40-150)'

DERMIS
1000-4000)'
DERMAL ABSORPTION

Fig. 4-3. Optical pathways of radiation within the skin.

The term direct is used here with transmission and reflection of a skin layer,
to separate these components from forward-scattered and back-scattered radia-
tion, respectively. Because measurements of the diffuse, direct reflection and
transmission of skin have not in practice been separated from back and forward
scatter, respectively, the general terms reflection and transmission are used here
to imply all radiation returning from or passin"g through a layer of skin. Direct
reflectance from the skin surface is due to the difference in refractive index of air
and stratum corneum, and is only about 5-10%, depending on the angle of inci-
dence. Total skin reflection in the visible spectrum may be as high as 75%. It is
important to remember, therefore, that what we call reflection from the skin is
largely radiation returning from within the turbid tissue.
Scattering is a non absorptive interaction of photons with matter in which the
direction of the radiation is altered by molecules or particles. When the particle
size is less than 1/100 of the wavelength (Rayleigh or molecular scattering), the
amount of scattering varies inversely as the fourth power of the wavelength. The
relationship between scattering and wavelength becomes less strongly inverse
when particle size and wavelength are of the same order of magnitude, and is
independent of wavelength for large particles (Mie scattering). More impor-
tantly, scattering is greatest when particle size and wavelength are on the same
order of magnitude. The spatial distribution of the scattered radiation relative to
the original path of radiation is also a function of wavelength and particle size,
mass, shape, and orientation.
The strong inverse relationship between molecular and small particle scat-
Optical Properties of the Skin and Eyes 65

tering and wavelength creates an effectively shallower depth of penetration for

shorter wavelengths in a scattering medium of this type than for longer
wavelengths. This is one mechanism accounting for the generally shallower
penetration of shorter-wavelength radiation in skin. Increased scattering also in-
creases the effective path length of radiation within a tissue layer. Because the
probability of a photon being absorbed in a medium is proportional to its path
length, scattering increases the absorption within a layer over that which would
be obtained if no scattering particles were present. This effect is similarly more
pronounced for shorter wavelengths.
In addition to the degree of scattering, the presence of optically absorbing
molecules (pigments) greatly affects the penetration of different wavelengths into
the skin. Transmission through a layer of tissue is less for wavelengths that are
absorbed by pigments within the layer. Therefore, excised skin layers show
spectral transmittance minima in regions where pigments within the sample show
absorbance maxima. Similarly, because what we have called reflection from skin
is largely from within the tissue, the absorption maxima of certain skin pigments
may be identified as minima in the spectral reflectance of skin in vivo. The con-
centration and location of skin pigments that absorb in the visible region largely
determine our perceptions of skin color.
Many organic molecules show some absorption of ultraviolet radiation. The
penetration and reflection of UV wavelengths are largely determined by the con-
centration of melanin in the epidermis, although other ultraviolet-absorbing pig-
ments may be discerned from ultraviolet spectral reflectance and transmittance
measurements. Furthermore, one may calculate the spectral absorption within an
excised skin layer by subtracting the sum of total spectral reflectance and total
spectral transmittance curves from 100%,6,7 but some difference from this calcu-
lated value occurs in vivo because of optical interactions between skin layers.
Because of the optical complexities of skin, a quantitative theoretical model
of the penetration and reflection of optical radiation in skin is lacking. However,
Atkins 8 has presented an interesting discussion of modified Kubelka-Munk dif-
fuse flux theory relevant to this problem. Many of the data necessary to develop a
rigorous quantitative model are lacking.

Measurements of the Penetration and Reflection of Optical

Radiation in Skin

Stratum Corneum and Epidermis

Many studies of the transmission of ultraviolet radiation through excised

human epidermis and stratum corneum have been reported since the original
work of Hasselbalch. 9 Much of the early work, however, failed to account accu-
rately for the diffuse nature of transmission through skin samples, largely be-
66 Chapter Four

cause of inadequate instrumentation. 9 - 13 Quantitative measurements of the diffuse

spectral transmission of skin samples must be made by collecting all the transmit-
ted radiation or by integrating measurements taken at many angles over a com-
plete hemisphere (goniometric measurements, see Fig. 4-4). Goniometric mea-
surements also yield precise information about the angular distribution of diffuse
transmission and reflection. Unfortunately, the only goniometric study of human
skin available 7 examined transmission of wavelengths 550 to 2400 nm but did
not examine ultraviolet wavelengths. However, in those studies that compare dif-
fuse versus direct (off-axis versus in-line optical path) transmission of epidermis,
the ratio of diffuse/direct transmissions does not appear to be wavelength depen-
dent. 6.13.14 This lack of dependence on wavelength suggests that the diffuse na-
ture of epidermal transmission of ultraviolet wavelengths is due more to irregular
refractive and reflective surfaces and large particle scattering than to small parti-
cle or molecular scattering within the epidermis. However, the nature and impor-
tance of scattering within epidermis remains to be determined.
Everett et al. 6 employed a Cary 15 double-beam spectrophotometer with an
integrating sphere to measure total diffuse spectral transmission and reflection of
Negro and Caucasian excised epidermis- and stratum corneum. The authors also

A.-_

INCIDENfr
RADIATION ~-­
IMONOCHROMATlC)

B DETECTOR

INCIDENT
RADIATION-------K"'""
IMONOCHROMATIC)

INTEGRATING SPHERE~
Optical Properties of the Skin and Eyes 67

c INCIDENT
RADIATION
(MONOCHROMATIC)

SKIN

PHOTOGRAPHIC PLATE

INCIDENT ,......._--, DETECTOR

RADIATION =1 I
(MONOCHROMATIC) SKIN

INCIDENT
RADIATION -------"F/--.~""re:::::-_~J SKIN SPECTROGRAPH OR
(NOT MONOCHROMATIC) SPECTRORADIOMETER

INCIDENT DETECTOR
RADIATlON--------t~
(MONOCHROMATIC)

Fig. 4-4. Spectroscopic methods employed for measurement of spectral transmittance of skin.
Methods A, B, and C measure total transmittance; methods D, E, and F fail to measure total transmit-
tance. (A) Goniometric spectrophotometer. (B) Integrating sphere spectrophotometer. (C) Contact
photographic plate. (D) In-line absorption spectrophotometer. (E) Spectrograph or spectroradiome-
ter. (F) Collecting lens.
68 Chapter Four

compared the ultraviolet spectral transmission of specimens obtained with a vari-

ety of separation techniques and found them to be remarkably similar. The total
diffuse transmission spectra of Caucasian stratum corneum samples appear in Fig
4-5. Transmission of Caucasian and Negro whole epidermis samples appear in
Fig. 4-6. Earlier studies of ultraviolet spectral transmission through skin show
general qualitative agreement with these data; the values for transmittance, how-
ever, are somewhat lower in many earlier studies, as would be expected when
only some portion of the diffuse transmittance is measured.
The data of Everett et al. 6 indicate that about 50% of incident UV -A radia-
tion is transmitted through the Caucasian stratum corneum to the viable cells of
the epidermis. The data also indicate that 35 to 50% of incident UV-A is trans-
mitted through Caucasian epidermis samples. The epidermis samples used in this
study did not include the basal cell layer. However, it is assumed that optical
extinction in the pigmented basal cell layer will not be much greater than for the
rest of the viable epidermis. It is therefore reasonable that UV -A exposure may
exert direct photobiologic effects on both epidermal (keratinocytes, melanocytes,
basal cells) and dermal (blood within capillaries, lymphocytes, nerves, fibro-
blasts) cells and structures. One might also hypothesize from these data that
UV -A would directly produce histologic changes in skin at greater depths than
would UV-B or UV-C radiation.
To demonstrate the significant epidermal transmission of wavelengths below
300 nm, Everett et al. exposed a volunteer to monochromatic radiation at 254,
280, and 297 nm with and without a specimen of excised epidermis in place over
the exposure area. The observed minimal delayed erythema dose with the speci-
men acting as a filter, as compared with control (no specimen) exposures, corres-
ponded well with that predicted by the transmission data of the specimen at each
wavelength. This offered some confirmation of the accuracy of their spectral
transmission data, done essentially by their use of the living human as an ul-
traviolet radiometer.
The minimum in spectral transmittance near 280 nm of stratum corneum
(Fig. 4-5) is due to absorption by small amounts of aromatic amino acid residues
in keratin, which is highly concentrated in the stratum corneum. This absorption
band has recently been shown to. shift toward longer wavelengths during matura-
tion of newborn rat skin, presumably as a consequence of some maturational
change in keratin of the stratum corneum. 15 This minimum of stratum corneum
transmittance at 280 nm is most likely the cause of a corresponding minimum in
the human ultraviolet erythema action spectrum. 16 No distinct minima or max-
ima of transmittance or reflectance are seen in the UV-A, but both UV-A trans-
mittance and reflectance are decreased with increasing melanin content. Thus,
while up to half of incident UV-A radiation may reach the basal cell layer in
whites, generally 5 to 15% reaches the same layer in Negro skin.
Optical Properties of the Skin and Eyes 69

A 100'
6 90'
in
en 80 ---,.. ..-. ,- :-. " ..-. :-.
:" :". :". :"
--------
..-. :-. :". :-. :". :"..": .-..-. ~................ .
~ •. ;..::.,; ....;_ioI
~ 70 .,.
.;
~ 60 I
I-
...J 50 ,:I
« ,:
,:
5l- 40 ,,:
I- 30
z
~ 20
CC
~ 10

25~0~50"""400 450 560 5~0 660 6~0 700

WAVELENGTH

B 100
80
60
z 40
0
in
en
~ 20
en
z
« 10
_ ----- -----
CC

,-- --
I- 8 ....

I-
z 6 ,
UJ ,
u 4
CC ,,
UJ ,,
0..
, ............. ---,'
2 ,,•
,,
"
250 300 350 400
WAVELENGTH

Fig. 4-5. (A) Stratum corneum: total transmission. Specimen 9 (sunburn)' •• ; specimen 10
(cantharidin)·- - - - - ; specimen II (cantharidin) - - . (B) Comparison of direct and total trans-
mission measurements for stratum corneum. Total transmission - - - ; direct
transmission - - - - - . (From Everett, M. A., Yeargers, E., Sayre, R. M., and Olson, R. L. Penetra-
tion of epidermis by ultraviolet rays. Photochem. Photobiol. 5 :533-542, 1966. Copyright, Pergamon
Press, 1966. Used with permission.)
70 Chapter Four

A
z 100
oen 90
en 80
:2:
~ 70
«
a: 60
I-
~ 50
bl- 40
I- 30
z
~ 20
ffi 10 .- .. -
0..
, ,
300 350 400 450 500 550 600 650 700
WAVELENGTH

B
100
80
60

z 40
0
en
en 20
:2:
en
z
« 10
a: 8
l-
I- 6
z
w , ,
u 4
,,,
a:
w , ,
0.. ,
2
,,,
,
,,
.
1~
250 300 "
350 400
WAVELENGTH

Fig. 4-6. (A) Intact epidermis: total transmission. Specimen I (stretch-heat) ° specimen 5 (water
0 0 ;

vacuum) - - - - - ; specimen I3 (Negro; water vacuum) _00_; specimen 14 (water

vacuum) - - - ; specimen 15 (stretch) _0_. (B) Comparison of direct and total transmission
measurements for whole epidermis. Total transmission - - - ; direct transmission - - - - - . (From
Everett, M. A. Yeargers, E., Sayre, R. M., and Olson, R. L. Penetration of epidermis by ultraviolet
rays. Photochem. Photobiol. 5:533-542, 1966. Copyright, Pergamon Press, 1966. Used with per-
mission.)
Optical Properties of the Skin and Eyes 71

Total spectral reflectance of excised stratum corneum, epidermis, whole

skin, and skin in vivo are given in Fig. 4-7. Comparison of the spectral reflec-
tance of whole skin with that of excised epidermis alone suggests that for
wavelengths shorter than 350 nm, the reflectance of skin is largely radiation re-
turning from within the epidermis. For wavelengths longer than 350 nm, reflec-
tance has an increasingly large dermal component throughout the visible light
region (400-700 nm).
The spectral absorption within excised layers may be calculated roughly by
subtracting the sum of spectral transmission and spectral reflectance from 100%.
Calculated spectral absorption curves of excised Caucasian stratum corneum and
epidermis are shown in Fig. 4-8; the absorption curve for whole Caucasian epi-
dermis presented in this figure agrees well with that determined much earlier by
Lucas. 5 The protective role of absorption within the stratum corneum is readily
apparent from the figure. This optical barrier function can be intentionally in-
creased through application of UV-absorbing sunscreens (see Chapter 11). The
optical barrier function can also be decreased for UV-B wavelengths by pro-
longed application of optically non-absorbing, aqueous media. 1 7 The slightly in-
creased sensitivity of skin to UV -B exposures following prolonged application of
water correlates quantitatively with slight increases in epidermal transmittance. 1 7

100

z 90
0 80
l-
e.>
w 70 ...... _.. _.. _.. -
-l
u.. 60 .- .....
w
II: 50
I-
Z 40
w
e.> 30 .... .......
II:
w
0- 2
10

250 300 350 400 450 SOO 550 600 650 700
WAVELENGTH

Fig. 4-7. Light reflected by skin. In vivo, dorsum hand, Caucasian - - ; in vitro, specimen 15
epidermis, S.Q. fat _00- ; specimen 15 epidermis - - - - - ; specimen II stratum corneum _0- ;

specimen 13 Negro epidermis, dermis, S.Q. fat ° 0 0 specimen 13 Negro epidermis __

; 0_- .

(From Everett, M. A., Yeargers, E., Sayre, R. M., and Olson, R. L. Penetration of epidermis by
ultraviolet rays. Photochem. Photobiol. 5:533-542, 1966. Copyright, Pergamon Press, 1966. Used
with permission.)
72 Chapter Four

100

90

80
..
'g

.a 70
0
'"tD
.a
60
C
.g 50
•
'g
:! 40
~
tD
30

I
~
u
c
'0 20 STRATUM CORNEUM (SAMPLE 111
11-
10

0
240 300 400 500
WAVELENGTH - nm

Fig. 4-8. Ultraviolet spectral absorption of Caucasian epidermis and stratum corneum (Calculated
from Everett, M. A., Yeargers, E., Sayre, R. M., and Olson, R. L. Penetration of epidermis by
ultraviolet rays. Photochem. Photobiol. 5:533-542, 1966.)

Dermis

Very little information is available on the ultraviolet optical properties of the

dermis, although several studies of spectral reflectance and transmission of ex-
cised dermis (washed to remove blood) have been made in the visible and in-
frared spectral regions. 7•18 The effective extinction coefficient (absorbance per
unit thickness) of the dermis is less than that of the epidermis in the visible re-
gion. 7 The dermal spectral reflectance data of Findlay 1 8 indicate an increase from
about 40% (at 400 nm) to 70% (at 700 nm) diffuse reflectance over the visible
spectrum, and these data also indicate that shorter wavelengths are effectively
scattered from shallower depths within the tissue. The visible region spectral
reflectance of various animal connective tissues resembles that of pure collagen
powder. 1 8 The qualitative picture that emerges for the visible optical properties
of the dermis is one of a relatively nonabsorbing but turbid, light-scattering tis-
sue. This would imply that UV wavelengths penetrate the dermis less deeply than
visible wavelengths.
The visible and near infrared goniometric studies of Hardy et ai. 7 tend to
support the qualitative picture of a high degree of scattering within the dermis.
For a given thickness of skin, including the upper dermis, scattering is more
complete, and the transmission less, at shorter wavelengths. The transmission of
Optical Properties of the Skin and Eyes 73

550 nm, the shortest wavelength measured (green visible), through a lo8-mm
thickness of dermis was about 0.5%. Near-infrared radiation between 700 and
1300 nm is highly penetrating compared to visible wavelengths,1·14.19.20 which in
tum are more penetrating than UV-A wavelengths. Infrared wavelengths greater
than about 3000 nm are absorbed by water and other molecules and, therefore, do
not penetrate as deeply as 700-3000 nm infrared radiation. Short-wavelength ul-
traviolet radiation does not penetrate signi ficantly through the 1-4 mm thick
human dermis; however, there may be some small transmission of UV-A
wavelengths to the subcutaneous tissues.
The penetration of optical radiation through Caucasian skin is summarized
diagrammatically in Fig. 4-9.

The Effects of Pigments

Characteristic spectral absorption bands of melanin, blood (oxygenated and

reduced hemoglobin), carotenes, water, and aromatic amino acid residues in
keratin may be identified with reflectance minima in the spectral reflectance
curves of living human skin19223 (Fig. 4-10). Melanin (epidermal) and blood
within the capillaries of the papillary dermis are the most important pigments that
may affect UV-A penetration into the skin, depending on their concentrations.
Carotenes present in epidermis and fatty tissue also influence UV -A penetration.
The epidermis may be viewed as a system that produces a protective pro-
duct, the stratum corneum. Part of this protection is the increased production of
melanin by melanocytes in the basal layer following exposure to ultraviolet radia-
tion. Thickening of the epidermis also results from increased cell proliferation
after UV exposure and plays some protective role. The melanocytes transfer
melanin granules (melanosomes) into the cells of the epidermis, which carry this
pigment until they are sloughed off as dead stratum corneum cells from the sur-
face. Although some melanosomes and melanin are degraded by the time a
keratinocyte reaches the stratum corneum, there may be a considerable concen-
tration of melanin granules in the stratum corneum. 2 4 The production of quantities
of melanin, the stability of one's melanosomes within keratinocytes, and the rela-
tive increase in melanin production following exposure (primarily UV-B radia-
tion) are largely genetically determined. However, if UV-A-photosensitizing
compounds, such as psoralens, are present, marked tanning and changes in
melanosome structure may follow relatively small UV-A exposure doses. 25
There is a large variation in melanin concentration between sun-exposed and
normally covered skin areas and between individuals. Melanin has a significant
effect in attenuating the epidermal transmission of radiation between 250 nm and
700 nm (Figs. 4-6A and 4-10), with greater attenuation at shorter wavelengths.
The degree of melanin pigmentation largely determines the penetration of all
wavelengths of ultraviolet radiation into skin. 6.7.22.24.26
 74 Chapter Four

(J)
:?!:?!
::::>::::> :?! C/)
I-W a:
«z
a: a:
w :?!
Cl a:
1-0 a.. w
(J)U w Cl

o
Ez
e:w
OW
It) a:
It)(!)

o
o

0
0
E« a:
e: I I-
0> >
~::::> z
0
0 z
0
I-
«
a:
I-
w
0 Z
w
a..
Ern l-
e: I z
0> w
g::::> U
a:
0 w
...
0 a..

EU
e: I
0>
~::::>
o
...
o
j I
o It) o
o o
wW'H.ld3a NI)IS
 700nm 1230nm 2200nm
DEEP RED N.I.R. IR.
I --. T ---.-, 'tI
Tl I ~ r I
Io
0.0
'i
....... STRATUM i-
s.
CORNEUM :f
CD
VI
:~
.... EPIDERMIS :ij'
III
::I
a.
E
E f
:c- "':\MI~
- II ~,
.....
0.. 0.5_
w
c DERMIS
z
~
(f)

- I

1.0 _
j r:{m.t.~"~""~[l
Fig. 4·9. General diagrammatic
summary of the penetration of radia.
100 o 100 o 100 o tion of different wavelengths into
Caucasian skin. Ui

PERCENT PENETRATION (IN VITRO)

 76 Chapter Four

HEMOGLOBIN
II)
!::
z
::J
w
U
z BETA-CAROTENE
«
CD
a:
o
II)
CD
«
w
~
«
~

....I
\AI
a:

200 300 400 500 600 700 800 900 1000 1100 1200
WAVELENGTH.nm

Fig. 4-10. Absorption spectra of skin pigments that absorb in the UV -A region. (After Edwards, E.,
and Duntley, S. Q. Spectrophotometry of living human skin, the ultraviolet range. 1. Invest. Der-
matol. 16:311, 1951.)

Although little is known about the penetration of UV-A into the dermis,
presumably the presence of blood exerts a considerable absorbing effect. The
facts that about half of incident UV"A radiation reaches the dermis of fair-
skinned Caucasians and that blood shows considerable absorption in the UV-A
region (Fig. 4-10) indicate that significant UV -A radiation is absorbed directly by
blood in the capillaries of the papillary dermis. This absorption may be of great
importance if, in the presence ofUV-A-photosensitizing compounds in the blood
or high UV-A exposure doses, the cells or plasma components of blood become
altered. Normally innocuous doses of solar radiation may damage cutaneous
blood vessels if endogenous photo sensitizers are present. 2 7 Absorption of UV-A
and visible light by the blood or by cells of noncutaneous origin present in the
dermis may also be of benefit in phototherapy and photochemotherapy aimed at
the treatment of certain systemic metabolic 28 or malignant 29 disease states. An
understanding of the factors affecting penetration of UV-A and other wave-
lengths through overlying skin layers is of obvious importance in the administra-
tion of such treatments.
The optical properties of the living skin are a summation of the properties of
Optical Properties of the Skin and Eyes 77

many living cell and tissue layers. For instance, over the UV -A and visible spect-
ral region, the cells of the epidermis are exposed to both incident radiation and
radiation reflected by the dermis. It should also be realized that the data presented
above are largely of nonliving excised tissues. In addition to gross absorption and
penetration of radiation through tissue, one must also consider each cell as a liv-
ing optical element. Many of the lethal and mutagenic effects of ultraviolet radia-
tion on cells have been associated with absorption of radiation by DNA or DNA
complexes with other molecules in the cell nucleus. In this sense, the absorption
of a given dose of radiation in a cell's nucleus may be of greater biologic impor-
tance than a similar exposure of the cytoplasm. For example, if cells are rounded
during UV -C exposure so that the nucleus is somewhat shielded by a thickness of
cytoplasm, less cell lethality occurs. It has been found in cultured human liver
cells that the cytoplasmic thickness for 50% optical extinction ("half-value
layer") at 254 nm is approximately 5.5 /Lm. 30 Although such data are lacking for
keratinocytes or other wavelengths, the intracellular distribution of melanin pig-
ment and the gradual flattening of these cells may affect the viability or response
of keratinocytes after UV exposure. In addition, those cells nearer the skin sur-
face may be exposed to greater incident radiation levels. Similar arguments may
apply to the scattered cells of the dermis. Exposures delivered to circulating ery-
throcytes, leukocytes, and lymphocytes also depend upon their transit time
through a region of exposure.

Summary of UV-A OptiCS of the Skin

Roughly 35-50% of incident UV-A radiation penetrates to the dermis of

Caucasians. This penetration may be modified by the concentration and distribu-
tion of melanin, carotenes, and topically applied substances. Increased epidermal
thickness and melanin production are induced following UV exposure and de-
crease the amount of UV radiation penetrating the epidermis. Although little is
known of the ultraviolet optical properties of dermis, it is clear that U V-A
reaches and is absorbed by blood and other constituents of the papillary dermis in
significant amounts. Other reviews of cutaneous optical properties are found in
references 31-33.

Ultraviolet Optics for the Eye

It is usual to consider the image-formation qualities of the eye when consid-

ering its optics; however, absorption, transmission, and reflection of ultraviolet
radiation are stressed in this section. The type and extent of damage to ocular
tissue depend on the energy absorbed, the wavelength of the radiation, and the
duration of the exposure. Because the ocular tissues affected by UV exposures
78 Chapter Four

are generally those in which the radiation is absorbed, the morphology of the eye
is also reviewed here.
Figure 4-11 presents a cross section of the human eye with the gross struc-
tures of the eye identified. Figure 4-12 provides a schematic representation of the
absorption characteristics of the eye for electromagnetic radiation ranging in
wavelength from gamma rays to microwaves. Gamma rays and X-rays of
wavelengths shorter than 0.01 nm mainly pass through the eye, but damage may
occur through primary and secondary ionization caused by the small fraction of
radiation that is absorbed. Soft X-rays and ultraviolet radiation in the 0.01-310
nm wavelength range are absorbed primarily by the cornea of the eye.
While ultraviolet radiation below 310 nm is absorbed principally by the
cornea, UV-A radiation is absorbed by the cornea and the lens, with the lens
absorbing more of the radiation at wavelengths approaching 400 nm. Some of the
ultraviolet radiation near 400 nm may reach the retina because there is some
transmission through the human lens at these wavelengths. Visible light (400-
700 nm) is refracted by the ocular media and forms the retinal image from which
the organism gains sensory information. Near-infrared radiation of wavelengths
from 740 nm to about 1300 nm is also focused on and absorbed by the retina but
does not stimulate vision. Middle- and far-infrared radiation (1300 nm to 100
/Lm) is absorbed by the cornea or the intraocular media before reaching the re-
tina. Microwave and radio frequency radiation is transmitted by the eye; how-
ever, certain wavelengths in the 1-10 cm wavelength range (microwave) are ab-
sorbed by the crystalline lens and are thought to pose a cataract hazard.
The biologic effects of wavelengths shorter than 310 nm are seen predomin-
antly in the corneal tissue. Kinsey 34 and Bachem 35 published ultraviolet trans-

OPTIC NERVE

CORNEA

Fig. 4-11. Structures of the human eye.

Optical Properties of the Skin and Eyes 79

GAMMA 6 X-RADIATION SHORT ULTRAVIOLET

LONG ULTRAVIOLET 6 NEAR INFRARED

VISIBLE

FAR INFRARED MICROMVES

Fig. 4-12. Generalized characteristics of the eye for electromagnetic radiation.

mission data for the rabbit cornea. Boettner and Wolter 36 provided ultraviolet
transmission data for the human cornea. A comparison of the rabbit and human
corneal transmittance of ultraviolet radiant energy is shown in Fig. 4-13. In each
instance, the data were obtained on instruments not specifically designed for
far-ultraviolet research and are therefore less reliable at shorter UV wavelengths.
The rabbit and human total corneal transmittance curves from Bachem and from
Boettner and Wolter, respectively, compare favorably at all wavelengths but dif-
fer materially from the total corneal transmittance found by Kinsey for
wavelengths longer than 310 nm. Nevertheless, the data confirm that little ul-
traviolet radiation shorter than 310 nm is transmitted through the cornea and that
most of this absorption occurs in the corneal epithelium. A comparison of the
human and the rabbit corneai spectral transmittance is shown in Fig. 4-14 for the
270 to 400 nm waveband.
These data clearly demonstrate that most of the ultraviolet radiation between
320 and 400 nm (UV-A) that enters the pupil is absorbed within the crystalline
 80 Chapter Four

lens of the eye. Therefore, the lens in particular may be subject to biologic effects
caused by UV-A radiation. The lens may also act as a protective UV-A-
absorbing filter for the retina. While most of the UV -A entering the eye is ab-
sorbed by the lens, the data of Boettner and Wolter 36 clearly show an 8-10%
transmission band centered at 320 nm of young human and rhesus monkey
lenses. Interestingly, this UV -A transmission band is reduced to less than 0.1 %
by age 22 in the human 36 which may be because of photochemical alterations or
maturation of the lens. Persons with aphakia can readily perceive UV-A radia-
tion; there is no apparent evidence, however, that those with aphakia or young
humans suffer retinal damage from normal environmental levels of UV -A.
When exposing the eye to optical radiations, it is difficult to determine the
total incident and absorbed radiant energy because of losses due to reflection,
largely at the surface of the cornea. These losses vary with the relative indices of
refraction of the media and with the angle of incidence. For large angles of inci-
dence, such as the marginal rays incident on the curved surface of the cornea,
reflectance may be more than 50% (see Fig. 4-13). The irradiance impinging on
the cornea is also reduced in direct proportion to the cosine of the angle of inci-
dence (see Fig. 3-1A). Because of these variations, quantitative experimental ex-
posures are generally confined to collimated irradiation at normal incidence to the
central region of the cornea. The central 4-mm-diameter area of the cornea re-

10

0.9

0.8
..J
~ 07
U
I&J
00.6
Z

~ 0.5
Z
t!
.... 0.4
~
~ 0.3
cS:
a::
.... 0.2

0.1

0.0 ........- ........-..-----.--Si

~~~~m~~~~~~~~~m*~~

WAVELENGTH IN NANOMETERS

Fig. 4-13. Transmission of ultraviolet radiant energy by the cornea. 0 = rabbit corneal epithelium
(after Kinsey 34); x = rabbit cornea (after Kinsey 34);. 0 = rabbit total cornea (after Bachem 3'); • =
human total cornea (after Boettner and Wolter 30 ).
Optical Properties of the Skin and Eyes 81

1.0

O.

<i
0.8

_0---
-0--
--<>
.- AI
'"
-- -
:I!
- y '"
07
U .0---
ILl
...- ..- ...-

--
C ..-~
0.6
~ ?"""
I ..r-
ILl 1
~ 0.5 ..-
~
~
0.4 1
/r
/
~ 1 1
rn 0.3 / 1
Z
« / 1
II:: / /
~ 02
11

----
1I
01 6:]1
1
0.0
210 280 290 300 310 320 330 340 350 360 310 380 390 400

WAVELENGTH IN NANOMETERS
Fig. 4-14. Comparison of rabbit and human transmittance of ultraviolet through the cornea, aqueous
humor, crystalline lens, and incident on the vitreous. 0 = incident on human aqueous; x = incident
on human lens; • = incident on human vitreous; D = incident on rabbit aqueous; ~ = incident on
rabbit lens; V = incident on rabbit vitreous. Most of the wavelengths below 300 nm are absorbed by
the cornea, while the wavelengths between 310 nm and 390 nm are absorbed by the lens. After Boett-
ner and Wolter 3 ., Kinsey", and Bachem".)

ceives more than 90% of such incident radiation. However, biologic effects are
certainly not limited to this central area when the eye is exposed to ultraviolet
radiation.

References

I. Parrish, J. A. Dermatology and Skin Care. McGraw-Hill, New York, 1975.

2. Montagna, W., and Parakkal, P. F. The Structure and Function o/Skin, 3rd ed. Academic Press,
New York, 1974.
3. Montagna, W., and Lobitz, W. The Epidermis. Academic Press, New York, 1964.
4. Fitzpatrick, T. B., Arndt, K. A., Clark, W. H., Jr., Eisen, A. Z., Van Scott, E. J., and Vau-
ghan, J. H. (Eds.). Dermatology in General Medicine. McGraw-Hill, New York, 1971.
5. Lucas, N. S. The permeability of human epidermis to ultraviolet radiation. Biochem. 1.25:57-
70, 1930.
6. Everett, M. A., Yeargers, E., Sayre, R. M., and Olson, R. L. Penetration of epidermis by ul-
traviolet rays. Photochem. Photobiol. 5:533-542, 1966.
7. Hardy, J. D., Hammell, H. T., and Murgatroyd, D. Spectral transmittance and reflectance of
excised human skin. 1. Appl. Physiol. 9:257-264, 1956.
82 Chapter Four

8. Atkins, J. T. Optical properties of turbid materials. In The Biologic Effects of Ultraviolet Radia-
tion (with Emphasis on the Skin) (F. Urbach, Ed.). Pergamon Press, Oxford, 1969, pp. 141-150.
9. Hasselbalch, K. A. Quantitative Untersuchungen iiber die Absorption der menschlichen Haut
von ultravioletten Strahlen. Skandinav. Arch. F. Physiol. 25:5-68, 1911.
10. Bachem, A. The ultraviolet transparency of the various layers of human skin. Am. J. Physiol.
91 :58-64, 1929.
II. Macht, D. I., Anderson, W. T., and Bell, F. K. The penetration of ultraviolet rays into live
animal tissues. J.A.M.A. 90:161-165, 1928.
12. Bachem, A., and Reed, C. I. The penetration of ultraviolet light through the human skin. Arch.
Phys. Therapy 11:49-56, 1930.
13. Kirby-Smith, J. S., Blum, H. F., and Grady, H. G. Penetration of ultraviolet radiation into skin
as a factor in carcinogenesis. J. Natl. Cane. Inst. 2:403-412, 1942.
14. Bachem, A., and Reed, C. I. The transparency oflive and dead animal tissue to ultraviolet light.
Am. J. Physiol. 90:600-606, 1929.
15. Rosencwaig A., and Pines, E. A photoacoustic study of newborn rat stratum corneum. Biochim.
Biophys. Acta 493:10-23, 1977.
16. Mitchell, J. S. The origin of the erythema curve and the pharmacological action of ultraviolet
radiation. Proc. R. Soc. Bioi. 126:241-246, 1938.
17. White, H. A. D., Anderson, R. R., Blank, I. H., and Parrish, I. A. Enhancement of transmission
of optical radiation through human epidermis by topical applications. Sixth Annual Meeting of
the American Society of Photobiologists, Vermont, June 1978, p. 68. (abstract).
18. Findlay, G. H. Blue skin. Br. J. Dermatol. 83: 127-134, 1970.
19. Jacquez, 1. A., Kuppenheim, H. F., Dimitroff, J. M., McKeehan, W., and Huss, J. Spectral
reflectance of human skin in the region 235-700 1IlJL. J. Appl. Physiol. 8:212-214, 1956.
20. Jacquez, J. A., Huss, J., McKeehan, W., Dimitroff, J. M., and Kuppenheim, H. F. Spectral
reflectance of human skin in the region 0.7-2.6 /L. J. Appl. Physiol. 8:297-299, 1956.
21. Goldzieher, J. W., Roberts. I. S., Rawls, W. B., and Goldzieher, M. A. "Chemical" analysis
of the intact skin by reflectance spectrophotometry. Arch. Dermatol. Syphilol. 64:533-548,
1951.
22. Edwards, E. A., and Duntley, S. Q. The pigments and color of human skin. Am. J. Anat. 65:1-
33, 1939.
23. Edwards, E. A., Finkelstein, N. A., and Duntley, S. Q. Spectrophotometry ofliving human skin
in ultraviolet range.J. invest. Dermatol. 16:311-321, 1951.
24. Kligman, A. Comments on the stratum corneum. In The Biologic Effects of Ultraviolet Radiation
(with Emphasis on the Skin). (F. Urbach, Ed.). Pergamon Press, Oxford, 1969, pp. 165-167.
25. Toda, K., Pathak, M. A., Parrish, J. A., Fitzpatrick, T. B., and Quevedo, W. C., Jr. Alteration
of racial differences in melanosome distribution in human epidermis after exposure to ultraviolet.
Nature [New BioI.1236:143-145, 1972.
26. Thomson, M. L. The relative efficiency of pigment and horny layer thickness in protecting the
skin of Europeans and Africans against solar ultraviolet radiation. J. Physiol. 127:236-246,
1955.
27. Mathews-Roth, M. M., Pathak, M. A., Fitzpatrick, T. B., Harber, L. C., and Kass, E. H. Beta
carotene as a photoprotective agent in erythropoietic protoporphyria. N. Engl. J. Med.
282:1231-1234, 1970.
28. Odell, G., Schaffer, R., and Simopoulos, A. (Eds.). Phototherapy in the Newborn, an Over-
view. National Academy of Sciences, Washington, D.C., 1974.
29. Gilchrest, B. A., Parrish, J. A., Tanenbaum, L., Haynes, H. A., and Fitzpatrick, T. B. Oral
methoxsalen photochemotherapy of mycosis fungoides. Cancer 38:683-689, 1976.
30. Williams, J. R. The survival of cultured human cells irradiated with ultraviolet light. Ph.D.
thesis, Harvard University, 1972.
Optical Properties of the-Skin and Eyes 83

31. Scheuplein, R. J. A survey of some fundamental aspects of the absorption and reflection of light
by tissue. J. Soc. Cosmet. Chern. 15:111-122, 1964.
32. Treagar, R. T. Physical Functions of the Skin. Academic Press, New York, 1966, pp. 96-107.
33. Magnus, I. A. Dermatological Photobiology. Blackwell Scientific, Oxford, 1976, pp. 11-22:
34. Kinsey, V. E. Spectral transmission of the eye to ultraviolet radiations. Arch. Ophthalmol.
39:508-513, 1948.
35. Bachem, A. Ophthalmic ultraviolet action spectrum. Am. J. Ophthalmol. 41 :969-975, 1956.
36. Boettner, E. A., and Wolter, J. R. Transmission of the ocular media. Invest. Ophthalmol.
1:776-783,1962.
CHAPTER 5

Effects of Ultraviolet Radiation on

Microorganisms and Animal Cells

Introduction

The photobiologic reactions of multicellular organisms depend upon what hap-

pens to individual living cells within their organ systems, which in tum is deter-
mined by individual intracellular photochemical reactions. Ultraviolet radiation
may inactivate enzymes in numerous biologic systems by producing alterations
in proteins. Peptide bonds may be split, photochemical oxidations may occur,
sulfide and disulfide bonds may be altered, and DNA may be photochemically
changed. In order for this photochemistry to occur, radiation must be absorbed,
providing the molecular activation energy to initiate photochemical reactions. All
absorption of radiant energy, however, does not lead to photochemistry, because
of competition by other mechanisms for dissipating the absorbed energy, such as
fluorescence, phosphorescence, or nonradiative deexcitation.
Absorption spectra and photochemical properties are specific for a given
molecule. A photon may be absorbed only when its energy corresponds to the
energy required for an allowed transition between quantized energy states of the
molecule. In the ultraviolet and visible wavelengths, the photon energies (E = hv
= he/>..; Table 5-1) are in the range associated with transitions of molecular elec-
trons into excited electronic states. Chemical bonding depends on the sharing be-
tween atoms of electrons in a given electronic energy state, and absorption of
ultraviolet radiation may therefore lead to the formation or breakage of chemical
bonds, that is, photochemical reactions. At longer wavelengths (infrared radia-
tion), the lower photon energies correspond to molecular rotational and lower
vibrational state transitions, expressed macroscopically as increased temperature.
Bimolecular chemical reactions are, in general, temperature-dependent, and ab-

85
86 Chapter Five

Table 5-1. The Energy Associated with Quanta of Visible and Ultraviolet Wavelengths

Color Wavelength (nm) Energy in kcal/mole

200 143
UV~l
UV.c
UV-B Invisible
250
280
114
102
UV-B 300 95
UV-A 360 79
UV -A/violet 400 72
Violet 420 68
Blue 470 60
Green 530 54
Yellow 600 47
Orange 630 45
Red 700 41

sorption of infrared radiation may therefore induce increased chemical reactions.

Many enzymes are heat-labile. It is important, however, to note that the
mechanism of infrared-induced chemical or biologic changes is not via electroni-
cally excited states, as is often the case in the ultraviolet region, but rather via
thermal effects.
Moreover, absorption of a given amount of radiant energy of any
wavelength causes nearly the same amount of heating. Thus, the biologic effects
of UV -A, for example, may involve both thermal and photochemical
mechanisms. Experimentally, one can discern these mechanisms distinctly.
Thermal effects are often immediate, and are always dependent upon the ir-
radiance of the source as well as the radiant exposure dose. Photochemical effects
generally exhibit reciprocity (for a given radiant exposure dose, the reaction is
independent of irradiance) over a wide range of irradiance. Furthermore, ther-
mally induced biologic effects generally show broad action spectra that relate to
the production of heat within the tissue and can be correlated with the spectral
absorbance of the tissue. On the other hand, the action spectra of photochemi-
cally induced biologic effects are usually more sharply defined and may corre-
spond to the absorption spectra of particular chromophores within the tissue. For
a basic review of the principles of photochemistry, see references 1-3.
There are several mechanisms of deexcitation "available to" a molecule in
an excited electronic state after absorption of a UV photon. In general, in an ex-
cited singlet state, a molecule has acquired additional energy through absorption,
yet still retains its original electronic spin. This state is short-lived (about 10- 9
sec), and the excited singlet-state molecule may return to the ground state
through emission of a photon ifluorescence) or through nonradiative deexcita-
Effects on Microorganisms and Animal Cells 87

tion. Molecules in a singlet excited state can also undergo internal chemical al-
teration or react with other molecules in the system. The extremely short lifetime
of the excited singlet state, however, may limit bimolecular photochemistry at
physiologic temperatures because of the comparatively long time required for
molecules to collide.
If, in addition to excitation, a change occurs in the excited electron's spin,
the excited state may become metastable, because the excited electron cannot
form an electron "pair" within its ground-state energy level until its spin is again
reversed. This excited metastable state is called the triplet state and may last as
long as several seconds. Radiative deexcitation of the excited triplet state results
in phosphorescence. The longer lifetime of the excited triplet state compared to
that of the excited singlet state accounts for the longer decay time observed for
phosphorescence than for fluorescence. Triplet-state excitation can also lead to
photochemical reactions and, because of the longer lifetime of the excited triplet
state than that of the excited singlet state, the triplet state may be more likely to
undergo allowed bimolecular photochemical reactions. Absorption of a UV
photon generally leads to an excited singlet state, which mayor may not then
undergo intersystem crossing (an electron spin reversal) to an excited triplet
state.
For photochemical reactions to occur, the energy of the photons absorbed
must be greater than or nearly equal to the activation energy required for the reac-
tion. That is, even in strongly exothermic photochemical reactions in which the
reaction products have lower chemical potential energy than the reactants, the
reaction may be dependent upon activation of one of the reactants to a higher
energy than that of its ground state. It is this activation energy that is supplied by
an absorbed photon. Many biologic photochemical reactions require activation
energies of 40-100 kcal/mole, which is why, with single photon absorption
processes, ultraviolet radiation is effective in causing photobiologic effects,
while visible radiation is less so, and infrared radiation essentially not at all (see
Table 5-1). As one approaches the more biologically active wavelengths of
UV-B and UV-C, the energy per photon increases to 100 kcal/mole or more.
The majority of phototoxicity reactions mediated via exogenous photosen-
sitizers in humans are activated by UV -A radiation. In this case, an exogenous
agent may absorb UV-A and initiate photochemical reactions, or it may act as a
UV-A sensitizer by complexing with another molecule (for example, DNA) in
such a way as to increase UV-A absorption or lower the activation energy re-
quired for a given photochemical alteration. Sensitization may also occur via
reactive photochemically produced intermediates formed in the presence of the
sensitizer. The intermediates, such as singlet oxygen, peroxide and other radi-
cals, or metastable excited-state molecules, then attack and alter important
molecules or organelles.
88 Chapter Five

Effect of Ultraviolet on Cells

Much of what we know about how radiation-induced molecular changes af-

fect living cells comes from research on microorganisms. Low doses of unfiltered
solar radiation kill a variety of organisms. 4.5 At the earth's surface, the most le-
thal component of sunlight appears to be UV_B. 6 • 7 In the UV-C and UV-B,
DNA is strongly implicated as the chromophore for many of the observed effects
of UV radiation on bacterial and mammalian cells, 8 and inactivation spectra for a
wide variety of cells correspond relatively closely to the absorption spectrum of
DNA.
It has been known for many years that the maximal bactericidal effective-
ness of ultraviolet radiation is in the UV -C region. The absorption spectra of
many bacteri-:> have a peak near 280 nm, where aromatic amino acid residues of
their proteins have maximum absorption. The bactericidal action spectrum in
most experimental conditions corresponds to the absorption spectrum of the nu-
cleic acids DNA and RNA, both peaking at about 260 nm. It has been concluded
that the primary event in UV -C and UV -B inactivation or killing of bacteria re-
sults from photochemical alteration of the cellular DNA. Furthermore, in many
experimental circumstances, DNA replication is inhibited by lower doses of ul-
traviolet radiation than is RNA synthesis or protein synthesis. For this reason,
research has been concentrated primarily on the effects of UV absorption by nu-
cleic acids and proteins occurring at wavelengths shorter than 300 nm.
In vitro and in vivo, in both prokaryotic and eukaryotic cells, one major
molecular consequence of exposing DNA to ultraviolet radiation is the formation
of pyrimidine dimers. Thymine dimers have received the most attention, perhaps
because DNA is easily labeled with rlH] thymine, and the dimer is an acid-stable
DNA photoproduct that may be easily isolated. In some systems, however, other
dimers, such as cytosine-cytosine and cytosine-thymine, make up the largest per-
centage of dimers. 9 Other DNA photoproducts are known to occur, including
other dimers, hydrated pyrimidines, and cross-links between two strands of
DNA, as well as between DNA and protein. 1 () DNA strand breakage, local dis-
ruption of hydrogen bonds, and changes in RNA also occur when cells are ir-
radiated. Changes in RNA similar to those in DNA certainly occur and may be
more numerous events. However, changes in RNA are not as evident or as easy
to measure as those in DNA, because multiple copies of each species of RNA
may exist within a given cell, whereas one, two, or, at most, a few copies of a
DNA sequence may be present in bacteria.
Cell protein structure may also be altered by UV irradiation. Such protein
changes would usually have to be extensive and numerous to affect the cell as
severely as do changes in DNA. However, many enzyme activities can be shown
to be decreased following UV irradiation. Proteins may be primarily affected and
enzymes may be either activated or inactivated. The effects on proteins may re-
Effects on Microorganisms and Animal Cells 89

sult from photochemical alterations of the individual amino acids of which the
proteins are composed. Decarboxylations, deaminations, and ring breakage oc-
cur. Among the more sensitive targets are the aromatic amino acids, especially
tryptophan. The disulfide bonds of cystine can be broken by ultraviolet radiation.
Ultraviolet-induced changes in macromolecular synthesis may have sig-
nificant effects on cellular metabolism. At radiation doses that have little or no
effect on oxygen consumption, there may be derangements in the synthetic
metabolism of the cell. These derangements affect DNA synthesis, RNA synthe-
sis, and therefore protein and enzyme synthesis. The eventual changes in the cell
may therefore be profound.
Because of DNA's essential role as the template for synthesis of RNA,
which in tum codes for protein synthesis, many of these other processes induced
by ultraviolet radiation are less important. Functional enzymes, however, are
necessary for the repair of damaged DNA. The effects of ultraviolet on animal
cells are complicated, and hundreds of UV-induced changes have been noted.
The effects usually mediated by wavelengths shorter than 320 nm have been
summarized by Giese 11 and Painter. 12

DNA Repair

Biologically important amounts of solar UV radiation reach the earth's sur-

face and have done so since the beginning of evolution. Mechanisms evolved
very early in biological time to protect cells and to aid in the recovery from the
damaging effect of photons.
In recent years, three major survival mechanisms have been described:
I. The damaged molecule or part of a molecule can be restored to its func-
tional state in situ. This restoration is accomplished either by enzymatic
mechanisms or by "decay" of the damage to some innocuous form.
2. The damaged part can be removed and replaced with undamaged mate-
rial to restore normal structure and sequence.
3. The damage may remain unrepaired, but during replication the cell may
be able temporarily to bypass or ignore the damage.
The type of repair as well as the extent of recovery depends on the nature of
the molecule that has been damaged. Because of the importance of the sequence
of events necessary for normal replication and function of DNA molecules, effec-
tive repair of damaged DNA must usually be completed within some narrowly
defined period of time before cell division or normal function may resume. A
number of different DNA repair mechanisms have been described to date, and
others are being discovered with increasing frequency as new methods for pro-
duction and analysis of repair are studied. An excellent review of known repair
90 Chapter Five

mechanisms can be found in Smith and Hanawalt I 3 and in the proceedings of a

symposium on molecular and cellular repair processes. 14 Only the most
thoroughly described processes are summarized in this text.

Photoreactivation (Fig. 5-1A)

Enzyme-catalyzed photoreactivation is a form of in situ repair in which ex-

posure to radiant energy facilitates the direct repair of UV-induced thymine di-
mers. It has been clearly shown that most nonmammalian celis contain a photoac-
tivated enzyme system that monomerizes pyrimidine dimers, thus restoring a
normal DNA strand in situ. The photoreactivating enzyme (PRE) binds spec-
ifically to the UV-induced pyrimidine dimers to form a complex that is stable in
the dark. If this complex is then exposed to radiation of wavelengths approxi-
mately 330 to 600 nm, energy transport occurs through the protein and causes a
separation into the active enzyme and a repaired DNA segment that can no longer
bind to the enzyme. Illuminating the enzyme or the damaged DNA prior to com-
plex formation has no effect on UV damage repair. IS,I6
The photoreactivation mechanism is of greatest importance for the survival
of certain microorganisms as well as for some plants and small animals (such as
insects) in the field, accounting for the capability of these organisms to survive in
tropical and mountainous areas. A photoreactivating enzyme has been isolated
from human leukocytes by Sutherland. I 7 Molecular weight of this protein is ap-
proximately 40,000, and the action spectrum is broad, extending through the
UV-A and up to 600 nm. The exact properties, media, requirements, and spec-
ificity of human PRE are not known and appear to be different from the ubiqui-
tous PRE of bacteria. The light-mediated monomerization of thymine dimers by
this human enzyme may be an incidental property of an enzyme whose major
function is not understood. Compounds of appropriate electronic properties could
absorb energy and, by electronic excitation transfer, monomerize nearby
pyrimidine dimers. Therefore, the exact role of PRE in higher organisms and in
the internal viscera of vertebrates is not clear.

Excision Repair (Fig. 5-18)

The studies of Setlow 18 provided the first experimental evidence leading to a

model for repair of UV damage in the dark. A repair mechanism was postulated
in which defective regions in one of the two DNA strands could be excised and
then subsequently replaced with normal nucleotides utilizing the complementary
base-pairing information in the intact strand. This mechanism, also known as
cut-and-patch repair, is of widespread significance for the repair of a variety of
structural defects of DNA.
Effects on Microorganisms and Animal Cells 91

Excision repair involves the following 13:

1. Recognition. This system is capable of recognizing a variety of struc-
tural defects in DNA, including those that do not involve pyrimidines
and those not due to UV effects (usually caused by alkylating agents,
etc.). The exact nature of the recognition mechanism is not known.
2. Incision. Following recognition of DNA damage, a single-strand break
near the damage point must be produced. This is a specific endo-
nuclease-mediated process.
3. Excision and resynthesis (repair replication, unscheduled DNA synthe-
sis, UDS). Enzymatic excision of a segment containing damaged nu-
cleotides and opposite strand-dependent synthesis may occur as sepa-
rate steps or concurrently.
4. Rejoining. Completion of the repair process requires rejoining of the
repaired segment to the continuous DNA strand. A polynucleotide
ligase is most likely the enzyme responsible for this step.
Evidence for excision repair mechanisms has been found in microor-
ganisms, and mammalian cells. This repair system responds to a variety of chem-
ical and radiation-induced DNA alterations. 19 Altered DNA nucleotides are
eliminated and apparently normal cell replication can follow.

Postreplication "Repair" (Fig. 5-1C)

The observation by Howard-Flanders 20 that double mutant strains of Es-

cherichia coli, deficient in both excision repair of DNA and recombination of
genetic loci, were more sensitive to UV than either of the single mutant strains
alone, suggested the existence of a DNA repair system, other than excision re-
pair, that might involve enzymes active in recombination. It is postulated that
some of the mechanisms and enzymes of genetic recombination are utilized in
these repair systems, especially in those "repairs" not completed until during or
after cell replication.
Newly synthesized segments of DNA in UV -irradiated cells are initially
smaller than segments synthesized in nonirradiated cells and gradually elongate
to control sizes. It is not clear how replication proceeds with UV damage in the
parental DNA, but it is assumed that gaps are left in the daughter strand opposite
damaged sites. At later times, some mechanism permits DNA synthesis in these
gaps. This gap-filling process has been termedpostreplication repair. 21,22 Since
the presence of a single pyrimidine dimer in a bacterial genome can be a lethal
condition if repair does not remove it, DNA polymerases probably cannot use the
damaged region as a template. Hence, synthesis of nucleotides in gaps opposite
such dimers during replication presents a special problem to the cell, since sub-
92 Chapter Five

------~,

PRE
c;:::]
I hY

-----/
1I

PRE (?
m

B
:-@'"'
I. Recognition
,"01.
II. Excision

III. Excision
..nJ
P'---7'-
IV. Degradation

,
V. Repair replication
~
..,

VI. Rejoining

Alternate Steps
III'. Repair replication -~
~
IV'. Excision

;0.. ___ -

V'. Degradation
Effects on Microorganisms and Animal Cells 93

C
a

b ~
~

--_._-,.

c 'III;
-
•
,
~ • • -
• ~ w
-
d
"'" ~
~.
~~

Fig. 5-1. (A) Schematic representation of photorepair mech"nism of UV -A-damaged DNA. I: UV-
induced pyrimidine dimer. II: Photoreactivation enzyme (F RE) complexes with dimer and absorbs
UV-A or visible photon. III: Cleavage of dimer (repair) and release of PRE. (B) Schematic represen-
tation of the postulated steps in the excision repair of damaged DNA. Steps 1- VI illustrate the cut-
and-patch sequence. An initial incision in the damaged strand is followed by local degradation before
the resynthesis of the region has begun. In the alternative patch-and-cut model, the resynthesis Step
III begins immediately after the incision Step II, and the excision of the damaged region occurs when
repair replication is complete. In either model, the final step (VI) involves a rejoining of the repaired
section to the contiguous DNA of the original parental strand. (From Smith, K. c., and Hanawalt,
P. C. Molecular Photobiology. Copyright, Academic Press, Inc., New York, 1969. U,sed with per-
mission.) (C) A model for postreplication repair of UV-damaged DNA. (a): Dots indicate radiation
lesions produced in DNA. (b): DNA synthesis proceeds past the lesions in the parental strands, leav-
ing gaps in the daughter strands. (c): Filling of the gaps in the daughter strands with material from the
parental strands by a recombinational process. (d): Repair of the gaps in the parental strands by repair
replication. (From Smith, K. C. The roles of genetic recombination and DNA polymerase in the re-
pair of damaged DNA. In Photuphysiology, Vol. 6, A. C. Giese, Ed. Copyright, Academic Press,
1969. Used with pennission.)
94 Chapter Five

sequent excision repair of residual damage requires a correct template. If postrep-

lication repair plays an active role in promoting survival of a cell line, it must be
assumed that the filling of "gaps" left opposite damaged DNA sites during repli-
cation must be an accurate process. That is, the nucleotide sequence inserted into
the gap opposite the pyrimidine dimers must be the correct one. Because the cor-
rect nucleotide sequence is present in the homologous sister duplex, speculations
on the mechanism of postreplication repair have postulated processes involving
interchange between the two sister helices.
At present, several possibilities for gap filling have been studied:
I. Filling by repair synthesis without exchange (which has been proposed
as a mechanism for UV mutagenesis but does not provide for accurate
repair)
2. Filling by sister exchange involving insertion of a newly synthesized
strand formed on a template of the sister duplex
3. Filling by a sister strand exchange involving preexisting DNA
Studies of radiosensitive bacterial mutants indicate that at least five genetic
loci are associated with gap filling in newly synthesized DNA. The details of the
mechanisms, however, for restoring altered DNA during and after replication are
unclear, both in bacteria and in mammalian cells. Some of these processes may
result in errors in nucleotide sequence and subsequently may lead to mutations
and possibly to carcinogenesis. While direct DNA damage by UV-A occurs to
some extent, an indirect damage via photoproducts formed from such naturally
existing chromophores as tryptophan 23 may involve other repair systems.
In bacterial systems, some enzymes involved in repair processes appear to
be induced by ultraviolet light. An inducible repair system, sometimes referred to
as SOS repair, initially postulated by Radman, 24 is thought to result either in the
direct induction of errors in nucleotide sequence or in a reduction in the fidelity of
all DNA synthetic systems, those involved both in repair and in normal replica-
tion.
All of the DNA repair mechanisms discussed above have been identified and
characterized in bacteria through the technique of UV -sensitive mutant strain
selection. Research into the DNA repair mechanisms in human cells and their
role in the repair of UV-induced damage has been greatly advanced by the iden-
tification of human beings whose cells are not only UV-C-sensitive but also
exhibit defects in DNA repair processes. Originally described by Cleaver, 25 cul-
tured cells from UV-sensitive, cancer-prone humans with xeroderma pigmen-
tosum show reduced levels of excision repair after UV-C irradiation. More
recently, it has been shown that cells from various individual patients can be
classified according to six complementation groups and a variant group, indicat-
ing that this DNA repair process may involve at least seven gene products. 26
Effects on Microorganisms and Animal Cells 95

Cells from three other human genetic conditions in which patients exhibit
higher cancer incidence have been shown to be sensitive to physical and chemical
agents, and reduced repair has been suggested. These are ataxia telangiectasia, 2 7
deletion-type retinoblastoma, 28 and Fanconi's anemia. 29 Cells from patients with
the first two conditions are sensitive to ionizing radiation, and cells from patients
with the third condition are sensitive to the chemical cross-linking agent,
mitomycin-C. Repair enzyme deficiencies in these diseases are reviewed in detail
elsewhere. 30

Effects of UV-A

Many of the experimental data evaluating the effects of UV -A on single-cell

or viral systems are difficult to interpret because of the wide variety of UV-A
sources used. This interpretation is further complicated by the fact that UV -A-
induced changes in cell function require as much as 10,000 times more energy
than those induced by UV-C or UV-B. Sources of UV-A that emit as much as a
fraction of a percent of their output as UV-B and UV-C radiation may markedly
alter experimental results. A similar problem exists for sources of UV-B or
UV-C that emit significant UV-A or visible radiation.
Another source of confusion in the literature is the frequent use of broad-
spectrum UV -A light sources that are rich in visible light. There are numerous
examples of inhibition of growth and respiration by visible light and UV-A in
prokaryotic and eukaryotic organisms. 31 Unfortunately, in these experiments,
possible UV -A effects are not separated from the widespread blue light
phenomena found in many one-celled organisms and lower plants and animals.
Finally, many of the published experiments do not utilize monochromatic light
sources. Therefore, interpretation of the data is difficult, because it is known that
UV-A can induce enzymes, alter enzyme function, and induce or inhibit repair
systems.
When radiation is limited to wavelengths longer than those at which DNA
maximally absorbs, photochemical reactions other than those occurring in DNA
may assume greater relative biologic importance. Although 1O:l_10 4 times more
energy may be required than at 260 nm, it is possible to kill bacteria with
longwave ultraviolet radiation or with visible light. UV -A and visible lethality
are very often partially oxygen-dependent and show survival curves that suggest
a greater multiplicity of targets than those of shortwave UV radiation. DNA is a
target molecule of UV -A, but it is not always clear in anyone experimental sys-
tem whether UV-A-induced alteration of DNA is direct (via minuscule but
measurable DNA absorption of UV-A), or indirect (via another chromophore,
such as the proteins surrounding DNA or an endogenous sensitizer such as tryp-
96 Chapter Five

tophan). It has been suggested that quinones, flavin, pyridine, porphyrins,

4-thiouracils of tRNA, pyridine nucleotides, or other molecules act as primary
chromophores in different specific UV-A-induced reactions.
Hollaender 32 first compared the effects of UV-A and UV-C radiation on
bacteria in 1943. Using E. coli, he found that 104 _10 5 times more energy of
UV -A (primarily 365 nm) was required to have the same killing effect as the
shorter wavelengths. Survival curves showed much larger shoulders, and the
temperature coefficient was twice that seen in the shorter-wavelength region. The
UV-A-irradiated cells became sensitive to physiologic saline immediately after
irradiation. He thought it likely that UV -A produced a toxic substance but also
considered the possibilities that UV -A destroyed some compound essential for
survival and multiplication of bacterial cells or that UV -A inactivation of bacteria
was a nor.<;pecific physiologic process. In 1946, Luckiesh confirmed that high
radiant exposures of UV-A and visible light could kill bacteria in the absence of
added photosensitizers. 33
Recently, expanded awareness and concern of biologic effects of UV -A has
led to a rapid accumulation of information about the effects of UV -A on microor-
ganisms, mainly E. coli. The recent interest in UV-A was preceded by the estab-
lishment of some of the basic knowledge of DNA repair systems and the de-
velopment of new techniques to induce, isolate, purify, and assay mutant cell
lines. Information is growing so rapidly that it is difficult to place it in proper
perspective relative to the data base of the better-known effects of UV -B and
UV-C on microorganisms. In general, chromophores for UV-A action are not
known. The lethal and mutagenic effects of UV -A radiation on microorganisms
have been thoroughly summarized by Webb. 34 Many of the exogenous photosen-
sitizing agents that affect microorganisms have action spectra in the UV-A range.
These effects have recently been summarized by several investigators. 35-38
UV -A lethality has been confirmed on a variety of bacterial strains that dif-
fer somewhat in survival curve shape, action spectrum, and dose require-
ments. 39- 41 Lethal effects of UV -A are strongly enhanced by oxygen in
carotenoid-deficient Sarcina lutea 42.43 and in recombination-deficient strains of
Bacillus subtilis, Salmonella typhimurium, andE. coli. 44-48 Control studies and
experience in other laboratories showed that this oxygen-dependent lethality is
clearly different from the relatively oxygen-independent inactivation by UV-C
and may suggest intracellular photodynamic processes with as yet unrecognized
endogenous photosensitizers. Certain repair-proficient strains of E. coli are more
sensitive to UV_C 49 - 51 or to UV_A52.53 during exponential growth than during
stationary phase. The relative increase in sensitivity during exponential phase is
much greater for UV-A than for UV-C. 53
DNA may act as the target molecule for UV -A lethality by way of direct
absorption of photons, by photosensitization via some other molecule or com-
plex, or by way of secondary damage or lack of repair by UV-A-induced
Effects on Microorganisms and Animal Cells 97

metabolic changes within the cell. It is certain that UV-A radiation results in al-
tered DNA that is susceptible to repair, but the relative importance of the variety
of UV-A-induced DNA or DNA repair system lesions is a matter of debate, and
depends largely on the system studied. UV-A can induce cyclobutane-type
pyrimidine dimers in bacterial DNA in vivo and in vitro, 54 that can be repaired by
photoreactivating enzyme. 55 The ratio of dimer yield at 365 nm to that at 254 nm
was 7.1 X 10 5, somewhat greater than ratios for the radiant exposure yielding
equal lethality at these wavelengths,55 and suggesting that damage other than
pyrimidine dimers is important. UV-A also produces oxygen-dependent single-
strand breaks or alkali-labile bonds in bacterial DNA, 56 and in intact and ex-
tracted phage DNA. 56 Single strand breaks (or alkali-labile bonds) appear to be
the most likely DNA lesion accounting for the strong oxygen dependence of
UV-A lethality of E. coli. 34.57 The chromophore for oxygen-dependent DNA le-
sions caused by UV-A may be a nucleoprotein. 58
UV -A irradiation of bacteria can induce several transient cellular effects at
exposures of one-tenth the lethal dose. One of the most general and sensitive re-
sponses of cells to UV-A is a 1-2 hr growth delay. According to Jagger, 59
UV-A-induced growth delay in E. coli, inhibition of capacity to support phage,
and inhibition of induction of tryptophanase all require the reI gene product,
which provides a stringent control of RNA synthesis by the cell. These effects
appear to be related to a U V-A -induced tRN A photo product. 59 The chromophore
strongly implicated is the 4-thiouracil of tRNAs. 60
Nonspecific membrane damage has also been reported in several organisms
after radiation with greater-than-lethal doses of UV-A or visible light. 32.61,62
UV -A and visible radiation in exposure doses causing little inactivation of
repair-proficient strains can, however, damage transport and metabolic systems
in bacteria and yeast. 59,63- 65 In microorganism systems, two separate leucine
transport systems can be inhibited 66 and respiration can be inhibited 67,68 by
365 nm radiation that does not kill the cell. Relatively low-irradiance UV-A also
inhibits induced formation of galactosidase 69 and tryptophanase. 70 In some bac-
teria, RNA synthesis may be more sensitive to UV-A irradiation than protein or
DNA synthesis. 71
DNA repair capability is an important factor in UV-A lethality in E.
coli, 39,40 and repair inhibitors have been shown to increase sensitivity of E. coli
to 365 nm radiation. 39,53 Elucidating the mechanisms for UV -A-induced lethal-
ity is complicated by the fact that UV-A can both cause DNA damage and inhibit
the function of DNA repair systems. 44,50,55,57,72 Both excision repair and recom-
bination repair systems are impaired by UV-A radiation. 72 UV-A radiation can
also inhibit the repair of :!1NA single-strand breaks induced by X-radiation of E.
coli. 57 The relative roles ofthese effects in producing cell lethality is explored by
Webb. 34
UV -A effects on microorganisms are further complicated by the fact that
98 Chapter Five

while the action spectrum for photoreactivation often extends from the shorter-
wavelength visible spectrum into the longwave ultraviolet, UV -A can also inac-
tivate E. coli photoreactivating enzyme in vivo and in vitro. 50 UV -A inactivation
of yeast photoreactivating enzyme is oxygen-dependent.44 The capacity of
photoreactivation of 254 nm lethality in exponential growth phase of E. coli was
inhibited by prior exposure to 365 nm radiation. 3.
The ability of bacteria to support the multiplication and liberation of phage
can be impaired by UV-A, 49,69,73,74 an effect long known to be a result ofUV-C
radiation. 75 Partially dehydrated bacteria may be more sensitive to ultraviolet
radiation than cells in liquid suspension. 76,77 This effect of relative humidity is
found to be greater for UV-A than for UV-C radiation. 78-80 Carotenoids may
partially protect against UV-induced lethality,81 an effect that is best shown
against UV-A-induced lethality. 34,82,83
Transforming DNA can be inactivated by UV-A 84-86 and shows a response
curve that differs from the response to UV-C radiation. Action spectrum studies
for inactivation of tryptophan and leucine markers of B. subtilis transforming
DNA suggest that the lesions responsible for marker inactivation are different for
UV-A and UV-C. 34 In contrast to a large photoreactivable component of UV-C
and UV-B inactivation of Haemophilus injluenzae DNA, no photoreactivation
was evident after 365 nm inactivation. 87 UV-A inactivation of H. injluenzae
transforming DNA is oxygen-dependent unless the transforming DNA is purified
before being added to the B. subtilis. 58 This lack of oxygen dependence after
purification strongly suggests that the chromophore for the oxygen-dependent
transforming DNA lesions induced by UV-A is some nucleoprotein associated
with DNA. Histidine protects against the UV-A inactivation of transforming
DNA of B. subtilis, 58,88 but offers no protection at wavelengths shorter than 300
nm. Clearly UV-A and UV-C produce different lesions in these experiments.
The mechanisms of UV -A inactivation of transforming DNA and the evidence
and mechanism for UV-A-induced bacteriophage lethality is discussed by
Webb. 34
In the absence of known exogenous photosensitizers, UV-A and visible
radiation are mutagenic. 89-91 The action spectrum for mutagenesis in continuous
cultures of E. coli Bir show~d a broad peak between 350 and 480 nm and demon-
strated a strong oxygen dependence at wavelengths longer than 320 nm. 34 The
demonstration of UV-A-induced mutation using broad band sources 92 - 94 should
be reexamined by the use of reliable monochromatic sources and exact
dosimetry.
It is not known to what extent these findings in microorganisms are applic-
able to ultraviolet-induced changes in animal cells. Some of the same molecular
alterations must occur in multicellular animals, but it is not known whether such
injury causes the same effect on the cell. Subtle changes that do not cause muta-
tion or cell death may, by altering the performance of specialized cells, have
Effects on Microorganisms and Animal Cells 99

physiologic implications on the affected cell, or on its neighbor cells or organs.

For example, cell permeability is affected by ultraviolet. Such a change may have
little effect on the growth of a bacterium but could lead to significant physiologic
changes in animals if increased excitability of membranes or changes in the water
or ion content of certain cells occurs as a result of UV -A radiation. Microor-
ganisms' DNA repair and other systems may repair or reverse ultraviolet damage
differently from those of animal cells. And while decreases in the rate of cell
division may temporarily retard the creation of new bacteria, the same effect can
interfere significantly with the function of a complex organ system and the survi-
val of the organism. Alternatively, the epidermal cells of animals are the most
exposed cells and are a population that is already programmed to die, and some
lethality of the differentiated cells may not be adverse. Presumably, mutagenicity
of the stem cells that supply such populations is of greater concern.
In organ systems, the results of radiation may be quite complicated. Sub-
stances within cells or on the cell surface may absorb UV -A and transfer the exci-
tation energy to nucleic acids, catalyzing or altering the action spectrum for ge-
netic damage caused by radiation. Pyridine nucleotides, melanin, nucleopro-
teins, flavins, or porphyrins in living cells may play such roles. Substances may
also intercalate with DNA and, upon absorption of UV -A, induce photochemical
reactions that ultimately alter the DNA. The presence of such endogenous or
exogenous substances may markedly reduce the amount of radiation needed to
cause biologically significant events. For instance, the exposure dose of UV-A
needed to produce skin erythema (redness) in the presence of photo active psora-
lens is much lower than that needed in the absence of psoralens. When 4,5' ,8-
trimethylpsoralen is applied topically in proper concentrations, the dose of UV-A
needed to cause delayed redness of human skin may approach the relatively small
doses of UV-B needed to cause erythema.
UV-A irradiation of tissue culture medium (Dulbecco's modified Eagle's
medium without serum or phenol red) can cause death of cells growing in that
medium. The irradiated medium remains lethal to cells added after UV-A expo-
sure, while similar UV-A exposure to cells in phosphate-buffered saline is not
lethal. The formation of toxic photoproducts within the medium via UV -A-
induced photodynamic action involving riboflavin, tryptophan, and tyrosine
seems the most likely mechanism. 95 Tryptophan photoproducts formed by expo-
sure to UV -A have been shown to be mutagenic to bacteria, influencing the ge-
netic recombination process. 47 UV-A irradiation OfL-tryptophan, in the presence
of oxygen, produces photoproducts that are toxic for bacteria 96 and mammalian
cells in tissue culture. 97 These products can also sensitize bacterial DNA to sub-
sequent 365 nm radiation to increase the yield of DNA strand breaks, 98 contrib-
ute to UV-A lethality when irradiating E. coli, 98 and inhibit replication gap clo-
sure inE. coli. 99
Furthermore, Glatzer et al. 23 have recently shown that the UV-A-induced
100 Chapter Five

tryptophan photoproducts bind to DNA and may induce lesions that interfere
with either DNA replication or repair. Such an observation is important not only
because it suggests mechanisms for biologic injury by UV -A but also because the
presence of photoproducts in tissue culture may lead to misinterpretation of the
effects of ultraviolet on in vitro systems. Not all biologic effects of radiation re-
sult from light-induced molecular changes within the cell. Irradiation of tissue
cultures in nonnutritive but isotonic media presumably eliminates effects due to
the formation of toxic photoproducts within the medium.
To avoid the production of toxic photoproducts that may occur in tissue cul-
ture medium, Danpure and Tyrrell I 00 irradiated Chinese hamster cells and HeLa
cells (human tumor cell line) in an inorganic buffer. Both cell lines were approx-
imately 10 4 times more resistant to inactivation by 365 nm radiation than by
254 nm radiation, and the UV -A inactivation curves had a large shoulder not
seen with the shorter-wavelength radiation. Both of these mammalian cell lines
were about 10 times more sensitive to UV -A lethality than exponential phase E.
coli. In further contrast to UV-C effects, UV-A lethality was strongly oxygen-
dependent, and UV-A exposure sensitized HeLa cells, but not hamster cells, to
subsequent X-irradiation. The authors hypothesized that the oxygen dependence
of UV-A-induced lethality in mammalian cells could indicate that a photo-
dynamic process is involved. Other possible explanations include damage to
DNA repair processes, induction of a separate oxygen-dependent class of DNA
lesions,44 alteration of oxygen metabolism, or other unknown mechanisms.
As with other wave bands, UV -A sensitivity may vary greatly depending on
cell type. A cumulative UV-A exposure dose of approximately 16 J/cm2 is re-
quired to produce a noticeable decrease in the number of total population doubl-
ings of human fibroblasts irradiated in saline. 101 Using the same source, human
lymphocyte viability in vitro is compromised by as little as 100 mJ/cm 2 , 102 and
shows a biphasic survival curve. More information is needed about the effect of
UV-A on mammalian cells.
Finally, UV-A cannot be considered a monomorphous entity. The arbitrary
nature of the separation of ultraviolet wave bands must be remembered. Many
different biologic phenomena may be induced by radiation wavelengths over the
320-400 nm region. Synergism, photoaddition, photoprotection, photoreactiva-
tion, and other competing or supportive photobiologic events may be occurring
simultaneously when polychromatic radiation is employed. 103 Repair enzymes
may be induced, inhibited, or inactivated by various spectral regions. Poly-
chromatic sources may produce different cellular responses than monochromatic
radiation, and irradiance may affect the biologic end point. Visible light emitted
by the experimental source and the environmental lighting of the laboratory must
not be considered as innocuous and should be eliminated when one is performing
precise experiments. Cool white fluorescent lamps have been shown to be
mutagenic and toxic to cultured mammalian cells in vitro, 104,105 and a host of
enzyme inductions or blue light effects may alter cell response to UV radiation.
Effects on Microorganisms and Animal Cells 101

Summary

It is well established that DNA damage and the cells' ability to repair this
damage are of major significance in the effects of UV-A, UV-B, UV-C, and
ionizing radiations on microorganisms and mammalian cells in tissue culture. A
variety of damage produced in DNA of various cells by UV -A exposure has been
recently observed. Non-oxygen-dependent damage to DNA after UV-A exposure
occurs in the form of pyrimidine dimers (photo- or excision-repairable) and
thymine glycols 34 (significance unknown, not oxygen-dependent). Oxygen-
dependent UV-A-induced DNA damage, which may be mediated by free-radical
intermediates, includes single-strand chain breaks or alkali-labile bonds and
other lesions. Endogenous and exogenous photosensitizing molecules also playa
role in UV-A-induced DNA damage; DNA photoproducts, DNA-protein cross-
linking, or DNA complexed with photosensitizing molecules may result from
such reactions, which mayor may not be oxygen-dependent.
In addition to DNA damage, UV-A may damage DNA repair mechanisms
in microorganisms. Photoreactivating enzyme is both destroyed and activated by
UV-A. Excision repair and single-strand break repair mechanisms may also be
inhibited. Presumably, the effects of UV -A on DNA and DNA repair interact in a
complex manner. UV-A (365 nm) appears to alter active transport processes,
proteins, and enzyme activities and to inhibit DNA repair. Unlike that of UV-C,
UV-A-induced mutagenesis may be highly irradiance-dependent and oxygen-
dependent in some cells. The effects of UV-A on single cells are just beginning
to be understood and appear to be highly variable, depending on the conditions of
exposure and the cells exposed. It is therefore difficult to generalize or form any
precise theory or model of these effects of UV-A. Finally, one must remember
that "UV_A" is a broad wavelength region, within which the many photo-
biologic effects produced may interact.

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30. Friedberg, E. C., Cook, K. H., Duncan, J., and Mortelmans, K. DNA repair enzymes in
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31. Epel, B. L. Inhibition of growth and respiration by visible and near-visible light. In Photo-
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Effects on Microorganisms and Animal Cells 103

32. Hollaender. A. Effect of long ultraviolet and short visible radiation (3500 to 2900 A) on Es-
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35. Spikes, J. D. Photodynamic action. In Photophysiology, Vol. 3 (A. C. Giese, Ed.). Academic
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37. Knowles, A. The effects of photodynamic action involving oxygen upon biological systems. In
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39. Webb, R. B., and Brown, M. S. Sensitivity of strains of Escherichia coli differing in repair
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365 nm in strains of E. coli differing in repair capability. Mutat. Res., in press, 1977.
41. MacKay, D., Eisenstark, A., Webb, R. B., and Brown, M. S. Action spectra for lethality in
recombinationless strains of Salmonella typhimurium and Escherichia coli. Photochem. Photo-
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42. Webb, R. B., and Lorenz, J. R. Oxygen dependence and repair of lethal effects of near ul-
traviolet and visible light. Photochem. Photobiol. 12:283-289, 1970.
43. Mathews, M. M., and Sistrom, W. R. Function of carotenoid pigments in non-photosynthetic
bacteria. Nature 184:1892-1893, 1959.
44. Tyrrell, R. M. RecA+-dependent synergism between 365 nm and ionizing radiation in log-
phase Escherichia coli: A model for oxygen-dependent near-UV inactivation by disruption of
DNA repair. Photochem. Photobiol. 23:13-20, 1976.
45. Eisenstark, A. Sensitivity of Salmonella typhimurium recombinationless (rec) mutants to visi-
ble and near-visible light. Mutat. Res. 10:1-6, 1970.
46. Eisenstark, A. Mutagenic and lethal effects of visible and near-ultraviolet light on bacterial
cells. In Advances in Genetics, Vol. 12 (E. W. Caspari, Ed.). Academic Press, New York,
1971, pp. 167-198.
47. Eisenstark, A. Tryptophan photoproduct as a genetic probe: Effects on bacteria. In Stadler
Genetics Symposium, Fifty (G. Kimber and G. P. R. Redei, Eds.). University of Missouri,
Columbia, 1973, pp. 49-60.
48. Ferron, W. L., Eisenstark, A., and Mackay, D. Distinction between far- and near-ultraviolet
light killing of recombinationless (recA) Salmonella typhimurium. Biochim. Biophys. Acta
277:651-658, 1972.
49. Hanawalt, P. C. The UV sensitivity of bacteria: Its relationship to the replication cycle. Photo-
chem. Photobiol. 5:1-12, 1966.
50. Tyrrell, R. M., Webb, R. B., and Brown, M. S. Destruction of photoreactivating enzyme by
365 nm radiation. Photochem. Photobiol. 18:249-254, 1973.
51. Morton, R. A., and Haynes, R. B. Changes in the ultraviolet sensitivity of Escherichia coli
during growth in batch cultures. J. Bacteriol. 97:1379-1385, 1969.
52. Harrison, A. P. Survival of bacteria: Harmful effects of light, with some comparisons with
other adverse physical agents. Ann. Rev. Microbiol. 2/:143-156,1967.
53. Peak, M. J. Some observations on the lethal effects of near-ultraviolet light on Escherichia
coli, compared with the lethal effects of far-ultraviolet light. Photochem. Photobiol. 12:1-8,
1970.
104 Chapter Five

54. Tyrrell, R. M. Lethal effects. Abstracts, Fifth Annual Meeting of the American Society for
Photobiology, San Juan, 1977, p. 14.
55. Tyrrell, R. M. Induction of pyrimidine dimers in bacterial DNA by 365 nm radiation. Photo-
chem. Photobiol. 17:69-73, 1973.
56. Tyrrell, R. M., Ley, R. D., and Webb, R. B. Induction of single-strand breaks (alkali-labile
bonds) in bacterial and phage DNA by near-UV (365 nm) radiation. Photochem. Photobiol.
20:395-398, 1974.
57. Tyrrell, R. M. The interaction of near-UV (365 nm) and X-radiations on wild-type and repair-
deficient strains of Escherichia coli K12: Physical and biological measurements. Int. J. Radiat.
Bioi. 25:373-390, 1974.
58. Peak, M. J., Peak, J. G., and Webb, R. B. Inactivation of transforming DNA by ultraviolet
light. II. Protection by histidine against near-UV irradiation: Action spectrum. Mutat. Res.
20:137-141, 1973.
59. Jagger, J. Effects and mechanisms of near-UV (UV-A) radiation actions on cells. Sublethal
effects. Abstracts, Fifth Annual Meeting of the American Society for Photobiology, San Juan,
1977, p. 1<.
60. Ramabhadran, T. V., and Jagger, J. Mechanism of growth delay induced in Escherichia coli
by near ultraviolet radiation. Proc. Natl. Acad. Sci. USA 73:59-63, 1976.
61. Bruce, A. K. Response of potassium retentivity and survival of yeast to far ultraviolet, near
ultraviolet visible, and x-radiation. J. Gen. Physiol. 41 :693-702, 1958.
62. Koch, A. L., Doyle, R. J., and Kubitschek, H. E. Inactivation of membrane transport in Es-
cherichia coli by black light. J. Bacteriol. 126:140-146, 1976.
63. Barran , L. R., Dooust, J. Y., Labelle, J. L., Martin, W. Cr., and Schneider, H. Differential
effects of visible light on active transport in E. coli. Biochem. Biophys. Res. Commun.
56:522-528, 1974.
64. Sprott, G. D., Dimock, K., Martin, W. G., and Schneider, H. Coupling of glycine and alanine
transport to respiration in cells of Escherichia coli. Can. J. Biochem. 53:262-268, 1975.
65. Doyle, R. J., and Kubit"hek, H. E. Near ultraviolet light inactivation of an energy-
independent membrane transport system in Saccharomyces cerevisiae. Photochem. Photobiol.
24:291-293, 1976.
66. Robb, F. T., Hauman, J. H., and Peak, M. J. Similar spectra for the inactivation by mono-
chromatic light of two distinct leucine transport systems in Escherichia coli. Photochem.
Photobiol., in press, 1977.
67. Kashket, E. R., and Brodie, A. F. Effects of near-ultraviolet irradiation on growth and oxida-
tive metabolism of bacteria. J. Bacteriol. 83: 1094-1100, 1962.
68. Jagger, 1. Growth delay and photoprotection induced by near-ultraviolet light. In Research
Progress in Organic, Biological and Medicinal Chemistry, Vol. 3, Part I (U. Gallo and L.
Santamaria, Eds.). American Elsevier, New York, 1972, pp. 383-401.
69. Webb, S. J., and Bhorjee, J. S. The effect of 3000-4000 Alight on the synthesis of B. galac-
tosidase and bacteriophages by Escherichia coli B. Can. J. Microbiol. 13:69-79, 1967.
70. Swenson, P. A., and Setlow, R. B. Inhibition of the induced formation of tryptophanase in
Escherichia coli by near-ultraviolet radiation. J. Bacteriol. 102:815-819, 1970.
71. Ramabhadran, T. V. Effects of near-ultraviolet and violet radiations (313-405 nm) on DNA,
RNA, and protein synthesis inE. coli B/r: Implications for growth delay. Photochem. Photo-
bioi. 22:117-123, 1975.
72. Tyrrell, R. M., and Webb, R. B. Reduced dimer excision following near ultraviolet (365 nm)
radiation. Mutat. Res. 19:361-364, 1973.
73. Hill, R. F. Effects of illumination on plaque formation by Escherichia coli infected with T1
bacteriophage. J. Bacteriol. 7/:231-235, 1956.
74. Day, R. S., and Muei, B. Ultraviolet inactivation of the ability of E. coli to support the growth
of phage T7: An action spectrum. Photochem. Photobiol. 20:95-102,1974.
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75. Anderson, T. F. The growth ofT2 virus on ultraviolet killed host cells. J. Bacteriol. 56:403-
410,1948.
76. Wells, W. F., and Wells, N. W. Air-borne infection. J. A. M. A. 107:1698-1703, 1936.
77. Kaplan, R. W., and Kaplan, C. Influence of water content on UV-induced S-mutation and kill-
ing in Serratia. Exp. Cell Res. 11:378-392, 1956.
78. Webb, S. J. The effect of relative humidity and light on air-dried organisms. J. Appl. Bacteriol.
26:307-313, 1963.
79. Webb, S. J., and Tai, C. C. Lethal and mutagenic action of 3200-4000 A light. Can. J. Mi-
crobiol. 14:727-735, 1968.
80. Webb, S. J., and Tai, C. C. Differential, lethal and mutagenic action of 254 nm and 320-400
nm radiation on semi-dried bacteria. Photochem. Photobiol. 12:119-143, 1970.
81. Sistrom, W. R., Griffiths, M., and Stanier, R. Y. The biology of photosynthetic bacterium
which lacks colored carotenoids. J. Cell Compo Physiol. 48:473-515, 1956.
82. Denniston, K. J., Webb, R. B., and Brown, M. S. Action spectrum for carotenoid protection
against lethal photo-oxidation of Sarcina lmea. Abstr. Am. Soc. Microbiol., 1972, p. 184.
83. Mathews-Roth, M. M., Wilson, T., Fujimori, E., and Krinsky, N. 1. Carotenoid chromophore
length and protection against photosensitization. Photochem. Photobiol. 19:217-222, 1974.
84. Szybalski, W., and Opara-Kubinska, L. Radiobiological and physiochemical properties of
5-bromodeoxyuridine-labelled transforming DNA as related to the nature of the critical
radiosensitive structures. In Cellular Radiation Biology. Williams & Wilkins, Baltimore, 1965,
pp. 223-240.
85. Peak, M. J., Peak, J. c., and Webb, R. B. Inactivation of transforming DNA by ultraviolet
light. I. Near-UV action spectrum for marker inactivation. Mutat. Res. 20:129-135, 1973.
86. Cabrera-Juarez, E., and Swenson, P. A. Action spectrum for the oxygen independent inactiva-
tion of Haemophilus injiuenzae transforming DNA with near-UV light. Photochem. Photobiol.
2/:193-196,1975.
87. Peak, M. J., Peak, J. G., and Webb, R. B. Inactivation of transforming DNA by ultraviolet
light. III. Further observations on the effects of 365 nm radiation. Mutat. Res. 20:143-148,
1973.
88. Peak, M. J., and Peak, J. G. Protection by histidine against the inactivation of DNA transform-
ing activity by near-ultraviolet light. Photochem. Photobiol. 18:525-527, 1973.
89. Kubitschek, H. E. Mutagenesis by near-visible light. Science /55:1545-1546, 1967.
90. Webb, R. B., and Malina, M. M. Mutagenesis in Escherichia coli by visible light. Science
156:1104-1105, 1956.
91. Webb, R. B. Photodynamic lethality and mutagenesis in the absence of added sensitizers. In
Organic, Biological and Medicinal Chemistry, Vol. 3, Part 2. (U. Galo and L. Santamaria,
Eds.). American Elsevier, New York, 1972, pp. 511-530.
92. Kaplan, R. W. Dose-effect curves of S-mutation and killing in Serratia marcescens. Arch.
Microbiol. 24:60-79, 1956.
93. Kelner, A., and Halle, S. Mutagenesis by visible light in a mutable strain of Escherichia coli.
Society of American Bacteriologists, Abstracts of 60th Annual Meeting, Philadelphia, May
1-5, 1960, p. 67.
94. Cabrera-Juarez, E., and Espinosa-Lara, M. Lethal and mutagenic action of black light (325-
400 nm) on Haemophilus injiuenzae in the presence of air. J. Bacteriol. 117:960-964, 1974.
95. Stoien, J. D., and Wang, R. J. Effect of near-ultraviolet and visible light on mammalian cells in
culture. II. Formation of toxic photoproducts in tissue culture medium by blacklight. Proc.
Natl. Acad. Sci. USA. 7/:3961-3965, 1974.
96. Yoakum, G., and Eisenstark, A. Toxicity of I.-tryptophan photoproduct on recombinationless
(rec) mutants of Salmonella typhimurium. J. Bacteriol. 112:653-655, 1972.
97. Wang, R. J., Stoien, J. D., and Landa, F. Lethal effect of near-ultraviolet irradiation on mam-
malian cells in culture. Nature 247:43-45, 1974.
106 Chapter Five

98. Yoakum, G. H. Tryptophan photoproducts(s): Sensitized induction of strand breaks (or alkali-
labile bonds) in bacterial deoxyribonucleic acid during near-ultraviolet irradiation. J. Bacteriol.
122: 199-205, 1975.
99. Yoakum, G., Ferron, W., Eisenstark, A., and Webb, R. B. Inhibition of replication gap closure
in Escherichia coli by near-ultraviolet light photoproducts of L-tryptophan. J. Bacteriol.
119:62-69, 1974.
100. Danpure, H. J., and Tyrrell, R. M. Oxygen dependence ofnear-UV (365 nm) lethality and the
interaction of near-UV and X-rays in two mammalian cell lines. Photochem. Photobiol.
23:171-177, 1976.
101. Gilchrest, B. A. Personal communication.
102. Morison, W. L. Personal communication.
103. Peak, M. J., Peak, J. G., and Webb, R. B. Synergism between different near-ultraviolet
wavelengths in the inactivation of transforming DNA. Photochem. Photobiol. 21:129-131,
1975.
104. Bradley, M. 0., and Sharkey, N. A. Mutagenicity and toxicity of visible fluorescent light to
cultured mammalian cells. Nature 266:724-726, 1977.
105. Ritter, M. A., and Williams, J. W. Endonuclease-sensitive sites induced by fluorescent light.
Radiation Research Society, May 1977, Abstract Ei-4.
CHAPTER 6

Immediate and Short-Term Biologic

Effects of Ultraviolet Radiation
on Normal Skin

Introduction

A host of biologic effects of ultraviolet exposure of normal human skin begin

immediately after absorption of ultraviolet photons within the tissue. Biophysical
and photochemical molecular events are followed by alterations in biochemistry
and subsequent changes in cell metabolism. Electron-microscopic and his-
tochemical studies demonstrate changes in cell structure and function occurring
in minutes to hours after UV exposure. Subsequently, products of photochemis-
try and altered metabolism lead to cellular changes recognizable by routine light
microscopy. Over hours to days, changes in blood flow, cell kinetics, and pig-
ment production cause grossly observable changes in the whole organ.
The only known beneficial effect of ultraviolet radiation on skin is the con-
version of 7-dehydrocholesterol to vitamin D;l in skin. Vitamin D;l affects cal-
cium absorption and calcification of bone, and insufficient solar UV-B exposure
is reported to cause rickets in children. 1 In a long-term controlled study using
artificial lighting, Neer et at. have shown increased calcium absorption in sub-
jects receiving greater UV-B exposures. 2 While vitamin D~ is produced within 3
the skin by photochemical alteration of 7-dehydrocholesterol, hydroxylation of
vitamin D;l to 25-0H cholecalciferol (25-0HCC) in the liver 4 and hydroxylation
of 25-0HCC to 1,25 dihydroxycholecalciferol (1 ,25-0HCC) in the kidney 4.5 are
largely responsible for calcium absorption and homeostasis. 6
This chapter outlines the recognizable immediate and short-term effects of
UV exposure of normal skin. The events discussed here may occur after asingle
adequate exposure. Biologic response in the presence of pathologic conditions or

107
108 Chapter Six

photosensitizing compounds and the chronic effects of multiple UV exposures

are reviewed in the following two chapters.
The skin's response to UV exposure is, in general, a reparative and protec-
tive reaction. Sunburn is an example of inflammation, a generalized protective
pathophysiologic response of skin. In this sense, many features of the skin's re-
sponse to UV exposure are similar to those caused by other irritating or toxic
agents.
The various components of inflammation are usually useful to the organ as a
whole. The vasodilation and increased capillary permeability associated with
inflammation allow increased transfer of proteins and other molecules across the
endothelial capillary lining. The larger-molecular-weight proteins that normally
are retained in the capillaries are allowed to enter the dermis. The resulting shift
of osmotic pressure between vessels and the tissues of the dermis draws excess
water into the tissue, causing edema. The altered capillary endothelium also
permits polymorphonuclear leukocytes, lymphocytes, and monocytes to pass
from vessels into the dermis and, to some extent, the epidermis. The numerous
polymorphonuclear leukocytes phagocytize foreign matter that may be present
when the skin has been broken and infected. Monocytes, also called mac-
rophages, can phagocytize larger objects, including dead polymorphonuclear
cells.
Thickening of the epidermis is a generalized protective response often as-
sociated with inflammation. The increased proliferation of keratinocytes follow-
ing UV exposure of skin leads to thickening of the epidermis and may be as-
sociated with a noticeable increase in desquamation or "peeling." Delayed tan-
ning of the skin may also result from a single UV exposure. This protective re-
sponse is caused by increased quantities of melanin produced by melanocytes in
the basal layer. Hyperpigmentation may also result from other causes of inflam-
mation. The degree of baseline pigmehtation and pigmentation response induced
by ultraviolet is genetically determined.
Inflammation is a primitive response designed to remove an injurious agent
that invades the skin. Radiant exposure is one cause of inflammation in which no
matter actually enters the tissue. This situation is also true of some thermal bums
or trauma. The delayed onset of the inflammation of sunburn indicates that
radiant exposure presents a special class of injury. The "injurious agents" of UV
exposure are certain photochemical products formed within the tissue, which
may include photochemically altered DNA, RNA, proteins, membranes, and
other molecules or structures. It is the task of the photobiologist to elucidate the
complicated pathways that lead from these initial photochemical alterations to the
observable short-term and chronic biologic effects of ultraviolet radiation. The
appearance of the delayed erythema component of ultraviolet-induced inflam-
matory response has been the most commonly used end point in dermatologic
photobiology. While much of our knowledge of the effects of ultraviolet radia-
Effects of Ultraviolet Radiation on Skin 109

tion of skin is obtained by the use of this end point, it must be remembered that
erythema is only one small part of the cutaneous response to ultraviolet exposure.

Erythema

Ultraviolet-induced erythema (redness) of skin has been studied by many

investigators and experienced by all but the most pigmented or protected persons.
Upon exposure to sun or artificial ultraviolet sources, a faint, transient redness
may begin within minutes. However, the major erythema response of skin to ul-
traviolet radiation is delayed in onset. This delayed redness may not appear for
several hours after UV exposure, gradually increases to reach its maximum at
12-24 hr after exposure, and then fades over several days. The delayed erythema
response is the reaction commonly referred to as sunburn. Exposed areas are
warm to the touch, are often tender, and can be blanched with pressure.
Erythema caused by ultraviolet radiation is grossly confined to exposed
areas and reflects blood vessel dilatation and increased quantities of blood in the
dermis. It is often assumed that the initial photochemical reaction is in the
epidermis, where photon absorption by keratinocytes may lead to liberation of
intracellular substances that diffuse into the papillary dermis to cause vasodila-
tion. This diffusion theory is supported by the existence of a latent period be-
tween exposure and erythema and by the fact that much of the radiant energy of
the erythemogenic wavelength region is absorbed by the epidermis. However,
there may also be direct injury to the vascular endothelium or to other sites in the
dermis.
In experimental animals, the vascular response to ultraviolet radiation is
biphasic. Transient immediate vasopermeability 7.8 is followed, after a latent
period of 2 to 8 hr, by a period of delayed, prolonged, increased vasopermeabil-
ity and vasodilation. In some animal models, the initial vasopermeability is ac-
companied by a faint erythema (vasodilation), which may begin during exposure.
This immediate erythema has been attributed to release of histamine, 9.1 0 possibly
because of a direct effect of photons on dermal mast cells. There is evidence that
serotonin 11 • 12 may also play some role. In rats and guinea pigs, serotonin and
histamine antagonists suppress the immediate phase of UV vascular responses. 13
In vivo studies of human skin have shown transient appearance of kinins within
minutes after ultraviolet radiation. 14 Kinins were not found after the onset of de-
layed erythema.
The mediators of the delayed phase of ultraviolet-induced vascular response
have been difficult to define. Anlihistamines do not suppress the delayed
erythema induced by ultraviolet radiation in guinea pigs, rats, or humans. 9 Ki-
nins have been stated to be absent ll or not elevated. 1o • 1fi Delayed erythema was
not suppressed by using various inhibitors of proteases, plasminogen activators,
110 Chapter Six

or kallikrein. '3 Serotonin has been found in urine following ultraviolet expo-
sure, 12 but the significance of this finding is not clear.
More recently, prostaglandins, a group of long-chain fatty acids with vas-
oactive properties, have been implicated as possible mediators of the delayed
phase of erythema. Prostaglandins are produced in human and animal skin 17,18
(paE and paF groups of prostaglandins), intradermal injection of prostaglandins
produces erythema 19 (paE mainly; paF group are much less active), and the
production of prostaglandins increases following UV exposure. 15,20 Indometha-
cin is a potent inhibitor of the conversion of arachidonic acid to active prosta-
glandin, a reaction catalyzed by the enzyme prostaglandin synthetase. Topical
indomethacin in both humans and guinea pigs produces a profound and pro-
longed blanching of UV-B-induced delayed erythema,21,22 and in humans in-
tradermal indomethacin has been shown to consistently decrease erythema from
UV_B 23 but not from all sites irradiated with UV_C 24 ; these effects appear to be
due to inhibition of prostaglandin synthetase. 23 Oral indomethacin in humans has
been shown to decrease the cutaneous blood flow peak that accompanies a
UV-B-induced erythematous reaction,23 but a visual assessment of UV-B-
induced delayed erythemal response did not show observable reduction of
erythema by oral indomethacin. 26
The fact that a transient tissue leukocytosis precedes the peak of the delayed
erythema reaction leads to a theoretical consideration of release of vasoactive fac-
tors from leukocytes. 27 However, ultraviolet-induced delayed erythema was the
same in guinea pigs made neutropenic with nitrogen mustard as in irradiated but
untreated controls. 7
It has been suggested that the complex delayed erythema reaction known as
sunburn may result from hydrolytic enzymes and possibly other substances re-
leased by lysosomes 28,29 within keratinocytes. The possibility that light could
rupture cutaneous lysosomes was first suggested by Weissman and Fell. 30 Irradia-
tion (300 nm) of fetal rat skin resulted in necrosis of epidermis and dermis. His-
tologic evidence of destruction and necrosis was decreased by prior addition of
hydrocortisone, and this decrease was felt to be due to stabilization of lysosomes
by hydrocortisone. In addition,. ultraviolet irradiation was shown to release acid
protease from isolated rat liver lysosomes. 30 Subsequent studies have confirmed
this observation and showed release of the enzyme to be dose-related. 31
Johnson 32 found that irradiation with wavelengths shorter than 320 nm reduced
the extractable acid phosphatase in mouse ears but did not deplete histochemi-
cally demonstrable succinic dehydrogenase. He felt this was evidence for
lysosomallabilization in the absence of nonspecific leakage due to cell death.
Histochemical studies of human skin exposed to ultraviolet radiation were
consistent with the theory that specific damage to lysosomal membranes caused
partial to complete rupture and release of enzymes. 28 At 4 hr after irradiation,
enzymes were leaking from the lysosomes, and at 6 hr there was evidence of
Effects of Ultraviolet Radiation on Skin 111

lysosomal rupture. Isolated cells subsequently showed more severe nuclear and
intercellular damage, presumably secondary to the action of the enzymes, and
these cells stained abnormally as the so-called sunburn cells. 33 Wilgram et al. 34
have shown ultraviolet-induced keratinosome disintegration or disappearance
from human skin. The structures described as keratinosomes contain acid phos-
phatases and have several features in common with lysosomes. Other studies
have demonstrated numerous vacuoles in epidermis 1 hr after ultraviolet irradia-
tion. 35 These vacuoles appear similar to the acid-phosphatase-containing au-
tophagic vacuoles seen in psoralen-induced phototoxic reactions. 36 Ultraviolet-
induced lysosomal labilization of human foreskin epidermis can be blocked by
the application of a sunscreen. 37
Lysosomal damage may not only cause the release of lysosomal enzymes
and subsequent damage of the keratinocytes but may also release enzymes, va-
sodilator substances, or subsequently formed cell-breakdown products into the
dermis, where they may firectly or indirectly lead to erythema. It has been
suggested that the immediate erythema that results from ultraviolet radiation re-
sults from disruption of lysosomes of the endothelial cells of blood vessels with
release of chemical mediators. 38 The delayed erythema could result from secon-
dary diffusion of proteinases from the epidermis secondary to lysosomal rup-
ture. 39 It is also possible that direct photon damage to the lysosomes of mast cells
of the dermis or endothelial cells of dermal blood vessels may play a role in the
delayed erythema response. 29 It has also been suggested that lysosomal leakage
or rupture may playa role in photosensitization. A number of exogenous photo-
sensitizing substances become concentrated within lysosomes, and subssquent
exposure to radiation of the appropriate wavelengths results in lysosomal damage
leading to cell death.4o.41 Lysosomal rupture appeared a primary event in cell
death resulting from photosensitization with acridine orange but not in cell death
resulting from metabolic poisons. 38
The central role of lysosomes in ultraviolet erythema remains conjectural. It
remains to be determined whether the effect of ultraviolet radiation on lysosomes
is a primary event in the chain of photochemical reactions that result in delayed
erythema. It is not known whether lysosomal membranes are damaged by direct
absorption of photons upon their surface or whether they are damaged secondar-
ily by production of free radicals or mediators produced at some other site. An
increase in free radicals, which are known to damage lipid membranes, have
been demonstrated in human skin following ultraviolet radiation.29.42.43
Lysosomal stabilizers given prior to radiation have been shown to alter the degree
of erythema produced by a given dose of ultraviolet radiation, 44 but sunburn can-
not be entirely prevented in this way. Furthermore, recent electron-microscopic
studies have shown that cytoplasmic and nuclear edema with vesiculation appear
within minutes after irradiation, before lysosomes are noted to be damaged, and
that the lysosomes in "sunburn cells" often appear completely normal. 45
112 Chapter Six

The mediators of UV-A erythema mayor may not be the same as those of
UV-B erythema. In a study of the release of bound hydrolases located in the
lysosomal particles of guinea-pig epidermis, it was noted that ultraviolet of
wavelengths longer than 330 nm was found to be effective in releasing acid
DNase. 46 On the other hand, recent studies have shown that topical intradermal
or oral indomethacin does not reduce UV-A-induced delayed erythema in hu-
mans. 26 In the same subjects, the same doses of indomethacin reduced UV-B-
induced delayed erythema. This result suggests that prostaglandins do not playas
important a role, if any, as mediators of the UV-A-induced delayed erythema
reaction.
Multiple ultraviolet chromophores may exist in skin, and radiation probably
leads to activation of a complicated cascade of mediators whose final end point is
erythema. Multiple pathways may exist. It is also possible that photons have ad-
ditional direct effects on blood vessels 47 or nerves. Ultraviolet radiation causes
dilation of isolated exposed dermal blood vessels. 48 Dermal proteins may be di-
rectly changed by radiation. 49.50
Using a mathematical approach utilizing diffusion theory, van der Leun 51
hypothesized that erythema induced by 300 nm sources results from the action of
radiation on the epidermis. A mediator substance, which according to calcula-
tions of the diffusion coefficient is not necessarily macromolecular, then diffuses
into the dermis to cause vasodilation. He quoted evidence of a slowly spreading
"diffusion flush" seen at the periphery of striking U V-induced erythema 5 2 and
provided quantitative evidence for diffusing substances liberated by 300 nm radi-
ation. On the other hand, van der Leun believes the erythema induced by 250 nm
radiation to be mediated by a direct effect of photons on the dermis. He is in
agreement with Rottier 53 - 55 that UV-B and UV-C have two independent
erythema reactions involving two separate chromophores. Van der Leun suggests
that the chromophore for UV-A erythema is the same as that for UV-C-induced
erythema and exists in the dermis.
The presence and degree of delayed erythema induced by exposure to ultra-
violet radiation are dependent upon exposure dose. For a given area, the expo-
sure dose equals the product of irradiance and exposure time. The erythema is
relative to the radiant energy delivered per unit area of skin surface and not to the
rate of delivery (irradiance) per se. Within extremely wide limits (10 7 -fold range)
delayed erythema response is independent of irradiance ("dose rate"); that is, the
influence of doubling the irradiance may be compensated for by a halving of the
exposure time. 56-59 This relationship, of a given exposure dose yielding a con-
stant biologic response, is termed the reciprocity law. Most photobiologic re-
sponses that follow the reciprocity law over a wide range of irradiances may be
assumed to derive from photochemical causes. That is, constant quantities of
some photochemical reaction products are produced per absorbed photon at given
wavelengths, and these products yield a constant biologic response. Thus, the
Effects of Ultraviolet Radiation on Skin 113

delayed erythema response to UV exposure is initiated mainly via photochemical

products. It is important to realize, however, that not all photochemically derived
biologic responses necessarily follow the reciprocity law. The complexities of
repair systems, inducible responses, factors affecting the transport, localization
and stability of photochemical products, and the complicated cascade of events
that ultimately lead to erythema or other gross responses may in essence obscure
the fact that the primary photochemical event itself follows a reciprocity relation-
ship. In addition, the products of photochemical reactions in which a significant
percentage of the absorbing molecules are excited at any given time need not
follow a reciprocity relationship.
Reciprocity over a wide range of irradiances is certainly not the case for
thermally induced photobiologic responses. As a site is exposed, heat transfer
away from the exposure area occurs until either some equilibrium temperature is
reached at the site during exposure or the exposure ceases. At equilibrium, the
temperature attained at the exposure site is proportional to the irradiance, and not
the exposure dose, and a deviation from the reciprocity law is seen when the
biologic response corresponds to the temperatures produced during exposure.
Many of the biologic effects of UV-A require a greater radiant exposure dose
than do those of shorter wavelengths. Therefore, higher irradiances are often
employed in experimental UV-A exposures, and one must be careful to separate
the biologic effects particular to UV -A from the biologic effects of radiant heat-
ing of tissue. Examination of the adherence of a given biologic effect to the reci-
procity law is a relatively simple way of making this separation. Another means
is to compare the effects of UV -A with infrared or other wavelengths that heat the
tissue similarly to UV -A.
The size of the exposure site may also affect photobiologic responses,
whether induced by heat or by photochemical reactions. At any given irradiance,
the total thermal energy delivered and the total quantity of a given photochemical
reaction product produced during exposure are proportional to the area of the ex-
posed site. If transfer of either of these agents from the exposure site is sig-
nificant, the biologic response caused by them will be partially dependent on the
area of the exposed site. In cases where a systemic effect results from light expo-
sures, such as in visible-light phototherapy for hyperbilirubinemia of newborns,
the total area exposed is a major factor determining the degree of the systemic
response. 6o ,61 Except for exposure areas a few millimeters in diameter, UV-
induced delayed skin erythema is independent of the size of the exposure site. 62
Variation of UV-A-induced delayed erythema with area of exposure has not been
reported.
The variation of UV-B erythema with angle of incidence of the radiation
appears to follow a cosine function (cf. Chapter 3).63 Because significant absorp-
tion of UV wavelengths occurs within the stratum corneum (cf. Chapter 4), one
would not expect this response function unless the stratum corneum also scatters
114 Chapter Six

UV wavelengths. In the absence of scattering, the transmittance of the stratum

corneum according to Beer's law would vary as e -Ed sec 0, where () is the angle of
incidence, d is the thickness of stratum corneum, and E is an extinction
coefficient. Since irradiance at the skin surface caused by a collimated beam is
de fined as £(0=0) cos (), exposure to the viable cells of the epidermis would vary
with angle of incidence as cos ()e-·dsec 8, which would predict a cosine-weighted
erythemal response function only for small values of () (near-normal incidence).
In the presence of a high degree of scattering (turbidity), however, the average
photon pathlength within the stratum corneum is much less dependent upon (),
and the viable epidermal exposure varies more nearly with the surface irradiance,
£(o=O)cos (). A cosine-weighted erythemal response would be expected even at
larger angles of incidence. Within the UV -C and UV -B wavelength regions, this
latter situation appears to be the case. The variation of erythema with angle of
incidence of UV-A remains to be documented, but one would expect a nearly
cosine response function for UV-A wavelengths as well because of the lesser ab-
sorption of UV-A wavelengths by the stratum corneum as compared with UV-B
wavelengths. The application of substances that reduce scattering or increase ab-
sorption within the stratum corneum may theoretically modify the variation of
erythemal response of skin with angle of incidence.
One obtains the action spectrum for delayed erythema by plotting the recip-
rocal of the exposure dose necessary to produce a certain predetermined grade of
redness versus wavelength. The classic erythema action spectrum of Hausser and
Vahle 64 (Fig. 6-1) has been more recently modified by the use of modem mono-
chromators and newer views regarding the end points for minimal erythema
dose 65 • 66 (Fig. 6-2). The exact shape of the erythema action spectrum is still dis-
puted and is affected by the degree of erythema that is being plotted, the time of
reading after irradiation, and other variables.
Evaluation of redness induced by ultraviolet radiation is difficult to quantify;
degrees of redness of skin are usually estimated by subjective visual evaluations,
which differ from one observer to the next. Therefore, in determining action
spectra, most observers have attempted to describe simply the presence or ab-
sence of redness. After irradiating adjacent sites of skin with increasing incre-
ments of UV exposure doses, the skin is observed to find which sites sub-
sequently become red as the result of such exposure. The lowest exposure dose
required to produce SUbsequent erythema is regarded as the threshold or break-
point dose and is called the minimal erythema dose (MED). It is usually the re-
ciprocal of this value, determined at various wavelengths, that is plotted as an
erythema action spectrum.
Even when the presence or absence of erythema is used as an end point,
exact determinations of erythema action spectra are impossible. The threshold or
break points establishing MED are usually not definite or easily agreed upon. The
Effects of Ultraviolet Radiation on Skin 115

en
en
w
Z .7
w
'#. >
>- I-
U u
z 10.0 w
w u..
u..
.5
u w
u.. w
u..
w 1.00 >
« I-
« .3
~
w ...J
w
J:
a:: .2
I- .100
>-
a:: .1
w

.0 1 0 L...L_----L---lI---.J_---.Ju......-:-'~
250 300 450 250 260 270 280 290 300 310
WAVELENGTH (nm) WAVELENGTH (nm)

Fig. 6- I. Erythema action spectrum for human
skin. Note logarithmic vertical axis. (Modified
from Hausser. K. W., and Vahle, W. Sunburn
and suntanning. In The Biologic Effects of Ul·
traviolet Radiation with Emphasis on the
Skin. F. Urbach, Ed. Pergamon Press, Ox-
ford, 1969, pp. 3-21.)
Fig. 6-2. Relative erythemal effectiveness of
various wavelengths. (From Everett, M. A.,
Sayre, R. M., and Olson, R. L. Physiologic
response of human skin to ultraviolet radia-
tion. In The Biologic Effects of Ultraviolet
Radiation with Emphasis on the Skin. F. Ur-
bach, Ed. Pergamon Press, Ltd., Oxford,
1969. Used with permission.)

presence of minimal erythema may be difficult to detect. While some observers

feel that any patch of redness anywhere in the exposed field is enough to establish
MED, others insist that the redness be definite and well defined and that the bor-
der of the redness clearly outline the radiation site. To this confusion over end
points must be added the radiometric difficulties of accurately and reproducibly
measuring and delivering ultraviolet radiation of known irradiance and spectral
distribution. In addition to these difficulties in determining MED, many other
variables must be considered when evaluating any erythema response of human
skin to ultraviolet radiation. The most important of these are listed in Table 6-1,
which also defines a commonly used visual grading system of erythema. The de-
gree of erythema, timing of the peak response, and duration of erythema vary
with ultraviolet dose 67 and may be influenced by other factors, such as skin
temperature, 68 wind exposure, 69 and ambient humidity. 70
116 Chapter Six

Table 6-1. Variables Affecting Erythema Evoked by Ultraviolet Irradiation

The optical source

Wavelengths emitted
Irradiance, size of aperture, degree of collimation
Exposure
Distance from source
Duration
Field size ± shadow effects
Angle of incidence
Environment (heat, humidity, wind)
The skin
Skin color (pigmentation)
Previous light exposure
Anatomic site (thickness of stratum corneum, degree of vascularity, amount of hair)
Abnormal reactions (rare)
Presence of topical or systemic substance that may affect response
The end point
Definition of end point
Time of reading
Orthostatic effect (posture of the subject) during reading of MEDa
Grading of erythema:

Symbol Abbreviation Definition

o NR No reaction; identical to surrounding nonirradiated skin

± MPE Minimal perceptible erythema; blotchy areas of faint
erythema confined to the irradiated site; borders
indefinite
+ 1+ (MED) Minimal erythema with four sharp borders
++ 2+ More pronounced, even bright, erythema without edema
+++ 3+ Marked erythema with edema
++++ 4+ Violaceous erythema with vesiculation

a MED (minimal erythema dose) is an exposure dose of radiation, not a grade of erythema,
and is expressed in units of radiant exposure (J/cm 2 ) or exposure duration for a light
source of known irradiance.

The comparison of the erythemogenesis of various wavebands of elec-

tromagnetic radiation is compounded by the fact that there exist definite differ-
ences in the quality of the erythema produced by various ultraviolet wavelength
regions. 65 ,66,71 For example, erythema produced by UV-C (germicidal lamps,
254 nm) appears sooner, reaches its peak more quickly, and is of shorter duration
than that of UV-B. When fair-skinned persons are irradiated with 254 nm
sources, erythema reaches its maximum intensity between 5 and 8 hr after expo-
Effects of Ultraviolet Radiation on Skin 117

sure. * It takes up to twice this time before peak erythema is observed after
297 nm irradiation. 53.65,66,71-75 The color of the erythema produced by 254 nm
is more pink than red. Less energy is required to produce a minimal perceptible
erythema with 254 nm wavelengths than with 297 nm wavelengths, but much
larger doses do not proportionally increase the redness; that is, the dose-response
curve is relatively flat. Therefore, even though the MED of 254 nm may be lower
than at 297 nm, it takes less energy to induce moderate-to-strong erythema with
297 nm than with 254 nm wavelengths. 72.76,77 It takes more than 30 multiples of
the MED of 254 nm radiation to produce a 2+ erythema, 78 while as little as 10
times the MED of 297 nm radiation may produce 3+ or 4+ erythematous reac-
tions. Five times the MED of midday sun, which is polychromatic UV-B,
UV-A, and longer wavelengths, produces painful sunburn and blistering. 79
Any reasonable criterion for erythema shows that the longer-wavelength ul-
traviolet irradiation (UV -A) is much less erythemogenic than wavelengths short-
er than 320 nm. 64-66 However, the literature contains little information about the
parameters of delayed erythema produced by UV-A radiation of human skin.
K. W. Hausser and Vahle 64 . 75 first described the erythemogenic property of
UV-A in 1921. They compared the erythemogenic effects of 297, 313, 334,
365, 405, and 436 nm radiation on the skin of normal individuals. They found
the erythemal effectiveness of 365 nm radiation to be approximately 1% of that
of the maximally effective wavelength (297 nm). On the basis of evidence pre-
sented in 1935, the International Commission on Illumination stated that the
erythemal effectiveness of 365 nm radiation was 0.1 % of that of 297 nm.80 I.
Hausser later observed that when equal degrees of redness were produced with
300 nm and 365 nm, the 365 nm radiation had a shorter latent time and a less
steep dose-response curve to increasing multiples of radiation than did 300
nm. 81 Meyer and Seitz 82 believed that the site of action of UV -A radiation was

• As noted in Chapter 2, low-pressure mercury vaporlamps emit primarily at the spectral line of253.7
nm. Such radiation is a line spectrum, not a broadband or polychromatic source, and is the most often
used source of UV-C. The reactions to such radiation are often spoken of as synonymous with the
reactions to all radiation of UV -C wavelengths. Although not completely accurate, this assumption is
usually acceptable because: (l) the cutaneous effect of radiation in other waveband regions within
UV-C is approximately the same; (2) most of what is known about cutaneous and other biologic
reactions to UV -C is derived from low-pressure mercury lamps generating largely 254 nm; and (3)
UV-C to which humans may be exposed is usually from low-pressure mercury vapor lamps.
Similarly, 297 nm radiation has often been used to be representative of UV-B. This usage may
be less acceptable because the erythema action spectrum may vary considerably throughout the UV-B
range, and the UV -B in our natural environment is variable. Other writers use 250 nm and 300 nm to
be representative of or synonymous with UV-C and UV-B, respectively. Using 365 nm (a
strong spectral line of high-pressure mercury lamps) to represent UV-A may be an even less safe
assumption.
118 Chapter Six

Q)
u
c:
IV
2.0
.
"C
IV

...
,\
IV 1.5
u
II)
Co
1.0

\
1/1

..
II)
.~
IV
I

II) 0.5
a:

250
)
350
~ 450
J
550
J\
650 750
Nanometers

Fig. 6·3. Spectral energy distribution of UV -A fluorescent lamp source (see text).

the dermis. Miescher 56 first noted that the UV-A radiation caused damage to
dermal capillaries but caused little alteration in the epidermis.
Van der Leun 83 compiled information from older literature 64, 79,82 to derive
a UV-A action spectrum that, when compared to the erythemal efficiency of
300 nm radiation, gradually falls from about I % at 320 nm to 0.1 % at 365 nm
and less than 0.05% at 400 nm. He suggested that the site of action of UV-A-
induced erythema is the dermis. According to van der Leun, ultraviolet radiation
exerts two distinct erythemal actions: (1) a direct dermal effect with a smooth
action spectrum with diminishing effect from 250 to 400 nm; (2) a more effec-
tive, and often overriding, effect on the epidermis with a narrow wavelength re-
gion action spectrum peaking at 300 nm. The latter is postulated to be due to a
vasodilating mediator substance, which diffuses to the dermis to cause erythema.
Parrish et al. 84,85 have found the MED to polychromatic UV-A (320-380
nm) to be 10-50 J/cm 2 for fair-skinned Caucasians. With a monochromatic
source (337.1 nm laser), MED ranges of 10-30 J/cm 2 were found. 86 When
Juhlin irradiated 60 persons with 30 J/cm 2 of 365-nm radiation, he observed min-
imal erythema at 24 hr in 10 of the persons, suggesting a similar range of human
MED.87 Using a glass-filtered, high-pressure mercury arc lamp with highest
UV-A output at 365 nm, Tanenbaum et al. 88 found fair-skinned Caucasians to
have an MED for UV-A of 25 to 35 J/cm 2 • Willis and Cylus 89 found the average
MED of 320-400 nm radiation from a 1600-W xenon solar simulator to be 91
J/cm 2 ; after filtering to produce 350-370 nm radiation, the average MED was 26
J/cm 2 •
When 7 fair-skinned Caucasians (skin types I and II) were exposed through
a Mylar filter 6 inches from a fluorescent UV-A source (spectral irradiance given
Effects of Ultraviolet Radiation on Skin 119

in Fig. 6-3), the MED was 36.0 ± 8 J/cm 2 of 320-380 nm radiation. With a
400-W high-pressure mercury lamp filtered through glass, the MED for delayed
erythema in 15 fair-skinned Caucasians was 29 ± 5.2 J/cm 2 • In the same sub-
jects, the ratio of the UV-A MED to the UV-B MED was consistently about
1000:1 (Table 6-2).
It is most likely inaccurate to discuss uV -A erythema as a single entity be-
cause the erythema action spectrum probably differs throughout the 320-400 nm
waveband region. The amount of energy needed to produce erythema at 365 nm
is somewhat greater than that needed at 337.1 nm. It is probable that even larger
amounts of radiant exposure are required to produce erythema with wavelengths
longer than 380 nm. Exact exposure requirements will not be known until more
sophisticated high-intensity UV -A sources permit accurate extension of the
erythema action spectrum.
Exposure to polychromatic UV-A (30 mW/cm2 , 320-400 nm, with major
output at 365 nm) produced by a filtered high-pressure xenon mercury arc lamp
resulted in maximum delayed erythema 8 to 10 hr after exposure. 90 The erythema
that results from threshold doses of 337.1 nm nitrogen laser radiation follows a
similar time course to UV-B erythema. 86 Redness begins several hours after ir-

Table 6-2. Minimal Erythema Doses to UV-A (Primarily 365 nm) and UV-8 (280-315 nm)

Subject UV-A MED J/cm 2D UV-8 MED mJ/cm 2 b UV-A MED/UV-8 MED

1 24 34.8 0.69 (X 10 3 )
2 30 21.7 1.38
3 36 21.7 1.66
4 21 26.1 0.80
5 30 26.1 1.15
6 27 39.1 0.69
7 36 21.7 1.66
8 36 47.8 0.75
9 24 26.1 0.92
10 36 30.4 1.18
11 30 39.1 0.77
12 24 43.5 0_55
13 30 21.7 1.38
14 24 26.1 0.92
15 27 21.7 1.24
Average: 29 ± 5.2 29.9 ± 8.8 1.05 ± 0.36 (X 10")

DUV-A source: 400-W high-pressure mercury arc filtered through glass. Exposure doses in
table are those of the 365 nm UV-A radiation, delivered at approximately 10 mW/cm'.
Visible and infrared radiation also present.
bUV-B light source: Westinghouse FS40 fluorescent sunlamp bulbs mounted 15 cm apart
and 4.5 cm in front of a planar specular aluminum reflector. These lamps have a con-
tinuous output spectrum between 275 nm and 380 nm, with a broad peak at 313 nm_
Exposure doses in this table are those in the band 280-315 nm.
120 Chapter Six

radiation and has a broad peak between 12 and 24 hr after exposure. However,
exposure doses delivered at high average laser irradiances may produce im-
mediate erythema that may persist for hours, to the extent of blending into the
delayed erythema response. The presence of striking immediate erythema de-
pends on the irradiance, suggesting that local heating of the skin during exposure
may be involved in its production, although other mechanisms may also be in-
volved. At high average irradiance levels, immediate erythema may be caused by
total exposure doses lower than those required to produce delayed erythema.
However, the relationship between immediate and delayed erythema thresholds
is not predictable. Once present, immediate erythsma may appear to last longer
than 24 hr, but it is possible that the two classic phases of U V-induced vascular
change run together. The production of immediate erythema is probably a func-
tion of UV-A wavelength as well, since the fraction of energy absorbed and its
depth of penetration into the skin varies over the UV-A region (cf. Chapter 4).
Irradiation with conventional medium-pressure mercury arc sources of UV-A
that have most of their output at 365 nm may also result in an immediate, as well
as a delayed, erythema. At sufficiently high UV -A irradiances, subjects may feel
pain at the site of irradiation during the exposure, and a local rise in skin tempera-
ture can be documented. 86
Bucher 91 stated that for wavelengths greater than about 340 nm, delayed
erythema cannot be distinguished from the erythema produced by heating of the
skin during the exposure. This statement is not entirely true, since delayed
erythema may be produced by 365-nm radiation with no immediate erythemal
response if sufficiently low irradiances on fair-skinned subjects are employed.
However, the immediate erythema often produced by intense UV-A or other
radiation appears to be induced primarily by radiant heating of the skin. Shorter
visible (blue) wavelengths are somewhat more effective in producing immediate
erythema than longer wavelengths, 91 an effect presumably related to the fact that
over the range of 250 to 1300 nm, skin absorbs shorter wavelengths more than
longer wavelengths (cf. Chapter 4). The exact relationships between immediate
erythema and penetration, absorption, and wavelength are not known, and it is
possible that other factors contribute to the immediate erythemal response. In any
situation of high-irradiance exposures, as is often the case in UV-A photo-
biologic research, thermal effects must be considered.
In a study attempting to demonstrate photorecovery phenomena to ultra-
violet erythema (300 nm) by longwave ultraviolet and visible radiation (320-500
nm), van der Leun and Stoop92 observed that when 300-nm irradiation was pre-
ceded by exposure to 5 hr of sunlight filtered through window glass (,\ > 315
nm), the sunlight-irradiated sites exhibited an increased sensitivity to 300-nm
radiation. These investigators believed that the long-wavelength ultraviolet com-
ponents of filtered sunlight had erythemogenic properties that could be additive
to the erythema produced by 300 nm. More recently, Willis et ai., 93 using clini-
Effects of Ultraviolet Radiation on Skin 121

cal observation and histologic and autoradiographic techniques, evaluated the re-
sponses of human skin to UV-A, UV-A plus UV-B, and UV-B alone. Their ob-
servations indicated that UV-A radiation had a striking augmentative effect on
sunburn damage caused by UV-B. Compared to sites that were exposed to UV-B
alone, preirradiation of skin with UV -A increased the redness that occurred from
UV -B. This response was interpreted by the authors as a striking synergistic ef-
fect between UV-A and UV-B.
The notion of photoaddition was first suggested by Adams et at. in 1931.94
Sayre et at. 66 showed evidence of photoaddition phenomenon of 254 nm and
297 nm ultraviolet radiation. More recently, Ying et at. 85 found that in the
threshold ranges, the erythemogenic properties of UV-A and of UV-B were not
augmentative but were linearly additive. They used fractions of predetermined
minimal erythema doses to show that various combinations of suberythemal ex-
posures to UV-A and UV-B could be added linearly to produce erythema. One-
half MED to UV -A and one-half MED to UV -B given in any order caused mini-
mal perceptible erythema. When various incremental fractions of UV-A and

10 0
......
- ........
'\. ./
" SOLAR SPECTRUM
ERYTHEMA
10- 1 \ /
ACTION
SPECTRUM \ /
1\
/ \
10- 2
(/)
/ \
~

z /
::::l
w 10- 3
~
~
«
...J
w
a:: 10- 4

10- 5

10-6L-~__~__~__~~__~__~__~~__~__~~
260 280 300 320 340 360 380
WAVELENGTH,nm

Fig. 6-4. Erythema action spectrum, noontime solar spectral irradiance, and resultant spectral
erythemal effectiveness.
122 Chapter Six

UV-B erythema were used, erythema resulted only when the sum of UV-A
dose/MEDA and UV-B dose/MEDB was 1 or greater. Therefore, disagreement
exists as to whether UV-A radiation augments or simply adds to the erythema
produced by UV-B radiation alone.
The question of photoaddition or photosynergism between various
wavelength bands has immediate practical implications. U.S. governmental reg-
ulatory agencies have assumed photoaddition to be the case when proposing
safety standards for UV exposure 9S (see Chapter 11). Photoaddition is often as-
sumed during the administration of UV phototherapy treatments as well.
Moreover, photoaddition is assumed whenever a photobiologic action spectrum
is multiplied by a spectral irradiance curve of some source of radiation to predict
or explain an observed response. An excellent example is the erythemogenesis
produced by sunlight. If one weights solar noontime spectral irradiance by an
erythema action spectrum, the most effective wavelength for sunburn is approx-
imately 306 nm. The contribution of noontime solar wavelengths away from 306
nm falls rapidly (Fig. 6-4).
Even though UV -A is 200 to 2000 times less erythemogenically efficient
than UV-B, the role of UV-A in producing sunburn reactions cannot be ignored
because of the high UV -A irradiance present in solar radiation and because of the
existence of additive or augmentative erythemogenesis between UV-A and
UV-B radiation. Considering the solar UV-A irradiance to be 5 mW/cm 2 at
noontime and considering that approximately 20 min of solar exposure induces
delayed erythema in Caucasians, UV -A may be responsible for approximately
15% of the erythemally effective radiation at noon. While solar UV -B and UV-A
both decrease at times other than noon, UV-B is attenuated more than UV-A.
UV-A may therefore playa somewhat greater role than 15% in producing mini-
mal erythema at times other than noon. This role is apparent in Table 6-3, in
which the erythemogenic percentage of UV -A for 1 MED is calculated as a func-
tion of solar angle. The sunburn reaction that one observes after 2 or more hours
of sunbathing is due to the combined effects of UV-B and UV-A radiation and
should not be ascribed to the effects of UV-B alone. In addition, several of the
diseases and chemical influences that cause man to be abnormally sensitive to
ultraviolet radiation are precipitated or made worse by 320-400 nm radiation.
Persons with such disorders may be sensitive to UV-B or to UV-A or to a wide
spectrum of ultraviolet and visible light.
Van der Leun and Stoop92 have reported that the delayed erythema reactions
to UV -C and UV -B exposures are lessened if the UV exposures are followed by
hours of exposure to sunlight filtered through window glass to remove solar
UV-B. This phenomenon was termed "photorecovery" by the authors. The
erythemal reactions to UV-C were unchanged by exposure to filtered sunlight
prior to UV exposure, but in the case of UV-B, prior exposure to filtered sunlight
resulted in greater erythemal response. The increased erythemal response seen
 Table 6-3. Approximate Relative Erythemogenesis of Solar UV-B and UV-A Components as a Function of Solar Zenith
i
2-
Angle /the Angle of the Sun from Directly Overhead)Q c:

Solar zenith angle ~

o
0° 30° 50° 70° i
:II

UV-A spectral irradiance (at 350 nm, in mW/cm2 • nm) 0.08

!.
0.06 0.03 0.005
~
UV-B spectral irradiance (at 300 nm, in mW/cm 2 • 'nm) 7 X 10 41 4 X 10"" 8 X 10~ 3 X 10-6 g
Approximate total UV-A irradiance (at 320-400 nm, g
in mW/cm2 ) 5.1 3.6 1.8 0.3 en
!!!:
::I
Spectral irradiance ratio UV-A (350 nm) I UV-B (300 nm) 110 150 375 1650
Tilne for MED from solar UV-A alone (30 J/cm2 ) 1.6 hr 2.3 hr 4.6 hr
Time for MED from solar UV-B alone 16 min 34 min 3hr
Time for MED from solar UV assuming additivity of
UV-B and UV-A erythemogenesis

=~) 13.7 min 27.3 min 1.8 hr

( MED fA + fB

Percent of 1 MED due to solar UV-A 14% 25% 39%

Q Because of the dependence of solar UV irradiances upon many factors other than solar zenith angle, the figures of this table may be con-
sidered only as approximation for clear-day exposures of fair-skinned individuals. Calculated from Bener, P. The diurnal and annual variation
of spectral intensity of ultraviolet sky and global radiation on cloudless days at Davos, 1590 m.a.s.l. Air Force Contract AF61 (052)-618.
Technical Note No.2, Davos, January 1963.

.-
tl
124 Chapter Six

when UV-B exposure was preceded by filtered-sunlight exposure may be due to

the additive or synergistic effects of the solar UV -A exposure and the artificially
produced UV-B alone. Interestingly, such addition or synergism was not noted
with UV-C exposures. The "photorecovery" noted by van der Leun and Stoop
produced a change in the minimal erythema dose of only 20 to 40%, which is on
the order of the errors involved in determining the MED itself. The action spec-
trum for photorecovery of delayed erythema is unknown but presumably includes
UV-A and visible wavelengths. The authors also suggested that the exposure to
UV-A and visible wavelengths required to produce maximum photorecovery is
very small, on the order of the UV-B or UV-C exposure required to elicit the
erythema. It is not known whether "photorecovery" plays any significant role in
determining the delayed erythemal response, or whether this possible effect is
related mechanistically to the action of a photoreactivating enzyme.
Thus, it is possible that UV-A may initiate recovery from, add to, or aug-
ment the delayed erythemal responses to UV-B and UV-C, depending on ex-
perimental conditions and exposure doses.

Histology

Most histologic studies of the ultraviolet erythemal response describe

changes induced by UV-B and UV-C. 96.97 The histologic response of normal
skin to erythemal 'doses of UV-B has been studied in rats, mice, and guinea
pigs. 8 Exposure doses used in these studies were considerably greater than the
minimal erythema dose. At 6 to 8 hr, scattered "sunburn cells" appear in the
epidermis. These cells, characterized by homogeneous eosinophilic cytoplasm
and unrecognizable or pyknotic nuclei, are usually surrounded by normal-
appearing cells. Although they are characteristically found in sunburn, similar
cells are seen in skin altered by other processes. At 14 to 20 hr after exposure,
foci of frank necrosis are present in the epidermis and are often invaded by
leukocytes. In the dermis, a perivascular infiltrate of lymphocytes and polymor-
phonuclear leukocytes can be detected as early as 30 min after UV-B exposure.
This inflammatory response reportedly peaks at 8 to 24 hr in these animals. At 30
hr, the repair process begins, manifested by mitoses in the basal layer. Human
skin demonstrates a somewhat similar sequence of changes. 67
The histologic effects of UV-A irradiation have not received much atten-
tion. In 1957, Miescher 56 first noted that UV -A radiation causes changes princi-
pally in the dermis, with evidence of capillary injury and endothelial cell ne-
crosis. The epidermal cytotoxic and disruptive changes noted with UV-B radia-
tion were much less evident with UV-A. Willis et al. 93 noted no histologic
changes in skin exposed to 30 min of UV-A from a xenon source, but the dose
was suberythemogenic. Stem 9 7 compared the histologic effects of U V-B, UV -C,
Effects of Ultraviolet Radiation on Skin 125

and UV-A and concluded that in all instances the common denominator was vas-
cular injury. However, he used nonerythemogenic doses of UV-A and unequal
erythema for UV-C and UV-B, and he biopsied only at 24 hr after exposure.
Willis and Cylus 89 compared histologic changes present in adult humans after
exposure to UV-B (xenon solar simulator 290-400 nm) and various wavebands
of UV -A (same light source with various filters). Sites irradiated with 2 to 4
times the measured MED were biopsied at 48 hr. The degree of redness was not
mentioned. Skin exposed to UV-B showed epidermal changes consisting of sun-
burn cells, vesiculation, and liquefaction degeneration of the basal layer. A
perivascular infiltrate of lymphocytes and histiocytes was present in the upper
dermis. The epidermis of the UV-A-irradiated sites remained normal, while the
dermis showed similar or more impressive perivascular lymphohistiocytic
in filtrate.
In comparing histologic changes at 24 hr postradiation of equally erythemo-
genic doses of UV-B and nitrogen laser (337 nm) UV-A in guinea-pig and
human skin, Parrish et al. 57 noted that the UV-A-induced changes occurring in
the dermis are similar to those occurring from U V-B but that there is considera-
bly less epidermal alteration. Individual keratinocyte dyskeratosis is less fre-
quent, and there is less evidence of cytotoxicity and disruption of orderly epi-
dermal cell proliferation.
Rosario et al. 98 compared the histologic changes occurring after radiation
with UV-A, UV-B, and UV-C by serial observations at 1,2,3, and 7 days after
exposure. UV-A exposures were studied with and without prior systemic ad-
ministration of the potent photosensitizer methoxsalen. By varying the exposure
dose of each source, equal ultraviolet-induced delayed redness (2+ by a visual
grading scale, Table 6-1) was achieved at each test site. Many histologic
parameters showed reproducible changes when correlated with time after expo-
sure, but because they were uniformly caused by exposure to all wavelengths,
they were not of great value in discriminating between changes induced by
UV-B, UV-C, and UV-A with and without methoxsalen. Within the epidermis,
hyperkeratosis, parakeratosis, and acanthosis appeared concurrently. These were
absent for all wavelengths at 24 and 48 hr and appeared in UV-B and UV-C at 72
hr and in methoxsalen plus UV-A at 7 days. They never appeared in UV-A and
were most striking in UV-c. The granular layer was focally absent with UV-B
and UV -C at 24 hr and 48 hr and focally increased with all wavelengths at 72 hr
and 7 days. Intracellular edema of moderate degree was noted for all wavelengths
at all time intervals. Vacuolization of the basal layer was noted in moderate de-
gree at all time intervals and wavelengths.
In general, the shorter UV wavelengths, UV-C and UV-B, affected the
epidermis in greater degree than the longer wavelengths. The latter affected the
dermis in greater degree at all times. Nucleoli of keratinocytes were normal 24 hr
after all exposures. They increased in size 48 to 72 hr after UV-B and UV-C
126 Chapter Six

exposure and returned to normal by 7 days. For UV-A, particularly after ad-
ministration of methoxsalen, nucleoli were enlarged at 7 days after exposure. At
all postexposure times, dyskeratosis was greater from UV-B and UV-C than
from UV-A with or without the photosensitizer. At 24 to 48 hr in UV-B- and
UV-C-irradiated sites and after 72 hr in UV-A-irradiated sites, dyskeratotic cells
("sunburn cells") were scattered throughout the midepidermis. UV-A elicited
the greatest degree and depth of dermal inflammatory cell infiltrate at 1, 2, and 3
days after exposure. At 7 days, UV-A plus methoxsalen.showed the most im-
pressive infiltrate.
Of the time intervals used, maximum histologic changes of both epidermis
and dermis were present at 48 to 72 hr for UV-A, UV-C, and UV-B, while for
UV-A plus methoxsalen, maximum changes were seen at 7 days. The time re-
quired for the skin to complete its histologic response to a single ultraviolet expo-
sure was least with UV-B and greatest with methoxsalen plus UV-A. For UV-A,
the time for completion is unknown as the histologic changes were still obvious
at 7 days.
Alterations of skin induced by single and repeated UVA exposures were
examined with the electron microscope by Kumakiri et al. 99 The superficial ves-
sels of the papillary dermis showed widely open endothelial gaps and extravasa-
tion of blood cells. Extravascular fibrin deposition was noted in the reticular
dermis. Multiple exposures led to the formation of cross-banded filamentous
aggregations in and around vessels throughout the dermis. Similar changes were
not produced by solar simulator or UV -B exposures. The presence of immediate
erytQema in the subjects and the high irradiance used make it impossible to rule
out heat effects influencing the ultrastructural changes.

UV and Epidermal Macromolecular Synthesis

Germinative cells in tissues such as the epidermis proliferate by synthesiz-

ing twice their normal complement of deoxyribonucleic acid (DNA) and other
cellular macromolecules and then dividing into two daughter cells (mitosis).
Either the daughter cells differentiate, mature, and die, or they reenter the ger-
minative cycle by resuming DNA synthesis before proceeding to mitosis. This
cycle has been divided into four phases: S, the period of active, scheduled nuc-
lear DNA synthesis; G2 , the interval from the end of DNA synthesis to the begin-
ning of mitosis; M, the period of mitosis; and G 1 , the interval from completion of
mitosis to the beginning of DNA synthesis (S phase). The term unscheduled DNA
synthesis (UDS) refers to autoradiographically detectable DNA synthesis occur-
ring apart from S phase and is thought to represent DNA repair processes.
UV radiation produces a biphasic response in the rate of DNA, RNA, and
protein synthesis by the cells of the epidermis. There is an initial transient de-
crease followed by a sustained increase above normal rates. The extent and tim-
Effects of Ultraviolet Radiation on Skin 127

ing of these changes vary with the wavelength of radiation, the system studied,
and the presence or absence of photosensitizers. Most studies of the effects of
UV radiation on cell kinetics have concerned UV_B100-104 and PUVA (Psoralen
plus UV-A, UV-A radiation of skin previously sensitized with Psoralen). 105-1 08
There are few data on the effects of UV-A alone.
The skin of albino mice was exposed to 450 mJ/crri 2 of UV-B and biopsied
at intervals up to 7 days after radiation. 104 Prior to each biopsy, mice were in-
jected with a tritium-labeled precursor of DNA (thymidine), RNA (cytidine), or
protein (histidine or methionine). The sectioned biopsy material was applied to
photographic emulsion for up to 4 weeks before staining and examination. In the
nucleic acid studies, the number of nuclei containing silver grains was expressed
as a fraction of the total number of nuclei. Nuclei containing more than 5 silver
grains after [3H] thymidine' injection were considered to represent cells undergo-
ing scheduled DNA synthesis (S phase). A quantitative assessment was not pos-
sible in the case ofthe amino acid precursors. By the use of this autoradiographic
technique, it was found that the rate of synthesis of nucleic acids and protein fell
within 3 hr after irradiation but by 24 hr had recovered, partially (RNA and pro-
tein) or completely (DNA). At 48 hr, scheduled DNA synthesis was 6 to 7 times
control levels, and at 7 days, a significant increase was still apparent. A similar
stimulation of RNA and protein synthesis was evident 72 hr after exposure.
With the use of colchicine to arrest dividing cells at metaphase, the mitotic
rate or number of cells entering mitosis per hour could be measured at intervals
after irradiation. The mitotic rate fell sharply by 1 hr, remained depressed for 7
hr, and returned to normal by 24 hr. By 72 hr, the rate was significantly increased
over un irradiated controls. Thus, two processes of the epidermal cell cycle are
interrupted soon after UV -B irradiation, DNA synthesis and mitosis. In addition,
0 1 and O 2 resting phases may be extended by depression of RNA and protein
synthesis, blocking entrance into S phase or mitosis. The subsequent acceleration
of mitotic and synthetic activity may account for the epidermal acanthosis and
hyperplasia seen several days after UV exposure. These observations in mice are
consistent with more limited data available from human subjects. 100.101, 103
Autoradiographic studies with PUVA, using topical 8-methoxypsoralen on
hairless mice, have, in general, furnished results with some similarities to those
from UV -B. 108 Scheduled DNA synthesis decreased within 2 hr after exposure,
but by 48 hr it was significantly higher than control levels and was still elevated
at 7 days. In contrast to the findings from UV-B, unscheduled DNA synthesis
(UDS), manifested by sparse autoradiographic labeling, was not seen in PUVA-
treated skin. At the doses used in this study, RNA synthesis was not inhibited by
PUVA, and protein synthesis was discontinued only in the upper epidermis be-
tween 36 and 48 hr after irradiation. In studies with human fibroblasts, scheduled
DNA synthesis was reduced to 10% of controls within minutes of UV -A irradia-
tion of methoxsalen-sensitized cells. 106 A small amount of UDS was noted.
Willis et al. 93 examined the clinical, histologic, and autoradiographic re-
128 Chapter Six

sponse of human skin to UV-B, UV-A, and the combination. UV-B with or
without UV -A produced a biphasic response of scheduled DNA synthesis, as de-
scribed above, and extensive UDS. UV-A alone was associated with a transient
increase in scheduled DNA synthesis at 20 and 48 hr and a minimal increase of
UDS. Protein synthesis, reduced by UV-B, was unaffected by UV-A. Willis et
al. ' s data are difficult to interpret because equally erythemogenic doses of UV-A
and UV-B were not used.

Effects of UV-A on Mucous Membrane

The literature on this subject is minimal. Payne I 09 irradiated the mucous

membrane of the lip with a xenon arc solar simulator and found that compared
with relatively untanned skin, the lip mucosa was slightly more sensitive to
UV-B. Parrish llo found that irradiation of the oral labial mucosa with a light
source emitting intense UV-A produced no erythema with doses of 30 to 40
J/cm 2 , the dose usually producing erythema on untanned human skin. Thus, it
appears clinically that the oral mucosa is not substantially more sensitive than
untanned skin to either UV-B or UV-A, in spite of the relative absence ofprotec-
tion due to the lack of the cutaneous stratum corneum and relative decrease in
melanin pigmentation.
Lampert and Loew III observed sequential histologic changes in the oral
mucosa of albino mice after exposure of the mucosa to unfiltered medium-
pressure mercury arc radiation. Hyperemia and extravasation of red blood cells
occurred within 24 hr after exposure, and hyperemia, edema, and vacuolization
of epithelial cells were evident 2 and 3 days after exposure, similar to the histol-
ogy of UV-exposed skin. Histologic appearance returned to normal by 7 days
after exposure. The source of radiation was a lamp from a dental-filling photo-
polymerization system (Nuva-Lite). Because the authors did not determine the
minimal erythemal dose of this source on human skin, it is difficult to evaluate
the exposures given to the mice in terms of human erythemal effectiveness. The
authors stated that there is no evidence of injury to the mucosa as the dental in-
strument is normally used (protective UV -A transmissive filter in place).

Tanning

Constitutive skin color in man is that "baseline" color that develops in the
absence of exposure to solar radiation or other environmental influences and that
results from genetically determined levels of melanin pigmentation. Facultative
skin color or "tan" is that increase of melanin pigmentation above the constitu-
tive level that is induced by ultraviolet radiation II 2 or by pituitary hormones such
 Table 64. Responses of Human Melanocytes to Ultraviolet Radiation a

UV-irradiated skin (single exposure)

Light Electron Unirradiated
microscopy microscopy skin Immediate tanning (IT) Delayed tanning (DT)

Dopa reaction Weak. No definite change. Markedly increased.

Number of melanocytes No change. Increased, but variable.
Perikaryon Small. Small. Markedly enlarged.
Dendrites Poorly developed. Well developed. Well developed.
Nucleus shape Oval, round. Indented. Markedly indented.
Chromatin Heterochromatin granules Increased heterochromatin Increased euchromatin.
in the periphery; sparse granules; no change in
euchromatin. euchromatin.
Nucleolus Few in number, small. No increase in number, but Increased in number and en-
slightly enlarged in size. larged in size; occasional
segreation of nucleolar
components.
Melanosomes Few. No change; possibly Markedly increased.
(number) decreased (?)
Stages of formation In pigmented skin, mostly Larger proportion of mel- Melanosomes in various stages,
of melanosomes melanized stages, but in anized melanosomes (?) particularly in melanized
light skin, unmelanized stages.
stages.
Distribution pattern Perinuclear. Dendrites (predominantly). Perinuclear areas and dendrites.
IOO-A filaments Dense, perinuclear Dense aggregation in Diffusely scattered in perikaryon
aggregation. dendrites, and melanosomes and dendrites.
interspersed between fila-
ments.
Continued
 Table 6-4. Continued

UV-irradiated skin (single exposure)

Light Electron Unirradiated
microscopy microscopy skin Immediate tanning IITI Delayed tanning (DT)

Golgi apparatus Poorly developed. Poorly developed. Well developed; marked increase
in size and number.
Rough endoplasmic Poorly developed. Poorly developed. Well developed.
reticulum
Free ribosomes A few. A few. Abundant.
Microtubules Perinuclear area. Dendrites. Perinuclear area and dendrites.
Autophagic vacuole Absent. Absent. Present.
Lipid droplet Absent. Absent. Present.
Basal lamina below Monolayered. Multilayered. Multilayered.
melanocyte

a FroID Quevedo etal., pp. 176_177. 112

Effects of Ultraviolet Radiation on Skin 131

as MSH or ACTH. The facultative tanning (darkening) response induced by solar

exposures can be divided into immediate tanning (IT), which occurs within min-
utes after exposure to sunlight, and delayed tanning (DT), which becomes evi-
dent several days after exposure (Table 6-4).

Immediate Tanning (IT)

IT, also called immediate pigment darkening (IPD) or the Meirowsky

phenomenon, is best seen in pigmented individuals. In general, the darker the
unexposed, baseline, inherited coloration (constitutive skin color), the greater the
ability to exhibit IT. Previously exposed or tanned (DT) skin usually yields more
impressive IT than does unexposed skin. Individual's capabilities to demonstrate
IT parallels that of DT: those persons genetically best able to suntan show the
most striking IT. Fair-skinned, easily sunburned individuals who suntan poorly
may have poor or absent IT.
Within 5 to 10 min of exposure to noonday sun, a gradual darkening is
noticed that is confined to exposed skin. If exposure continues, darkening in-
creases until it reaches its maximum, at about I hr. The color is light brown to
dark brown or may be gray-brown to black in the more deeply pigmented races.
Brief exposures to sunlight may lead to slight to moderate IT, which begins to
fade within one-half hour after discontinuation of sun exposure and is barely
noticeable after 3 to 8 hr. Prolonged sun exposure or high-intensity artificial
longwave ultraviolet radiation sources can lead to striking IT that lasts longer
than 36 hr.
IT is optimally produced by UV -A but is also produced by visible
light. 113.114 The function, if any, of IT is not certain. Although protection from
ultraviolet irradiation appears the most likely reason for darkening immediate-
ly after sun exposure, studies to document such protection have not been con-
vincing.
The mechanism of IT reaction is believed to involve:
1. Photooxidation of preformed melanin. 43 ,115 The result is increased
melanization of melanosomes. Studies with electron spin resonance
spectroscopy115 have shown that IT is most likely an oxidation reaction
that involves the generation of unstable semiquinonelike free radicals in
melanin. Radiation from 320 to 700 nm elicits such radicals. 43
2. Changes in distribution pattern of melanosomes within basal and sup-
rabasal keratinocytes.
3. Changes in the distribution pattern of micro filaments and microtubules
of melanocytes. Absorption of 340-640 nm radiation causes shifting
and dispersion of micro filaments and microtubules from the perinuclear
region of the melanocyte to the periphery of the cytoplasm and leads to
prominence of dendritic processes. 116
 4. Movement of melanosomes from perikaryon to dendrites.
5. Increase in transfer of melanosomes from melanocytes to keratinocytes.
No changes in the size of melanosomes or in the number of melanocytes
have been noted to accompany IT. 115.11 6

Delayed Tanning (DT)

Delayed tanning (DT), also referred to as melanogenesis, may be noticeable

beginning about 72 hr after exposure to ultraviolet radiation, although electron-
microscopic studies show ultrastructural evidence of melanogenesis beginning
much earlier. The major action spectrum of DT is the same as that for sunburn
(UV-B), but DT can also follow exposure to UV-C 7I • 117 and to UV-A and blue
visible light. 67 Melanogenesis is the formation of new melanosomes and mela-
nin. The mechanisms involve the following, any or all of which may be pres-
ent l15 :
1. Increase in the number of functioning melanocytes; this may result
from proliferation of existing pigment cells or from activation of previ-
ously undetectable dormant melanocytes.
2. Increase in size of melanocytes and increased arborization of dendrites.
3. Increased melanosome synthesis within the melanocytes.
4. Increased melanization of melanosomes.
5. Increase in size of melanosomes.
6. Increased enzyme (tyrosinase) activity due both to an increase in
amount of enzyme synthesized in melanocytes and possibly to enzyme
activation as an effect of radiation of sulfhydryl compounds that may
inhibit tyrosinase.
7. Increased transfer of melanosomes from melanocytes to keratino-
cytes. 112
Dendritic processes of the melanocytes, within the basal layer of the
epidermis project between epidermal cells, and pigmented organelles called
meianosomes, produced within the melanocytes, are transferred from these den-
dritic processes into the keratinocytes. One melanocyte transfers melanosomes to
approximately 36 keratinocytes in its vicinity. 112 Melanosomes may occur as
discrete, dark particles or as aggregates of three or more particles within a mem-
brane analogous to a lysosome. This state of aggregation of melanosomes is at
least partly dependent upon racial (genetic) factors (Fig. 6-5).
Ultraviolet irradiation can modify the distribution pattern of melanosomes in
keratinocytes. In Mongoloids, repeated sunlight exposures result in a predomi-
nance of nonaggregated, single melanosomes within keratinocytes, whereas in
the keratinocytes of habitually unexposed skin, there is a predominance of aggre-
Effects of Ultraviolet Radiation on Skin 133

CAUCASOID NEGROID
KERATINOCYTE KERATINOCYTE

(4)
MELANOSOME

DEGRADATION

:~
,.
III
II

• MELANOSOME
(1)

0 FORMATION

EPIDERMAL EPIDERMAL
MELANOCYTE MELANOCYTE

Fig. 6-5. Biologic processes underlying melanin pigmentation and racial differences in aggregation
state of melanosomes. Nonaggregated melanosomes (right. Negroid) are more stable than aggregated
melanosomes (left, Caucasoid). (From Quevedo, W. C., Jr., Fitzpatrick, T. B., Pathak, M. A., and
Jimbow, K. Light and skin color. In Sunlight and Man: Normal and Abnormal Photobiologic Re-
sponses, M. A. Pathak, L. C. Harber, M. Seiji, and A. Kukita, Eds.; T. B. Fitzpatrick, Consulting
Ed. Copyright, University of Tokyo Press, Tokyo, 1974. Used with permission.)

gated melanosomes. 112 The interpretations of variations in melanosome "pack-

aging" within the epidermis must take into account a history of exposure of indi-
viduals to solar radiation II 2.11 8 and other factors. 119 In Caucasians, alteration of
melanosome aggregation may be induced by UV-A in the presence of a photo-
sensitizer. Application of 4,5' ,8-trimethylpsoralen to the unexposed buttocks of
a Caucasoid subject and a single exposure to UV-A resulted in the appearance of
nonaggregated or single melanosomes in keratinocytes, 120 while the adjacent un-
exposed buttock skin of the same subject showed aggregated melanosomes.
Epidermal melanin absorbs over a broad spectral region (see Chapter 4),
thus decreasing the amount of ultraviolet radiation that reaches the lower portions
of the skin. The presence of melanin in the stratum corneum may significantly
134 Chapter Six

K
DT

IT

I~~

j - I I I 11111 I I 11111
1.0 10 100

2
J/em laser exposure dose: 337.1 nm nitrogen pulsed laser

Fig. 6-6. Radiant exposure dose ranges for biologic phenomena in II fair-skinned Caucasians ex-
posed to 337.I-nm pulsed nitrogen gas laser. A = Average values;~= range of values; DT =
delayed tanning; IT = immediate tanning ("immediate pigment darkening"). (Modified from Par-
rish, J. A., Anderson, R. R., Ying, C. Y., and Pathak, M. A. Cutaneous effects of pulsed nitrogen
gas laser irradiation. J.lnvest. Dermatol. 67:603-608, 1976.)

decrease the amount of UV -B and UV -A that reaches the viable keratinocytes or

penetrates into the dermis to strike blood vessels (Chapter 4). As constitutive or
facultative pigmentation increases, the MED to UV-B increases.
UV-A induces DT but, as with erythema, it takes much more energy with
UV-A than with UV-B. IT, on the other hand, is caused by UV-A but apparently
not by UV-B. Within the UV-A spectral region, slightly less energy may be re-
quired to cause DT or IT than is required to cause erythema, but the amount re-
quired varies from subject to subject and is also dependent on constitutive and
facultative pigmentation. Darker persons may consistently show IT or DT with
much less energy than is required to produce delayed erythema. Part of this ap-
parent difference in relative energy requirements may be because erythema is
difficult to appreciate through darkly pigmented skin. In fair-skinned persons, the
differences are smaller (Fig. 6-6).

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Effects of Ultraviolet Radiation on Skin 137

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69. Owens, D. W., Knox, J. M., Hudson, H. T., and Troll, D. Influence of wind on ultraviolet
injury. Arch. Dermatol. 109:200-201, 1974.
70. Owens, D. W., Knox, J. M., Hudson, H. T., and Troll, D. Influence of humidity on ul-
traviolet injury. J. Invest. Dermatol. 64:250-252, 1975.
71. Breit, R., and Kligman, A. M. Measurement of erythemal and pigmentary responses to ul-
traviolet radiution of different spectral qualities. In The Biologic Effects of Ultraviolet Radiation
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72. Everett, M. A., Olson, R. L., and Sayre, R. M. Ultraviolet erythema. Arch. Dermatol.
92:713-719,1965.
73. Bachem, A. Time factors of erythema and pigmentation, produced by ultraviolet rays of dif-
ferent wavelengths. J. Invest. Dermatol. 25:215-218, 1955.
74. Olson, R. L., Sayre, R. M., and Everett, M. A. Effect of anatomic location and time on ul-
traviolet erythema. Arch. Dermatol. 93:211-215, 1966.
75. Hausser, K. W., and Vahle, W. Die Abhangigkeit des Lichterythems und der Pigmentbildung
von der Schwingungszahl (Wellenlange) der erregenden Strahlung. Strahlentherapie 13 :41-71,
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138 Chapter Six

76. Rottier, P. B. The ratio of the 260/300 nm MED's in the judgement of skin sensitivity to short
wave ultraviolet radiation. In The Biologic Effects of Ultraviolet Radiation (with Emphasis on
the Skin) (F. Urbach, Ed.). Pergamon, Oxford, 1969, pp. 187-195.
77. Adams, E. Q., Barnes, B. T., and Forsythe, W. E. fIber die Erythemwirksamkeit ultraviolet-
ten Lichtes. Strahlentherapie 48:235-249, 1933.
78. Parrish, J. A. Expanding the clinical spectrum of sunscreen testing. Cutis 8:498-506,1971.
79. Luckiesh, M. Application of Germicidal Erythemal and Infrared Energy. Van Nostrand, New
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81. Hausser, I. Uber spezifische Wirkungen des langwelligen ultravioletten Lichts auf die
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82. Meyer, A. E. H., and Seitz, E. O. Ultraviolette Strahlen. de Gruyter, Berlin, 1949, pp. 227-
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83. van der Leun, J. C. Theory of ultraviolet erythema. Photochem. Photobiol. 4:453-458, 1965.
84. Parrish, J. A., Ying, C Y., Pathak, M. A., and Fitzpatrick, T. B. Erythemogenic properties
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86. Parrish, J. A., Anderson, R. R., Ying, C. Y., and Pathak, M. A. Cutaneous effects of pulsed
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88. Tanenbaum, L., Parrish, J. A., Pathak, M. A., Anderson, R. R., and Fitzpatrick, T. B. Tar
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90. Rosario, R., and Shea, S. Unpublished observations.
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93. Willis, I., Kligman, A., and Epstein, J. Effects of long ultraviolet rays on human skin: Photo-
protective or photoaugmentative? J. Invest. Dermatol. 59:416-420, 1972.
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95. Criteria for Recommended Standard-Occupational Exposure to Ultraviolet Radiation. US-
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96. Stem, W. K. Anatomic localization of the response to ultraviolet radiation in human skin.
Dermatologica 145:361-370, 1972.
97. Stem, W. K., and Urbach, F. The diagnostic significance of minimal erythemal dose. Arch.
Dermatol. 105:387-393, 1972.
98. Rosario, R., Mark, G. J., Parrish, J. A., and Mihm, M. C., Jr. Histologic changes produced
in skin by equally erythemogenic doses of UV-A, UV-B, UV-C, and UV-A with psoralens.
Submitted for publication.
99. Kumakiri, M., Hashimoto, K., and Willis, I. Biologic changes due to long-wave ultraviolet
irradiation on human skin: Ultrastructural study. J. Invest. Dermatol. 69:392-400, 1977.
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100. Baden, H. P., and Pearlman, C. The effect of ultraviolet light on protein and nucleic acid
synthesis in the epidermis. J. Invest. Dermatol. 48:71-75, 1964.
101. Fukuyama, K., Epstein, W. L., and Epstein, J. H. Effect of ultraviolet light on RNA and pro-
tein synthesis in differentiated epidermal cells. Nature 216: 1031-1032, 1967.
102. Epstein, J. H., Fukuyama, K., and Epstein, W. L. UVL induced stimulation of DNA synthesis
in hairless mouse epidermis. J. Invest. Dermatol. 51 :445-453, 1968.
103. Epstein, W. L., Fukuyama, K., and Epstein, J. H. Early effects of ultraviolet light on DNA
synthesis in human skin in vivo. Arch. Dermatol. 100:84-89, 1969.
104. Epstein, J. H., Fukuyama, K., and Fye, K. Effects of ultraviolet radiation on the mitotic cycle
and DNA, RNA and protein synthesis in mammalian epidermis in vivo. Photochem. Photobiol.
12:57-65, 1970.
105. Trosko, J. E., and Isoun, M. Photosensitizing effects of trisoralen on DNA synthesis in human
cells grown in vitro. Int. J. Radiat. BioI. 19:87-92, 1971.
106. Baden, H. P., Parrington, J. M., Delhanty, J. D. A., and Pathak, M. A. DNA synthesis in
normal and xeroderma pigmentosum fibroblasts following treatment with 8-methoxypsoralen
and longwave ultraviolet light. Biochem. Biophys. Acta 262:247-255, 1972.
107. Walter, J. F., Voorhees, J. J., Kelsey, W. H., and Duell, E. A. Psoralen plus black light in-
hibits epidermal DNA synthesis. Arch. Dermatol. /07:861-865, 1973.
108. Epstein, J. H., and Fukuyama, K. Effects of 8-methoxypsoralen-induced phototoxic effects on
mammalian epidermal macromolecular synthesis in vivo. Photochem. Photobiol. 21 :325-330,
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109. Payne, F. T. An evaluation of actinic blocking agents for the protection of lip mucosa. J. Am.
Dent. Assoc. 92:409-411, 1976.
110. Parrish, J. A. Unpublished observations, 1974.
III. Lampert, F., and Loew. R. Physicalische und biologische Pnifung des Nuva-Lite-Genites.
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112. Quevedo, W. C., Jr., Fitzpatrick, T. B., Pathak, M. A., and Jimbow, K. Light and skin color.
In Sunlight and Man: Normal and Abnormal Photobiologic Responses (M. A. Pathak, L. C.
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113. Pathak, M. A., Riley, F. J., Fitzpatrick, T. B., and Curwen, W. L. Melanin formation in
human skin induced by long-wave ultraviolet and visible light. Nature /93:148-150, 1962.
114. Pathak, M. A. Photobiology of melanogenesis: Biophysical aspects. In Advances in Biology of
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tern of micro filaments and microtubules and on the nucleus in human melanocytes. Yale J. BioI.
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117. Parrish, J. A., Pathak, M. A., and Fitzpatrick, T. B. Topical protection against germicidal
radiation. Arch. Surg. 104:276-283, 1972.
118. Konrad, K., and Wolff, K. Hyperpigmentation, melanosome size, and distribution patterns of
melanosomes. Arch. Dermatol. 107:853-860, 1973.
119. Flaxman, B. A., Sosis, A. c., and Van Scott, E. J. Changes in melanosome distribution in
Caucasoid skin following topical application of nitrogen mustard. J. Invest. Dermato!'
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tion of racial differences in melanosome distribution in human epidermis after exposure to ul-
traviolet light. Nature [New Biol.l236:143-145, 1972.
CHAPTER 7

Adverse Cutaneous Reactions to UV-A

Introduction

Defining the action spectrum of light-induced or light-aggravated disease has

theoretical importance for at least two reasons. First, this information, by indicat-
ing likely chromophores, may lead to an understanding of etiology and
pathogenesis. A classical example of such a sequence was the identification of
erythropoietic protoporphyria 1 and its separation from the polymorphous light
eruption group. Second, photoprotective agents ideally are selected by matching
the action spectrum of the skin disease with the absorption spectrum of the sun-
screen.
Currently, however, a thorough classi fication of photodermatoses according
to action spectrum is impossible because the abnormal reaction may be provoked
by a wide and variable spectral range that often does not clearly correspond to a
particular chromophore. Clinical and histologic appearance of photodermatoses
is generally the means of classification and diagnosis.
The abnormal cutaneous reactions in which UV-A may playa part are:
Chemical photosensitivity (phototoxicity and photoallergy)
Persistent light reaction
Actinic reticuloid
Polymorphous light eruption
Solar urticaria
Porphyrias and other endogenous photosensitization syndromes
Melasma
Ephelides

Chemical Photosensitivity

This term embraces all forms of photosensitivity resulting from excitation of

an identified chemical by electromagnetic radiation. There are many mechanisms
141
142 Chapter Seven

by which compounds may cause photosensitivity to various wavelengths of radia-

tion. A wide variety of photosensitizing chemicals of therapeutic, industrial, ag-
ricultural, or other origin may reach the skin directly or via the bloodstream, 2
each having its own pattern of absorption, metabolism, and binding to skin com-
ponents. In general, these compounds possess highly resonant structures with a
molecular weight of less than 500 and absorb radiation in the ultraviolet and visi-
ble range. Table 7-1 lists the more common compounds. Penetration of radiation
into the skin depends largely on the wavelengths of radiation and on epidermal
thickness and melanization (see Chapter 4). In addition, radiation and chemicals
may have direct effects when acting alone. Host factors compound all these vari-
ables.
Chemical photosensitivity has been traditionally divided into two
clinicopathological classes, phototoxicity and photoallergy. The features distin-
guishing these two classes are summarized in Table 7-2 and Fig. 7-1. In practice,
however, there is often considerable difficulty in categorizing an individual
chemical photosensitization reaction on human skin. Although chemical photo-
sensitization is a subject of great theoretical interest, it occupies a very small part
of the day-to-day clinical experience of the practicing dermatologist.

Phototoxicity

Light-induced damage that is not dependent on an allergic mechanism may

be considered phototoxic in nature. Theoretically, these reactions will occur in
everybody if the skin is exposed to enough radiant energy of the proper
wavelengths and if enough of the photosensitizing molecules are present.

Table 7-1. Common Chemical Photosensitizers

Antimicrobial agents
Tetracyclines, especially demeclocycline
Sulfonamides, especially sulfanilamide
Griseofulvin
Halogenated salicylanilides

Other drugs
Phenothiazines, especially chlorpromazine
Thiazides
Psoralens
Sulfonylureas

Others
Sunscreens
Tar
Cosmetics (due to presence of eosin, psoralens, or antimicrobial agents)
Adverse Cutaneous Reactions to UV-A 143

Table 7-2. Characteristics of Drug-Induced Photosensitivity a

Reaction Phototoxic Photoallergic

Reaction possible on first exposure. Yes No

Incubation period necessary after
first exposure. No Yes
Chemical alteration of photosensitizer. No Yes
Covalent binding with macromolecule. Yes Yes
Clinical changes. Usually like sunburn Varied morphology
(Le., erythema, edema,
vesiculation)
"Flares" at distant, previously
involved sites possible. No Yes
Can persistent light reaction develop? No Yes
Cross-reactions to structurally related
agents. Infrequent Frequent
Broadening of cross-reactions follow-
ing repeated photopatch testing. No Possible
Relative concentration of drug High Low
necessary for reaction.
Incidence. Usually relatively high Usually very low (but
(theoretically 100%) theoretically could
reach 100%)
Action spectrum. Usually similar to Usually higher wave-
absorption spectrum length than absorption
spectrum
Passive transfer. No Possible
Lymphocyte stimulation test. No Possible
Macrophage migration inhibition test. No Possible

a From Harber, L. C., and Baer, R. L. Pathogenic mechanisms of drug·induced photo-

sensitivity. J. Invest. De;'matol. 58:327-339, 1972. Copyright, The Williams & Wilkins
Company, Baltimore, 1972. Used with permission.

The term photodynamic action was introduced to describe the in vitro

oxygen-dependent killing of paramecia by the synergistic action of acridine and
light. 3 The term was adopted by clinicians and was used by some as synonymous
with phototoxicity 4 but by others to refer to a particular clinical pattern of photo-
toxicity characterized by immediate wheal formation. The term is best dropped
from discussions of clinical aspects of photosensitivity and is currently used by
photobiologists to describe effects (usually destructive) that require a photosen-
sitizer, optical radiation, and oxygen.
144 Chapter Seven

PHOTOSENSITIZER + LIGHT
ENERGY ABSORPTION
EXCITATION

PHOTOTOXIC PHOTOALLERGIC

ENERGY FREE RADICAL FORMATION

TRANSFER AND/OR AND/OR OXIDATION

FREE RADICAL, NEW HAPTENE FORMATION

PEROXIDE AND
HEAT FORMATION
HAPTENE-PROTEIN COMBINATION

NUCLEAR SENSITIZATION VIA IMMUNO-

CYTOPLASM OR COMPETENT CELLS (ICC)
CELL MEMBRANE
ALTERATION PHOTOANTIGEN-ICC INT[RACTION

~ CELL DAMAGE - - - - - - - -
Fig. 7-\. Classification of phototoxic and photo allergic reactions. (From Harber, L. c., and Baer,
R. L. Pathogenic mechanisms of drug-induced photosensitivity. J. Invest. Dermatol. 58:327-339,
1972. Copyright, The Williams & Wilkins Company, Baltimore, 1972. Used with permission.)

A phototoxic response generally has the clinical appearance of sunburn.

Depending on the severity of the reaction, erythema, edema, and vesiculation,
followed by desquamation and hyperpigmentation, are seen in the exposed areas.
The reaction occurs upon simultaneous exposure to a phototoxic chemical and
radiation of the appropriate wavelength. The latent interval (time between expo-
sure and beginning of reaction) may be brief (hours), but phototoxic erythema
may not peak for 2 to 3 days after exposure. In theory, 100% of persons exposed
to the proper combination of chemical and radiution can react, and the severity of
the reaction is in proportion to the dose of both chemical and radiation. Histolog-
ically, the reaction is predominantly epidermal, showing intracellular edema and
death of keratinocytes. Compared to the histologic pattern seen in photoallergy,
the dermis appears to be relatively spared. The phototoxic effect in a given tissue
layer is dependent upon both the penetration and absorption of radiation within
the layer and the distribution of the specific phototoxic chemical. Nearly all
phototoxic reactions are produced by UV-A. An exception is vinblastine, in
which radiation in the UV-B range evokes the response. 5
Adverse Cutaneous Reactions to UV-A 145

Mechanisms of Phototoxicity

Radiation may be absorbed by the phototoxic molecule itself, by some

metabolite of this substance, or by complexes that these may form with cellular
molecules or organelles. The electronic excited singlet- and triplet-state
molecules or complexes may then undergo photochemical reactions with DNA,
RNA, proteins, or membranes. Free radicals may be formed that attack other
molecules, resulting in damage to lysosomal or cell membranes and nuclear and
mitochondrial injury. Peroxides may be formed that then oxidize the substrate.
Other possibilities include transfer of energy from the excited chemical to a
biologic substrate, which may then become oxidized, or the activated chemical
may be able to accept electrons, resulting in direct oxidation of the substrate. The
photosensitizing molecules may return to the ground state structurally unchanged
and induce further photoreactions as long as they are associated with the sub-
strate. Alternatively, some photosensitizing molecules or complexes may be
bound to the substrate or photochemically altered so that they are no longer
photosensitizing.
Differences in the clinical patterns of response to various photo toxic agents
may reflect differences in sites of primary damage and pathogenetic mechanisms.
However, the complexity of the chain of events that begins with photochemical
excitation and alteration of some molecules and leads to an observable response
often makes it hard to interpret the response in terms of the mechanisms or sites
of the initial photochemical damage.

Psoralen Phototoxicity (PUVA)

Two groups of UV -A phototoxic compounds, psoralens and chlor-

promazine, have been studied in some detail in an attempt to understand
mechanisms of phototoxicity. Psoralens in particular have been subjected to con-
siderable study because of their marked photoxicity and because of both the
harmful (phytophotodermatitis, berloque dermatitis) and beneficial (photo-
chemotherapy) effects of psoralen photo toxicity . The primary mechanism for
psoralen phototoxicity is thought to be a direct photochemical reaction of psora-
len with DNA, producing binding of psoralen and thymine, the formation of in-
terstrand crosslinks, 6- 8 and subsequent inhibition of DNA synthesis. 9-11 The ac-
tion spectrum for psoralen phototoxicity has been shown to lie in the UV-A
range. 12-15 The exposure dose necessary to elicit phototoxicity manifested by
delayed erythema depends on the particular psoralen compound and the route of
administration. The mechanisms and uses of psoralen photosensitization in
photochemotherapy of skin disorders are described in Chapter 10.
146 Chapter Seven

Chlorpromazine (CPZ) Phototoxicity

Phototoxicity has been studied in vitro using the photohemolysis system and
in the killing of mouse peritoneal macrophages. 16 When a medium containing
CPZ was irradiated and added to these systems, hemolysis of red cells and death
of macrophages resulted without further irradiation. It was proposed that a stable
photoproduct of CPZ was cytotoxic on the plasma membrane. There was no evi-
dence of damage to lysosomal membranes.
Tritium-labeled CPZ has been shown to form stable photoproducts with nu-
cleic acids, purines, and pyrimidines after irradiation with UV-A.17 CPZ-
nucleic acid photoproducts were fonned much more rapidly with single-stranded
than with double-stranded nucleic acid. It was suggested, therefore, that CPZ
photosensitization involved interaction with RNA rather than with DNA. Photo-
products were fonned on irradiation with adenine, thymine, uracil, and cytosine,
but not with guanine.
Using singlet oxygen quenchers and the deuterium effect in the photo-
hemolysis system, Nilsson et al. I B concluded that CPZ photosensitization, al-
though oxygen-dependent, did not act by a singlet oxygen pathway. Phototoxic-
ity has been quantified in vivo using the weight gain of tails of mice following
treatment with radiation and chemicals. 19 The tissue edema that followed expo-
sure caused measurable weight gain of the tails, which correlated with the sever-
ity of the phototoxic reaction. In addition to CPZ itself, five metabolites (includ-
ing two demethylated derivatives, a sulfoxide and a hydroxylate) and two photo-
products (the sulfoxide and the promazine) were studied. All were found to be
phototoxic, and the two demethylated derivatives were more so than the parent
CPZ.

Other Chemical Photosensitizers

Anthracene, porphyrin, neutral red, and acridine were shown to be taken up

by lysosomes in living cells in vitro. 20 When irradiated with 360-400 nm light,
lysosomal damage was demonstratable by acid phosphatase staining. After a
further interval, microscopic evidence of cell damage and death was apparent,
with cytoplasmic vacuolation and pyknotic nuclei. The same chemicals produce
photodynamic damage to cell membranes, as demonstrated in the photo-
hemolysis system.

Quantification of Phototoxic Response

The minimum exposure dose required to produce a photo toxic reaction

under a given set of conditions may be used to quantify and compare the poten-
cies of phototoxic compounds. The end point of erythema produced at the time of
Adverse Cutaneous Reactions to UV-A 147

maximum response is generally used, and the lowest exposure dose necessary to
elicit this erythema is called the minimum photo toxic dose (MPD). The MPD is
used to quantify the variables involved in phototoxicity reactions:
1. Concentration of phototoxic compound
2. Method of administration or application
3. Time between administration and radiant exposure
4. Methods of exposure (wavelength, size of exposure site, irradiance,
and exposure dose)
5. Variations among individuals due to pigmentation and other variables
Understanding these variables is the key to use of phototoxic reactions in photo-
chemotherapy (see Chapter 10) and to consideration of UV-A exposure safety
criteria in the presence of phototoxic compounds (Chapter 11). The MPD is
meaningful only when adequate information is supplied. MPD should be a spec-
trally defined exposure dose determined when all of the other variables listed
above are constant.
The ratio between MPD and MED (minimal erythema dose, without ad-
ministration of phototoxic compounds) is called the photo toxic index (PI) and is a
useful parameter for comparing the potencies of photo toxic compounds. 21 The PI
is used in order to minimize the effects of individual differences between sub-
jects. For example, comparison of the phototoxicity and action spectra of topical
tar preparations used in phototherapy of psoriasis has been compared using the
PI.
The advantage of using erythema as a clinical end point is that it is a nonin-
vasive, readily observed response. However, it is nonspecific and is produced by
a variety of mediators or stimuli (see Chapter 6). The degree of erythema pro-
duced in photo toxic reactions often correlates with damage seen histologically,
but absolute correlation cannot be assumed. It is therefore wise to combine clini-
cal, histologic, metabolic, and other means of evaluating phototoxic reactions.

Photoallergy

Photoallergy is photosensitivity mediated by immunologic pathways. Clini-

cally, it resembles allergic contact dermatitis and may have an urticarial, papular,
or eczematous morphology. The distribution, although primarily on light-
exposed areas, may extend onto covered areas of skin. An incubation period after
exposure to a chemical and subsequent repeated exposures to UV radiation or
light are required before a response is elicited. The response is not linearly dose-
related. The histologic pattern is not specific but differs from that seen in photo-
toxicity. Epidermal spongiosis and vesicle formation may accompany an ec-
zematous clinical response. Characteristically, there is a dense perivascular
round-cell infiltrate in the dermis.
148 Chapter Seven

Agents that have been reported to cause photoallergy 22 include most of

those listed above as phototoxins: sulfonamides, sulfonylureas, thiazides, and
phenothiazines. Others are the halogenated salicylanilides, antifungal agents,
cyclamates, and substituted benzoic acids. Methoxsalen has been reported to
cause photoallergy,23 but the phenomenon must be extremely rare in view of
subsequent infrequent reports. As with phototoxic agents, these chemicals are
resonating molecules. If administered in adequate amounts, all photoallergens
may produce a phototoxic response.
Many mechanisms have been proposed to account for the photo allergic re-
sponse 22 . 24 :
1. Radiation may alter the photosensitizing molecule. The photoproduct
(hapten) then combines with a protein carrier to form an antigen. If the
photoproduct is unstable, antigens may not be formed unless the carrier
protein is in apposition to the photosensitizer at the time of irradiation.
2. Radiation may alter a tissue protein, enabling it to act as a carrier for
either the photosensitizer itself or its photoproduct. This carrier-hapten
complex is then antigenic.
3. Radiant energy in the presence of a photosensitizer alters a tissue com-
ponent in such a way that the latter becomes a tissue antigen.
The halogenated salicylanilides, used as antibacterial agents in soaps and
cosmetics, were the cause of an epidemic of photo allergic reactions in the
1960s. 25 - 27 One member of this group of chemicals, 3,3',4' ,5-tetrachloro-
salicylanilide (TCSA), has been intensively studied as an experimental
model. 28,29 Longwave ultraviolet radiation of salicylanilides in solution resulted
in changes in absorption spectra and promoted firm binding to skin and other pro-
teins. Modified immunodiffusion and immunofluorescent techniques detected no
humoral immunoglobulin antibodies against either the salicylanilide or its
hapten-protein complex, and no contact or photocontact sensitivity could be in-
duced in recipients by transfer of serum from TCSA-sensitive animals. The mac-
rophage migration inhibition factor and the lymphocyte transformation test were
used as in vitro methods to demonstrate a cell-mediated immune mechanism re-
sponsible for the salicylanilide photocontact or photoallergic dermatitis. Evi-
dence has also been presented for the role of TCSA as both a contact and a photo-
contact sensitizer and for the necessity of both hapten and carrier protein in anti-
gen function.
Kochevar and Harber29 studied the photoreactions of TCSA with human
serum and bovine globulin. In the dark, TCSA anion binds noncovalently with
albumin but not with globulin. Subsequent irradiation with UV-A yields stable
covalent TCSA-albumin photoproducts. Both TCSA and albumin were altered
in this photochemical reaction, TCSA by sequential dechlorination (to TriCSA
Adverse Cutaneous Reactions to UV-A 149

and DiCSA) and albumin by a 15% reduction of its histidine content. From this
model, the following conclusions and speculations were made:
1. Not all proteins can act as the carrier for TCSA to form a complete anti-
gen.
2. Noncovalent binding in the dark is a necessary preliminary step, since
the photoexcited state of TCSA is very short-lived, and close proximity
of TCSA with protein is essential for TCSA-protein covalent photo ad-
duct formation.
3. Sequential dechlorination of TCSA, resulting in salicylanilide deriva-
tives having 3, 2, or 1 chlorines may account for the clinical observa-
tion of cross-reactivity between members of this group of compounds.
4. Alteration of a tissue protein may playa role in the development of per-
sistent light reactivity (see below).
The action spectrum of almost all of the photoallergenic compounds studied
lies in the UV-A range. 3U Two exceptions, in which the response appears to be
UV -B-activated, are photoallergy to dephenhydramine and triacetyldiphenolisa-
tin.31

Persistent Light Reactivity

A small proportion of patients suffering chemical photosensitivity reactions

remain photosensitive despite careful avoidance of the offending compound. The
term persistent light reactor was coined 32 to describe such individuals. Retention
in the dermis of small amounts of the photosensitizer has been proposed as the
mechanism for this phenomenon. 33 However, in some patients a different
pathogenesis is likely, since the action spectrum is considerably wider 34 ,35 than
is typically seen in photoallergy and extends from 290 nm through UV-A to visi-
ble light.

Actinic Reticuloid

This is a condition 36 of elderly males who commonly have a long history of

recurrent dermatoses (atopic, seborrheic, or contact dermatitis). A relationship to
persistent light reactivity has been proposed. Morphologically, the eruption is
that of a chronic lichenified dermatitis associated with plaques and papules in
light-exposed areas and commonly on the back of the neck. The eruption may
extend onto the covered areas. Photosensitivity may be exquisite, but the con-
tribution of light to the disease may not be appreciated by the patient or his physi-
150 Chapter Seven

cian because of the indirect relationship between exposure and symptoms. His-
tologically, the appearance is that of a chronic dermatitis in some parts of which a
dense dermal lymphomatoid infiltrate may be present. Progression to reticulum
cell sarcoma or mycosis fungoides has been described.
The action spectrum has not been accurately defined but includes the UV-A
and visible portions of the spectrum. 36 •37

Polymorphous Light Eruption

This common photodermatitis of unknown etiology presents, as the name

suggests, a variety of morphological apperances (erythema, eczema, papules,
plaques, blisters) on light-exposed areas. The onset is usually in adolescence or
early adult life and first appears 4 to 24 hr after sun exposure in spring. The erup-
tion subsides within 2 to 8 days of avoiding sunlight. The severity of sun-induced
eruptions tends to subside during summer ("hardening") but recurs each spring
with little tendency to remit. The action spectrum in most cases lies in the 290 to
320 nm range. UV-A is, however, responsible for a small minority of cases. 38
Indeed, there is evidence suggesting that some polymorphous light eruption is
not wavelength-dependent. Possibly any radiation (including X-ray and infrared)
that produces a delayed erythema response can reproduce the eruption. 39.40

Solar Urticaria

This condition consists of wheals appearing within seconds or minutes of

exposure to sunlight. Solar urticaria is evidently a syndrome that may result from
several different pathogenic mechanisms. Several classifications have been of-
fered 41.42 based on action spectra and ability to transfer the phenomenon with the
patient's serum to a normal individual. The latter procedure is now considered
unacceptable because of the danger of transferring viral or other potentially
harmful biologic matter to the recipient. This factor and the relative rarity of solar
urticaria result in no generally satisfactory classification of the disease. Ive et
al. 42 divide solar urticaria into four groups according to action spectrum: (1)
UV-B, (2) UV-A and visible, (3) visible, and (4) broadband (250-700 nm).

Porphyrias and Other Endogenous Photosensitization

Syndromes

Cutaneous photosensitization may be a feature of all the porphyrias except

acute intermittent porphyria. These syndromes are the only well-known exam-
Adverse Cutaneous Reactions to UV-A 151

pIes of photosensitization by identified endogenous chemicals (protoporphyrin,

uroporphyrin I, coproporphyrin I). The action spectrum 43, 44 conforms closely to
the absorption spectrum of porphyrins (380-440 nm, with a sharp peak near 400
nm, the Soret band).
Photosensitivity is a feature of several syndromes in which an absolute or
relative deficiency of niacin appears to be the central biochemical feature. These
syndromes appear as pellagra, as Hartnup syndrome, as carcinoid, and as an oc-
casional complication of Isoniazid therapy. The action spectrum and pathogene-
sis are, however, unknown. 45

Melasma and Ephelides 46

These are extremely common entities of circumscribed hyperpigmentation

in which UV-A as well as UV-B initiates and maintains excessive melanocyte
activity. Melasma. also known as chloasma, is a sharply outlined, confluent,
brown hyperpigmentation of the forehead, cheeks, upper lip, and chin, usually
occurring in women and most commonly associated with pregnancy or the use of
oral estrogens. It is more common and severe in dark-skinned individuals.
Ephelides are freckles. The importance of UV-A and visible light in the
pathogenesis of these conditions is evident when treatment is undertaken with
hydroquinone, a topical depigmenting agent. Unless appropriate sunscreens are
used, progress is negligible, since minimal sunlight exposure sustains the en-
hanced melanocyte activity. UV-B sunscreens are ineffective, and sunscreens
that block UV -B, UV -A, and visible light (titanium dioxide and the ben-
zophenones) are an essential aspect of therapy.

Management of UV-A-Induced Dermatoses

In broad terms, diagnosis involves identification of the action spectrum and

of possible photosensitizing chemicals, endogenous (porphyrins) or exogenous.

History

The most useful single point to be gained from the history is whether the
eruption can be produced by sunlight that has passed through window glass. If
so, one may conclude that the action spectrum includes UV-A or visible
wavelengths. A common presentation is that of a patient who observes that his
eruption evolved on sun-exposed areas while he was driving with all car windows
closed.
The history may be pathognomonic (as in erythropoietic protoporphyria). In
152 Chapter Seven

other cases, the timing of eruption with respect to light exposure may have diag-
nostic value, as in solar urticaria (minutes) or the phototoxic reaction (hours).

Examination

The eruption may have diagnostic morphologic features, as in porphyria

cutanea tarda. If, as is commonly the case, the rash has subsided, provocation by
exposure to sun or artificial sources designed for "phototesting" may be attemp-
ted.

Laboratory Findings

Microscopic examination and immunofluorescent studies of histologic

material from involved skin should be performed. Such tests are seldom diagnos-
tic but may be helpful in ruling out, for example, lupus erythematosus or in dis-
tinguishing between polymorphous light eruption and a photoallergic reaction,
each of which has characteristic if not diagnostic features. A complete blood
count, an antinuclear factor assay, and a urine or blood examination for presence
of abnormal amounts or types of porphyrins are essential.

Phototesting

Photopatch testing 47 for photoallergy involves placing suspected photocon-

tactants on the skin in a suitable concentration and vehicle. These areas are then
irradiated separately with UV-A and UV-B or simulated sunlight. Control sites
with test chemicals are shielded from irradiation, and untreated areas are also
irradiated as controls. The test areas are examined at 24 hr, at 48 hr, and, ideally,
1 week after irradiation. A positive response (eczematous) in an irradiated area is
diagnostic of photoallergy. Controls include negative reactions on sites exposed
to chemical alone and to light alone.
In theory, photodermatoses can be reproduced by exposure of the patient's
skin to an artificial light source that emits light of the appropriate wavelengths. In
practice, this procedure is not always rewarding, and reliance is most often
placed on the history or response to sunlight filtered through window glass.
The reason for difficulty in reproducing lesions using artificial lamps is un-
clear. Theoretically, the skin should respond similarly to photons of given
wavelengths whether from the sun or from an artificial source. The variables of
exposure site, site size, irradiance (dose-rate) as well as exposure dose, and
synergism between various wavelengths should be considered. Psychological
variables, cyclic responses, or environmental factors may also be involved. One
valuable approach is to use a large-area solar simulator, from which various
spectral regions such as UV-B, UV-A, or visible light may also be selected using
Adverse Cutaneous Reactions to UV·A 153

optical filters. Even so, it is sometimes impossible to reproduce the abnormal

response using artificially produced "sunlight" over a large area in a region of
the body that normally receives sunlight exposure. A monochromator is generally
not required for phototesting but is a valuable research tool for determining ac-
tion spectra of photodermatoses.

Treatment

Identified exogenous photo sensitizers are eliminated if possible. Exposure

to UV-A is minimized by appropriate clothing, timing of activities to avoid the
midday sun, and, if necessary, changing artificial light sources 35 at home or at
work. Topical sunscreens that absorb the harmful wavelengths may also be used.
There is currently no completely satisfactory sunscreen that protects against
UV-A and the blue-violet end of the visible spectrum. The benzophenones ab-
sorb well into the UV-A range 48 but suffer the common defect of most sun-
screens, namely, lack of substantivity (binding power to the skin). Thus, protec-
tion is lost when the sunscreen is washed off by normal activities involving
sweating or swimming; frequent reapplication is necessary. Opaque sunscreens
(titanium dioxide, zinc oxide) are effective, but few patients can be persuaded to
apply them with any regularity even when they are formulated with appropriate
skin tones.
The use of oral beta carotene to diminish photosensitivity in erythropoietic
protoporphyria 4 4 has provided impetus to the search for other oral sunscreens.
Beta carotene has been used empirically in other UV -A-provoked diseases
(polymorphous light eruption) with some success. Psoralens plus UV-A have
proved effective in controlling the symptoms in some patients with polymor-
phous light eruption.

References

I. Magnus, I. A., Jarrett, A., Prankero, T. A., and Rimington, C. Erythropoietic protoporphyria:
A new porphyria syndrome with solar urticaria due to protoporphyrinaemia. Lancet 2 :448-451,
1961.
2. Pathak, M. A., and Epstein, j. h. light plus exogenous agent. In Dermatology in General
Medicine (T. B. Fitzpatrick, K. A. Arndt, W. H. Clark, Jr., A. Z. Eisen, E. J. Van Scott, and
J. H. Vaughan, Eds.). McGraw-Hili, New York. 1971, pp. 1009-1021.
3. Raab, O. Z. Uber die Wirkung fluorescierender Stoffe auf infusorien. Z. Bioi. 39:524-546,
1900.
4. Epstein, S. Photoallergy and primary phototoxicity to sulfanilamide. J. Invest. Dermatol.
2:43-51, 1939.
5. Breza, T. S., Halprin, K. M., and Taylor, J. R. Photosensitivity reaction to vinblastine. Arch.
Dermatol. 111:1168-1170, 1975.
154 Chapter Seven

6. Pathak, M. A., and Kramer, D. M. Photosensitization of skin in vivo by furocoumarins (psora-

lens). Biochim. Biophys. Acta 195:197-206, 1969.
7. Cole, R. S. Light-induced cross-linking of DNA in the presence of a furocoumarin (psoralen).
Biochim. Biophys. Acta 217:30-39, 1970.
8. Dall' Acqua, F., Marciani, S., Ciavatta, L., and Rodighiero, G. Formation of interstrand cross-
links in the photoreactions between furocoumarins and DNA. Z. Naturforsch. Teil B 26:561-
569, 1971.
9. Baden, H. P., Parrington, J. M., Delhanty, J. D. A., and Pathak, M. A. DNA synthesis in
normal and xeroderma pigmentosum fibroblasts following treatment with 8-methoxypsoralen
and longwave ultraviolet light. Biochim. Biophys. Acta 262:249-255, 1972.
10. Epstein, J. H., and Fukuyama, K. A study of 8-methoxypsoralen-induced phototoxic effects on
mammalian epidermal macromolecule synthesis in vivo. Photochem. Photobiol. 21 :325-330,
1975.
II. Walter, J. F., Voorhees, J. J., Kelsey, W. H., and Duell, E. A. Psoralen plus black light in-
hibits epidermal DNA synthesis. Arch. Dermatol. 107:861-865, 1973.
12. Owens, D. W., Glicksman, J. M., Freeman, R. G., and Carnes, R. Biologic action spectra of
8-methoxypsoralen determined by monochromatic light. J. Invest. Dermatol. 51 :435-440,
1968.
13. Pathak, M. A. Mechanism of psoralen photosensitization and in vivo biological action spectrum
of 8-methoxypsoraien. J.lnvest. Dermatol. 37:397-407, 1961.
14. Nakayama, Y., Morikawa, F., Fukuda, M., Hamano, M., Toda, K., and Pathak, M. A. Mono-
chromatic radiation and its application-Laboratory studies on the mechanism of erythema and
pigmentation induced by psoralens. In Sunlight and Man: Normal and Abnormal Photobiologic
Responses (M. A. Pathak, L. C. Harber, M. Seiji, and A. Kukita, Eds.; T. B. Fitzpatrick, Con-
sulting Ed.). University of Tokyo Press, Tokyo, 1974, pp. 591-611.
15. Buck, H. W., Magnus, I. A., and Porter, A. D. The action spectrum of 8-methoxypsoralen for
erythema in human skin. Preliminary studies with a monochromator. Br. J. Dermatol. 72:249-
255, 1960.
16. Johnson, B. E. Cellular mechanisms of chlorpromazine photosensitivity. Proc. R. Soc. Med.
67:871-873, 1974.
17. Kahn, G., and Davis, B. P.ln vitro studies of longwave ultraviolet light-dependent reactions of
the skin photosensitizer chlorpromazine with nucleic acids, purines, and pyrimidines. J. Invest.
Dermatol. 55:47-52, 1970.
18. Nilsson, R., Swanbeck, G., and Wennersten, G. Primary mechanisms of erythrocyte photolysis
induced by biologic sensitizers and phototoxic drugs. Photochem. Photohiol. 22:183-186,
1975.
19. Ljunggren, B., and Muller, H. Phenothiazine phototoxicity. An experimental study on chlor-
promazine and its metabolites. J. Invest. Dermatol. 68:313-317, 1977.
20. Allison, A. C., Magnus, I. A., and Young, M. R. Role of Iysosomes and of cell membrane in
photosensitization. Nature 209:874-878, 1966.
21. Tanenbaum, L., Parrish, J. A., Pathak, M. A., Anderson, R. R., and Fitzpatrick, T. B. Tar
phototoxicity and phototherapy for psoriasis. Arch. Dermatol. 111 :467-470, 1975.
22. Harber, L. c., and Baer, R. L. Pathogenic mechanisms of drug-induced photosensitivity. 1. In-
vest. Dermatol. 58:327-341, 1972.
23. Fulton, J. E., and Willis, I. Photoallergy to methoxsalen. Arch. Dermatol. 98:445-450, 1968.
24. Storck, H. Photoallergy and photosensitivity. Arch. Dermatol. 9/:469-482,1965.
25. Wilkinson, D. S. Photodermatitis due to tetrachlorosalicylanilide. Br. J. Dermatol. 73:213-
219, 1961.
26. Calnan, C. D., Harman, R. R., and Wells, G. C. Photodermatitis from soaps. Br. Med. J.
5262:1266, 1961.
Adverse Cutaneous Reactions to UV·A 155

27. Harber, L. C., Targovnik, S. F., and Baer, R. L. Contact photosensitivity patterns to haloge-
nated salicylanilides in man and guinea pigs. Arch. Dermatol. 96:646-656, 1967.
28. Herman, P. S., and Sams, W. M. Soap Photodermatitis. Charles C Thomas, Springfield, Il-
linois, 1972.
29. Kochevar, I. E., and Harber, L. D. Phmtoreactions of 3,3' ,4' ,5-tetrachlorosalicylanilide with
proteins. J.Invest. Dermatol. 68:151-156, 1977.
30. Epstein, J. H. Phototoxicity and photoallergy: Clinical syndromes. In Sunlight and Man: Nor-
mal and Abnormal Photobiologic Responses. (M. A. Pathak, L. C. Harber, M. Seiji, and A.,
Kukita, Eds.; T. B. Fitzpatrick, Consulting Ed.). University of Tokyo Press, Tokyo, 1974, pp.
459-478.
31. Emmett, E. A. Diphenhydramine photoallergy. Arch. Dermatol. 110:249-252, 1974.
32. Jillson, O. F., and Baughman, R. D. Contact photodermatitis from bithionol. Arch. Dermatol.
88:409-418, 1963.
33. Willis, I., and Kligman, A. M. The mechanism of the persistent light reactor. J. !nvest. Der-
matol. 51:385-394,1968.
34. Epstein, J. H., Wuepper, K. D., and Maibach, H. I. Photocontact dermatitis to halogenated
salicylanilides and related compounds. Arch. Dermatol. 97:236-244, 1968.
35. Brown, S., Lane, P. R., and Magnus, I. A. Skin photosensitivity from fluorescent lighting. Br.
J. Dermatol. 81:420-428,1969.
36. Ive, F. A., Magnus, I. A., Warin, R. P., and Wilson Jones, E. "Actinic reticuloid": A chronic
dermatosis associated with severe photosensitivity and the histological resemblance to lym-
phoma. Br. J. Dermatol. 81:469-485,1969.
37. Menter, M. A., McKerron, R. A., and Amos, H. E. Actinic reticuloid: An immunological in-
vestigation providing evidence of basement membrane damage. Br. J. Dermatol. 90:507-515,
1974.
38. Frain-Bell, W., Dickson, A., Herd, J., and Sturrock, I. The action spectrum in polymorphous
light eruption. Br. J. Dermatol. 89:243-249, 1973.
39. Epstein, S. Studies on abnormal human sensitivity to light. II. Light sensitivity in prurigo aes-
tivalis, eczema solare, and urticaria photogenica. J. Invest. Dermatol. 5:225-241, 194.2..
40. Epstein, J. H. Polymorphous light eruptions: Wavelength dependency and energy studies. Arch.
Dermatol. 85:82-88, 1962.
41. Epstein, J. H. Adverse cutaneous ceactions to the sun. In Yearbook of Dermatology (F. D. Mal-
kinson und R. W. Pearson, Eds.). Year Book Medical Publishers, Chicago, 1971, pp. 5-43.
42. Ive, H., Lloyd, J., and Magnus, I. A. Action spectra in idiopathic solar urticaria. br. J. Der-
matol. 77:229-248, 1965.
43. Magnus, I. A., Porter, A. D., and Rimington, C. The action spectrum for skin lesions in por-
phyria cutanea tarda. Lancet 1 :912, 1959.
44. Mathews-Roth, M., Pathak, M. A., Fitzpatrick, T. B., Harber, L. C., and Kass, E. H. Beta-
carotene as a photoprotective agent in erythropoietic protoporphyria. N. Engl. J. Med.
282:1231-1234, 1970.
45. Wiskemann, A. Abnormal action spectra. In The Biologic Effects of Ultraviolet Radiation (with
Emphasis on the Skin) (F. Urbach, Ed.). Pergamon Press, Oxford, 1969, p. 197.
46. Pathak, M. A., and Epstein, J. H. Freckles (ephelides). In Dermatology in General Medicine
(T. B. Fitzpatrick, K. A. Arndt, W. H. Clark, Jr., A. Z. Eisen, E. J. Van Scott, andJ. H. Vau-
ghan, Eds.). McGraw-Hill, New York, 1971, p. 999.
47. Harber, L. c., Harris, H., and Baer, R. L. Photoallergic contact dermatitis due to halogenated
salicylanilides and related compounds. Arch Dermato!' 94:255-262, 1966.
48. Parrish, J. A., Pathak, M. A., and Fitzpatrick, T. B. Prevention of unintentional overexposure
in topical psoralen treatment of vitiligo. Arch. Dermatol. /04:281-283, 1971.
CHAPTER 8

Skin Aging and Carcinogenesis

due to Ultraviolet Radiation

Introduction

Skin cancers are unquestionably the most common malignant tumors of man.
They are also among the least studied spontaneous human neoplasms, in part
because the most frequently detected types rarely lead to death. The major types
of skin cancer are basal cell carcinoma, squamous cell carcinoma, and malignant
melanoma.
Basal cell carcinoma is an epithelial tumor of the skin that usually arises
from cells of the basal cell layer of the epidermis or the hair follicles. It is most
commonly seen clinically as a small, waxy, smooth nodule that tends to ulcerate
in the center and often shows a few small telangiectatic vessels on its surface. It
invades locally but rarely metastasizes.
Squamous cell carcinoma is derived from the prickle or squamous cells of
the epidermis and is more commonly seen as a shallow ulcer surrounded by a
wide, elevated, and indurated border. Sometimes the lesion is not ulcerated and
may be seen as keratotic or verrucous nodules. It may metastasize to regional
lymph nodes and from there to the other organs, but metastasis occurs late and in
less than 5% of squamous cell carcinomas.
Malignant melanoma is derived from the cutaneous melanocyte, the pig-
ment cell of the epidermis. In appearance, it is usually a pigmented (black, gray,
blue, brown, or red) lesion, which may be flat or elevated, eroded or ulcerated.
Not infrequently, "satellite" lesions are present around the periphery or a white
halo may be seen. It may metastasize widely through lymph and blood vessels to
other organs of the body.
Blum (1959),1 Urbach et al. (1959, 1972),2,3 and most recently Emmett
(1973) I have reviewed the evidence supporting the role of sunlight in the de-

157
158 Chapter Eight

velopment of basal cell carcinoma and squamous cell carcinoma in humans.

Briefly, the main arguments are:
1. It is clearly established that superficial skin cancers occur most fre-
quently on the head, neck, arms, and hands-parts of the body habitu-
ally exposed to sunlight. 4,5
2. Pigmented races sunburn much less readily than people with white skin
and have much less skin cancer. In pigmented races, skin cancer affects
areas not exposed to sunlight more frequently than exposed sites. 5
3. Among Caucasians, there appears to be much greater incidence of skin
cancer in those who spend more time outdoors than in those who work
predominantly indoors.'
4. Skin cancer is more common in white-skinned people living in areas
where sun exposure is greater. 5
5. Genetic diseases that increase skin sensitivity to the effects of solar
UV-B radiation are also associated with marked increases and prema-
ture skin cancer development (albinism, xeroderma pigmentosum). 5
6. Superficial skin cancers, particularly squamous cell carcinoma of the
skin, occur predominantly on the areas receiving the maximum
amounts of solar UV radiation, where histologic changes of chronic
UV damage are most severe. 3
7. Skin cancer can be produced readily on the skin of mice and rats with
repeated doses of UV radiation, and the upper wavelength limit of the
effective cancer-producing radiation is about 320 nm, which is the same
spectral range that produces delayed erythema in human skin. 1

Incidence of Skin Cancers in Man

Surveys of the incidence of skin cancer have been performed with varying
success in the recent past. The major surveys have been those performed by the
National Cancer Institute, National Institutes of Health, in 1947-1948 (Ten City
SurveY),6 the recent Third National Cancer Survey (1971-1972),7 and the M. D.
Anderson Institute Texas Survey (1962-1972).8 Data are also available from
Iowa (1950),9 Minnesota (1963),10 and several prevalence studies carried out in
Australia (1960-1970).11,12
The enumeration of skin cancer incidence in a population is made very
difficult by the relative benignity of the disease, which allows for curative treat-
ment in physicians' offices rather than in hospitals. Surveying all physicians in
anyone area is difficult and expensive, so that most skin cancer incidence studies
have seriously underestimated actual incidences. 7
In a comparison of the older and the more recent studies, there is a strong
suggestion that skin cancer has increased in the past several decades. For in-
Skin Aging and Carcinogenesis due to Ultraviolet Radiation 159

stance, for Minnesota, comparing the metropolitan areas, an apparent twofold

increase has occurred since 1963, perhaps a reflection of a trend for people to
receive more sun exposure over more of the body surface as life-styles, clothing,
and leisure time have changed.
Skin cancer, like most other forms of cancer, is a disease that occurs more
frequently in older age groups. However, unlike those for other types of cancer,
the rates for skin cancer are significantly higher even in the middle age groups,
that is, 35-54 years.
Probably the most intense and accurate surveys have been those of Scotto et
al. (Third National Cancer Survey, National Cancer Institute, 1971-1972r and
of McDonald (Texas only, 1960-1970).8 In both these surveys, and in data from
other studies, an inverse relationship with latitude is apparent (Fig. 8-1). The best
estimate for present annual incidence of nonmelanoma skin cancer in the United
States (in Caucasians) is 165/100,000 population. This means that at present
about 300,000 cases of skin cancer develop in the United States each year or that
about one-third to one-half of all cancers of all sites arise in the skin. It is indeed
fortunate that the cure rate for skin cancer exceeds 95%.7 The third Cancer Sur-
vey shows that the annual incidence rates for skin cancer (excluding malignant
melanoma) vary from 379/100,000 in Dallas-Fort Worth to 124/100,000 in
Iowa.
The best available estimate of current incidence rates of melanoma come
from the Third National Cancer Survey, 13 based on counts during the years
1969-1971. The average annual incidence rate per 100,000 was 4.1. It is six to
seven times more common in whites than in blacks.
According to the National Cancer Institute's End Results in Cancer 14 and
"Trends in Survival Rates of Patients with Cancer," 15 the 5-year adjusted sur-
vival rate is 67% for malignant melanoma patients diagnosed between 1965 and
1969. Melanomas made up 1.4% of all cases of cancer diagnosed between 1969
and 1971 and about 1% of all cancer deaths in 1970.

Epidemiologic Evidence Supporting the Role of Sunlight

There is excellent epidemiologic evidence supporting the role of sunlight in

three types of skin cancers: basal cell carcinomas, squamous cell carcinomas,
and malignant melanomas.

Basal Cell Carcinoma

Basal cell carcinoma is the most common skin cancer that occurs in Cauca-
sians. Extensive epidemiologic studies indicate that these tumors are found
primarily on the head and the neck-that is, in sun-exposed areas. They are ex-
 160 Chapter Eight

1000 NON ME lANOMA

500

----------------
--"k-
z --..0
-- ---------------
" - --....---
100
Q
>-
<t

i - -- ~Meial(~) i
..J
ir
~ 50 ITO'tohl;-
1 -., MeIcnJma oncldence m
1---. (NCI) t
- A Non-Me!cn:lmo oncld m
1 -------£ (NCI) t
1 ~0 Non - Melanoma oncod m
I - - -0 (Texas) f I

I IWV\NVV\. N(Jl- Melanoma prewlence I

\ (age I-i'!!, HANES, NCHSI)
...... _ - - - - - ---
Incidence G

o
w MELANOMA
>- 5
is"'-
w
0::

Mortality

05
25 30 35 40 45
LATITUDE

Fig. 8-1. Reported skin cancer rates among whites as a function of latitude. Sources: melanoma mor-
tality from Mason and McKay '2; melanoma incidence from National Cancer Institute"; non-
melanoma skin cancer incidence (NCI) from Scotto et al. '; nonmelanoma skin cancer incidence
(Texas) from Macdonald'3; and prevalence of nonmelanoma skin cancer based on preliminary data
from the Health and Nutrition Examination Survey of the National Center for Health Statistics
(McDowell). 74 (From Environmental Impact of Stratospheric Flight. Climatic Impact Committee,
National Academy of Sciences, Washington, D. C., 1975, p. 38. Used with permission.)
Skin Aging and Carcinogenesis due to Ultraviolet Radiation 161

ceedingly rare in deeply pigmented races. There is a statistically significant ten-

dency for patients with basal cell carcinomas to have light eyes, light hair, and
light complexions, to sunburn easily, and to spend more hours outdoors than a
matched control group of people wihhout such tumors. These findings add
further support to the importance of sunlight in basal cell carcinoma formation.
However, the distribution of this type of cancer on the head and the neck does
not entirely correspond with sites of greatest UV exposure: that is, they are con-
centrated around the eye, behind the ear, and in other relatively protected sites.
Thus, factors in addition to sun exposure must playa role in the production of
this type of skin cancer.

Squamous Cell Carcinoma

Squamous cell carcinoma is the second most common skin cancer in Cauca-
sians and is found more often than basal cell carcinomas in the pigmented races. 5
The evidence for the role of sunlight in producing squamous cell carcinomas
in Caucasians, albinos, and patients with xeroderma pigmentosum is even more
convincing than it is for basal cell carcinoma formation. 3 These growths are dis-
tributed primarily over the head and the neck and, to a lesser extent, the exposed
areas of the upper extremities. As noted for basal cell carcinomas, squamous cell
carcinomas are found most abundantly in men who work outdoors in areas of the
highest isolation. In addition, though both squamous cell carcinomas and basal
cell epitheliomas are more prevalent in geographic areas of high sun exposure,
there is a relatively greater increase in squamous cell carcinoma incidence with
decreasing latitude and increasing sun exposure. 3
The geometry of sunlight exposure on the body of erect man further sup-
ports the association of sunlight with basal cell and squamous cell cancer. The
areas receiving most UV when the sun is high are the head (and since the top of
the head is usually covered with hair and thus protected, the exposed areas are the
rim of the ear, the nose, the forehead, the cheeks, the lower lip, and the chin), the
shoulders, the back of the neck, and the upper arms, and to lesser extent the outer
surfaces of the arms and hands. The chest, back, abdomen, and legs are exposed
only when the sun is low and little UV-B is present in the solar radiation. This
concept gains additional support from experimental studies in which squamous
cell carcinomas to the exclusion of other types of malignancies, can be produced
in appropriate animals by UV irradiation. 4

Malignant Melanoma

Malignant melanoma is a serious life-threatening tumor because of its poten-

tial for biologic aggressiveness and metastases. It is as common as a primary
162 Chapter Eight

malignant brain tumor. Recent statistical studies 15 show that only two-thirds of
new melanoma patients survived for 5 years-about the same fraction as for
breast cancer.
The influence of latitude of residence on the incidence of mortality from
melanoma in whites is the original and strongest evidence of the importance of
exposure to sunlight as a cause of malignant melanoma. Ths gradient of death
rates from malignant melanoma are rising in all countries in which they have
cancers, but it is nonetheless substantial. 1 7 Both the incidence and the mortality
rates from malignant melanoma are rising in all countries in which they have
been studied. Mortality rates are rising by around 3 to 9% per year, so that the
rates may have approximately doubled in the last 15 years. It has been suggested
that this increase in incidence is related to increased sun-exposure habits. The
skin areas receiving sun exposure vary between the sexes because of conven-
tional dress and hair styles (e.g., ears and neck exposed more in males; lower
limbs exposed more in females). These sites more commonly exposed in one sex
show a higher incidence of melanoma than do the corresponding sites in the op-
posite sex (Table 8-1). While no substantial study of the influence of occupation
on melanoma incidence has yet been made, patients with malignant melanoma, in
general, have a history of greater sun exposure and are less pigmented than are
those in a control group. 16
There is a close correlation between the death rate from squamous cell car-
cinoma and malignant melanoma at various geographic areas within the United
States and the Canadian provinces, suggesting that a common environmental fac-
tor operates. If these tumors are all caused mainly by sun exposure, one would
expect the distribution of malignant melanomas over the body to be the same as
that of squamous and basal cell carcinomas, which correspond well with habitu-
ally sun-exposed areas. However, malignant melanomas are not concentrated on

Table 8-1. Percentage of Skin Cancers by Anatomic Location in

Males and Females

Percent

Head and Upper Lower Multiple

neck Trunk limb limb sites Total

Malignant melanoma
Male 32 25 9 22 12 100
Female 24 19 14 33 10 100
Other skin cancers
Male 81 7 4 6 2 100
Female 78 7 8 4 3 100
Skin Aging and Carcinogenesis due to Ultraviolet Radiation 163

the face and the neck, in obvious contrast to squamous and basal cell car-
cinomas. 17 Therefore, the paradox exists of, on the one hand, a relationship of
the incidence of malignant melanoma to sun exposure (correlation of incidence
with latitude, differentially exposed sites between the sexes, and occupational
exposure to the sun) and, on the other hand, a lack of concentration of
melanomas on exposed sites. This paradox has been most clearly presented in
Australian clinical studies. Direct questioning of Australian patients with malig-
nant melanoma fails to elicit a history of particular exposure on the site of the
primary lesion.
In spite of the improvements in treatment in the last generation, the inci-
dence and death rate from malignant melanoma are rising rapidly in many de-
veloped societies. The increase in death rates from malignant melanoma in the
younger age groups has been balanced by the decline in death rates from squa-
mous cell carcinomas in the elderly. As a result, the death rate from all forms of
skin cancer combined has shown no decline in recent years.
Although other factors may be present, it seems reasonable to postulate that
the increasing melanoma rate is related to the general increase in sun exposure
because of recent changes in clothing styles, life-styles, and customs.
The real possibility that human activity may alter the concentration of ozone
(0 3 ) in the stratosphere has caused concern about a concomitant increase in the
incidence of skin cancer in the future. A thin layer of ozone formed in the upper
stratosphere absorbs all the short (UV-C) and most of the mid-range (UV-B)
radiation. Ozone thus serves as the primary ultraviolet "shield" for living or-
ganisms on earth, and it is probable that life as we know it could not have
evolved without the development of this shield.
The effluents of supersonic aircraft engines, unbridled discharge of
chloroftuoromethanes, and even the huge increase in the use of nitrogenous fer-
tilizers may reduce the stratospheric ozone thickness. Such a reduction could in-
evitably result in an increase in UV-B radiation at the earth's surface and eventu-
ally in an increase in the incidence of skin cancer in susceptible individuals. The
~st available present calculations suggest that a 5% decrease in ozone could
eventually (at a new equilibrium-in 60-120 years) result in an increase of ap-
proximately 20% in skin cancer. Other biologic and ecologic effects of an in-
crease in UV -B reaching the earth are a matter of debate. 1 7

Animal Studies

Because of obvious moral and sociological objections, and because of the

long time to tumor induction in humans, the skin of man is not an appropriate
experimental model for the study of skin cancer. Therefore, the experimental
production of tumors has been confined to the laboratory animals that develop
skin tumors more rapidly, particularly mice. I If we are to interpret the action
164 Chapter Eight

spectrum for UV -induced carcinogenesis and exposure-incidence (dose-

response) relationships, photobiological models relating radiant energy to its
eventual biologic effects in living organisms are needed. In general, the fewer
variables involved, the simpler the model may become. However, the complex-
ity of theoretical models that attempt to explain experimental photocarcinogene-
sis data suggests that photocarcinogenesis is not a simple process. Such models
are known moderately well for mice but much less well for humans, for whom
evidence must be laboriously gathered from epidemiologic data.
Beginning in 1940, Blum and his co-workers 19 initiated and carried out a
series of remarkable experiments concerning the dose-response, reciprocity, and
tumor induction-time relationships in photocarcinogenesis. These studies remain
as guideposts in the field of experimental UV carcinogenesis. The authors
utilized a UV source monitored by a stable photoelectric cell and an integrating
meter, and they confined their observations to tumor formation on the ears of an
inbred strain of albino mice. The reproducibility of their results attests to the sig-
nificance of their findings. In addition, they used very large numbers of mice,
further enhancing the validity of their results.
Blum et al. found that, above a certain minimum daily exposure dose and
over a threefold range, reciprocity was observed for tumor induction. That is, a
given daily exposure dose, whether delivered at higher irradiance for a short time
or at lower irradiances for longer periods, yields a given carcinogenic response.
For exposure doses below that required for reciprocity, less efficiency in produc-
ing tumors was observed, suggesting the presence of active repair processes dur-
ing the time of these exposures. Continuous irradiation of the mice was not
studied, and the fluxes examined covered only a threefold range.
In subsequent studies, using an irradiance that conformed to the region of
reciprocity, Blum et al. reported that increasing the exposure dose and/or short-
ening the interval between doses accelerated tumor formation but did not alter the
shape of the incidence curve. In addition, if sufficient energy had been applied
over a long enough period of time, discontinuing exposures to animals prior to
the appearance of the first visible tumor still resulted in eventual cancer produc-
tion. 19
Recently, Forbes et al. i~ have completed new experiments investigating the
limits of reciprocity for photocarcinogenesis. For these experiments, a xenon-arc
solar simulator simulating the whole optical spectrum contained in sunlight was
used, and the animals irradiated were hairless mice. Giving the same daily
radiant exposure dose of simulated sunlight in 5, 50, or 500 min, Forbes et al.
showed that protraction of the exposure duration-that is, giving each dose at a
lower irradiance-caused a marked increase in the photocarcinogenic effect of
spectrally identical and equal total doses of UV radiation. Animals receiving the
UV dose in 500 min developed many more skin cancers much sooner than those
receiving the same dose in 5 or 50 min. Thus, reciprocity failed when doses of
Skin Aging and Carcinogenesis due to Ultraviolet Radiation 165

UV were given at irradiances 10 or 100 times lower than doses given the con-
trols. This observation is of considerable importance in comparisons of the
photocarcinogenic effect of UV sources with significantly differing irradiance
and for the understanding of the dose-rate effects expected from solar UV -B ir-
radiation, which normally varies greatly with time of day and season.
Based on many experiments on hundreds of mice, Blum concluded that the
development time of UV-induced skin tumors was proportional to the logarithm
of the number of doses. From this basic experimental data, Blum derived a model
of UV -B photocarcinogenesis in mice that states that cancerization is a continu-
ous, cumulative process, which begins with the initial exposure and is acceler-
ated by each subsequent exposure. 1 Whether this theoretical model is applicable
to carcinogenesis by other UV wavelengths, or in humans, is unknown.

Mechanisms of UV Carcinogenesis

The pathogenetic mechanism of UV carcinogenesis on the cellular level re-

mains obscure. The recent discovery of excision repair deficiencies in cancer-
prone humans with xeroderma pigmentosum 20 resulted in considerable research
activity because of the implication of unrepaired DNA damage in cancer produc-
tion. However, xeroderma pigmentosum variants were soon discovered in which
excision repair systems were apparently normal, although the patients were as
exquisitely cancer-prone as those showing little or no DNA repair. 21
Excision repair is now known to occur after damage in the skin of men and
mice, 22 rat liver and kidney, rabbit brain, UV -induced squamous cell carcinoma
of hairless mouse skin, 23 and human tumor cell suspensions 24 produced by both
carcinogenic and noncarcinogenic agents. 25 It is thus clear that a great variety of
mammalian cells and at least some malignant cells have excision repair capacity.
In addition to the well-documented capability of UV radiation to induce
cancer of the skin in man and mice, Setlow and Hart 26 have been able to show
that fish liver cells UV -irradiated in vitro and reinjected into isogenic recipients
give rise to tumors. The tumor induction is UV -dose-dependent, and illumina-
tion of the irradiated cells with visible light before injection markedly reduces
tumor production. Since fish cells possess the photoreactivating enzyme, these
data support the concept that pyrimidine dimers induced in cellular DNA by UV
are related to the development of the tumors.
The available evidence suggests that injury to DNA is somehow related to
carcinogenesis. In view of the evidence that DNA damage is related to
mutagenesis in cells, this is a tenable assumption. However, in mouse skin and in
most cancer patients, the DNA repair systems seem to be capable of repairing
most UV damage; thus, absence of DNA repair may not be the basis of most skin
cancers. An elegant experiment of Zajdela and Latarjet 27 suggests a possible
166 Chapter Eight

reason. They painted a solution of caffeine, a potent inhibitor of postreplication

DNA repair, on the skin of mice during irradiation with UV. The caffeine-treated
skin developed fewer skin cancers than an unpainted control area on the same
animal. Epstein et al. 22 and Zajdela and Latarjet 27 suggest that the production of
skin cancer by UV is initiated by repair of DNA, allowing the cell to survive, yet
leaving in place or even favoring subsequent errors in DNA replication, resulting
in a greater potential for malignant change. The recent discovery of additional
error-prone DNA-repair mechanisms such as recombination repair and the com-
plicated system repairing interstrand links, plus the increasing evidence for the
association of error-prone DNA-repair systems with mutagenesis, makes these
hypotheses progressively more attractive. 28
Recently, Kripke 29 - 31 has demonstrated that chronic UV irradiation (280-
340 nm) has a profound effect on the immune response to skin tumors induced by
UV light in C3Hf mice. Many of these UV -induced tumors are immunologically
rejected by normal syngeneic mice, but they grow progressively when trans-
planted to UV-irradiated hosts. The mechanism of UV light interference with
immunologic rejection may be related to induction of a population of suppressor
lymphoid cells, which prevents an immune response against the UV -transformed
tumor cells. Also, the growth of some non-U V-induced tumors is enhanced in
UV-irradiated recipients. UV light may be acting as a potentiating agent for the
growth of certain tumors of diverse etiology.
. Not enough is known about the carcinogenic effects of UV-A exposure and
the augmentation and possible diminution of UV-B carcinogenesis by UV-A,
and only the most general statements can be made about mechanisms associated
with UV-A carcinogenesis. Similarities between the DNA absorption spectrum,
carcinogenesis, and delayed erythema action spectra (including the UV -A re-
gion) suggest that direct DNA damage and alteration of DNA-repair systems are
probably involved in UV-A effects upon photocarcinogenesis. On the other
hand, biologic exposures to UV-A may alter a host of cellular molecules, en-
zymes, repair enzymes, or other components (see Chapter 5), and there is little
reason to assume that direct alteration of DNA or DNA repair by absorption of
UV-A photons is the only mechanism involved in augmentation or production of
photocarcinogenesis.

Action Spectrum of Animal Photocarcinogenesis and the

Carcinogenic Effects of UV-A

Since the original work of Findlay, 32 many investigators have studied non-
melanoma skin cancers in experimental animals using 250-320 nm radiation.
Precise determination of the action spectrum for tumor induction has not been
accomplished because of the problems associated with methodology. However,
Skin Aging and Carcinogenesis due to Ultraviolet Radiation 167

the action spectrum for photocarcinogenesis between 250 and 320 nm has been
tested with various broadband sources 33 - 37 and appears somewhat similar to that
for induction of delayed erythema. Freeman et al., 38 using a monochromator as a
light source, were able to show that the relative effectiveness of wavelengths of
300, 310, and 320 nm for skin cancer production in hairless mice varied in ap-
proximately the same way as for production of skin erythema in man. However,
unlike erythema production, UV-B is more efficient than UV-C in producing
tumors.33
Bain and Rusch 39 compared the carcinogenesis produced in albino mice by
a specially filtered mercury source (280-340 nm) with "whole spectrum" from
the same source (essentially the same UV-B, but more UV-A plus visible radia-
tion) and noted somewhat increased tumorigenesis with "whole spectrum" radia-
tion. However, UV-A had been considered ineffective in producing or augment-
ing photocarcinogenesis until the work of Urbach et al. 75 Groups of hairless mice
were exposed 5 days a week to UV radiation from each of two sources with equal
amounts of UV-B but differing in the content of UV-A. The daily dose from both
light sources was adjusted to produce the same amount of acute skin erythema in
the animals. The animals treated with the source containing UV-B and a large
amount of UV-A developed many more tumors than those treated with the same
amount of UV -B and very little UV -A. However, the spectral energy distribution
and irradiances of the two light sources used (FS40 fluorescent lamps and a
long-arc xenon solar simulator) were significantly different. It is likely that be-
cause of the enhancing effect of dose protraction on photocarcinogenesis, 18 these
results were due to differences in irradiance between the sources, not differences
in spectrum. Subsequently, Forbes has found by using these same two light
sources, and at comparable irradiances, that no enhancing effect of UV-A and
visible radiation on U V-B carcinogenesis in hairless mouse skin is evident. 18
Thus, it appears that UV -A, in doses of the same order of magnitude as that exist-
ing in sunlight, is not carcinogenic for mouse skin when given intermittently.
Pathak et al. 41 also were unable to observe an enhancement by UV-A of
UV-B-induced carcinogenesis in groups of 20 albino Swiss mice exposed to
daily doses of UV-A, UV-B, and UV-A plus UV-B, for a period of 39 weeks.
Special fluorescent lamps and filters were used to obtain a source of high ir-
radiance UV-A completely free of radiation below 320 nm. FS40 fluorescent
lamps were the UV-B source. Two exposure dose ranges of UV-A, UV-B, and
combination exposures were studied. The daily UV-A doses were as high as 200
J/cm 2 and the UV-B as high as 100 mJ/cm 2 , and yet mice receiving daily UV-A
plus UV-B showed no significant increase in tumor production or time of onset
over those receiving UV-B alone.
Forbes (personal communication) found that chronic irradiation of hairless
mice with UV-A alone, carefully filtered to remove UV-B, does not lead to
tumor formation provided that exposures are given daily in 8 hr or less. When the
168 Chapter Eight

animals were exposed to continuous UV-A radiation, erythema, inflammation,

and some skin cancer were noted. Continuous exposure at the irradiances
employed, without a daily dark period, introduced a set of conditions in the ani-
mals that was not at all comparable to a "normal" daily light cycle. Undet con-
tinuous exposure, dark-repair mechanisms may theoretically be unable to oper-
ate, and the response of a primarily nocturnal animal may also be altered. Thus,
this experiment showed simply that UV-A alone can produce tumors under cer-
tain conditions.
Pathak 41 was unable to produce any tumors in albino Swiss mice exposed
for 45 weeks to a daily UV-A dose of 200 J/cm 2 from fluorescent lamps filtered
with Mylar to remove essentially all wavelengths less than 320 nm and delivered
at 10 mW/cm 2 in less than 8 hr. Zigman and Vaughan,42 in studying ocular ef-
fects of chronic (fluorescent) UV -A exposure of albino mice, noted tumor forma-
tion primarily in the animals' tails. The system used by these authors employed a
somewhat lower UV-A irradiance (3 mW/cm 2), but unfiltered UV-A fluorescent
lamps, which emit a small but significant UV-B component, were used. The
tumorigenesis noted here was probably due to UV-B emitted by the unfiltered
fluorescent lamps.
In summary, the present status of a variety of experiments with miGe con-
cerning the possible photocarcinogenic effect of UV-A shows the following:
l. UV -A given intermittently in doses equivalent to those occurring in na-
ture is not carcinogenic.
2. UV-A given in the ratio to UV-B that exists in noontime sunlight does
not augment or add to the carcinogenic effect of UV-B.
3. UV-A alone either (a) in doses equal in erythemogenic effect to UV-B
(i.e., a 1000:1 ratio) or (b) given continuously without a rest period,
may be carcinogenic. More data are needed.

Other Factors Influencing or Associated with

Development of Skin Cancer

Prolonged Erythema

Amount of pigmentation and amount of sun exposure are clearly related to

the development of skin cancer. The Celtic races, possessing a low level of pig-
mentation, are more susceptible to skin cancer, sunburn, and "aging"; thus,
"Ceiticity," the number of grandparents of Celtic origin, is sometimes used as an
indicator of a person's presumed skin cancer susceptibility. 43 In an attempt to
further define that popUlation susceptibl~ to skin cancer, Tanenbaum et al. 44
studied the duration of UV -induced delayed erythema in skin cancer patients ver-
Skin Aging and Carcinogenesis due to Ultraviolet Radiation 169

sus a normal control group. Erythema was prolonged in a significantly higher

percentage of the skin cancer patients than in control subjects, lasting 2 to 3
weeks after single exposures of six to eight times the patients' minimal erythema
dose (MED). No significant correlation was seen between prolonged erythema
and Celticity, fairness of skin or eyes, tanning, or other established risk charac-
teristics for cutaneous carcinoma. The small number of patients involved and
other factors make it hard to state clearly at present that prolonged erythema is
related to skin cancer risk; but this idea merits further investigation.

Heat, Wind, Humidity

The enhancing effect of heat on the degree of cutaneous injury and intensity
of erythema response to UV energy has been documented by Bain et aI., 45 and
more recently Freeman and Knox 46 demonstrated that increased temperature
present at the time of UV exposure accelerates tumor production. Utilizing en-
vironmental chambers to irradiate experimental animals, Owens and Donald 47
found that the influence of wind blown through the chamber was that more ani-
mals exposed to ultraviolet light and wind developed tumors than those animals
receiving the same dose of ultraviolet light alone. In other animal groups, it was
noted that animals maintained at high humidity developed tumors more rapidly
than those maintained at low humidity.

Chemical Influences on UV-Induced Carcinogenesis

The presence of carcinogenic and tumor-promoting chemicals in our envi-

ronment has made the experimental evaluation of chemical influences on UV
carcinogenesis of practical importance. At the current rate of introduction of new
compounds into the environment, it has become important to determine whether
a particular property-for example, phototoxicity-can be used to predict which
compounds could enhance photocarcinogenesis. The additive cancer-producing
effect of two separate chemical carcinogens has been well established. 48 Recent
studies have demonstrated that UV and chemical carcinogenic stimuli are also
additive in nature. 49 In addition, repeated applications of croton oil (a tumor-
promoting noncarcinogenic chemical)51 following a single UV exposure result in
significant cancer formation. These experimental findings suggest that environ-
mental chemicals may playa role in the development of skin tumors. 51,52
Equally significant is photoinduced carcinogenesis following application of
agents to the skin that are phototoxic but not in themselves carcinogenic. The
literature is brief: Biingeler 53 reported that subcutaneous injection of eosin,
hematoporphyrin, and a tar solution enhanced the ability of light to produce skin
tumors. His experiments are difficult to interpret, since the mortality in each of
the experimental groups was very high long before tumors appeared.
170 Chapter Eight

Miescher 54 did not succeed in producing tumors in mice exposed five times
weekly to a clinically phototoxic combination of UV-A and anthracene. He con-
cluded that there were no reasons for viewing photodynamic action as "a new,
specific carcinogenic agent." Heller,55 in an attempt to reproduce Miescher's re-
sults, produced necrotizing phototoxic skin changes with UV-A and anthracene
in mice, which developed neoplasia at the borders of the phototoxically produced
skin ulcerations. Heller's conclusion was diametrically opposed to that of
Miescher-he concluded in fact, that photodynamic reactions "should be recog-
nized as a new carcinogenic principle."
The introduction of 8-methoxypsoralen (8-MOP) as a therapeutic agent for
certain human skin diseases was followed by reports that it could enhance photo-
carcinogenesis in mice. 2 ,37 These demonstrations had the experimental advan-
tage of newer and more convenient lamps, which produced less stress on the
animals, limited skin ulceration, and improved animal survival. Evidence indi-
cated that carcinogenesis could result from the interacting effects of a compound
and irradiation with a particular waveband of light, neither of these agents being
a primary carcinogen in the doses used.
Forbes et al. 56 recently investigated the relative enhancing effect of two
widely recognized photoactive compounds (8-MOP and anthracene) on photo-
carcinogenesis. These were selected because they were available in 99% pure
form, leaving no doubt as to the identity of the active ingredient, because they
represented different classes of chemicals inducing similar but not identical
physiologic responses, and because neither was a chemical carcinogen. The re-
sults of the study are pertinent to the opposing conclusions of Miescher and Hel-
ler. The development of more appropriate light sources (solar simulators), ani-
mals (hairless mice), and test procedures (subacute phototoxicity), has in fact,
made possible a type of study envisaged by Miescher.
The experiments utilized genetically hairless mice pretreated three times
weekly with application df 40 Jl.. I of a 0.1 gm /liter solution of either 8-MOP or
anthracene dissolved in reagent-grade methanol. This volume covers about 20
cm 2 of skin on the back of a mouse. Approximately one-half hour later, the ani-
mals were exposed to a dose of simulated sunlight producing 30 mJ/cm 2 UV-B
(equivalent to about 1 human ME D) from a xenon-arc solar simulator. Chemical
and UV treatment continued for 38 weeks.
Compared with the vehicle alone, the 8-MOP solution, but not the an-
thracene solution, markedly enhanced photocarcinogenesis to simulated sunlight.
The times to 50% tumor prevalence for the vehicle-treated and anthracene groups
were 27.2 weeks and 28.2 weeks, respectively. These values were not sig-
nificantly different from each other. The time to 50% tumor prevalence for the
8-MOP groups differed significantly from each of the other two groups. Simi-
larly, tumor prevalence was significantly greater in the 8-MOP group. Beginning
with week 18, tumor yield was significantly greater in the 8-MOP group than in
Skin Aging and Carcinogenesis due to Ultraviolet Radiation 171

either of the other groups. Under the conditions of the experiment, both test
compounds were phototoxic, but only 8-MOP enhanced photocarcinogenesis.
Although methodological difficulties affect interpretations, studies using hairless
or albino mice 37 ,57-61 have demonstrated that oral administration .(either in the
diet or by oral intubation) of 8-MOP is, at the least, much less carcinogenic than
by topical application or intraperitoneal administration. This finding may be re-
lated to the dose of the drug or the total dose of light or to the metabolic biotrans-
formation of the drug. Daily ot'al administration of 8-MOP to hairless mice in
doses up to 50 times that used to treat humans followed by UV -A irradiation does
not cause carcinogenesis. 61 At present, there are no data available for comparing
the effective cutaneous dose for photocarcinogenesis after topical, oral, and in-
traperitoneal administration of 8- MOP.

Ultraviolet Radiation and Aging

The appearance of the exposed skin of Caucasian individuals is often re-

ferred to as either actinic damage (sunlight-induced) or premature aging. This
term is used because most of the features-for example, wrinkling and
yellowing-are similar to those observed in the process of "natural aging." It
should be noted that chronic sun-induced damage produces changes, such as
solar keratosis and solar elastosis, that are not necessarily seen in the process of
natural aging. However, the difference may be one of degree, since there is evi-
dence that actinic damage and normal aging share a common pathogenesis.
Susceptibility to actinic damage is genetically determined, light-skinned
Caucasians being most prone while blacks are most resistant. While skin color
and ability to produce melanin in response to sunlight exposure are the main fac-
tors involved, other genetic factors, such as the ability to repair the light-induced
damage, may be operative also. In Caucasians, the so-called farmer's skin or
sailor's skin is the end result of prolonged exposure over many years to solar UV
radiation. The picture includes dryness and cracking of the skin, senile len-
tigines, solar keratoses, a loss of elasticity, telangiectasias, and ecchymoses.
These changes reflect alterations in the epidermis, the dermis, and the adnexal
structures.
Histochemical and biochemical investigations have characterized the
sunlight-induced dermal changes as a decrease in insoluble collagen and in-
creases in elastin and ground substance. 62 A striking microscopic change occurs
in the dermis and is termed solar elastosis. This is a continuum of changes in the
elastin fibers 63 from curling and fragmentation to simple hyperplasia, then hyper-
trophy, and finally culminating in massive degeneration. Solar elastosis, which is
found only in sun-exposed skin, is most marked in the papillary dermis but can
involve the full thickness of the dermis.
172 Chapter Eight

Attempts to induce solar elastosis experimentally in humans have not been

successful. 64 Changes compatible with solar elastosis have been produced in al-
bino mice 65 and in rats 66 using a polychromatic UV radiation source, but this
approach leaves the question open as to whether UV-B (290-320 nm), UV-A
(320-400 nm), or both are responsible for the changes. UV-B is, in equivalent
doses, much more inflammatory or erythemogenic than UV-A, and that fact,
coupled with the suggestion that solar elastosis is a postinflammatory degenera-
tion,67 would favor UV -B as the causative portion of the UV spectrum. Klig-
man 6S has suggested that the deeper-penetrating UV-A is responsible for the de-
generation in the deeper layers of the dermis but that UV-B, which is more
energetic but less penetrating, produces the changes in the upper dermis. The
action spectrum of solar elastosis in human skin has not been ascertained because
of the need for chronic exposures.
The cross-linkage hypothesis of aging has recently been put forward. 69,70
Bjorksten 69 has proposed that intermolecular and intramolecular cross-linking
reactions are primarily responsible for age-dependent changes. The number of
potential cross-linking materials within cells is large, and a gradual accumulation
of cross-linking of many types may be unavoidable. The presence and biological
effects of cross-linking within the cell, particularly in chromatin, 70 are beginning
to attract interest. It is postulated that DNA adducts in chromatin and cross-links
between DNA and protein molecules are constantly being formed and repaired
but that the repair process may not always be complete. Probably of greatest im-
portance in terms of aging would be the formation and failure to repair DNA
adducts. This suggestion is supported by the correlation found by Hart and Set-
low 71 between the extent of excision repair of UV-radiation-induced DNA dam-
age and the aging rate for a number of different mammalian species.
Ultraviolet radiation is one agent that may produce cross-linking in DNA.
This possibility has been suggested by (1) the reduced extractability of protein-
free DNA; (2) the altered template capacity of chromatin; and (3) the identifica-
tion of specific DNA adduct products after ultraviolet irradiation. 70 It may be that
natural aging is related to the accumulation of DNA adducts by faulty DNA re-
pair. Thus, ultraviolet radiation may be simply accelerating the natural aging
process of skin. The presence and biological importance of cross-linking of DNA
by ultraviolet radiation and DNA-repair mechanisms in the process of aging of
human skin demands further study.

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26. Setlow, R. B., and Hart, R. W. Direct evidence that damaged DNA results in neoplastic trans-
formation: A fish story. Radiat. Res. 59:73-74, 1974.
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29. Kripke, M. L. Antigenicity of murine skin tumors induced by ultraviolet light. J. Natl. Cancer
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31. Kripke, M. L. Target organ for a systemic effect of UV radiation. Photochem. Photobiol.
24:599-600, 1976.
32. Findlay, G. N. Ultraviolet light and skin cancer. Lancet 2:1070-1073, 1928.
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3400 A. Cancer Res. 3:425-450, 1943.
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Meeting of the American Society for Photobiology, Sarasota, florida, 1973, p. 136.
41. Pathak, M. A., Parrish, J. A., and Anderson, R. R. Unpublished observations.
42. Zigman, S., and Vaughan, T. Near ultraviolet light effects on the lenses and retinas of mice.
Invest. Ophthalmol. 13:462-465, 1974.
43. Lane-Brown, M. M., and Melis, D. F. A genetic diathesis to skin cancer. J. Invest. Dermatol.
61:39-41, 1973.
44. Tanenbaum, L., Parrish, J. A., Haynes, H. A., Fitzpatrick T. B., and Pathak, M. A. Prolonged
UV-induced erythema and the cutaneous carcinoma phenotype. J. Invest. Dermatol. 67:513-
517, 1976.
45. Bain, J. A., Rusch, H. P., and Kline, B. E. The effect of temperature upon ultraviolet car-
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46. Freeman, R. G., and Knox, J. M. Recent experience with skin cancer. Arch. Dermatol.
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47. Owens, D., and Donald, W. The influence ofheat, wind, and humidity on UV injury (abstract).
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21-23, 1977.
48. Saffiotti, U., and Shubik, P. The effects of low concentrations of carcinogen in epidermal car-
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Academic Press, New York, 1970, pp. 2~5-273.
50. Epstein, J. H., and Roth, H. L. Experimental ultraviolet light carcinogenesis: A study of croton
oil promoting effects. J. Invest. Dermatol. 50:387-389, 1968.
51. Berenblum, I., and Shubik, P. The persistence oflatent tumor cells induced in the mouse's skin
by a single application of 9: lO-dimethyl-l:2-benzanthracene. Br. J. Cancer 3:384-386, 1949.
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52. Pound, A. W. Induced cell proliferation and the initiation of skin tumor formation in mice by
ultraviolet light. Pathology 2:269-275, 1970.
53. Biingeler, W. Uber den Einfluss photosensibilisierender Substanzen auf die Enstehung von
Hautgeschwulsten. Z. Krebsforsch. 46: 130-167, 1937.
54. Miescher, G. Experimentelle Untersuchungen iiber Krebsenzeugung durch Photosen-
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56. Forbes, P. A., Davies, R. E., and Urbach, F. Phototoxicity and photocarcinogenesis: Compara-
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57. Hakim, R. E., Griffin, A. C., and Knox, J. M. Erythema and tumor formation in methoxsalen-
treated mice exposed to fluorescent light. Arch. Dermatol. 82:572-577, 1960.
58. Pathak, M. A., Daniels, F., Hopkins, C. E., and Fitzpatrick, T. B. Ultraviolet carcinogenesis in
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59. Griffin, A. C. Methoxsalen in ultraviolet carcinogenesis in the mouse. J. Invest. Dermatol.
32:367-372, 1959.
60. O'Neal, M. A., and Griffin, A. C. The effect of oxypsoralen upon ultraviolet carcinogenesis in
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61. Langner, A., Wolska, H., Tarzabek-Chorzelska, M., and Pawinska, M. Dermal toxicity of
8-methoxypsoralen in hairless mice irradiated with long wave UV. J. Invest. Dermatol.
64:279-280, 1975.
62. Smith, J. G., Davidson, E. A., Sams, W. M., Jr., and Clark, R. D. Alterations in human dermal
connective tissue with age and chronic sun damage. J. Invest. Dermatol. 39:347-350, 1962.
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27:183-190,1957.
64. Shellow, W. V. R., and Kligman, A. M. An attempt to produce elastosis in aged human skin by
means of ultraviolet irradiation. J. Invest. Dermatol. 50:225-226, 1968.
65. Sams, W. M., Jr., Smith, J. G., and Burk, P. G. The experimental production of elastosis with
ultraviolet light. J. Invest. Dermatol. 43:467-471, 1964.
66. Nakamura, K., and Johnson, W. C. Ultraviolet light induced connective tissue changes in rat
skin: A histopathologic and histochemical study. J. Invest. Dermatol. 5/:253-258,1968.
67. Lovell, W. W. Ultraviolet irradiation of dermal collagen in vivo. I. Single doses of radiation.
Trans. St. John's Hosp. Dermatol. Soc. 59:166-174,1973.
68. Kligman, A. M. Solar elastosis in relation to pigmentation. In Sunlight and Man: Normal and
Abnormal Photobiologic Responses (M. A. Pathak, L. C. Harber, M. Seiji, and A. Kukita, Eds.;
T. B. Fitzpatrick, Consulting Ed.). University of Tokyo Press, Tokyo, 1974, pp. 157-163.
69. Bjorksten, J. The crosslinkage theory of aging. J. Am. Geriat. Soc. 16:408-427, 1968.
70. Cutler, R. G. Crosslinkage hypothesis of aging: DNA adducts in chromatin as a primary aging
process. In Aging, Carcinogenesis and Radiation Biology (K. C. Smith, Ed.). Plenum Press,
New York, 1975,p. 443.
71. Hart, R. W., and Setlow, R. B. Correlation between DNA excision repair and life-span in a
number of mammalian species. Proc. Natl. Acad. Sci. USA. 71:2169-2173, 1974.
72. Mason, T. J., and McKay, F. W. U.S. Cancer Mortality by County: 1950-1969. DHEW Pub.
No. (NIH) 74-615. U.S. Government Printing Office, Washington, D. C., 1974.
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74. McDowell, A. National Center for Health Statistics. Personal communication, 1974.
75. Urbach, F., Epstein, J. H., and Forbes, P. D. Ultraviolet carcinogenesis: Experimental, global,
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versity of Tokyo Press, Tokyo, 1974, pp. 259-283.
CHAPTER 9

Effects of Ultraviolet Radiation on the

Eye

Introduction

In humans, the eye has evolved into an incredibly sophisticated organ whose
neurophysiologic responses to photons in a certain portion of the electromagnetic
spectrum provide a constant detailed map of our immediate environment. The
action spectrum for this response lies primarily within the 400-700 nm
wavelength range, which has therefore been labeled the visible spectrum, or
"light. " The maximum of the eye's spectral response corresponds roughly to the
maximum of solar spectral irradiance. Because solar UV radiation is present dur-
ing most of the daylight hours, the eye may be exposed daily to some amount of
solar ultraviolet radiation throughout life.
The ultraviolet optical properties of the human eye are reviewed in Chapter
4. Wavelengths shorter than approximately 290 nm are partially or completely
absorbed within the cornea and conjunctiva (see Chapter 4). The acute effects of
exposure to these wavelengths are primarily those of conjunctivitis and a corneal
inflammation reaction known as photokeratitis. The inflammatory reaction of the
outermost layer of the eye to UV-C and UV-B radiation is similar to that of the
skin in many respects.
The ordinary clinical picture of photo keratitis follows a characteristic
course. After exposure, there is a period of latency varying somewhat inversely
with the amount of exposure. The latent period may be as short as 30 min or as
long as 24 hr, but it is typically 6 to 12 hr. Conjunctivitis, often accompanied by
an erythema of the skin surrounding the eyelids, is associated with the sensation
of a foreign body or "sand" in the eyes, varying degrees of photophobia (intoler-
ance to light), lacrimation (tearing), and blepharospasm (spasm of lid muscles).
Corneal pain can be very severe. The individual is usually incapacitated for some
period oftime. These acute symptoms usually last from 6 to 24 hr, and almost all

177
178 Chapter Nine

discomfort disappears within 48 hr. Very rarely does exposure result in perma-
nent damage. Unlike the skin, the ocular system does not develop tolerance to
repeated ultraviolet exposure. Swelling or shrinking of groups of corneal epithe-
lial cells leads to visibly recognizable stippling or irregular mosaic granulation of
the corneal surface. With UV doses greater than the threshold for photokeratitis,
surface epithelial cells show nuclear fragmentation, mid-epithelial cells show
vacuole formation, basal cells show inhibition of mitosis, and clouding of the
corneal stroma occurs. Inflammation is also present in the conjunctiva where
vasodilation, edema, and inflammatory cell infiltrate is followed by desquama-
tion.
Because wavelengths longer than 290 nm are largely transmitted by the
cornea, the underlying lens and iris are exposed to UV-A. The lens absorbs es-
sentially all of the UV -A striking it and may therefore be the ocular tissue espe-
cially susceptible to alteration by UV -A exposure of the eye. The possible pro-
duction of lenticular cataracts in humans by U V-A exposure is therefore of major
concern.
Any alterations of the lens or its capsule that result in apparent decreased
transmission or increased scattering of visible light may be called a cataract.
Minimal alterations, although detectable by careful biomicroscopic examination,
cause no change in routine visual acuity, but more marked alterations of light
transmission may impair or eliminate vision. The term cataract is often reserved
for this symptomatic decrease in vision. Cataracts may result from:
I. Injury to the epithelial cells that differentiate into lens fibers. In this in-
stance, fibers made after injury may scatter light more than normal
fibers. This type of alteration may not become evident for months be-
cause the lens fibers differentiate slowly. Corticosteroid-induced
cataracts are thought to result from changes in epithelial cell fiber
synthesis.
2. Acute osmotic imbalance. In this instance, water is drawn into the lens
and alters the light-scattering properties of the lens because of changes
in refractive indices. An example of osmotic cataract is the sugar
cataract, but similar changes may follow a traumatic, chemical, or
thermal injury to the lens capsule.
3. Alteration Of lens metabolism resulting in changes in water, calcium,
sodium, potassium, and phosphate content and the decreased transmis-
sion of visible light.
4. Alteration of the lens proteins so that high-molecular-weight protein
aggregates occur and scatter light. Osmotic swelling and then water in-
hibition lead to development of lakes of low density, low-refractive-
index material interspersed among areas of high-density and high-
refractive-index material. This is thought to be the mechanism of the
aging cataract. 1,2
Ultraviolet Radiation Effects on the Eye 179

Relatively little is known about the basic mechanisms or fundamental

changes that result in the cataractogenic process.
In persons with aphakia, the retina is exposed to UV -A because of the lack
of the lens in their dioptric system. Theoretically, UV-A-induced photochemical
alteration of the retina is possible in these persons. Another special case of in-
creased ocular sensitivity to UV-A exposure may be introduced when photosen-
sitizing compounds are present.

Morphology and Histology of the Cornea and the Lens

The gross morphology of the eyes is summarized in Fig. 9-1. Ultraviolet

radiation under various conditions of exposure has been reported to affect the
tissues of the cornea, the aqueous humor, the iris, the lens, and the retina.
The cornea is the first and principal absorber of ultraviolet radiation below
310 nm, the spectral region of maximum (lowest threshold) UV-induced injury
of the eye. Much of the research has been concentrated on this spectral region.
Because most of the research on the ocular effects of ultraviolet radiation has
been on rabbits, some of the anatomic features of the rabbit cornea are outlined
here. Histologically, the rabbit cornea has four tissue layers. From the outer sur-
face to the inner limit, the tissue layers are the epithelium, the stroma, Des-
cemet's membrane, and the endothelium (Fig. 9-2). The major difference be-
tween the rabbit, the human, and the primate cornea is that Bowman's layer,
separating epithelium from stroma, is not prominent in the rabbit cornea. This
layer is known as the basement membrane for the rabbit.

OPTIC NERVE

CORNEA

Fig. 9-1. Sagittal section of the human eye.

180 Chapter Nine

----Stratified squamous
epithelial layers

Stroma

.. Descemet's Membrane
~ Endothelium
Fig. 9-2. Histology of the normal rabbit cornea. Formalin-fixed, paraffin-embedded, 6-JL-thick sec-
tion (Gomori trichrome stain, x 104). Reduced 23% for reproduction.

The external layer of the epithelium is composed of five to six layers of strat-
ified squamous epithelial cells. These cells and their oval, pale-staining, poorly
outlined nuclei have their long axis parallel to the surface of the cornea and, in
most histochemical preparations, have a much lighter appearance than the
polyhedral and basal cells (Fig. 9-3). Organelles remain distinguishable even in
the outermost squamous cells, where there is an increase in small vesicular struc-
tures in the perinuclear region. One or two cell layers of polyhedral-shaped cells
are seen just beneath the stratified squamous cells. These cells contain nuclei that
are generally spherical and have irregularities in their profiles. The innermost
layer of the epithelium is composed of basal cells that are columnar-shaped and
have oval nuclei, usually located in the superficial portion of the cell. These large
ellipsoidal to spherical nuclei are oriented with the axis of the cell and perpen-
dicular to the corneal surface. Distinct, irregular, lobed nucleoli are frequently
found in the nuclei of the epithelial cells and are composed of finely granular,
moderately dense material.
Mitochondria, Golgi apparatus, and endoplasmic reticulum are seen in cells
of all three layers of the epithelium but are most abundant in the basal layer,
where they are most often found in perinuclear areas (Fig. 9-4). Mitochondria
appear to be long and filamentous, to have small diameters, and to possess few
cristae. The randomly distributed ribosomes observed in all epithelial cells are
most numerous in the basal cells. Membrane-bound dense bodies are also ob-
served in the cytoplasm.
The cytoplasmic matrix of all three zones is composed of densely packed
tonofibrils, smme of which appear to attach to desmosomes. The tonofibrils are
the most prominent in polyhedral and superficial squamous cells. Numerous
desmosomes are present along opposing membranes of adjacent cells of all
layers. Adjacent to the basement membrane, dense, half-desmosomelike struc-
 Ultraviolet Radiation Effects on the Eye 181

tures are located on the basal epithelial cell borders. The cell membranes of adja-
cent cells of all layers form projections that interdigitate with each other and give
the cell borders a convoluted appearance.
There are no actual or artifactual intercellular spaces visible by light micros-
copy since all of the cells in the epithelium are closely knit. The normal
epithelium contains no blood vessels or leukocytes. Occasionally, mitotic cells
are found in the basal epithelial layer of the epithelium. The basal epithelial cells
rest on a very thin basement membrane, which is difficult to visualize by light
microscopy. Bare nerve fibers are observed occasionally in the basal zone of the
epithelium. Nerve fibers are not distinguishable in the superficial layers.
Ths human and the primate corneas demonstrate a much more prominent
basement membrane, which is called Bowman's layer. 3 The Bowman's layer of
the human and primate corneas is composed mainiy of collagen. The anterior
surface of Bowman's layer is smooth and blends with the basal layer of the
epithelium. Posteriorly, Bowman's layer becomes intertwined with the anterior
lamellae of the stroma, making the transition less distinct.
Abutting the basement membrane of the rabbit cornea is the stroma layer or
substantia propria (Figs. 9-2 and 9-4), which is avascular and is composed of
collagen fibers. Interspaced between the collagen fibers are fibrocytes or kerato-
cytes that have long filamentous nuclei and indistinguishable cell borders. The
stroma comprises approximately 85% of the corneal thickness. The stroma con-
tains nonmyelinated nerve fibers from which minute branches penetrate into the
overlying epithelium. The fine nerve fibers can be demonstrated only with special
histologic techniques.
Descemefs membrane is positioned subadjacent to the stromal layer. Des-
cemefs membrane is relatively uniform in thickness, 8-10 /L, and is composed

Superficial flattened
epithelial cells
~ ~~~~~J
Polyhedral epithelial c e l l s <

Columnar b a s a l <
epithelial cells

Basement membrane .-
Stroma~

Fig. 9-3. Corneal epithelium of a nonnal rabbit cornea as seen by oil-immersion light microscopy in
1-IL-thick, Ep<m-embedded sections (paragon stain, x 1260). Reduced 30% for reproduction.
182 Chapter Nine

Fig. 9-4. Low-magnification electron micrograph of the normal rabbit corneal epithelium. Note the
columnar-shaped basal cells (BC), the polyhedral cells (PC) in the midportion of the epithelium, and
the outer squamous cells (SC). Cell organelles are not numerous, but cell borders are distinct. A
portion of the stromal layer (S) is visible in the lower left comer (X 4040).

of an amorphous PAS-positive material. The final layer of the cornea is formed

by a delicate single layer of cuboidal endothelial cells situated on the internal
aspect of Descemet's membrane. The endothelial cells line the corneal surface of
the anterior chamber of the eye.
The human crystalline lens has been selected for description because its
anatomy is well established and most mammalian lenses are similar to the human
lens. The crystalline lens is a biconvex structure situated just posterior to the iris
and anterior to the vitreous body. The human crystalline lens is 9-10 mm in
Ultraviolet Radiation Effects on the Eye 183

diameter and 4-5 mm thick, and its anterior surface is less convex (9-mm radius)
than the posterior surface (5.5-mm radius). The equator is the border where the
anterior and posterior surfaces meet. The anterior pole lies at the center of the
anterior surface, and the posterior pole is located at the center of the posterior
surface. The crystalline lens consists of the capsule, the anterior epithelium, the
lens fibers, and the cement substance.
The capsule is a transparent, membranous envelope that encases the entire
lens. It is highly elastic, being thinner at the anterior and posterior portions and
thicker at the periphery and the equator (Fig. 9-5).
The anterior epithelium consists of a single layer of cuboidal cells that lie
just underneath or posterior to the anterior capsule and cover the entire front sur-
face of the lens (Fig. 9-5). There is no posterior epithelium because the posterior
epithelial cells fill the central cavity of the lens vesicle during the embryological
development of the lens. The anterior epithelial cells gradually become columnar
and elongate into fibers as they approach the equator. As the anterior epithelial
cells pass beyond the equator, that portion of the epithelial cell in contact with the
capsule becomes the posterior portion of the fiber, and the anterior part of the cell

ANTERIOR CAPSULE

CORTEX

_. -- _.. ---------------
--"- .... -
..... -....,-"
ADULT NUCLEUS

FETAL NUCLEUS
-._-._-'\.,
"""i
,.
,,
;

..
.....
,
',EMBRYONAL
, H.,'
, " ,,
'.....' "

-. -------- ----- ---------'-'

""

----------'-'
---
_............ ---

-
POSTERIOR CAPSULE

Fig. 9-5. The capsule, the anterior epithelium, the cortex, and the nuclear zones of the adult lens are
schematically shown. Note the variations in the thickness of the capsule and the elongation of the
anterior epithelium at the equator, where lens fibers originate. (After Hogan, M. J., Alvarado, J. A.,
and Waddell, J. E. Histology of the Human Eye, An Atlas and Textbook. W. B. Saunders, Philadel-
phia, 1971.)
184 Chapter Nine

becomes the anterior fiber. At the lens equator, the nuclei of the epithelial cells
form an S-shaped zone ofnuciei throughout the entire circumference ofthe lens.
The cement substance holds the lens materials together and is not visible in
ordinary histological sections. The cement substance lies beneath the capsule,
deep to the anterior epithelium, and forms the central strand. The central strand
runs somewhat like an axle from the posterior pole to the anterior pole of the
lens. In addition, the axial cement substance projects three strands or rays to-
ward the equator, which divides the lens into at least three sectors. Under the
biomicroscope, the strands of the cement substance form a Y, with each arm

Fig . 9-6. The embryonal and adult lens. A: Embryonal nucleus with the anterior Y at a and the pos-
terior Y at b. The equatorial lens cells project their fibers to the tip of the Y suture at one surface of the
lens and to the suture at the pole of the other surface of the lens. This relationship is maintained
throughout the entire length of each suture. B: Adult lens cortex with a more complex suture organi-
zation. The fibers that arise from the tip of a branch of the suture insert farther anteriorly or pos-
teriorly into the suture at the posterior pole . (After Hogan, M. J., Alvarado, J. A., and Waddell, J .E.
Histology o/the Human Eye, An Atlas and Textbook. W. B. Saunders, Philadelphia, 1971.)
Ultraviolet Radiation Effects on the Eye 185

being the same length, separated by 120 and extending from the pole toward the
0

equator. The anterior Y is vertical and the posterior Y is inverted (Fig. 9-6A).
The Ys are known as the anterior and posterior lens sutures. In the adult, the
rays of the central strand may be much more complicated, with as many as six
primary and additional secondary lens sutures (Fig. 9-6B). In spite of these com-
plications, the fetal or original Ys persist throughout life in front of and behind
the embryonic nucleus.
The lens fibers are long, prismatic, 6-sided albuminoid material that taper as
they reach the anterior or posterior sutures from the equator. The lens fibers are
formed from the anterior epithelial cells located near the equator and radiate an-
teriorly and posteriorly, and their ends or tips insert into the rays or the strands of
the cement substance. Newer fibers are formed externally to the old fibers and
force the old fibers toward the center of the lens in concentric layers as a part of
the aging process. The older fibers lose their nuclei as they are pushed centrally.
The unique growth process of the lens fibers causes the lens to take on the ap-
pearance of two zones. The external zone, consisting of the relatively new nu-
cleated fibers, is called the cortex. The internal zone, consisting of the old
nonnucleated denser fibers, is called the nucleus (Fig. 9-5).
Any injury, osmotic imbalance, alteration in lens metabolism, or alteration
in lens proteins may alter the lens structure and result in cataracts. The rabbit has
been used for experimental cataractogenesis. There is one major difference be-
tween the crystalline lens of the rabbit and of the human. The anterior suture of
the rabbit lens is a single, almost vertical, whitish strand that is canted slightly
caudally superiorly and cranially inferiorly. The posterior suture of the rabbit
lens is a single whitish horizontal strand. Both sutures of the rabbit lens extend
almost the entire length of the lens diameter.
Figure 9-7 shows a biomicroscopic photograph of the anterior segment of
the normal rabbit eye. The optical beam passes from right to left through the an-
terior chamber of the cornea and the crystalline lens. The different components of
the lens are readily seen in this figure. Of particular interest is the posterior sub-
capsular opacity seen at G, which projects a filament anteriorly to the nuclear
opacity seen at D. The cortex, E, is clear except for the filament at G. The rec-
tangular white spot just posterior to the anterior capsule and the anterior
epithelium is an artifact from the photographic flash.

Histologic Effects of Ultraviolet Radiation

Ultraviolet radiation is invisible to humans, and damaging exposures of UV

radiation can result before the recipient is aware of the potential danger. Numer-
ous cases of photokeratitis and cataracts have been reported to result from UV
exposure. 4 ,5 The sources involved were welding arcs, high-pressure pulsed
186 Chapter Nine

Fig. 9-7. A biomicroscopic photograph of the optical section of the cornea, the anterior chamber and
the crystalline lens of the normal rabbit eye. The optical beam passes from right to left through the
following components of the eye: A: cornea; B: anterior chamber; C: anterior capsule and anterior
epithelium of the lens; D: lens nucleus; E: lens cortex; F: posterior lens capsule; G: lenticular opacity.
See text for detailed description. (After Pins, D. G., and Cullen, A. P. Ocular ultraviolet effects
from 295 to 335 nm in the rabbit eye. A preliminary report, DHEW-NIOSH Publication No. 77-
130. National Institute of Occupational Safety and Health, Division of Biomedical and Behavioral
Science, Cincinnati, Ohio.)
Ultraviolet Radiation Effects on the Eye 187

lamps, and reflection of solar radiation from natural terrain (snow, desert, and
water). These sources all contain UV radiation below 310 nm. Cataracts have
been proved to result from UV radiation in animals. Ultraviolet cataracts in hu-
mans have not yet been demonstrated by controlled scientific observations.
Verhoeff et ai., 6 Buchanan et ai., 7 and Christner et ai. 8 have provided ex-
tensive reviews of the literature on the ocular effects of 200-300 nm ultraviolet
radiation. a few key references are reviewed here to summarize the research on
the effects of ultraviolet on the eye, with emphasis on research of more recent
years.
Verhoeff et ai. 6 (1916), studying rabbit eyes, formulated some of the basic
postulates relating to the ocular damage caused by radiation. Exposure doses that
were below the threshold of any observable effects were repeated at intervals of a
few minutes to an hour. The abiotic effects of each exposure were additive within
24 hr; that is, the total cumulative exposure was responsible for the effects. An
exposure of one-sixth the threshold dose repeated every 24 hr failed to produce
photophthalmia, but an exposure of one-half the threshold dose repeated within
14 hr produced the same effect as a single, threshold-dose exposure.
Duke-Elder 9 provided a summary of ocular photobiology in 1929. He ex-
posed rabbit corneas to an unfiltered medium-pressure quartz mercury lamp,
using the unexposed eye as the control. His source contained UV-C, UV-B,
UV-A, and visible radiation. Rabbits were sacrificed, and the eyes were studied
histologically at various periods of time ranging from 2 hr to 10 days after expo-
sure. The sequence of his observations of histologic changes observed in the
cornea is summarized below:
I. At 4 hr after exposure, the effects consisted mainly of occasional swell-
ing of a squamous or basal epithelial cell. The endothelium and stroma
were normal.
2. After 6 hr, a large number of epethelial cell nuclei stained red with
eosin, and the basal cells were widely spaced, indicating edema. The
most superficial cells of the stratified squamous epithelial layer became
irregular in arrangement.
3. These changes progressed and became most noticeable at 12 hr. In
some epithelial cells, granules appeared to fill the entire nucleus, which
was usually surrounded by a vacuolelike space. In contrast, other cells
remained normal. Subsequent desquamation occurred in the central
area of the cornea. The nuclei of the stroma or keratocytes stained
deeply with methylene blue and began to fragment.
4. These effects continued to increase in severity through 16 hr after ex-
posure, at which time swelling of the lamellae or collagen fibers of the
stroma was apparent.
5. At 24 hr, lamellae in the stroma were swollen, and the endothelium
showed some abnormal staining similar to that of the epithelium
(above) but did not exfoliate.
188 Chapter Nine

6. There were few additional changes from the 24-hr stage through to 36
hr after exposure.
7. The reparative process began at 130 hr after exposure. Epithelial cells
began to take on an orderly arrangement, and the changes progressed
slowly until 7 days after exposure, when the cornea was essentially
normal.
In addition to corneal changes, Duke-Elder reported changes in the aqueous
humor, the iris, the lens, and the retina. The aqueous humor showed a marked
increase in proteins and a slight increase in sugars but a decrease in the chlorides.
The iris appeared congested after 12 to 16 hr and showed a disturbance of the
iritic pigment. All iris conditions returned to normal within 80 hr. The lens
showed intercellular changes in the pupillary area of the anterior epithelium just
beneath the capsule beginning about 13 to 36 hr after exposure and returning to
normal within 10 days after exposure. The fibers of the lens stroma and nuclei
were swollen and lost their orderly arrangement.
Retinal damage was restricted to the region of the posterior pole of the eye.
The retinal changes consisted of a disintegration and chromatin bleaching of the
ganglion cells and a swelling of the nuclei of the inner nuclear layer. Slight, if
any, changes occurred in the outer nuclear layer, and the rod and cone layer re-
mained normal. The most marked retinal changes were found from 8 hr to 20 hr
after exposure. The retina returned to normal within 50 hr to 60 hr after exposure.
Although the ocular reactions reported by Duke-Elder were severe, they ap-
peared to be reversible. It is difficult to compare his data with those of other
researchers because of his broadband source, and because irradiance measure-
ments were not reported. However, his report does describe the destructive and
reparative processes that result from exposure to full-spectrum ultraviolet radia-
tion.
Buschke et al. 1 0 observed histologic changes and changes in the mitotic ac-
tivity of corneal epithelial cells after exposure to ultraviolet radiation. Their re-
search emphasized the destructive effects of ultraviolet on the nucleus of corneal
epithelial cells, the loss of adhesion of the epithelial cell to Bowman's mem-
brane, and the inhibitory effects of ultraviolet radiation on the healing process of
the cell after exposure.

Action Spectrum of Ocular Effects of Ultraviolet Radiation

The many authors attempting to define the ultraviolet ocular action spectrum
have used a variety of monochromatic radiation sources and experimental ani-
mals. Different criteria for measuring insult to the eye and for establishing a
threshold response have also been employed. In general, corneal damage is the
Ultraviolet Radiation Effects on the Eye 189

end point for most studies of wavelengths shorter than 320 nm, although recently
UV-A lasers have been shown to produce acute alterations of the cornea. Altera-
tions of the lens are, in general, used to study the effects of wavelengths between
290 and 400 nm. Threshold action spectra for corneal effects and lenticular ef-
fects are presented here separately.

Cornea

Using the rabbit cornea, Cogan and Kinseyll provided reliable quantitative
threshold exposure data for the spectral region of wavelengths shorter than 320
nm. Using a granular appearance seen within the corneal epithelium with the
biomicroscopic examination as an end point, they were unable to produce corneal
effects with radiation of wavelengths longer than 313 nm. The radiant exposure
threshold for corneal damage in the rabbit was 15 mJ/cm 2 , with the maximum of
sensitivity noted at 288 nm. Sherashov l2 reported an action spectrum using slight
corneal haze as the end point. Maximum sensitivity was observed at the 289.4
nm and 253.7 nm wavelengths, and wavelengths above 300 nm were reported
not to cause observable effects at the doses used. Sherashov l2 and Cogan and
Kinseyll both used mercury arc sources and quartz prism monochromators. Be-
cause mercury sources provide very strong emission lines at irregular intervals in
the ultraviolet spectrum, exposures at equal wavelength intervals across the ul-
traviolet spectrum cannot be readily achieved. Therefore, the number of points
introduced may not be adequate to provide sufficient resolution of the action
spectrum. The technique of Pitts et al. 13 - 15 using a xenon or argon arc lamp and
grating monochromator avoids this problem.
Pitts and Kay l6 also established a corneal action spectrum for the rabbit. A
40-kW argon gas forced-transpiration arc and grating monochromator were used
to produce 10 nm wavebands. A total 238 rabbit eyes were exposed at spectral
intervals of 10 nm from 200 nm to 350 nm. Granules, epithelial haze, and photo-
phobia were the criteria used to establish the rabbit corneal threshold response, as
observed with a biomicroscope. Figure 9-8 compares the data of Pitts and Kay
with those of Cogan and Kinsey. The rabbit corneal ultraviolet action spectrum
for photokeratitis ranges from 210 nm to 310 nm. The most effective wavelength
in producing photokeratitis was 270 nm, with an exposure dose of 5.0 mJ/cm 2 •
Using the same instrumentation, Pitts 13 and Pitts and Tredicj14 reported the
ultraviolet action spectrum for 83 rhesus-monkey eyes and compared the primate
and rabbit data (Table 9-1). Four criteria were used to establish threshold: epithe-
lial debris, epithelial haze, epithelial granules, and photophobia. The abiotic ac-
tion spectrum for the primate cornea covers the wavelength range from 210 nm to
320 nmwith a minimum threshold exposure dose of 4 mJ/cm 2 at 270 nm. Expo-
sures at wavelengths longer than 250 nm usually resulted in relatively few de-
monstrable surface cells in the tear layer on the cornea. Surface debris increased
190 . Chapter Nine

600
400 fj. Rabbit threshold
(after Cogan and Kinsey)
200
100.00 o Rabbit threshold
u
J: (after Pitts et al.l
80.0
...
CD

...o
:::l
60.0
c.,..
)(
CD ...
0
40.0
"0 ..x
... E 20.0
.t:'
10.0
~ U
.... .....
.t:
.... 8.0
:c.!l
til
a:: 6.0
4.0
2.0
0
220 240 260 280 300 320 340
Wavelength in nanometers

Fig. 9-8. Comparison of the rabbit ultraviolet action spectrum of Pitts and Kay'· with that of Cogan
and Kinsey. 11

markedly with shorter wavelengths in the region 210-250 nm. The dead epithe-
lial cells on the corneal surface were so numerous at times that it was difficult to
observe the granules deep in the epithelium. Figure 9-9 provides the human ul-
traviolet action spectrum curve and compares the human data with the primate
and rabbit data. A summary of the data of Pitts et al. 13-15 on human, primate,
and rabbit thresholds producing photo keratitis is listed in Table 9-1. The effect
of these threshold exposures on human visual acuity is shown in Table 9_2.'5
Pitts '5 found that the action spectrum for the human cornea-judged by the
same four criteria (above) and, in addition, subjective symptoms, visual acuity,
and corneal light scatter-ranges from 220 nm to 310 nm. The human threshold
curve is not materially different from that of the primate for wavelengths 260
nm or longer. However, for wavelengths 250 nm or shorter, the threshold curve
for the human eye shows radiant exposure values less than those (more sensitive)
of either the primate or the rabbit. The human threshold data show a rather shal-
low curve with a minimum at 270 nm of 4.0 mJ/cm 2 • Exposures at wavelengths
longer than 310 nm were not administered to humans because of the theoretical
possibility of the development of future lenticular anomalies caused by the ex-
perimental exposures.
Like those of the primate cornea, the reactions of the human cornea to
Ultraviolet Radiation Effects on the Eye 191

wavelengths of 250 nm and shorter were qualitatively different than for the
longer wavelengths studied.!5 The biomicroscopic signs and subjective
symptoms occurred much earlier after exposure to wavebands at 250 nm and
shorter wavelengths. Visual acuity decreased quickly and returned to normal
within 6 hr. Subjective symptoms always disappeared within 4 hr to 6 hr. For
exposures at wavelengths longer than 250 nm, the signs and symptoms were
more delayed in onset. Reduced visual acuity caused by wavelengths longer than
250 nm was not usually found until about 4 hr to 10 hr after exposure and re-
mained subnormal for 24 hr after exposure. Corneal light scatter was investigated
in humans so that an objective method for determining the effects of ultraviolet
on the eye could be established.! 7 The light scatter data were not materially dif-
ferent from the biomicroscopically established thresholds.

Mechanisms of Ultraviolet-Induced Corneal Damage

The photochemical and biochemical mechanisms underlying the response of

the cornea to wavelengths less than 300 nm are a matter of debate. Cogan and
Kinsey!! concluded that the action spectrum for the rabbit cornea corresponded
more closely to the absorption curve of albumin and globulin than to that of the
nucleoproteins; however, their ocular action spectrum peaked at 289 nm rather
than at the 270 nm maximum of sensitivity observed by Pitts (Fig. 9-9). Soll-
ner! 8 reported that the ultraviolet absorption of protein-free corneal extracts was

Table 9-1. Summary of Photo keratitis Thresholds for Rabbits, Primates, and
Humans l 3-1 S for the 210 to 320 nm Wavelength Range

Rabbit Primate Human

Radiant Radiant Radiant

Wave band exposure threshold exposure threshold exposure threshold
(nm) (mJ/cm' ) (mJ/cm' ) (mJ/cm' )

205-215 700.0 327.0

215-225 46.0 21.0 10.0
225-235 30.0 22.0 13.0
235-245 33.0 12.0 7.6
245-255 41.0 20.0 8.0
255-265 18.0 1l.0 7.5
265-275 5.0 4.0 4.0
275-285 11.0 6.0 5.9
285-295 12.0 7.0 7.0
295-305 50.0 1l.0 7.0
305-315 55.0 20.0 14.0
315-325 7250.0 350.0
192 Chapter Nine

70 • Human threshold
50
'"
o Rabbit threshold
0 30
x 10 X Primate threshold
~ 5.0
E
...,
(J

I
.= 4.0 I
(J

..
J:
Q)

I
I
"
en
0
3.0
a.
x I
Q)

31 2.0
0
j
..
~

~
en
Q)

I- 1.0
~

0.0 . 220. 240. i

260
i
280
i
300 320
,
Wavelength in nanometers

Fig. 9-9. Comparison of the radiant exposure thresholds forthe comeaofthe human, the primate, and
the rabbit. The data were established by the exposure of 238 rabbit eyes, 83 primate eyes, and 39
human eyes to ultraviolet radiation in IO-nm increments. (After Pitts, D. G. The human ultraviolet
action spectrum. Am. J. Optom. Physiol. Opt. 51 :946-960, 1974.)

due to the nucleopeptides and that the absorption peaks were found at 220 nm and
260 nm. It is a reasonable hypothesis that UV absorption by nucleoproteins is
involved in producing the observed corneal effects.
Hamerski und Zajaczkowska 19 used electrophoretic techniques to investi-
gate the changes in the corneal epithelium protein that take place during the 6 hr
to 8 hr asymptomatic period prior to photokeratitis. The electrophoretic changes
included a reduction in the .a-globulins and a simultaneous increase in the
'Y-globulins. Changes in the protein pattern began prior to 4 hr after exposure,
reached peak intensity at 8 hr, and began to regress 12 hr after exposure. In each
instance, the electrophoretic pattern changes preceded the appearance of photo-
keratitis. In another investigation, Hamerski 20 reported that a cysteine solution
instilled into the eye prior to exposure prolonged the symptomless period and
reduced the severity of the symptoms, while a glycerol solution prevented the
signs of photophthalmia. The histochemical changes during the 6-hr latency
period involved extensive changes in the -SH groups and S-bonds. Less ex-
tensive, but nevertheless distinct, changes were found in the activity of alkaline
phosphatase, acid phosphatase, ATPase, and 5-nucleotidase. It appears that
Ultraviolet Radiation Effects on the Eye 193

many biochemic~ changes were taking place during the latency period some of
which are related to the severity of the reaction.
Tapaszto and Vass 21 studied mucopolysaccharides of the tears and corneal
epithelium of albino rabbits after exposure to UV -C radiation. The rabbit cor-
neas received 1 J/cm2 of radiant exposure from a low-pressure mercury lamp
(primarily 254 nm). They found three proteins, two neutral mucopolysac-
charides, three acid mucopolysaccharides, and a lipoprotein fraction in the nor-
mal rabbit tear layer. After exposure, the three protein components decreased to
one, and the lipoprotein fraction increased. Concentration of the two neutral
mucopolysaccharide components decreased after 5 min of exposure but showed
an increase if the exposure lasted more than 10 mill. The first changes in the
corneal epithelium occurred as early as 3 hr after irradiation, but the most signi-
ficant changes were found 8-16 hr after exposure. Epithelial lipoid fraction was
increased but the mucopolysaccharides remained constant. Tapaszto and Vass
hypothesized that photokeratitis from ultraviolet might be negated if the chemical
alteration of the tear layer could be prevented.

Corneal and Lenticular Effects of Longwave Ultraviolet

Radiation

There are relatively fewer quantitative research data to establish the ocular
effects of exposure to wavelengths between 300 and 400 nm. Much of the early
work in this wavelength range employed broadband UV sources or sources for
which the emission spectra and irradiances were not accurately measured. The

Table 9-2. Visual Acuity Responses for Threshold Radiant Exposures

in 10 Human Subjects

Wave band Time after

(nm) Decimal acuity exposure (hr)

215-225 0.6 4
225-235 0.6 4
235-245 0.6 9
245-255 0.5 10
255-265 1.0 8
265-275 0.8 4
275-285 0.5 4
285-295 0.6 7
295-305 0.8 5
305-315 0.6 4
194 Chapter Nine

recent availability of UV-A lasers has added to our knowledge of the exposure
thresholds for acute effects of UV-A. However, the very high irradiance may
compromise the practical significance of these data.
Verhoeff et al. 6 used a medium-pressure mercury discharge lamp and water
filter system, and a magnetic iron-electrode arc lamp, both in combination with
glass cut-off filters and Uviol (visible-absorbing, UV -transmitting) glass. All
mercury lamp exposures of 5 min or longer for wavelengths longer than 295 nm
produced changes in the anterior lens epithelium which were maximum 48-72 hr
after exposure, and included swelling of cells, a persistent alteration of all nuclei
size and shape, appearance of cytoplasmic granules, and formation of a deeply
staining ring of cells near the periphery of the exposure area. Some transient
changes were reported in the lens substance, but only to a depth of 20 JLm. Two
quartz lenses were used to focus an image of the magnetic iron-electrode arc di-
rectly onto the eye and hence the exposure doses actually delivered are not
known. An often ignored finding of this early study was that the corneal en-
dothelium could be destroyed by wavelengths longer than 295 nm. Loss of the
endothelium was accompanied by a marked swelling of the corneal stroma to as
much as twice its original thickness.
The very broad spectral region used by Verhoeff et al. and other early inves-
tigators makes it difficult to interpret observations quantitatively regarding the
specific wavelengths that were responsible for the various observed effects.
Triimpy22 exposed rabbit eyes to wavelengths from 313 to 435 nm but also did
not provide detailed radiometric data on his source. He qualitatively described
lenticular changes that could not be compared to previous research. Van der
Hoeve 23 criticized the research of Triimpy and attempted to compare Triimpy's
work with previous data.
Fischer et al. 24 used narrow-bandwidth UV from 250 to 350 nm and ob-
served a change in the reflex image (specular reflection of an image) of the
cornea. They established 450 mJ/cm 2 as the threshold at 350 nm for this effect in
the rabbit cornea. The source was either a carbon arc or a tungsten strip-lamp
focused through a double monochromator. The irradiance as measured by a
thermopile varied from 200 to 2000 JL W/ cm 2 , and the duration of exposures
varied from 2 to 60 min. Fischer et al. did not use a biomicroscope to observe
lenticular and aqueous changes.
Bachem 25 concluded that repeated exposures of longer UV wavelengths can
cause cataracts through cumulative effects. He reported that the action spectrum
for cataracts begins abruptly between 293 and 297 nm, reaches a peak near 297
nm, and falls abruptly near 313 nm. Minimal effects exist through the remainder
of the UV -A. In both the rabbit and the guinea pig, reversible lenticular blurring
occurred 5 to 10 days after exposure. With repeated exposures to a waveband
from 297 to 365 nm, irreversible lenticular opacities occurred after a latency
period that varied between 2.5 and 15 months. Bachem was unable to produce
Ultraviolet Radiation Effects on the Eye 195

Table 9-3. Lenticular Damage Thresholds to Broadband UV (J/cm 2 a

Containing Only above

Reaction 297-289+ nm 302nm 334-365 nm only

Rabbit lens, opacity 2.0 X 10-' *1.5 X 10' *5.0 X 10 3

Guinea-pig lens, opacity 1.0 X 10-' *1.0 X 10' *5.0 X 10 3
Guinea-pig lens, cataracts 3.0 X 10-' *4.0 X 10' *3.0 X 10'
a After Bachem.2S
*Cumulative from repeated applications.

lenticular opacities using 254 nm radiation; however, exposure of 0.2 J/cm 2 at

297 nm and longer wavelengths did produce opacities in the rabbit lens. For simi-
lar lenticular opacity in the rabbit, cumulative exposure doses of 15 J/cm 2 were
required for wavelengths 302 nm and longer, and 5000 J/cm 2 were required using
334-365 nm radiation. Bachem's threshold radiant exposures in J/cm 2 are shown
in Table 9-3. Because daylight contains no UV-C or far infrared, and because
both visible and near infrared are freely transmitted by the lens, Bachem con-
cluded that solar ultraviolet (290-400 nm) was responsible for sunlight-induced
cataracts.
Pitts and Cullen 26 have studied the effects of the 300 to 400 nm
wavelength range on the rabbit eye in vivo. These data are reviewed here in de-
tail. A 5000-W Xe-Hg source and a double monochromator were used to produce
6.6 nm full band-pass ultraviolet radiation. Pigmented rabbit eyes were exposed
to the 6.6 nm band-pass ultraviolet radiant energy in 5 nm increments from 295
nm to 320 nm and at some wavelengths longer than 320 nm.
Prior to exposure, each eye was examined with the biomicroscope, and rab-
bits with anomalies of the anterior eye (cornea, anterior chamber, iris, or lens)
were rejected. The criteria used to determine corneal damage were epithelial de-
bris, epithelial stippling, epithelial granules, epithelial haze, epithelial exfoliation,
stromal haze, stromal opacities, and endothelial disturbances. Criteria for altera-
tion of the crystalline lens were subcapsular opacities, capsular and stromal
haze, stromal opacities, and increased prominence of the anterior suture. Anterior
chamber signs included aqueous flare and cellular debris. Criteria for UV-
induced change of the iris included the presence of the anterior chamber signs,
changes in clarity of the iris stroma, and a sluggish pupillary response.
Table 9-4 shows the threshold radiant exposures for the rabbit cornea and
the rabbit lens in the 295 to 395 nm wavelength range. * At 300 nm and longer
wavelengths, the lens and cornea curves are relatively parallel up to about 320

"Data from 295 to 335 nm are from Pitts and Cullen'"; data from 335 to 395 nm have not been
previously published.
196 Chapter Nine

Table 9-4. Ultraviolet Threshold Data for the Rabbit Cornea and Lens (Rabbit)

Corneal radiant Lens radiant

Wavelength exposure threshold exposure threshold
(nm) (J/cm 2 ) (J/cm 2 )

290 0.012 >3.00

295 0.02 0.75
300 0.05 0.15
305 0.07 0.30
310 0.055 0.75
315 2.25 4.50
320 7.25 12.60
325 18.00 >50.00
335 30.00 >60.00
345 50.00 >50.00
355 50.00 >70.00
365 65.00 >162.00
375 >58.80 >58.80
385 >28.20 >28.20
395 >23.50 >23.50

nm. At wavelengths longer than 320 nm, although corneal threshold increased to
50 J/cm 2 , no lenticular damage was produced by the highest radiant exposures
used (60-162 1/cm2 ).
The action spectrum for corneal threshold extends to 365 nm for high ex-
posure doses. The action spectrum for lenticular effects extends from 295 nm to
at least 325 nm; however, it appears that the most effective wavelength range for
producing lenticular opacities is from 295 nm to 315 nm, which agrees roughly
with Bachem's data. The most surprising finding is the relatively low radiant ex-
posures in the 295 to 315 nm wavelength range that are required to produce len-
ticular opacities. The long exposure times used and the sharp transitions in len-
ticular action spectrum indicate that thermal effects are not responsible for the
opacities noted.
Figure 9-10 compares corneal and lenticular damage in the 290 to 400 nm
wavelength range to the previous corneal data of Pitts et al. for the rabbit, pri-
mate, and human corneal thresholds. Limited data for wavelengths longer than
300 nm for the human eye do not allow a detailed comparison; however, the
human corneal threshold was considerably below that for either the rabbit or the
primate. The primate corneal threshold was somewhat below the rabbit corneal
threshold at wavelengths shorter than 320 nm.
Almost complete absorption of the ultraviolet radiation by the cornea oc-
curs at 290 nm and less, causing the sharp rise noted in the lens threshold below
300 nm. The radiant exposure required to produce a threshold response at 295
 Ultraviolet Radiation Effects on the Eye 197

nm was 5 times the threshold value at 300 nm (750 mJ/cm2 versus 150 mJ/cm2 ),
which suggests that the shorter wavelength limit of the action spectrum is at ap-
proximately 290 nm for lenticular opacities. Exposures of 3 J/cm 2 at 290 nm did
not produce lenticular damage in rabbits.
In general, the severity of corneal changes increased with increasing expo-
sure dose. The amount of epithelial debris increased steadily with increasing ex-

100.0
a
~ a
a
, ,o--~ •
N
Ie
u
-,
•
•
RABBIT CORNEA
RABBIT LENS
/'
~

I
10.0 V PRIMATE CORNEA
~ • HUMAN CORNEA

""a::en::> 5.0
a
0
n.
x
""
1.0
• •

V
~
Z 0.5
•
<l V
0
<l
a::
0
...J 0.1

.
0
• /'J
".-
:I:
en
/
0.05

""a::..... .-~
:I:
V-V'" / . . 11

0.01

0.005 ~~
210 220 no 240 250 210 270 280 210 300 310 520 330 S40 S50 !eO 370

WAVELENGTH IN NANOMETERS

Fig. 9-10. Comparison of rabbit corneal and lenticular thresholds at wavelength bands longer than
290 nm" with previous rabbit, primate, and human thresholds. The open squares represent the high-
est radiant exposures attained. No lenticular damage was produced at these exposure levels. The open
circle represents a single exposure at 345 nm which was judged to be corneal threshold. The data
points refer to a wavelength band rather than to discrete wavelengths. (After Pitts, D. 0., and Cullen,
A. P. Ocular ultraviolet effects from 295 to 335 nm in the rabbit eye. A preliminary report, DHEW
(NIOSH) Publication No. 77-130. National Institute of Occupational Safety and Health, Division of
Biomedical and Behavioral Science, Cincinnati, Ohio.)
198 Chapter Nine

posures beyond the threshold dose. At high radiant exposures of wavelengths

between 250 nm and 300 nm, the granules coalesced to form a network. Epithe-
lial damage, stippling, and haze were detected within 2 hr following such expo-
sures; stromal damage, haze, and opacities appeared within 24 hr; and endothe-
lial changes were noted within 4.5 hr after exposure. Exposures of twice the
corneal threshold usually resulted in irreversible damage to the cornea.
Severe corneal reactions were accompanied by secondary anterior uveitis
characterized by ciliary injection, aqueous flare, the membranous inflammatory
by-products deposited on the corneal endothelium (Fig. 9-11). Such changes
made it difficult to observe the lens or its capsule. Anterior uveitis usually re-
gressed spontaneously within approximately two days. Table 9-5 provides infor-
mation on the wavelength, exposure dose, time of onset (latent period), and time
for recovery from anterior uveitis. It is apparent that no anterior uveitis was in-
duced by ultraviolet radiant exposures of wavelengths longer than 315 nm.

Fig. 9-11. An optical section of the anterior segment of the eye that demonstrates anterior uveitis. A:
cornea; B: membranous inflammatory by-products on the corneal endothelium; C: aqueous flare.
Animal H080R, 305 nm, radiant exposure 0.5 J/cm 2 , 27 hr after exposure . (After Pitts, D. G., and
Cullen, A. P.Ocular ultraviolet effects from 295 to 335 nm in the rabbit eye. A preliminary report,
DHEW-NIOSH Publication No. 77-130. National Institute of Occupational Safety and Health, Di-
vision of Biomedical and Behavioral Science, Cincinnati, Ohio.)
Ultraviolet Radiation Effects on the Eye 199

Table 9-5. Anterior Uveitis (Rabbit)

Radiant exposure Time of Time for

Wavelength (J/cm 2 ) onset (hr) recovery (hr)

295 0.75 1 24
300 0.50 24 48
305 0.30 24 48
310 1.00 2 24
315
320
335
345
355 No anterior uveitis found at the highest exposure level
365
375
385
395

Because the whole intact eye is exposed in vivo from outside the cornea, the
optical properties of each layer and the relative thresholds for various changes
must be considered together. A few examples of rabbit eye damage induced by
various UV wavebands illustrate this point:
At 295 nm, the radiant exposure required to produce lenticular alterations
also produced an immediate corneal reaction manifested by epithelial haze,
granule formation, stippling, and anterior stromal haze over the entire irradiated
area. The epithelium stained extensively with sodium fluorescein, confirming the
immediate response. The severity of the reaction increased to complete exfolia-
tion of the irradiated area at 20 hr postexposure. The anterior stromal haze of the
cornea also increased as the radiant exposure increased, so that the anterior
chamber, iris, and lens were difficult to see at I hr postexposure with a radiant
exposure of 0.75 J/cm 2 • Recovery was complete within 24 hr after exposure.
At 300 nm, radiant exposures above 0.3 J/cm 2 (twice the threshold exposure
for the lens) resulted in immediate corneal damage. The animals displayed ex-
treme photophobia. Permanent damage to the corneal epithelium, stroma, and
endothelium resulted. The iris was swollen, with a sluggish pupillary response.
The anterior chamber demonstrated a slight flare and a few cells were found in
the aqueous. Minor anterior uveal changes were found at 0.2 J/cm 2 radiant expo-
sure, which returned to normal within 3 days. Below 0.2 J/cm 2 , anterior uveal
changes were not found.
At 305 nm, radiant exposures above 0.3 J/cm 2 resulted in granules,
opacities, stippling, and fluorescein staining of the corneal epithelium. In addi-
tion, the stroma of the cornea was hazy and developed opaque striae after about 8
200 Chapter Nine

days. Within 24 hr, there were severe fibrinous endothelial deposits. An aqueous
flare was noted within 4 hr postex"posure but was reduced in severity within 24 hr.
All signs of anterior uveal inflammation disappeared within 8 days. There was an
exfoliation of iritic tissue within 24 hr after exposure at the 1.0 J/cm 2 radiant
exposure. Below 0.3 J/cm 2 , no anterior uveal changes were found at 305 nm.
There was a general increase in corneal damage as the radiant exposure was in-
creased above the corneal radiant threshold of 0.04 J/cm 2 •
At 310 nm, radiant exposures of 1.5 J/cm 2 (twice the lens threshold) pro-
duced a very minor aqueous flare, which disappeared within 48 hr (Fig. 9-12).
Permanent lenticular opacities and stromal opacities were also found at this' level
of radiant exposure. The endothelial disturbance resulted in a slight area of ex-
foliation. There was an increase in corneal involvement as the radiant exposure
exceeded the corneal threshold value of 0.047 J/cm 2 •

Fig. 9·12. Ultraviolet lenticular damage. The optical section demonstrates: A: the cornea; B: dotlike,
discrete anterior subcapsular opacities; and C: the anterior suture line, a vertical whitish line bisecting
the pupil. Animal HOCR, 310 nm, and 1.5 J/cm 2 radiant exposure, 24 hr postexposure (2 x Qd.
(After Pitts, D. G., and Cullen, A. P. Ocular ultraviolet effects from 295 to 335 nm in the rabbit eye.
A preliminary report, DHEW-NIOSH Publication No. 77-130. National Institute of Occupational
Safety and Health, Division of Biomedical and Behavioral Science, Cincinnati, Ohio.)
Ultraviolet Radiation Effects on the Eye 201

Table 9-6. Transient Lenticular Opacities (Rabbit)

Wavelength Radiant exposure Appearance of Disappearance of

(nm) (J/cm 2 ) lens opacities (h r) lens opacities

295 0.75 2 2 days

300 0.15 12 3 days
305 0.30 24 7 days
310 0.75 24 2 weeks
315 4.50 48 1 week
320 12.60 24 3 days
325 50.00
335 60.00
345 50.00
355 70.00 Transient opacities not achieved
365 162.00
375 58.80
385 28.20
395 23.50

At 315 nm, no anterior uveal or aqueous changes were found with radiant
exposures up to 7.0 J/cm 2 • The general pattern of epithelial granules, epithelial
haze, and stromal haze increased in severity as the radiant exposure increased.
Fluorescein showed a generalized, diffuse staining of the epithelium.
At 325 nm, 18 J/cm2 was required to produce corneal damage. At this
wavelength, no anterior uveitis was seen and no transient or permanent lenticular
opacities were noted at exposure doses as high as 50 J/cm 2 • Corneal threshold
continues to rise at wavelengths longer than 325 nm but appears to remain below
the lens threshold (see Tables 9-4, 9-5, 9-6).
The first biomicroscopic signs of lenticular damage in the Pitts study 26
were: (1) loss or reduction of "orange-peel" appearance of the anterior capsules
and (2) an increased prominence of the anterior suture line. These two biomicro-
scopic signs regressed to normal within 24 hr postexposure. As the radiant expo-
sure approached threshold, many small, discrete white dots appeared in the an-
terior subcapsular epithelium of the lens (Fig. 9-12). These anterior subcapsular
opacities appeared similar to the corneal epithelial granules. Table 9-6 presents
the wavelength; the radiant exposure necessary to produce the transient, granular
lenticular opacities; the time of appearance of these opacities after exposure; and
the time of disappearance of these opacities after exposure.
Exposures greater than the lens threshold resulted in permanent lenticular
opacities (cataracts). The change from the small, discrete white anterior subcap-
sular granules into the permanent opacities developed as follows:
202 Chapter Nine

Table 9-7. Permanent Lenticular Opacities (Rabbit)

Wavelength Radiant exposure Appearance of

(nm) (J/cm 2 ) lens opacities (hd '

295 1.0 60
300 0.5 24
305 0.5 24
310 1.5 24
315 6.0 24
320 15.5 24
325 50.0
335 60.0
345 50.0
355 70.0 Permanent opacities not achieved
365 162.0
375 58.8
385 28.2
395 23.5

1. The fine opacities organized into fewer, more dense opacities.

2. The fine discrete opacities migrated toward the anterior suture line,
then posteriorly. into the anterior cortex of the lens.
3. A concurrent increase in the haze of the lens cortex was detected.
The permanent opacities usually developed in proximity to the anterior su-
ture line. In addition to the permanent opacities, occasionally the presence of a
vacuole was seen. An animal followed for 3 months postexposure did not show
nuclear or posterior subcapsular opacities. Table 9-7 gives the wavelength, the
radiant exposure, and the time after exposure for the appearance of the permanent
lenticular opacities and indicates that, in general, the threshold dose producing
permanent lenticular opacities was approximutely twice the acute lenticular
threshold dose.
Using single UV exposures, the threshold for corneal damage appears to be
lower than that for lens damage at all wavelengths. It is not possible to produce
cataract without also producing photokeratitis. This may not be true for repeated
subthreshold exposures, cumulative alterations, or damage induced by various
combinations of ultraviolet spectral bands. Furthermore, the appearance of the
characteristic UV-induced lens changes without and prior to the anterior uveitis
indicates that the lenticular effects are essentially independent of anterior uveitis.
The suggestion is that the lens is directly altered by its absorption of radiant
energy.
Ultraviolet Radiation Effects on the Eye 203

UV-A Laser Radiation Effects on the Eye

Ultraviolet lasers have recently been used to produce corneal and lenticular
damage. Ebbers and Sears 27 exposed 100 rhesus-monkey eyes to a helium-
cadmium laser with monochromatic output at 325 nm. This laser produced con-
tinuous output of approximately 30 mW over a 1. 78-mm-diameter field (1200
mW/cm 2 ). The end point for corneal damage was a well-defined corneal lesion
observed by biomicroscopic examination in 50% of the subjects (ED5o). An
ED50 of 0.8 J (32 J/cm 2 ) was established and the corneal damage was completely
reversible within 24-48 hr. The investigators noted that permanent lenticular
cataracts were produced with exposures greater than 6.5 J (260 J/cm 2 ) or approx-
imately 8 times the radiant exposure required to produce ~orneal damage.
MacKeen et al. 2 8 exposed weanling rabbits to the same model He-Cd laser
used by Ebbers and Sears, operating at 325 nm. The reported power density was
approximately 600 mW/cm 2 for the 1.5-mm-diameter beam. Exposure durations
of 4, 6, 8, 16, or 32 min to the continuous beam were administered to the anes-
thetized rabbits' eyes. The initial response observed was an edema of the corneal
epithelium that occurred within the first few hours and was reversible. All 32-
min-duration exposures (1150 J/cm 2 ) showed localized anterior lenticular sub-
capsular changes after 3 days, which did not spread laterally with age. The final
appearance was a division of the initial subcapsular cataract into a small subcap-
sular opacity and a larger anterior cortical opacity. Adult rabbits of 9-11 kg
body weight developed deep anterior cortical cataractous changes after a period
of 6 months.
Zuclich and Connolly29 reported on corneal and lenticular damage in the
rhesus monkey from a krypton-ion laser with output at 350.7 nm and 356.4 nm
(irradiance ratio 350.7 nm/356.4 nm = 3), an argon-ion laser with continuous
output at 351.1 nm and 363.8 nm (irradiance ratio of 1.0), and a pulsed nitrogen
laser emitting at"337.1 nm with a lO-ns pulse width, maximum repetition rate of
50 Hz, and peak power of 1 MW. The krypton-ion laser (350.7 nm and 356.4 nm)
produced reversible corneal epithelial lesions that developed within 12-24 hr
after exposure and returned to a normal appearance within 48 hr. Multiple expo-
sures with pulse widths varying from 250 JLsec to 1 sec and a pulse train of 30 sec
all gave a corneal damage threshold at 67 J/ cm 2. This reciprocity relationship
was interpreted as evidence that a single-photon photochemical process was re-
sponsible for the corneal lesions. The argon-ion laser (351.1 nm and 363.8 nm)
gave a corneal threshold of 82 ± 3.3 J/cm 2 with a continuous 30-sec exposure.
However, the nitrogen laser (337.1 nm) gave a corneal radiant exposure
threshold of 8.4 ± 3.3 J/cm 2 for trains of lO-nsec pulses. Zuclich and Connolly
suggested, therefore, that although the corneal damage observed following expo-
sure to the krypton and argon-ion lasers was the result of a single-photon photo-
chemical process, the nitrogen laser, with its much higher peak power (l06 W)
204 Chapter Nine

induced corneal damage that was the result of some other mechanism, perhaps a
multiple-photon absorption or a partially thermal process.
Lenticular clouding was produced with a single lO-nsec pulse of the nitro-
gen laser (337 nm) with a radiant exposure of 1.1 J/cm2 • Immediately visible
cataracts were produced with two or more pulses. The immediate formation of
nitrogen-laser-induced cataracts again suggests a thermal mechanism for these
particular experimental exposures. The threshold for lenticular cataracts with the
argon-ion laser was 76 J/cm2 if exposed for 4 sec and 19 J/cm2 if exposed for 1
sec. Both ofthese thresholds amount to an irradiance of 19 W/cm2 ; the lenticular
damage noted with the argon-ion laser may therefore have been more a function
of irradiance than of exposure dose, again suggesting thermally induced lenticu-
lar cataracts. Retinal damage was reported with the krypton and argon laser, but
threshold determinations could not be made because of the variability between
animals.
Subsequent work by Zuclich ,and Kurtin 30 demonstrated that corneal dam-
age induced by argon-ion laser wavelengths (351 nm and 363 nm) in rhesus
monkeys is somewhat oxygen-dependent. The authors obtained average ~orneal
damage thresholds of 82 J/cm 2 in air, 66 J/cm 2 after the eyes had been exposed to
O 2 for 15 min prior to exposure, and 133 J/cm 2 after exposure of the eyes to 15
min of N2 • The authors took these results as evidence of a photodynamic effect of
UV-A on corneal tissue. When exposure duration was varied, the threshold ir-
radiance correspondingly changed in such a way that the total energy dose re-
mained relatively constant and the effects of repetitive exposures were cumula-
tive. These observations were felt to indicate that corneal damage was initiated
by a single-photon photochemical process.
Pitts and Cullen 2 6 assumed that both the corneal and the lenticular damage
noted in their studies was photochemical in origin. Their study employed much
lower irradiances (0.1 mWtcm 2 ), larger exposure areas (1.6 cm x 1.8 cm), and
longer exposure durations than did the laser studies cited above, and it is there-
fore unlikely that direct thermal damage to the lens was the cause of cataracts in
the studies of Pitts and Cullen. A thermally induced lenticular damage
mechanism is, however, a possibility for the laser exposure data. The laser beam
exposures were coherent radiation, highly collimated, with a small beam diame-
ter. Tne radiant exposure levels necessary to produce damage were achieved in
less than minutes with the lasers, while the radiant exposure for the high-
pressure arc system of Pitts and Cullen took hours. Zuclich and Connolly 2 9 report
that their data is consistent with a thermally induced lenticular damage
mechanism. These differences may make the comparison of the ocular effects of
laser exposures and of broadband incoherent radiation exposures questionable.
The average worker in an industrial environment would more likely be exposed to
low-power, large-diameter, uncollimated, incoherent UV sources. The laser data
do indicate that researchers who use UV-wavelength lasers or who are exposed
Ultraviolet Radiation Effects on the Eye 205

to high irradiances of essentially any radiation absorbed within the human lens
must use extreme caution.
It is well known that the thermal effects caused by infrared exposure of the
human eye include cataract formation. 31 There is also evidence that any radiant
energy that is absorbed by the iris causes an increased ocular temperature in the
region of the lens, leading to thermally induced cataracts. 32 The facts that
cataracts may be induced thermally, that UV -A reaches and is strongly absorbed
by both the iris and lens, and that reciprocity is not always observed in the pro-
duction of cataracts by pulsed UV-A lasers suggest that a thermal mechanism
may playa role in cataractogenesis by high-irradiance UV-A exposure of the
eye. This thermal-mechanism hypothesis does not exclude UV-A-induced
photochemical alteration of the crystalline lens resulting in cataracts, as dis-
cussed below.

Solar Radiation

The incidence of cataracts has been stated to be increased in the temperate

zone. 33 The incidence of cataract at all ages is greater in Israel than in Oxford,
England. 34 In the United States, geographic areas with greater sunlight duration
had a higher incidence of cataracts than those areas of shorter sunlight duration. 35
In the Philippines, the incidence of brunescent cataracts is twice as common in
persons who work outdoors when compared with indoor workers. 36 Such cir-
cumstantial evidence does not prove a relationship between sunlight exposure
and human cataracts, but this evidence and the experimental data in animals
make a causal relationship a likely possibility, especially for cataracts involving
lens pigmentation.

Biochemical Studies and Suggested Mechanisms of Ultraviolet-Induced

Lens Damage

The normal lens has fluorescent emission in the blue region, primarily from
lens "pigments" which absorb in the UV-A region. There are probably many
ultraviolet absorbing pigments present in the lens. The major ones which have
been identified are oxidation products of tryptophan. It is likely that the lens
serves some protective role in filtering UV-A radiation to prevent it from
reaching the retina. Aphakic persons can see wavelengths much shorter than 400
nm und perceive UV-A as violet. As the human lens ages, there is an increase in
the amount of ultraviolet-absorbing substances. Unfortunately, this increasing
pigmentation may lead to alterations in the optical properties of the lens which
result in decreased transmission of visible light. Human lens fluorescence in-
creases with age and the fluorescent emission shifts from blue to blue-green.
206 Chapter Nine

In the lens, biochemical alterations occur with aging, cataract formation,

and ultraviolet exposure. The interrelationships, sequence, and possible causal
relationships of these changes are the subject of study in several laboratories.
Clark et al. 37 and Lerman 38 reported that insoluble protein albuminoid in normal
human crystalline lens increases from a minimum of about 3% below the age of
10 to about 40% at the age of 80-89 (Table 9-8). There were also a large de-
crease in the concentration of the smaller ,),-crystalline protein, a small decrease
in ,8-crystalline protein, and an increase in the large a-crystalline protein (Table
9-9). With advanced brunescent and nigrescent nuclear cataracts, the level of in-
soluble protein is about 70% in human lens nucleus.
Specific amino acids of lens protein and their photooxidation products may
be involved in the cataractogenic process, or lens protein may become linked to
metabolically derived chromophores that absorb UV-A and act as photosensitiz-
ers. Free aromatic amino acids may be photooxidized into pigmented compounds
with strong binding affinities for lens protein. Yellow-brown coloration of lens
material may playa role in the formation of brunescent cataracts. UV-A-
absorbing pigmented and fluorescent substances have been found to be cova-
lently linked to specific peptides of lens protein. 39
A number of studies have suggested that tryptophan plays a pivotal role in
UV-A-induced changes in the lens. N-formylkynurenine, a photooxidation prod-
uct of tryptophan, 40,41 is present in human lens 42 and can photosensitize further
tryptophan oxidation by UV_A.40,41 Tryptophan is photolysed by UV-A into
many unidentified pigmented and fluorescent compounds,43.44 some of which
bind to human and animal lens protein in vitro, altering the physical and chemical
properties of these proteins. 45

Table 9-8. Age-Related Soluble and Insoluble Protein Fractions in Human Lensesa

Soluble Insoluble Average weight

Age protein protein of lens
(years) (%) (%) (g)

0-9 96.7 3.3 0.1167

10-19 97.1 2.9 0.1484
40-49 96.7 3.3 0.2130
50-59 90.9 9.1 0.2096
60-69 84.3 15.7 0.1840
70-79 82.6 17.4 0.2246
80-89 60.1 39.9 0.2328
Brunescent and nigrescent cataracts
70-90 29.0 71.0 0.2255

a After Clark et al. 37 and Lerman. 38

Ultraviolet Radiation Effects on the Eye 207

Table 9-9. Age·Related Changes in a-, {3-, and 'Y-Crystalline Concentrations a

a-Crystalline {3 -Crystall ine 'Y-Crystalline

Species and age (%) (%) (%)

Human
10-19 years 37 38 25
60-{i9 years 55 33.6 11.4
80-89 years 57.6 33.7 8.7

Rat
5-{i weeks 20 20 60
6 months 38 42 20
11 months 41 46 13

a After Clark et al. 27 and Lerman. 38

Pirie 42 and van Heyningen 46 feel that lens proteins are photooxidized by de-
struction of tryptophan groups in the protein. Free fluorescent substances that can
sensitize the photooxidation of lens protein are found in human lens and appear
to be derived from tryptophan metabolism. 46 Tryptophan oxidation products have
also been located in human lens by ESR spectroscopy. 46,48 Fluorescent pig-
mented photoproducts may link the brown insoluble protein to lens protein in the
formation of human cataracts. 42 A portion of the fluorescent substances of lens
protein is derived from tryptophan. 49 ,50 Fluorescence and phosphorescence
studies of whole lens 51 ,52 suggest that tryptophan photoproducts are bound to
human cataractous lens protein, and that the deeply pigmented lenses, especially
brunescent cataractous lenses, result from a UV-A-induced depletion of
protein-bound tryptophan and replacement by tryptophan photooxidation prod-
ucts. Photoproducts of tryptophan may be cytotoxic to lens epithelium,48 in-
hibit or inactivate enzymes necessary for formation or maintenance of proteins, 53
or cause structural protein aggregation from direct or photosensitized UV-A-
induced cross-linking. 54 As lens tissue ages, tryptophan residues become
oxidized and convert to kynurenine-type compounds. The subsequent alteration
of proteins leads to aggregation to form albuminoid. These "aging" effects ap-
pear to be accelerated by UV-A.
Zigman et al. 54 studied the chemical effects of UV-A-induced tryptophan
photoproducts on human crystalline lenses. The source was a medium-pressure
mercury arc lamp filtered to produce a wavelength range from 340 to 380 nm
with an irradiance of 3.0 mW/cm 2 and a maximum emission at 365 nm. They
soaked the human lenses in a tryptophan solution and found that, after several
hours of exposure to near UV, chromatic photoproducts are produced that bind
to and alter the solubility of lens proteins. Human lens material without added
tryptophan did not show chromatic changes upon exposure to UV-A until after
208 Chapter Nine

48 hr of exposure. Tryptophan has an excitation wavelength at 278 nm and a

fluorescent emission at 330 nm. However, following UV-A exposure of tryp-
tophan, the investigators observed an additional 360 nm excitation and 440 nm
fluorescence similar to that found in the brunescent human cataract lenses, and
they suggested that tryptophan photoproducts bound to lens proteins are involved
in UV-A-induced cataractogenesis.
In an attempt to determine the role of UV -A in the production of cataracts in
vivo, Zigman et al. 55 reared albino mice in a continuous UV-A environment. The
UV-A irradiance varied from 200 JLW/cm 2 to 600 JLW/cm2 during the course of
the experiment. At 6- to 7-week intervals, the mice were sacrificed for histologic
and biochemical study. The in vivo effect ofUV-A in this study was to inhibit the
accumulation of soluble proteins in the crystalline lens. This inhibition could
have resulted fwm a direct effect on the protein-synthesizing system or from the
direct blockage of amino acid uptake as a result of lens capsule and fiber altera-
tions. The exposed mouse lenses also showed a marked increase in insoluble pro-
tein between 16 weeks and 43 weeks. Thus, it appears that UV-A may have ac-
celerated the "aging" process of the lens in mice by changing soluble crystallines
into insoluble crystallines. These authors also studied biochemical effects of
UV-A exposure of freshly enucleated dogfish lenses in vitro. 56 An increase in
insoluble lens proteins and a decrease in soluble lens proteins was observed after
24 hr exposure to 3 mW/cm 2 UV-A (320-380 nm, fluorescent lamps). These
changes were inhibited by the addition of ascorbic acid.
Zigman et al. 43,56,57 studied mouse eye tissue damage after intermittent (12
hr per day) UV-A exposures (fluorescent lamps) of the mice up to 90 weeks. The
lens epithelial cells seemed to lose their ability to differentiate into fiber cells
after 35 weeks of exposure. Anterior and posterior cataracts developed at 50
weeks. The retinal photoreceptors of these mice became thin, then were invaded
by phagocytizing wandering cells and destroyed. Photoreceptor thinning was
noted by 14 weeks and the total loss of the photoreceptors occurred by 70 weeks.
Little or no corneal damage was observed, although the exposed mice had both
cataracts and extensive retinal injury.
The action spectrum for UV-induced cataracts ends relatively abruptly
about 290 nm because corneal absorption increast:s greatly at wavelengths
shorter than 300 nm. However, it extends into the UV-A and this longer
wavelength "tail" coincides with the phosphorescence excitation spectrum of
tryptophan. 58 Other amino acid photoproducts, cytochromes, or the presence of
unidentified photosensitizers may also explain the UV-A portion of the action
spectrum but present evidence supports a role of tryptophan. Kurzel et al. 58 have
summarized the evidence for UV induction of cataracts and proposed a chemical
model describing the pivotal role of tryptophan. At the peak of the cataract action
spectrum, most of the radiation is absorbed by tryptophan residues. Because of
Ultraviolet Radiation Effects on the Eye 209

tryptophan's low-lying first triplet state, excitation energy from other excited
species can also be transferred to tryptophan. The action spectrum of UV-
induced cataracts may therefore result from lens protein damage mediated by two
photochemical processes which proceed via the first excited singlet state of tryp-
tophan. The major reaction, maximal at 293-313 nm, is electron photoejection.
This photoionization appears to be the primary process resulting in the decom-
position of tryptophan. The second process results in cleavage of the N-H bond
of tryptophan and indole ring cleavage. Because the N-H bond has a bond
strength of only 75-80 Kcal/mol, this reaction can result from the less energetic
300-365 nm radiation and may explain the UV -A tail of the cataract action spec-
trum.58
Fluorescent material with 360 nm excitation and 440 nm emission similar to
that which accumulates in nonnally aging lenses can be generated photochemi-
cally in cultured rat and human lenses and in mouse lenses in vivo. 59 When ir-
radiated with UV, the fluorescence intensity of incubated rat lens increases
linearly with time, suggesting the photochemical reaction is not inhibited by its
own photoproducts. The action spectrum for production of this 360/440 fluoro-
gen has been detennined and shows little action at 360-320 nm, increases
sharply at 300 nm, and remains constant in the 300 to 280 nm range, and then
exhibits a further gradual rise from 270 to 250 nm.59 If lens absorption is taken
into account, this action spectrum closely parallels the action spectrum for chem-
ical destruction of tryptophan in solution. 60 In whole-lens homogenates, the
presence of tryptophan photoproducts has been shown to further intensify the in-
duction by UV-A of 360/440 fluorescence, suggesting that photooxidized try'p-
tophan may also act as a photosensitizer.
All of these studies support the hypothesis that light absorption by aromatic
amino acids in lens protein is the initial event in UV -induced lens damage. Al-
though many investigators support the thesis that brown nuclear cataracts or "ag-
ing" cataracts result from solar photooxidation of lens protein,47,51.52.56.57.62,63
the idea has not been proved. Arguments against this etiologic connection have
been stated. 64 Although maximum ultraviolet absorption in brown nuclear
cataract takes place at the front portion of the lens, it is the nucleus that is pig-
mented. The absence of any pigmentary change in the anterior lens could not be
explained by focusing effects or by difference in susceptibility of photooxidation.
Proteins from the cortex of nonnal human lens are photooxidized in vitro at the
same rate as those from the nucleus. 65 Also, although in vitro solar photooxida-
tion of lens protein destroys tryptophan, 66 the tryptophan content of protein from
brown cataract nucleus is the same as that found in the nonnallens. 65 These ar-
guments are less valid if one a~sumes that in vivo in man it may take from years
to decades for the UV-initiated photochemical events to alter the physical and op-
tical properties of the lens protein and that these altered proteins are therefore
210 Chapter Nine

among the oldest and most central proteins which are altered. Also, in vivo pro-
teins of certain age and solubility properties may be more susceptible to
ultraviolet-induced changes.
Using metameric color matching to estimate lens pigmentation, and ques-
tionnaires to estimate sun exposure, Girgus et ai. 67 found no difference between
estimates of pigmentation density and estimates of UV exposure in spectacle
wearers compared to normal persons. They concluded that solar UV exposure
did not cause pigmentation of human lens in vivo. However, because the subjects
were only 18-25 years old, because many spectacles transmit significant
amounts ofUV-A, and because sun exposure histories are often unreliable, this
type of study needs more convincing confirmation.
Another likely target for UV insult to lens tissue is DNA. Repair of lens
epithelial DNA following ultraviolet irradiation damage has been demonstrated
in cultured rat lens 68 and in vivo in frog lens. 69 After 254 nm radiation, un-
scheduled DNA synthesis was noted in the nuclei of lens epithelial cells. Repair
process was also present in those cells which had begun to differentiate into
fibers. In other cell systems, repairable DNA damage has also been caused by
longer wavelength radiation (see Chapter 5). Although UV-A is much less
efficient at causing lesions in DNA, the repair may be different. A variety of
agents, such as x_ray 70 and alkylating agents, 71 which damage DNA, also cause
cataracts.
The exact role of DNA damage and repair in cataractogenesis is not known.
As with the skin, UV radiation can cause tumors of the cornea. 72 Lens pigments
may also playa role in DNA protection. 58 Indole derivatives including tryp-
tophan protect DNA from UV radiation 73 by energy transfer mechanisms involv-
ing triplet-triplet energy transfer from nuclei acid bases to the lower triplet
energy level of tryptophan. 74
Careful study of the action spectra and threshold studies for UV damage to
the eye shows that, using conventional continuous sources, at any wavelength
studied, the threshold for photo keratitis is less than that for cataract induction. It
is tempting to conclude that any cataractogenic ultraviolet insult would be known
because symptomatic photokeratitis would result. It must be remembered, how-
ever, that most dose-response information available is derived from single expo-
sure experiments and that chronic or intermittent low intensity UV-A exposure
which is well below photokeratitis threshold may carry a cataract risk because of
differences in repair capabilities between the cornea and lens. Also, the lens does
not routinely exclude protein or cells as does the cornea. Any minor protein alter-
ations are cumulative because the nature of the lens metabolism is such that its
macromolecular contents remain within the capsule for a lifetime. Finally, it has
been argued that because the UV component of solar radiation is small and be-
cause a large portion of this radiation is absorbed by the cornea, UV induction of
human cataract in vivo is unlikely. This argument is difficult to support when one
Ultraviolet Radiation Effects on the Eye 211

considers that significant photochemical changes may be caused by very little ex-
posure, and that the same small fraction of sunlight affects the skin, which pos-
sesses the protective filter of a pigmented stratum corneum. The lens absorbs
UV-A more than does the cornea and contains pigments which can act as photo-
sensitizers.
Extrapolations from experimental animal data to humans must be qualified;
human cataractous lenses do show increased insoluble proteins and/or the pres-
ence of brown pigmented material with many of the same characteristics as the
tryptophan photoproducts. Additionally, the human lens does absorb most of the
UV-A radiation striking the eye at an appropriate angle. UV-induced cataracts
have been experimentally produced in mice, rabbits, monkeys, and guinea pigs
and, after both single and multiple daily exposures, over a wide range of UV-A
irradiances. Thus, UV-A is implicated as a potential cause of cataracts in hu-
mans.

Effects of UV-A on the Retina

Absorption of radiation by the cornea and lens of the human eye is such that
very little radiation of wavelengths shorter than 390 nm reaches the retina. In
persons with no lens, much of the UV -A striking the eye reaches the retina.
UV-A irradiation of normal phakic animals with and without the presence of
UV-A-sensitizing compounds produces damage to the retina. Multiple animal
models have been used to study biochemical alteration in the retina by UV-A.
Free-radical scavenger studies suggest that UV-A damages turkey retina via lipid
peroxidation. 75 Protein and RNA synthesis in dogfish retinal rods are suppressed
by irradiation at 340-380 nm 76 but in protein extracts the rat retina protein syn-
thesis was suppressed by 320 nm radiation and not by 340 or by 360 nm expo-
sure.
UV-A-induced morphological and histological changes in the retina have
been studied by exposing the intact whole eye. Thinning of the photoreceptor
outer segments of mouse retina was noted 10 weeks after exposure to 365 nm
radiation. 62 The animals were exposed for 12 hr each day to UV-A fluorescent
lamps (450 JLW/cm2). By 16 weeks, outer rod segments were further destroyed
and remnants were partially digested by phagocytic cells. Similar changes have
been seen after exposure to visible light. 77 ,7H Laser radiation in the region of
350-365 nm also causes damage to outer segments of rhesus monkey retina. 29
Chronic exposure of rats to high intensity UV-A over 3 years led to atrophy of
the first neuron, partial degeneration of the second neuron, and destruction of
retinal structure. 79
The recent data of Ham et at. 80 in rhesus monkeys clearly demonstrate mar-
kedly increased retinal damage by exposure to the shorter visible wavelengths.
212 Chapter Nine

Blue laser light caused retinal lesions in the monkeys at approximately 1/1000
the irradiance necessary to produce thermally induced retinal lesions with red
laser light. The action spectrum for such retinal lesions may include the UV-A
region around 400 nm. There is reason to believe that UV-A could photochemi-
cally induce retinal lesions if the ocular spectral transmission of the species in
question allows UV-A to reach the retina.

The Ocular Effects of UV-A Exposure in the Presence of

Photosensitizing Compounds (Psoralens)

Dramatic increases in the sensitivity of skin to UV-A and other wavebands

are induced bv a variety of compounds (see Chapters 6 and 7). The most potent
UV-A-photosensitizing compounds known for skin are certain furocoumarins
called psoralens. Some experimental animal studies indicate that
8-methoxypsoralen, one of the more phototoxic psoralens when applied to skin
or taken orally, also sensitizes the eyes of certain species to UV -A exposure. It is
not yet known how this sensitization relates to the use of psoralens in photo-
chemotherapy of humans.
Griffin 8 ! examined the erythemal and carcinogenic response associated with
oral (dietary, 0.5 g/kg diet) and intraperitoneal (0.4 mg/mouse/day, I hr before
UV exposure) administration of methoxsalen and subsequent exposure of albino
mice to UV-A radiation (A > 320 nm). The mice were irradiated daily for a
period of 6 to 12 weeks. Scar tissue formation around the ears, eyes, and face of
the mice was observed. The eyes of almost all the animals showed corneal opac-
ity and cataract formation.
Cloud et al. 82 studied the effects of oral 80 mg/kg of 8-methoxypsoralen
plus UV-A radiation in albino and pigmented guinea pigs. The animals were ex-
posed continuously for 24 hr. Extensive damage involved the ears, lids, cornea,
iris, and lenses. Less damage was seen in the pigmented animals as compared to
the albinos.
Cloud et al. 83 performed another experiment in which albino mice received
10 min/day of UV exposure 1 hr after intraperitoneal injection of 4 mg (160
mg/kg) of 8-methoxypsoralen. The animals received the drug and were exposed
to light 6 days a week for 5 months and were observed for an additional 5
months. The radiation source was Sylvania (BLB) black light blue tubes that
emitted strongly between 320 and 400 nm; the irradiance was not given. Reac-
tions in the mice were severe. Of the mice receiving 8-methoxypsoralen, 50%
died, whereas 13% of the controls not receiving the drug died. The ears of all
mice receiving psoralen plus UV-A had damage, which ranged from shriveling
of the edges of the ear to the loss of the entire external ear. Atypical leukoma with
or without vascularization and punctate keratitis were found in the cornea. Of the
Ultraviolet Radiation Effects on the Eye 213

28 mice that lived 10 weeks, 89% developed cataracts, 64% of which were an-
terior cortical cataracts. There is no doubt that at the extreme drug doses given,
8-methoxypsoralen increased ocular damage in the mice.
Freeman and Tro1l 84 investigated the action spectrum for eye damage in
guinea pigs receiving oral 8-methoxypsoralen (88 mg/kg). The animals' eyes
were exposed to various single-exposure doses of 5 nm half-bandwidth radiation
from a xenon arc and grating monochromator system. Exposures were given 1 hr
after oral administration of the 8-methoxypsoralen and at 10 nm spectral inter-
vals from 300 to 390 nm. The animals were observed for evidence of eye injury
for 72 hr postirradiation. Special attention was given to corneal injury, clouding
ofthe anterior chamber, and conjunctival hyperemia. The maximum efficie~cy of
photosensitization of the eyes was found between 320 and 340 nm, and no effect
of 8-methoxypsoralen was noted for wavelengths longer than 380 nm. Freeman
and Troll also found that guinea pigs were more susceptible to photosensitizing
injury with psoralens and UV-A than were rabbits.
When guinea pigs were given repeated doses of 8-methoxypsoralen and
UV-A comparable to photochemotherapeutic doses in man, no eye injury was
detected. 84 8-Methoxypsoralen was administered intraperitoneally (0.5 mg/kg)
to albino guinea pigs daily for 13 months. Animals were exposed daily to 10 hr of
fluorescent UV-A (F40 BLB) lamps at a distance of 10 in. This long-term study
revealed no gross, ophthalmoscopic, slit lamp, or histologic manifestations of
ocular injury. These results suggest that the ocular photosensitizing effects of
8-methoxypsoralen may therefore be limited to single exposures above some
threshold value.
Phosphorescence spectra of intact rat lenses revealed that 8-methoxypsora-
len could be detected and quantitatively measured 21h hr 85 but not 24 hr 86 after a
single intraperitoneal injection of 4-6 mg/kg 8-methoxypsoralen in dimethyl
sulfoxide. The characteristic 8-methoxypsoralen phosphorescent emission
showed a significant loss if lenses are exposed to UV or ambient lighting after
8-methoxypsoralen injection. Exposure of normal unsensitized rat lens to UV or
ambient light causes an increase in fluorescence at 440 nm. This UV-induced
increased fluorescence is enhanced by previous intraperitoneal injection of
8-methoxypsoralen, suggesting a photosensitizing effect of 8-methoxypsora-
len. 86
Egyed et al. H7 fed 16 two-to-three-week-old ducklings the seeds of
Ammi majus, the plant from which 8-methoxypsoralen is extracted. The animals
were exposed to the sun for 4-5 hr per day. Acute conjunctivitis was observed in
2-3 days. The animals developed mydriasis (dilation of pupils) and severe pig-
mentary retinopathy after 1 month. No cataracts were observed. A control group
of ducklings exposed to the sun but without the ingestion of plant seeds contain-
ing 8-methoxypsoralen showed relatively few ocular changes. Of the control
ducklings, 20% showed areas of retinal hyperpigmentation similar to those that
214 Chapter Nine

developed in the experimental animals of the study; however, in the control

group, these areas were less dense than in the seed-treated group. The work of
Egyed et al. has recently been confirmed by that of Barishak et al. ss who re-
ported pigmentary retinopathy and histologic changes of the choroid in ducklings
photosensitized by force-feeding of Ammi majus seeds and subsequent exposure
to sunlight.
Although humans have used 8-methoxypsoralen therapeutically for de-
cades s9 - 91 no cataracts have been reported. 92 However, it seems wise to limit the
use of psoralen photochemotherapy to those with significant skin disease (see
Chapter 10) and to use adequate UV -A eye protection during the course of
therapy (see Chapter 11). After ingesting psoralens, patients should protect their
eyes at least the remainder of that day.
Chronic administration of chlorpromazine and ultraviolet exposure has led
to cataracts in guinea pigs. 93 Corneal and lens opacities were observed in beagle
dogs given 2,6-dichloro-4-nitroaniline, a fungicide, by mouth and subsequently
exposed to sunlight. No lesions developed in the eye of each dog which was sewn
shut to act as a control. 94 Phototoxic keratoconjunctivitis has been seen in roofers
exposed simultaneously to coal-tar pitch volatiles and to sunlight. The same sub-
stances also produced keratoconjunctivitis in rabbits subsequently exposed to
UV-A from fluorescent lamps. 95

Summary: UV-A Exposure of the Eye

Experimental exposures of animals and humans indicate that for

wavelengths shorter than approximately 310 nm, keratitis and alteration of the
cornea are major ocular hazards. These reactions and effects are usually painful
but reversible. UV -induced alteration of the cornea appears to follow a photo-
chemical mechanism.
Ultraviolet wavelengths longer than 290 nm may reach the lens, the iris, and
the aqueous humor (anterior chamber). Experimental evidence in rabbits and
monkeys indicates that permanent lenticular cataracts may be produced by
single, high-irradiance or long-exposure durations to UV-B or UV-A. This ob-
served damage may be induced by thermal and/or photochemical mechanisms in
the lens. Furthermore, in albino mice, cataracts may be produced by multiple
daily UV-A exposures that are below the single-exposure threshold dose of ob-
servable corneal damage.
Biochemical studies in human and other lenses indicate that UV-A may
conceivably induce lenticular cataracts via alteration of lens crystalline proteins
from soluble, lower-molecular-weight crystallines to insoluble, higher-
molecular-weight crystallines, which may cause light scattering within the lens
Ultraviolet Radiation Effects on the Eye 215

(cataract). There is evidence that long-tenn daily UV-A exposure destroys the
retinas of mice.
Although some epidemiologic studies suggest that human cataracts may be
related to sun exposure, a clear demonstration that UV-A is involved in the de-
velopment of the brunescent and other human cataracts is lacking. Extrapolation
of experimental animal data to exposure of humans to UV-A may be highly un-
certain. UV-A may induce cataractous changes by photooxidation of tryptophan.
In the presence of 8-methoxypsoralen, increased ocular damage is observed
following UV-A exposures of experimental animals. In the albino guinea pig,
maximum ocular sensitization by 8-methoxypsoralen occurs over the range of
320 to 340 nm, with no effect observed at wavelengths longer than 380 nm. In
ducklings, pigmentary retinopathy develops following ingestion of psoralen-
containing seeds and daily solar exposure. At extremely high doses of oral
8-methoxypsoralen and subsequent UV-A exposure of guinea pigs, severe ocular
sensitization occurs. Pigmented guinea pigs have shown somewhat less ocular
damage than have albino guinea pigs. Severe ocular effects of 8-methoxypsora-
len and daily UV-A exposure have also been shown in albino mice. The
mechanism of ocular photosensitization by 8-methoxypsoralen is uncertain, but
presumably the same theories applicable to skin sensitization (see Chapter 6) may
hold for the eye.
An attempt to mimic the doses of 8-methoxypsoralen and UV-A employed
in photochemotherapy of humans produced no observable ocular changes in
guinea pigs after daily treatment for 13 months. The ocular hazard attributable to
oral psoralens plus UV-A in humans is not known. The need for verification and
elucidation of the ocular effects produced by UV -A alone and by UV -A in com-
bination with photosensitizing drugs is apparent.

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Uses of UV-A Involving Exposure of Humans 219

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and electron microscopic study. Am. J. Anat. 139:503-518, 1974.
79. Weisse, I., and Stotzer, H. Age and light dependent changes in the rat eye. Virchows Arch.
Pathol. Anat. Physiol. 362:145-156, 1974.
80. Ham, W. T., Mueller, H. A., and Clarke, A. M. Retinal sensitivity to damage from short
wavelength light. In Symposium on Biologic Effects and Measurement of Light Sources. U. S.
Dept. HEW Publication (FDA) 77-8002. BRH Rockville, Maryland, 1977, pp. 37-47.
81. Griffin, A. C. Methoxsalen in ultraviolet carcinogenesis in the mouse. J. Invest. Dermatol.
32:367-372, 1959.
82. Cloud, T. M., Hakim, R., and Griffin, A. C. Photosensitization of the eye with methoxsalen. I.
Acute effect. Arch. Ophthalmol. 64:346-351, 1960.
83. Cloud, T. M., Hakim, R., and Griffin, A. C. Photosensitization of the eye with methoxsalen. II.
Chronic effects. Arch. Ophthalmol. 66:689-694, 1961.
84. Freeman, R. G., and Troll, D. Photosensitization of the eye by 8-methoxypsoralcn. J. Invest.
Dermatol. 53:449-453, 1969.
85. Lerman, S. A method for detecting 8-methoxypsoralen in the ocular lens. Science 197:1287-
1288, 1977.
86. Lerman, S., Jocoy, M., and Borkman, R. F. Photosensitization of the lens by
8-methoxypsoralen.lnvest. Ophthalmol. Visual Sci. 16:1065-1068, 1977.
87. Egyed, M. N., Singer, L., Eilat, A., and Shlosberg, A. Eye lesions in ducklings fed Ammi
majus seeds. Zentralbl. Veterinaermed. Reihe A 22 :764-768, 1975.
88. Barishak, Y. R., Beemer, A. M., Egyed, M. N., and Eilat, A. Histology of the retina and
choroid in ducklings photosensitized by feeding Ammi majus seeds. Ophthalmic Res. 8:169-
178, 1976.
89. Parrish, J. A., Fitzpatrick, T. B., Tanenbaum, L., and Pathak, M. A. Photochemotherapy of
psoriasis with oral methoxsalen and longwave ultraviolet. N. Engl. J. Med. 291:1207-1212,
1974.
90. Parrish, J. A., Fitzpatrick, T. B., Shea, C., and Pathak, M. A. Photochemotherapy of vitiligo
with oral psoralen and a new high-intensity longwave ultraviolet light (UV-A) system. Arch.
Dermatol. 112:1531-1534, 1976.
91. Pathak, M. A., Kramer, D. M., and Fitzpatrick, T. B. Photobiology and photochemistry of
furocoumarins. In Sunlight and Man: Normal and Abnormal Photobiologic Responses (M. A.
Pathak, L. C. Harber, M. Seiji, and A. Kukita, Eds.; T. B. Fitzpatrick, Consulting Ed.). Uni-
versity of Tokyo Press, Tokyo, 1974, pp. 335-368.
92. Melski, J., Tanenbaum, L., Parrish, J. A., Fitzpatrick, T. B., Bleich, H. L., and 28 participat-
ing investigators. Oral methoxsalen photochemotherapy for the treatment of psoriasis: A
cooperative clinical trial. J. Invest. Dermatol. 68:328-335, 1977.
93. McDonald, C. J., Snell, R. S., and Lerner, A. B. The effect of chlorpromazine on oculocutane-
ous pigmentation in the guinea pig. J. Invest. Dermatol. 49:39-42, 1967.
94. Bernstein, H. N., Curtis, J., Earl, F. L., and Kuwabara, T. Phototoxic corneal and lens
opacities. Arch. Ophthalmol. 83:336-348, 1970.
95. Emmett, E. A., Stetzer, L., and Taphorn, B. Phototoxic keratoconjunctivitis from coal-tar pitch
volatiles. Science 198:841-842, 1977.
CHAPTER 10

Uses of UV-A Involving Exposure of

Humans

Introduction

The uses of UV-A in medicine and industry are expanding. Such applications
generally employ UV-A-induced fluorescence as a diagnostic tool, or useful
UV-A photochemical or photobiologic reactions. In industry, UV-A-induced
fluorescence is used to detect flaws in metal castings. Industrial use of UV-A
photochemical reactions includes the curing of coating materials, inks, and adhe-
sives and the reprogramming of programmable integrated circuits in the elec-
tronics industry.
Medical uses of UV-A include:
1. Phototherapy and photochemotherapy of skin disorders
2. Photopolymerization (hardening) of plastic dental fillings
3. Hardening of body casts
4. Dermatologic and dental diagnosis

Therapeutic Uses of Ultraviolet Radiation

Around the tum of the century, N. R. Finsen utilized the apparent bacterici-
dal action of ultraviolet radiation to treat tuberculosis of the skin. The later dis-
covery of the role of ultraviolet radiation in the cutaneous conversion of
7-dehydrocholesterol to vitamin D;l reinforced the ancient belief that a "robust"
color was a sign of health. In addition, the social attributes of healthy glow and a
tan have led to popular use of sunlamps (artificial sources of UV-B) and sunbath-
ing.
The use of UV radiation from solar or artificial sources to treat disease is

221
222 Chapter Ten

now usually confined to the therapy of certain skin diseases. Acne vulgaris,
mycosis fungoides, psoriasis, and some forms of chronic eczema are frequently
treated with exposure to UV radiation. The spectral power distribution of most
dermatologic treatment sources has been such that the therapeutic range is usu-
ally within the UV-B region. UV-B has also been used successfully to treat the
pruritus associated with chronic renal failure. 1 Ultraviolet radiation has been
used in combination with topically applied crude coal tar or its derivatives to treat
psoriasis and eczema or with other topical and oral photo active drugs to treat
psoriasis and vitiligo. Certain of these treatment systems are designed to emit
primarily UV-A.
Neonatal hyperbilirubinemia is treated by exposing the infant to visible
(blue) light. 2 The emission of light sources used in the past has included sig-
nificant amounts of ultraviolet radiation. 3 However, while most phototherapy
systems now have eliminated the UV-B and UV-C wavelengths, measurable
UV-A radiation often reaches the infant. These exposures to UV-A are less than
those required to produce erythema in adult Caucasian skin but may be of sig-
nificance in newborns. Occlusive eye patches and glass or plastic filters that ab-
sorb ultraviolet radiation are recommended. 4

Ultraviolet Treatment of Psoriasis

Psoriasis is a disease of unknown etiology that is characterized on both clin-

ical and laboratory grounds as a disorder of epidermal cell proliferation. It is a
common chronic intractable skin disease that affects 1-5% of the world's popula-
tion. Medical costs related to the treatment of psoriasis reach more than $1 billion
a year in the United States alone.
Onset is most often in early life, but psoriasis may begin at any age. Once
manifest, the course of the disease is unpredictable. It may remain localized to a
few sites or become generalized: it may be present either intermittently or con-
tinuously. The lesions of psoriasis are elevated, erythematous, sharply cir-
cumscribed, scaling plaques, which tend to occur symmetrically over bony prom-
inences, such as elbows and knees, but may occur in any site. The tendency to
have psoriasis is inherited.
Microscopically, there is marked epidermal thickening with a regular elon-
gation of rete ridges and elongation of dermal papillae that contain dilated capil-
laries. Within the lower layers of the thickened epidermis, there is an increased
number of reproducing cells, an increased number of mitoses, and acceleration of
the cell cycle. The increased number of skin cells and rapid cell kinetics that
characterize the disorder lead to the formation of lesions that shed scales. The
outer cell layers of skin are manufactured too rapidly. It follows that the most
Uses of UV-A Involving Exposure of Humans 223

likely mechanism by which various treatments of psoriasis are effective is that of

cytotoxicity. Most treatments that are useful lead to transient retardation or inter-
ference with the reproduction and growth of epidermal cells. Ultraviolet irradia-
tion, with or without added photosensitizers, is one such agent.
It has been known for over 100 years that psoriasis often improves during
the summer months. Most patients with psoriasis are improved after repeated sun
exposures. Likewise, UV-B from artificial sources leads to improvement in most
cases of psoriasis. 5 ,6 The therapeutic exposure doses are within the range re-
quired to produce erythema of the uninvolved normal skin. Exposure doses must
be increased as patients tan or mobilize other cutaneous defenses against ul-
traviolet radiation. It has been shown that UV-A alone in large doses also leads to
improvement of psoriasis. 7 Because the UV-A exposure dose required is within
the range needed to produce erythema, treatment of large areas is impractical:
light sources of adequate intensity are not available and the long-term risks are
unknown. While UV-B treatment doses range from 10 to 500 mJ/cm 2 given as a
single daily dose, the daily exposure dose of UV-A must be 30-300 J/cm 2 or
greater.
In 1925, Goeckerman 8 • 9 introduced tar and UV irradiation therapy for
psoriasis. After evaluating other photosensitizing agents, he settled on crude coal
tar, which he believed would enhance the beneficial effects of UV irradiation.
Two subsequent studies claimed that tar alone was as effective as tar plus ul-
traviolet radiation, 1 0,11 but neither study gives ultraviolet dosimetry and it is not
clear whether ultraviolet-induced erythema was achieved. Studies using
erythema-producing doses of ultraviolet showed dithranol plus ultraviolet 12 or tar
plus ultraviolet 13 to be superior to either alone.
It has been shown for coal tar pitch, 14 coal tar, 15 and several commercially
available tars 16 that the action spectrum for delayed erythema induced in skin by
ultraviolet exposure in the presence of these compounds is within the UV-A re
gion. Although the phenomenon of phototoxicity is offered as the explanation for
the therapeutic benefit of the Goeckerman program, the light sources used have
been primarily artificial UV-B sources. Two studies compared the effectiveness
of UV-A plus tar with that of UV-B plus tar in psoriasis therapy and found the
UV-B-tar combination therapy to be more effective. 12,13 However, the exposure
doses of UV -A in both these studies were probably insufficient to elicit a tar
phototoxicity reaction. 16
When using exposure doses adequate to cause cutaneous phototoxicity
(manifest by delayed erythema), Parrish et at. 17 found UV-A and UV-B to be
equally therapeutic to tar-treated psoriatic lesions. It was felt that the UV-B
therapeutic effect is independent of tar photo toxicity, because of the exposure
dose of UV-B required to cause delayed erythema was the same with or without
the tar. TAR-UV-A phototoxicity was demonstrated by the fact that less UV-A
224 Chapter Ten

was required to elicit delayed erythema in the presence of tar than when skin was
irradiated without tar application. Even so, the dose of UV-A required to benefit
tar-treated skin is large enough to be impractical with currently available light
sources.
After application of crude coal tar or tar products to the skin, subsequent
exposure to UV -A may lead to an unpleasant, burning, painful sensation that has
been referred to as the smarting reaction. 14,16,17 Smarting begins relatively ab-
ruptly at some point during UV-A exposure, is relieved simply by discontinuing
the exposure, and may return when exposure is resumed. Smarting usually oc-
curs at doses of UV -A slightly less than that needed to produce delayed erythema
in tar-treated skin, but considerable individual variation exists. The action spec-
trum appears to include the UV-A but may extend into visible wavelengths. The
mechanism of smarting is not known. Cutaneous nerVe endings may be directly
affected by some photochemical event, or they may be indirectly affected via
damage of other cells or components in the skin.
The Goeckerman regimen (crude coal tar product applied topically and sub-
sequent ultraviolet exposure) has become a standard treatment for severe
generalized psoriasis. Its overall effectiveness is without question. Differences in
reported efficacy are probably due to variation in ultraviolet doses, the tar com-
pounds used, the duration and frequency of treatments, and the level of en-
thusiasm and fastidiousness of the therapist. The relative value of the various
components of the Goeckerman regimen are often debated. In summary:
1. The literature is contradictory, but there exists a widespread clinical be-
lief that tar plus UV-B clears psoriasis better than tar or UV-B alone.
Of the two agents, UV-B in daily erythemogenic doses appears to be
the most effective.
2. The action spectrum for erythema associated with tar phototoxicity is
within the UV-A waveband but not the UV-B waveband.
3. If adequate UV-A exposure doses are used, tar plus UV-A is therapeu-
tic. Such treatment is not currently practical.
Considering the spectral power distribution of treatment sources most often
used and the erythema action spectrum of tar-treated skin, the major therapeutic
component of the popularly used Goeckerman treatment is probably UV-B.
Other factors, such as tar, hospitalization, and patient-physician and patient-
nurse interactions, also play an important role. The vehicle used to apply the tar
and the ointments used to treat scaling, dryness, and itching may also be
therapeutic. These lubricants remove scale and also probably alter the optical
properties of the stratum corneum to facilitate the transmission of ultraviolet
radiation. Patient motivation is necessary for toleration of the inconvenience and
messiness of the treatment and is also a component of the therapy.
Uses of UV-A Involving Exposure of Humans 225

Photochemotherapy

Principles of Therapy

Photosensitization reactions (see Chapter 7) are usually regarded as harm-

ful, but it is possible to take therapeutic advantage of this phenomenon. Photo-
chemotherapy is a term defining the combination of light and drug to bring about
a beneficial effect. Usually, in the doses used, neither drug nor radiation alone
has any significant biologic activity: it is only the combination that is therapeutic.
Hematoporphyrin has been used to selectively photosensitize malignant
cells to visible and U V-A radiation. Cell culture studies and in vivo studies in rats
showed that prior infusion of hematoporphyrin led to selective killing of tumor
cells with radiation, while drug alone and radiation alone had no such effect. 18 A
hematoporphyrin derivative (probably purified hematoporphyrin) in combination
with red light has been used to treat human cancer patients. 19 The chemical was
given intravenously, and I to 8 days later the tumors were exposed to visible
light. Complete or partial tumor reduction correlated with histologic tumor ne-
crosis without necrosis of overlying skin. The hematoporphyrin derivative is
thought to remain in malignant tumors longer than in normal tissue. 19 Acridine
orange has been used to photosensitize epithelial tumor cells in mice so that the
tumors could be destroyed by subsequent argon laser exposure. 20
PUVA (Psoralen and UV-A) is a specific form of photochemotherapy. It de-
scribes the oral administration of psoralen and subsequent exposure to UV-A.
Psoralens are naturally occurring, tricyclic, furocoumarinlike compounds, some
of which can be activated by UV-A to produce phototoxic reactions. When
psoralens are added to intact cells or partial cell systems, absorption of energy
from photons within the 320-380 nm waveband results in the formation of
thymine-psoralen photoproducts and leads to transient inhibition of DNA syn-
thesis. Photo active psoralens can be delivered to the skin by either topical or sys-
temic administration. Subsequent exposure to UV-A results in erythema and
pigmentation, which are delayed in onset, occurring hours to days after expo-
sure.

Oral Psoralen Photochemotherapy

PUVA Erythema

The PUVA erythema reaction may range from a barely perceptible redness
to a severe inflammatory reaction accompanied by swelling, tenderness, blister-
ing, and pain. The possibility of a severe phototoxic reaction is the limiting factor
226 Chapter Ten

in using the PUVA reaction for treatment. The occurrence and degree of redness,
however, are related to the dose of both drug and optical radiation and are pre-
dictable. After a fixed amount of psoralen is given, the minimal amount of UV-A
energy subsequently required to produce delayed erythema can be measured
(MPD or minimal phototoxic dose). Increases of UV-A exposure dose beyond
the MPD increase the degree of delayed erythema. UV-A exposures more than
several times the MPD may cause blistering. Baseline melanization (constitutive
melanization), the sunburn history, the number of previous sun or PUVA expo-
sures, and the thickness of the skin influence the amount of UV -A energy needed
to penetrate into the skin to cause photobiologic reactions.
The erythema that results from PUVA differs from sunburn in its time
course: the onset and maximum are later, and the duration is longer. PUVA red-
ness may be absent or just beginning at 12 to 24 hr after ultraviolet exposure
(when UV-B or sunburn erythema is normally at its peak) and may not peak until
48 to 72 hr later. Therefore, PUVA treatments given as frequently as every 24 hr
carry some risk because a second treatment occurs before the extent of the
erythema resulting from the previous treatment is known. In addition, cumulative
phototoxic responses may result from daily exposures. Because skin diseases can
be treated with drug-plus-ultraviolet-exposure doses that are less than the doses
causing severe redness, PUVA treatments are relatively safe when administered
in combination with careful dosimetry.

PUVA and Pigmentation

It has been known for decades that topical application of certain plant de-
rivatives containing psoralens could cause erythema and hyperpigmentation. A
variety of natural products, such as citrus oils or bergamot, may lead to hyper-
pigmentation without noticeable preceding erythema. In 1953, Lerner et al. 21 re-
ported the melanogenic effect of topical psoralen in a worker who accidentally
contacted psoralen and subsequently exposed his skin to ultraviolet radiation. In
the same report, it was observed that vitiligo patients and albinos treated with
oral psoralens showed decreased sensitivity to sunlight. Several years later,
Lerner 22 and Fitzpatrick et al. 23 reported that oral ingestion of
8-methoxypsoralen followed by exposure to sunlight markedly increases
melanogenesis. Further investigations 24 - 26 supported these observations and
found that the sun-protective effects of oral psoralens were not immediate but
were delayed and paralleled the augmentation of solar-induced melanogenesis. In
contrast, shortly after the ingestion of psoralens, the skin becomes highly sensi-
tive to sunlight,23 a consequence of UV -A-induced psoralen phototoxicity. Ar-
tificiallight sources can also be used to induce and augment melanogenesis after
psoralen ingestion. Stegmaier 27 attempted to use repeated exposure to "black
light" fluorescent bulbs, but the inadequate UV -A irradiance of his lamps led to
unimpressive results.
Uses of UV-A Involving Exposure of Humans 227

Histologically and morphologically, the pigmentation that results from

PUVA appears similar to normal melanogenesis (delayed tanning, see Chapter
6). Pigmentation appears to reach a maximum at about 7 to 14 days after PUVA
exposure and lasts weeks to months. As with solar or UV-B-induced tanning, the
individual's tanning response to PUVA is dictated by genetic factors. However,
multiple erythemogenic PUVA exposures can stimulate melanogenesis more than
equivalently erythemogenic UV-B or sun exposures. PUVA has been used to in-
duce tanning in outdoor workers or persons with sun-sensitive skin disorders in
order to increase their tolerance to sun. Careful dosimetry is necessary to prevent
painful reactions and yet induce maximum melanization. While it may be possi-
ble to take advantage of this easily induced melanogenesis to protect persons who
are abnormally sensitive to UV-B radiation, persons abnormally sensitive to
UV-A may not tolerate enough UV-A to permit PUVA treatments.

PUVA Treatment of Vitiligo

Vitiligo is an acquired cutaneous amelanosis of unknown etiology. It is

asymptomatic and, except for the hazard of sunburn of depigmented lesions, is a
benign cosmetic problem. However, the disfigurement produced can be devastat-
ing psychologically and socially, especially for dark-skinned people in certain
cultures.
In Egypt, the plant Ammi majus Linn has been used orally and topically to
treat vitiligo for centuries. 21 .28 EI-Mofty29 found that when certain crystalline
constituents of the plant were ingested prior to sun exposure, repigmentation was
achieved in over 75% of vitiligo patients. Psoralens have been isolated from and
identified as the major photoactive substances of Ammi majus Linn. 3()-32 Certain
of these photo active psoralens 28 have proved to be of benefit in treating vitiligo.
Several investigators have treated vitiligo with topical psoralens and artificial
UV-A sources.21.34-39 Oral psoralen photochemotherapy requires larger UV-A
exposure doses than those required with topical therapy. Many commercially
available artificial sources of UV-A are not of adequate intensities or appropriate
spectral distribution to make therapy of vitiligo with oral psoralens practical, but
if long exposures and large numbers of fluorescent black lights are used, such
treatments are effective. 40 A high-intensity source of UV-A has recently been
developed to meet the requirements of indoor oral psoralen therapy of vitiligo. 41
Significant spontaneous repigmentation of vitiligo occurs in less than 5% of
patients with vitiligo and is usually limited to minimal perifollicular spotty pig-
mentation. In contrast, about 70% of patients treated with oral trimethylpsoralen
and sunlight repigmented when the medication was taken 2 hr prior to sun expo-
sure and repeated at least twice weekly for more than 12 months. 34 Progress is
slow. Significant repigmentation of vitiliginous skin may require 100 to 300
photochemotherapy treatments over a period of months to years. If patients dis-
228 Chapter Ten

continue therapy, they may again lose pigmentation. Total repigmentation is un-
usual and about 30% of patients do not respond at all despite months of therapy.
Methoxsalen (8-methoxypsoralen) may be used in the same doses as those used
for treatment of psoriasis (see below). Orally administered trimethylpsoralen
(0.5-1.0 mg/kg) appears effective in repigmentation of vitiligo 42 even though it
is much less erythemogenic and melanogenic when given orally than is
8-methoxypsoralen. 43

PUVA Treatment of Psoriasis

The concept of using photosensitizing agents to treat psoriasis is not new.

Goeckerman 8 ,9 formalized and popularized this approach in 1925 with the use of
coal tar and ultraviolet radiation. More recently, psoriasis has been successfully
treated with topically applied psoralens and subsequent exposure to UV_A.44-50
This topical treatment of generalized psoriasis is, however, laborious and time-
consuming and sometimes results in tender erythema or blistering reactions. In
addition, intense irregular hyperpigmentation may result at the site of treated
plaques. Some of these side effects have been reduced by immersing patients into
baths containing small amounts of psoralens and subsequently exposing the pa-
tient to ultraviolet sources containing UVA. 51-53 But with the use oflarger expo-
sure doses of UV-A, it is possible to treat psoriasis with oral psoralens. This
treatment is effective 5 ,6,54 and the hyperpigmentation is generalized and uniform.
Careful drug and UV-A dosimetry also makes it possible to obtain therapeutic
results without painful erythema. Bilateral comparison studies showed PUVA to
be superior to the use of ultraviolet radiation. 5,6

Table 10-1. Requirements for Clearing Psoriasis by Skin Typeo,b

Mean J/cm 2 at
Skin type Mean total Mean no. of Mean no. last clearing
(no. of patients) J/cm 2 treatments of weeks treatment

I (48) 191 23.6 12.5 9.6

II (214) 187 21.7 11.0 12.3
III (571) 253 23.6 11.6 14.4
IV (126) 296 24.2 12.3 16.4
V (17) 410 31.7 15.9 19.2
VI (16) 469 22.4 9.6 21.1
p < 0.01 n.s. (p > 0.05) n.s. (p > 0.05) P < 0.01

aFrom Melski, J., Tanenbaum, L., Parrish, J. A., Fitzpatrick, T. B., Bleich, H. L., and 28
Participating Investigators. Oral methoxsalen photochemotherapy for the treatment of
psoriasis: A cooperative clinical trial. J. Invest. Dermatol. 68: 328-335, 1977. Copyright,
The Williams & Wilkins Company, Baltimore, 1977. Used with permission.
bBased on the mean values of 992 of 1005 patients who cleared and for whom complete
data were available. P-values for column trends are for one-way analyses of variance for
unequal sample sizes.
Uses of UV-A Involving Exposure of Humans 229

Table 10-2. Skin Types, Criteria, 320-380 nm UV-A Exposures at the First
Treatment, and Percentage of Patients in the Cooperative Clinical Trial a

Percentage of
Skin type Criteria b J/cm 2 patients

Always bum, never tan 1.5 5.4

II Always bum, sometimes tan 2.5 22.1
III Sometimes bum, always tan 3.5 56.3
IV Never bum, always tan 4.5 12.7
V Moderately pigmented patients 5.5 1.4
VI Heavily pigmented patients 6.5 2.0

aFrom Melski, J., Tanenbaum, L., Parrish, J. A., Fitzpatrick, T. B., Bleich, H. L., and 28
Participating Investigators. Oral methoxsalen photochemotherapy for the treatment of
psoriasis: A cooperative clinical trial.J. Invest. Dermatol. 68:328-335, 1977. Copyright,
The Williams & Wilkins Company, Baltimore, 1977. Used with permission.
bThe criteria for patients with skin types I, II, III and IV, generally of European extraction,
are based on the history of the usual reaction to the first hour of full sun exposure in early
summer. Skin type V generally includes patients of Asiatic, American Indian, Mexican, and
Puerto Rican extraction. Skin type VI generally includes patients of African extraction.

The rationale for the use of PUVA to treat psoriasis is inhibition of the in-
creased DNA synthesis within keratinocytes of the psoriatic lesions. 55 This inhi-
bition is accomplished by a specific interaction of psoralen and DNA molecules
that involves absorption of radiant energy in the UV-A range (320-400
nm).56-58 Absorption of photons in the UV-A spectrum by psoralen molecules
generates photoexcited species (singlet -excited and triplet -excited states). This
activation leads to photoconjugation of psoralen with the pyrimidines of DNA,
leading to the formation of monofunctional single-strand photoadducts with
thymine bases and interstrand cross-links (bifunctional adducts) between a single
psoralen molecule and opposite pyrimidine bases. 59-61 Presumably, this process
leads to an inhibition of DNA synthesis, and thus cell division, within the rapidly
dividing psoriatic epidermis. This may, however, not be the only mechanism re-
sponsible for the observed regression of psoriatic lesions caused by PUVA
therapy. Direct effects of PUVA on the dermis and systemic effects are other pos-.
sible mechanisms for successful PUVA therapy of psoriatic skin.
Patients are exposed to UV-A 2 hr after ingestion of 0.6 mg/kg of
8-methoxypsoralen. The initial UV-A exposure dose (1.0-5.0 1/cm2 ) depends
not only on the spectral energy distribution of the treatment source but also on the
patient's degree of melanization and sunburn history. Because absorption by
melanin in the epidermis reduces the UV-A radiation reaching the level of the
dividing cells, the darker-skinned subjects require greater UV -A exposure doses
(Table 10-1, 10_2).62 Exposure doses must be increased as tanning occurs.
Ideal PUVA treatment sources are those that produce high UV-A irradiances
in the absence of other wavelengths, especially the highly erythemogenic UV-B
230 Chapter Ten

or UV-C regions. The design should allow the entire body surface to be
uniformly treated at one time. Safety devices and reliable methods of measuring
and delivering exact UV -A exposure doses are essential.
The major advantages of PUVA are its ease of administration, the use of
oral instead of topical medication, the relative speed with which remissions are
obtained, and the fact that psoriatic lesions often disappear completely without
trace. This treatment may be effective when other treatments have failed. Be-
cause photoactivation of the drug is confined to the skin, systemic toxicity may
be avoided. Patients can be maintained free of lesions by intermittent mainte-
nance treatment. In one series, 85% of patients were kept in remission,6 a per-
centage that compares quite favorably with the common experience with conven-
tional therapy. 63,64 Disadvantages include the inconvenience of travel to a treat-
ment center, the necessity of avoiding excess sun for 6 to 8 hr after ingestion of
psoralen, and the fact that the scalp is not treated because hair blocks the UV-A.
In addition, some patients are bothered by nausea or itching.

PUVA Treatment of Mycosis Fungoides

Mycosis fungoides (MF) is an uncommon malignant lymphoma that is

characterized by a polymorphous cellular infiltrate. It appears to originate in the
skin but may disseminate and involve visceral organs and cause death. While in
the late stages, characteristic or diagnostic histologic changes occur in the skin,
early in the course of the disease it is difficult to confirm the diagnosis. 65-67
Management is also complicated by the fact that occasionally the light-
microscopic 68 and electron-microscopic 69 - 71 appearance of benign dermatologic
disorders is indistinguishable from MF.
Treatments of MF are many and include topical steroids 72 or nitrogen mus-
tard,73 X-irradiation,74,75 electron-beam irradiation,76-69 and systemic
chemotherapy. 66,74,80- 85 Because of the difficulties in making an early definitive
diagnosis, proposals to use potentially toxic agents early in the course of the dis-
ease in order to achieve possible cure 73,77 carry the risk oftreating some patients
who do not have MF, Because no one therapeutic regimen has been clearly
shown to alter the life expectancy of patients with MF,66,81 it seems reasonable to
use the therapy that has least morbidity, toxicity, or side effects. Sunlight may be
beneficial to early stages of MF. 86,87
PUVA has been used to treat MF. 88 Its ease of administration and apparent
safety make it an appealing alternative, but reports are preliminary and the course
of MF is long and unpredictable. Because the only recognized hazards of PUVA
to date are painful erythema resulting from errors in dosimetry and actinic
changes resulting from prolonged therapy, there is no apparent contraindication
to its use in early cases of MF in which the diagnosis may be somewhat uncertain
and in which treatments with more definite morbidity and toxicity are less appeal-
ing. Considerations of theoretical risk-benefit ratio suggest that UV -A radiation
Uses of UV-A Involving Exposure of Humans 231

of patients with MF receiving topical or oral photo sensitizers merits further in-
vestigation. MF may not arise in the skin but may represent abnormallympho-
cytes with skin tropism. It may be that if skin lesions are cleared, the absolute
tumor mass is decreased, thus permitting the host defenses to eradicate the dis-
ease.

PUVA Treatment of Eczema

Atopic dermatitis (a common genetic form of eczema) may be successfully

treated with oral 8-methoxypsoralen plus UV-A photochemotherapy.89 Atopic
eczema is disabling in its severe form because of the cosmetic appearance, the
severe pruritus, and the occurrence of secondary infections associated with the
disease. Treatment with oral or topical corticosteroids may produce marked side
effects. Reports are preliminary, but the use of PUVA for clearing eczema is po-
tentially superior to existing therapy in severe cases. Further careful study of this
use of PUVA is required in a larger number of patients.

Immunological Effects of PUVA Therapy

A suggested advantage of PUVA over other systemic therapies for skin dis-
ease is the apparent limitation of its effect to the skin. However, the skin is not a
separate and distinct organ but instead contains important elements of other organ
systems. The immune system is well represented in the skin with antibodies,
lymphocytes, and polymorphonuclear leukocytes in the blood vessels and lym-
phatics, plus lymphocytes, mast cells, and macrophages in the tissues of the
dermis. Langerhans' cells in the epidermis may also be involved in immune reac-
tions. Therefore, both the epidermis and dermis are involved in immune func-
tion. Up to 50% of incident UVA radiation penetrates the upper dermis, and there
is evidence to suggest that noncutaneous cells take up psoralens. It is, therefore,
likely that circulating and tissue-fixed cells which form part of the immune sys-
tem could be affected by PUVA, resulting in an alteration in their function and
viability.
Support for this concept comes from reports of a beneficial effect of PUVA
in mycosis fungoides. In this condition, abnormallymphoreticular cells are pres-
ent in the dermis, and these cells share a common origin with normal cells of the
immune system. PUVA can eliminate these cells from the dermis, and this
suggests that at least abnormal lymphoreticular cells can take up psoralens and
that the dose of UVA used is sufficient to influence their viability. Furthermore,
PUVA has been found to be beneficial in atopic eczema, a condition that proba-
bly has an immune pathogenesis. Whether PUVA in this instance is acting via an
effect on cell viability, via mediator production, or via some other means is un-
known.
A number of studies have recently appeared in which the effect of PUVA,
232 Chapter Ten

applied both in vitro and in vivo, on lymphocyte viability and function has been
studied. In vitro irradiation of human mononuclear cells with UVA in the pres-
ence of 8-methoxypsoralen decreases cell viability and function in a dose-related
manner. 90-93 This is not surprising as a similar response oflymphocytes to UVC
radiation has been reported,94 and PUVA has been shown to affect viability in
many cell systems. The relevance of these in vitro effects to in vivo events during
PUVA therapy is not clear. A decrease in the percentage of circulating T lym-
phocytes following PUVA treatments has been reported. 95 ,96 Depression of
thymidine incorporation by mononuclear cells, immediately following PUVA
treatment, has also been seen. 97
These studies are of interest as they may give further insight into the
mechanism of action of PUVA in various diseases. They are too preliminary to
give any clear indication as to whether impairment of immune function should be
added to the list of potential risks of PUVA therapy. However, as yet, there is no
clinical evidence of any sustained immune suppression in patients undergoing
treatment for up to three years.

Risks of PUVA Therapy

Concerns about possible long-term side effects of psoralen photo-

chemotherapy are based on animal experiments and on the known properties of
psoralens and UV-A. Topical psoralen in the presence of UV-A is a potent car-
cinogen in certain laboratory animals. Mice injected intraperitoneally with high
doses of psoralens and subsequently exposed to ultraviolet have developed skin
cancer. 98-102 The same striking carcinogenic effect is not seen after oral ad-
ministration, 28,1 03 but the presence of a low incidence of tumors in one study of
mice fed methoxsalen 104 suggests that more information is needed. Skin cancer
in psoralen-treated patients has not been reported despite 25 years of experience
using oral psoralen and sun exposure to treat pigment disorders. Indeed, some
investigators have considered that the pigmentation produced by PUVA may pro-
tect against sun-induced skin cancer. While evidence for this protection is lack-
ing, one study showed that 2 years of daily use of psoralens did not increase skin
cancer in persons exposed to Texas sunlight. 105 Since PUVA may be used for
decades to treat chronic skin diseases, careful observation for the development of
skin cancer must continue. Repeated phototoxic reactions are also likely to accel-
erate "aging" (see Chapter 8) of the skin especially in persons who tan poorly.
UV-A penetrates the cornea and is absorbed by the lens (see Chapter 4).
Various animal species chronically exposed to high-intensity UV radiation in
laboratory settings have developed cataracts, an effect that is augmented by large
doses of psoralens. 98 ,99,106,107 It is not known how these animal data relate to
humans. Cataracts known to be related to psoralens have not been reported in
humans. 108 However, it is wise to use protective goggles during treatment and to
Uses of UV-A Involving Exposure of Humans 233

avoid prolonged unprotected sun exposure for several hours after psoralen inges-
tion.
Psoralen-UV-A-induced mutagenicity in one-celled organisms has been
demonstrated in vitro. 109.110 Carter et al. 111 have shown that PUVA promotes
sister chromatid exchanges (SCE) in human lymphocytes in vitro. A more recent
study by Wolff-Schreiner et al. 112 using peripheral blood lymphocytes from
psoriatic patients undergoing PUVA therapy could demonstrate no increased fre-
quency of SCE in vivo, both in the short term and in the relatively long term (2
years). The significance of SCE is not yet fully understood but is believed to
reflect chromosome damage and/or repair. 113 Wolff114 has shown that the in-
crease in SCE produced in vitro can be correlated with cross-link excision. SCE
may, however, result from different lesions than those that cause other chromo-
some aberrations. 115 Thus, while the latter are correlated with cell death, SCE
are associated with cell survival and mutagenesis. Information about repair of
psoralen-thymine photoproducts is insufficient. Despite these concerns, after
centuries of use in Egypt and India and 25 years of use in this country,
documented reports of toxic effects of psoralen in humans are lacking. Prospec-
tive studies of PUVA patients and more research information is needed.

UV-A-Activated Polymerization of Resinous Dental

Restorations

Uses of ultraviolet radiation in dental phototherapy duri.ng the first three dec-
ades of this century included treatment of pyorrhea alveolaris, "dental neural-
gia:' pulpitis, and stimulation of the epithelium of the mouth. 116 These uses all
involved primarily the effects of UV-B and UV-C, although radiation measure-
ments and dosimetry were seldom accurate in this early period. Current use of
ultraviolet radiation in dentistry is limited to UV -A photopolymerization of plas-
tic resinous fillings, restorations, and sealants, and use of UV-A fluorescence as
a diagnostic tool.
Resinous restorations are often less traumatic and more aesthetically accept-
able to patients than are restorations using conventional materials. The protection
afforded by resinous sealants against caries initiated at sites of pits and fissures on
the teeth is significant. 117.11S Autopolymerizing resins, once mixed, harden over
a given length of time. UV-A-activated polymerization has the practical advan-
tage over autopolymerizing resins of allowing the dentist to control the time of
hardening of the plastic materials.
The possible biologic hazards associated with this use of UV -A are a matter
of debate. The interior of the mouth normally receives very little exposure and
lacks a protective stratum corneum. The oral mucosa appears to be only slightly
more sensitive to single ultraviolet exposures than is the skin of fair-skinned in-
234 Chapter Ten

Table 10-3. Skin Fluorescence under Wood's lamp

Color observed
with
Clinical setting Fluorescent substance Wood's lamp

Normal skin Dermal and epidermal protein Blue-white

Tinea capitis Pteridine Blue-green
(e.g., Microsporum)
Erythrasma Porphyrin Coral-red
(Corynebacterium minutissimum)
Pseudomonas infection Fluorescein Yellow-green
Porphyria cutanea tarda Uroporphyrin Orange-pink or
pink-orange

dividuals. 119 Current dental uses of UV-A involve several brief suberythemal
exposures to small areas of the mouth, unlike the chronic wide-area and occa-
sionally erythemogenic UV exposures to which skin is subjected daily.

Diagnostic Uses of UV-A

In 1903, Wood 12 0 developed a source of UV-A emitting very little visible

light, consisting of a medium-pressure mercury arc and a nickel oxide glass filter.
Modern "Wood's lamps" vary somewhat but are all sources ofUV-A with min-
imal visible radiation. UV-A is absorbed by the material under examination, and
light in the visible spectrum (400-700 nm) is emitted (fluorescence and phos-
phorescence). Practicing dermatologists have used Wood's lamp to diagnose
such disorders as tinea capitis, erythrasma and some pseudomonas infections,
and porphyria cutanea tarda by noting the characteristic fluorescence of hair
shafts, skin, and urine, respectively (Table 10_3).121
Blue-white fluorescence of both epidermal and dermal proteins occurs under
Wood's lamp illumination. UV-A penetrates the skin less deeply than visible
light and is absorbed more than visible light by epidermal melanin. Therefore,
variations in epidermal melanin pigmentation are more apparent under Wood's
lamp than under visible light. For example, areas of vitiligo, which completely
lack epidermal pigmentation, are much more apparent when viewed under
Wood's lamp than under standard visible light. 122 Because of the lesser penetra-
tion of UV-A as compared with visible light, variations in dermal (pathologic)
pigmentation are much less apparent under Wood's lamp than under visible light.
For example, blue nevi, a dermal melanocytoma, are frequently unseen under
Wood's lamp, whereas under visible illumination the blue-gray lesions are easily
Uses of UV-A Involving Exposure of Humans 235

seen. The lamp may therefore be used as one means to determine the localization
of pigmented lesions.
A similar diagnostic procedure is useful in dentistry. UV -A fluorescence is
altered by the degree of calcification of teeth and bones. 103 Localized changes in
UV -A fluorescence of teeth may also aid in diagnosis of early dental caries, and
tetracycline incorporation into the teeth may be readily observed. 124 Plaque, cal-
culus, and some stains are also more apparent when viewed (as fluorescence)
under UV-A than under visible light.

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87. Van Scott, E. J., and Haynes, H. A. Cutaneous lymphomas. In Dermatology in General
Medicine (T. B. Fitzpatrick, K. A. Arndt, W. H. Clark, A. Z. Eisen, E. J. Van Scott, and
J. H. Vaughan, Eds.). McGraw-Hill, New York, 1971, pp. 557-567.
88. Gilchrest, B. A., Parrish, J. A., Tanenbaum, L., Haynes, H. A., and Fitzpatrick, T. B. Oral
methoxsalen photochemotherapy of mycosis fungoides. Cancer 38:683-689, 1976.
89. Morison, W. L., Parrish, J. A., and Fitzpatrick, T. B. Oral psoralen photochemotherapy of
atopic eczema. 1. Invest. Dermatol. 67:561, 1976.
90. Scherer, R. The human peripheral Iymphocyte-a model system for studying the combined
effect of psoralen plus black light. Klin. Wochenschr. 55:137-140,1977.
91. Scherer, R., Kern, B., and Braun-Falco, O. UVA-induced inhibition of proliferation of PHA-
stimulated lymphocytes from humans treated with 8-methoxypsoralen. Br. 1. Dermatol.
97:519-527, 1977.
92. Lischka, G., Bohnert, E., Bachtold, G., and Jung, E. G. Effects of 8-methoxypsoralen (8-
MOP) and UVA on human lymphocytes. 1. Invest. Dermatol. (Abstract) 68(4):245, 1977.
93. Edelson, R., Jacobs, D., Brin, M., and Kochevar, L. Lymphocyte sensitivity to
8-methoxypsoralen (8-MOP) and ultraviolet-A (UVA). Clin. Res. 25:281A, 1977.
94. Horowitz, S., Cripps, D., and Hong, R. Selective T cell killing of human lymphocytes by ul-
traviolet radiation. Cell. Immunol. 14:80-86, 1974.
95. Cormane, R. H., Hamerlinck, F., Simon, M., and Siddiqui, A. H. Photoimmunology in
psoriasis. 1. Invest. Dermatol. (Abstract) 68(4):253, 1977.
96. Ortonne, J. P., Claudy, A. L., Alario, A., and Thivolet, J. Decreased circulating E rosette
forming cells in psoralen UVA treated patients. Arch. Dermatol. Res. 258:305-306, 1977.
97. Kraemer, K. H., and Weinstein, G. D. Decreased thymidine incorporation in circulating
leukocytes after treatment of psoriasis with psoralen and long-wave ultraviolet light. 1. Invest.
Dermatol. 69(2):211-214, 1977.
98. Clark, D. H. Photosensitization by 8-methoxypsoralen. 1. Invest. Dermatol. 37:171-174,
1961.
99. Griffin, A. C. Methoxsalen in ultraviolet carcinogenesis in the mouse. 1. Invest. Dermatol.
32(Suppl.):367-372, 1959.
100. O'Neal, M., and Griffin, A. C. The effect of oxypsoralen upon ultraviolet carcinogenecity in
albino mice. Cancer Res. 17:911-916, 1957.
101. Hakim, R. E., Griffin, A. C., and Knox, J. M. Erythema and tumor formation in
methoxsalen-treated mice exposed to fluorescent light. Arch. Dermatol. 82:572-577, 1960.
102. Urbach, F. Modification of ultraviolet carcinogenesis by photoactive agents. 1. Invest. Der-
matol. 32(Suppl.):373-378, 1959.
103. Langner, A., Wolska, H., Jarzabek-Chorzilska, M., and Pawinska, M. Dermal toxicity of
8-methoxypsoralen in hairless mice. 1. Invest. Dermatol. 64:279-280, 1975.
240 Chapter Eleven

104. Griffin, A. c., Hakim, R. E., and Knox J. The wavelength effect upon erythema and car-
cinogenic response in psoralen-treated mice. J. Invest. Dermatol. 31 :289-295, 1958.
105. MacDonald, E., Griffin, A. C., Hopkins, c., Smith, L., Garrett, H., and Black, G. L. Psora-
len prophylaxis against skin cancer: Report of clinical trial. J. Invest. Dermatol. 41 :213-223,
1963.
106. Cloud, T. M., Hakim, R., and Griffin, A. C. Photosensitization of the eye with methoxsalen.
Arch. Ophthalmol. 64:346-351, 1960.
107. Cloud, T. M., Hakim, R., and Griffin, A. C. Photosensitization of the eye with methoxsalen.
II. Chronic effects. Arch. Ophthalmol. 66:689-694, 1961.
108. Cogan, D. Photosensitization and cataracts. Arch. Ophthalmol. 66:28-29, 1961.
109. Igali, S., Bridges, B., Ashwood-Smith, M., and Scott, B. Mutagenesis in Escherichia coli.
Mutat. Res. 9:21-30, 1970.
110. Swanbeck, G., and Thyresson, M. Induction of respiration-deficient mutants in yeast by psora-
len and light. J. Invest. Dermatol. 63:242-244, 1974.
111. Carter, D. M., Wolff, K., and Schnedl, W. 8-Methoxypsoralen and UVA promote sister-
chromatid exchanges. J.lnvest. Dermatol. 67:548-551, 1976.
112. Wolff-Schreiner, E. C., Carter, M., Schwarzacher, H. G., and Wolff, K. Sister chromatid ex-
changes in photochemotherapy. J.lnvest. Dermatol., 69:387-391, 1977.
113. Latt, S. A. Sister chromatid exchanges, indices of human chromosome damage and repair: De-
tection by fluorescence and induction by mitomycin C. Proc. Natl. Acad. Sci. USA. 7/:3162-
3166, 1974.
114. Wolff, K. Oral methoxsalen photochemotherapy of psoriasis: Results-Follow-up and pathol-
ogy. In Proceedings of the Second International Symposium on Psoriasis, Stanford University,
Stanford, California (E. M. Farber and A. J. Cox, Eds.). Dun-Donnelley, Yorke Medical
Books, New York, 1977, pp. 300-309.
115. Wolff, S., Rodin, B., and Cleaver, J. E. Sister chromatid exchanges induced by mutagenic
carcinogens in normal and xeroderma pigmentosum cells. Nature 265:347-349, 1977.
116. Christen, A. G. Ultraviolet radiation and fluorescence in diagnosis, therapy, and research. J.
Oral Ther. Pharmacol. 3:62-79, 1966.
117. Silverstone, L. M. Fissure sealants. Caries Res. 8:2-26, 1974.
118. Buonocore, M. G. Caries prevention in pits and fissures sealed with an adhesive resin
polymerized by ultraviolet light: A two-year study of a single adhesive application. J. Am.
Dent. Assoc. 82:1090-1093, 1971.
119. Payne, T. F. An evaluation of actinic blocking agents for the protection of lip mucosa. J. Am.
Dent. Assoc. 92:409-411, 1976.
120. Wood, R. W. Secret communications concerning light rays. J. Phys. 5e Serie, T. IX, March
1919.
121. Caplan, R. M. Medical uses of the Wood's lamp. J.A.M.A. 202:1035-1038, 1967.
122. Gilchrest, B. A., Fitzpatrick, T. B., Anderson, R. R., and Parrish, J. A. Localization of
melanin pigmentation in the skin with Wood's lamp. Br. J. Dermatol. 96:245-248, 1977.
123. Benedict, H. The fluorescence of teeth as another method of attack on the problem of dental
caries. J. Dent. Res. 9:274, 1929.
124. Hefferren, J., Cooley, R., Hall, J., Olsen, N., and Lyon, H. Use of ultraviolet illumination in
oral diagnosis. J. Am. Dent. Assoc. 82:1353-1360, 1971.
CHAPTER 11

Safety Measures and Protection

against Ultraviolet Exposure

Introduction

There are potential biologic hazards associated with exposure to essentially every
spectral region of the continuum of electromagnetic radiation. Exposure to mi-
crowave radiation is associated with heating of tissues, and possibly cataract
formation and deleterious effects upon. the central nerv.ous system. Infrared
energy may also cause destructive heating of the skin and eyes and cataract for-
mation ("glass blower's cataract"). Near infrared radiation and visible light fo-
cused on the retina may produce retinal bums.~The shorter wavelengths of the
visible light spectrum produce retinal damage at much lower irradiances than the
longer visible and near-infrared wavelengths. I The biologic effects and hazards
of ultraviolet radiation on the skin and eyes of humans have been well recorded in
the literature. Ionizing radiation (X rays and gamma rays) can cause tissue de-
struction and mutagenesis and can penetrate deeply into the body.
It is known that radiant energy in the visible region of the electromagnetic
spectrum may produce a wide variety of biologic injuries, including cell death
and bacterial mutagenicity. 2,3 Visible light may be mutagenic, lethal, or car-
cinogenic to animals that have been exposed to certain photosensitizing sub-
stances. 4 - 9 Visible light also affects numerous neuroendocrine functions in
mammals. 10-14 Despite the state of incompleteness of existing information, the
obvious beneficial effects of visible light to man cause us to accept some risks of
these deleterious effects. The absence of such obvious benefits of ultraviolet radi-
ation engenders a less permissive ,md more defensive attitude toward this spectral
region.
Chronic and acute exposure of humans to electromagnetic radiation, includ-
ing solar or artificial ultraviolet radiation, may entail both benefits and hazards,
depending upon the wavelengths present, the exposure dose and duration, the
241
242 Chapter Eleven

areas exposed, and the condition of the individual. For instance, the amount of
solar UV -B exposure may be largely responsible for fluctuations in vitamin D3
levels in humans. Vitamin D levels affect calcium absorption rates, and in-
sufficient UV exposure can result in rickets in children. 15 There is some evidence
that the ultraviolet emission of environmental lighting affects calcium absorp-
tion. I6 UV-A may also playa role in the photorepair of DNA damage associated
with UV-B and UV-C exposures. Chapter 10 reviews beneficial industrial, diag-
nostic, and therapeutic uses of UV-A. Other unknown or less documented bene-
ficial effects of UV exposure probably exist. However, chronic UV exposures
may potentially lead to carcinogenesis and actinic keratosis of skin 17,18 and pos-
sibly to cataract formation. Acute exposure hazards include painful sunburn and
keratoconjunctivitis, depending on the exposure and wavelengths. Clearly, it is
not beneficial to live either in total darkness or under daily intense radiation ex-
posure. However, the definition of the beneficial region between these extremes
is not clear in the case of UV exposure, especially for the UV -A spectral region.

Ultraviolet Exposure Safety Standards

In the past 25 years, the number of commonly used ultraviolet sources has
increased steadily, extending into consumer, medical, industrial, and military
use and necessitating the development of safety standards for ultraviolet expo-
sure. 19-23 The setting of such standards is complicated because value judgments
based on inadequate experimental data are often involved. Considerable latitude
therefore exists for the expression of arbitrary personal opinions on the healthi-
ness or hazards of UV exposure.
The most accepted ultraviolet exposure standard for conventional (nonlaser)
UV sources is that proposed by the National Institute for Occupational Safety and
Health (NIOSH),24 although other standards have been proposed.I 9,25 The
NIOSH-proposed standard is based on an envelope action spectrum developed by
Sliney 26 (Fig. 11-1), combining the action spectra for eye photokeratitis and
Caucasian skin erythema over the wavelength range of high sensitivity, 200 nm
to 315 nm. The envelope curve was defined as a smooth curve somewhat below
the energies required for development of observable eye or skin reactions. There-
fore, the criterion used in defining "safe" in this standard is any exposure that is
below that causing observable acute effects in most people. The standard is in-
tended to apply to environmental or occupational exposures to artificial sources
during an eight-hour working day.
The proposed envelope action spectrum (Fig. 11-1) shows maximum sen-
sitivity at a wavelength of 270 nm, corresponding to the eye photokeratitis
maximum, and defines 3.0 mJ/cm 2 as the maximum safe exposure to 270 nm
radiation. The maximum allowable exposure for monochromatic sources in this
wavelength region can be read directly from the curve. Maximum allowable ex-
Safety Measures and Protection against Ultraviolet Exposure 243

1 .0r--..--.--.,---,----r--nr----=I

.E
--....,.
u

>-
f-
C/)
Z
w
o
>-
t:l 0.01
0:
w
Z
w

o.00 1~-...1_--L.----I---'--....J....-___L...--...J
200 300 320 340
WAVELENGTH, nm

Fig. II-I. Envelope action spectrum used to define the ultraviolet exposure standard recommended
by NIOSH and accepted by ACGIH. (From Occupational Exposure to Ultraviolet Radiation. US-
DHEW, National Institute for Occupational Safety and Health (NIOSH), HSM 73-11009,
Washington, D.C., 1972.)

posures to broadband ultraviolet sources must be calculated by weighting the

source's spectral irradiance of wavelengths less than 320 nm relative to 270 nm.
The weighting factors, SA, are given in Table 11-1. The summation of the weigh-
ted spectral irradiance from 200 to 315 nm is the effective hazardous irradiance,
E eff , typically in p,W/cm 2 • The permissible effective exposure dose, determined
by Eeff x exposure duration in seconds, is 3.0 mJ/cm 2. The method of summa-
tion of the effectiveness of wavelengths less than 315 nm assumes both that there
is additivity of the hazards presented by these wavelengths and that no significant
synergistic or protective interactions exist between wavelengths; it is unlikely
that either of these assumptions is true. However, summation of hazard effec-
tiveness by wavelength is a practical means of defining a standard until more data
are available.
More explicitly, the effective hazardous irradiance relative to 270 nm is
313nm
Eeff (W/cm2) =I EASALlA
200 nm

where E A = spectral irradiance in W / cm 2 • nm, SA = relative spectral effective-

ness (weighting factors, Table 11-1), LlA = spectral bandwidth used for the
244 Chapter Eleven

Table 11-1. N IOSH Total Permissible Eight-Hour Exposure Doses

for Monochromatic Sources, and Relative Effectiveness S A (Weighting Factors)

NIOSH permissible NIOSH

Wavelength 8-hr exposure relative spectral
(nm) (mJ/cm 2 ) effectiveness SA

200 100.0 0.03

210 40.0 0.075
220 25.0 0.12
230 16.0 0.19
240 10.0 0.30
250 7.0 0.43
254 6.0 0.50
260 4.6 0.65
270 3.0 1.00
280 3.4 0.88
290 4.7 0.64
300 10.0 0.30
305 50.0 0.06
310 200.0 0.D15
315 1000.0 0.003

summation, in nm. The maximum permissible exposure time, t max in seconds is

3 X 10- 3 (J/cm2)
t max =
Eeff (W/cm 2 )

Three examples of the application of the NIOSH 200-315 nm exposure

standard are presented below. The first is of a monochromatic source, the second
is of typical noontime solar radiation, and the third is of cool-white fluorescent
lamps. Experimentally determined thresholds for skin erythema are presented for
these examples and compared with the hazard calculations.
Example I: A low-pressure mercury discharge lamp (germicidal lamp) is used to maintain
sterile eonditions within a tissue culture hood. This lamp emits over 90% of its radiant output at
a wavelength of 254 nm. The maximum permissible 8-hr exposure dose at this wavelength is 6.0
mJ/cm 2 (6.0 x 10- 3 J/cm 2 ). If the irradiance at the working surface of the tissue culture hood is
30 p.W/cm2 (3 x 10- 5 W/cm 2), then the maximum permissible exposure time is

{max =
6 X 10- 3 J/cm 2
3 X 10- 5 W/cm 2

or 200 sec. That is, in the course of a working day, the user of the hood should receive a cumula-
tive exposure duration of less than 200 sec. This calculation oft max is applicable only within the
hood, and for a specified working distance from the lamp, at which point the UV-C irradiance
measurement was taken. In practice, the total UV exposure dose is accumulated by the user at
Safety Measures and Protection against Ultraviolet Exposure 245

different times, at a variety of distances, and also depends on variables such as relative shape and
orientation of the hood and the user and the presence and type of clothing. While it may be
possible to estimate the range of UV-C exposure doses that a typical user might receive, one
should realize the impossibility of precision in such estimations. The minimal erythemal dose
(MED) of fair Caucasian skin to this source is 6- IO mJ/cm 2 , essentially equal to the NIOSH
maximum permissible exposure.
Example 2: Typical noontime solar UV spectral irradiances, E A, at selected wavelengths
less than 315 nm, are given in Table 11-2. The NIOSH weighting factors, SA, are also given for
these wavelengths. The product of E A and SA is then multiplied by the wavelength interval, in
this case 5 nm, and the results are then summed to arrive at E eff, the total effective hazardous
irradiance defined by the proposed standard. In this example, Eeff is 3.5 /LW/cm2, and the
maximum allowable exposure time for solar wavelengths less than 315 nm is 14 min 20 sec
(Table 11-2). The MED of this sunlight is approximately 20 min in fair-skinned Caucasians.
Example 3: One of the most commonly used indoor light sources is the cool-white fluores-
cent lamp. These lamps have a soda-lime glass envelope that strongly attenuates UV -C and
UV-B emission lines of the lamp's internal mercury discharge, but does allow some transmis-
sion of 297, 302, 313 nm, and longer wavelength spectral emission lines. Calculation of the
maximum allowable exposure to a bank of 4 of th~se lamps at close distance is presented in
Table 11-3. The NIOSH-proposed standard would in this example set an exposure duration limit
of 77 min. The minimal erythema dose was measured 24 hr after exposing the untanned backs of
two fair-skinned Caucasian subjects to these lamps. For this source, the proposed standard's
limit is well below the lowest exposure causing delayed erythema. Using I-hr increments and
continuous exposure at a distance of 30 cm, the MED for both subjects was 8 hr. Delayed
erythema was followed by tanning. It is interesting that grossly observable biologic effects in
humans could be seen after exposure to this very common source of light but the exposure dis-
tance was only 30 cm and the time required was a full 8 hr.
In the 320-400 nm (UV-A) region, a maximum total exposure of 1.0 J/cm 2
for periods less than 1000 sec (about 17 min) and a maximum UV-A irradiance
of 1.0 mW/cm 2 for periods greater than 1000 sec are allowed by the standard.
For example, noontime solar UV-A irradiance is approximately 5.0 mW/cm 2 ,
therefore exceeding the 1.0 mW/cm 2 limit for long exposures and producing the
maximum allowed exposure dose of 1.0 J/cm 2 in about 3.5 min. In this case, the
NIOSH recommended standard for UV-A appears very conservative. However,
for a source emitting 1.0 mW/cm 2 UV-A irradiance, the maximum allowable ex-
posure would be (1.0 mW/cm 2 ) x (8 hr) x (3600 sec/hr), or 28.8 J/cm 2 • This
UV-A exposure dose may be sufficient to cause delayed skin erythema in some
fair-skinned individuals (see Chapter 6). In any case, the UV-A exposure stan-
dard was recommended by NIOSH in the absence of adequate biologic data and
therefore reflects some uncertainty.
It is important to note that the NIOSH-proposed standard is drawn below the
threshold of observable effects and that the standard is based on monochromatic
experimental exposures, mainly of the eyes. The eyes are often naturally shielded
from radiant exposure, depending upon the subject's position and the source in
question. If, for instance, the ultraviolet source is overhead, as is roughly the
case with sunlight, certain skin areas are likely to receive much more UV expo-
sure than the eyes. In such situations, a direct application of the recommended
standard is overly conservative.
 Table 11-2. Application of the N IOSH-Recommended UV Exposure Standard
to Noontime Solar UV Wavelengths Less than 315 nm a

NIOSH effective
NIOSH rel;;.tive irradiance,
Solar spectral irradiance effectiveness Eeff (W/cm') over 5 nm,
Wavelength, nm E"A (W/cm' • nm) b weighting factor, ~ = E"AS"Ab."A

290 0.0 0.0

295 1.0 X 10- 7 0.47 2.4 X 10- 7
300 5.4 X 10- 7 0.30 8.1 X 10- 7
305 4.4 X 10- 6 0.06 1.3 X 10- 6
310 1.1 X 10- 5 0.Ql5 8.3 X 10- 7
315 2.3 X 10- 5 0.003 3.4 X 10- 7

Total Eeff 3.5 X 10- 6

fYI/em' )

Maximum permissible 3 X 10-3 J/cm' 860 sec, or

t max (sec)
exposure time, tmax Eeff W / cm ' 14 min 20 sec

aThe NIOSH maximum permissible exposure time for noontime solar UV-A is about 3.S min (see text).
bSolar spectral irradiance data is for summertime conditions, solar zenith angle of 2So, at Davos, Switzerland. (Modified from Bener, P. The diurnal
and annual variations of the spectral intenSity of ultraviolet sky and global radiation. Contract AF61(OSw)-618, Technical Note 2, 1963.)
 VI

j
Table 11-3. Application of NIOSH-Recommended UV Exposure Standard to Nearby Bank of Four Cool-White Fluorescent Lamps 3:
m
III
r::::
Cool-white fluorescent NIOSH relative NIOSH effective iil
III
lampb spectral irradiance,c effectiveness weighting irradiance, Eeff 1\1
~
Wavelength, nmil FA (W/cm 2 • nm) factor, SA (W/cm2) = EASAD.A Q.
'tI

254 <1.0 X 10- 8 0.50 <0.50 X 10-" ~

297 6.2 X 10- 7 0.40 2.5 X 10- 7
302 9.1 X 10- 7 0.20 1.8 X 10- 7
o·~
~

313 2.8 X 10- 5 0.008 2.2 X 10- 7 1\1

CQ
334 2.8 X 10-' 1\1

365 4.3 X 10- 5

:i"
!!l.
Total Eeff (W/em 2 ) 6.5 X 10- 7 ...~
1\1

3 X 10-3 J/em 2
o·<
Maximum permissible i
t max (see) 460 see (77 min)
exposure time, t max Eeff W/em 2 m
-g><
Exposure time for minimal III
r::::
erythema dose (MED) in two very 480 min (8 hrs) iil
fair-skinned volunteers

a UV emission spectrum consists of Hg spectral lines.

bPlanar array of four G.E. Mainliner F40 CW lamps spaced IS cm apart, at a distance of 30 cm from detector. Illuminance was 1300 fc.
cSpectral irradiance expressed for an emission line bandwidth of 1 nm.

~
-.j
248 Chapter Eleven

Another complication in the enforcement of an exposure standard is that of

accurately monitoring spectral irradiances reaching humans in each application
of a source of ultraviolet radiation. Some portable radiometers are available with
a spectral response that approximates that of the recommended standard for
wavelengths less than 320 nm, and UV-A may be measured separately. Another
approach to hazard determination has been the wearing of photosensitive film
badges which mimic the skin's erythemal response. Given the complexities of
accurate UV measurements and the uncertainties involved in the setting of the
standard itself, one is tempted to suggest that the threshold delayed erythema re-
sponse of willing fair-skinned Caucasians be the final test used to assess biologic
hazard levels. Even so, it cannot be assumed that possible hazards of chronic
exposures correlate with delayed erythema or eye photokeratitis responses, both
of which are apparently reversible.
The need for more photobiologic research relevant to human exposure is
both evident and pressing. We are just beginning to understand the complicated
biologic effects of optical radiation. And yet there already exists a wide array of
strong opinions, from those who insist that daily ultraviolet exposure is necessary
for emotional health to those who insist that minimal ultraviolet exposure from
any artificial source is a menace to health.
It is important that groups involved in the manufacture, use, and regulation
of sources emitting ultraviolet radiation carefully consider the positions they
take. Industry should not find comfort in the public's willingness to accept risks,
the natural occurrence and unregulatability of sunlight, and the equation of "min-
imal skin erythema dose" with "safety envelopes." Regulatory agencies should
not allow the comfort of conservatism to become the policy of regulation and
assume dictatorial power. Finally, the scientist must be swayed by neither group
and must be willing to admit that being asked to be an authority does not, by
definition, make one qualified to be so or increase one's base of information.

Sunscreens

Ultraviolet-radiation protective agents are substances applied topically,

given systemically, or produced endogenously that alter adverse ultraviolet-
induced reactions of the skin by altering optical or photochemical pathways. *
Most of the agents employed as solar ultraviolet sunscreens are topically applied
chemicals that absorb UV-B radiation. These agents include para-aminobenzoic
acid, esters of para-aminobenzoic acid, cinnamates, and salicylates. It has been
shown that para-aminobenzoic acid in 70% alcohol is a very effective sunscreen
that retains protection for a prolonged period of time under a variety of conditions

• Modified from the unofficial working definition adapted by the sunscreen session of the American
Society of Photobiology, Denver, Colorado, 1976.
Safety Measures and Protection against Ultraviolet Exposure 249

of use. 27 It has been suggested that this protection and substantivity result from a
chemical reaction between the ultraviolet absorber and certain components of the
stratum corneum. 27 The effectiveness of para-aminobenzoic acid in alcohol, es-
ters or para-aminobenzoic acid,28 or cinnamates in combination with ben-
zophenones,29 when properly used, permits exposures of more than 10 times the
minimal erythema dose before any delayed erythema may occur. If it requires 12
to 30 min of solar radiation exposure to cause delayed erythema on Caucasian
skin, some of the better sunscreens allow 2 to 5 hr of sun exposure before delayed
erythema is produced.
However, many sunscreens that absorb UV-B wavelengths absorb relatively
little UV-A. Their use therefore increases solar UV-A exposure to the user by
allowing extended sunbathing. Although solar UV-A is much less effective than
solar UV-B in causing erythema and other biologic effects in human skin, sun
exposure in the presence of sunscreens may be of long enough duration to allow
UV-A erythemogenesis or sufficient UV-A exposure to be additive to the
erythemogenesis of UV-B.
Many sunscreen manufacturers consider transmission of UV-A desirable
because of the known role of UV-A in inducing the immediate tanning reaction
and in augmenting delayed tanning reaction or melanogenesis. UV-A is thought
to stimulate melanogenesis with less relative erythemogenesis than that induced
by UV-B. It is true that melanin pigmentation can be induced by UV-A alone. It
is also true that in some individuals the UV -A melanogenesis dose is less than the
UV-A erythema dose, but this is not necessarily unique to UV-A. A concept that
has been exploited commercially in the sale of sunscreens involves an erroneous
belief that sunscreens actually stimulate tanning reaction. This belief is abso-
lutely incorrect. The pigment-producing cells of the skin are activated by the ab-
sorbed energy in the UV-B as well as in the UV-A spectrum and not by any
ingredient of the sunscreen preparations. If a sunscreen is effective in blocking all
of the erythemogenic rays of both UV-B and UV-A, no tanning (delayed
melanogenesis) occurs.
Ideally, an effective sunscreen agent should block the major portion of the
impinging UV-B radiation that causes the sunburn reaction. It should be substan-
tive to the skin and should remain under varying conditions, including sweating,
immersion of skin in water, and high and low humidity. It should be cosmetically
acceptable. The final testing of sunscreens should be conducted under field condi-
tions. 27 ,30,31 Some investigators have explored the effectiveness of nonopaque
ultraviolet absorbers as protectants against UV _C 32 and against UV -A. 33,34
Some of the newer approaches to sunscreen formulation combine compounds
such as benzophenones with cinnamates or other UV-B blockers to obtain a
broader spectrum of protection. 35
Benzophenones and benzotriazoles absorb over a wide spectral range, in-
cluding some portions of UV-A and UV-C. The absorption curves 36 - 38 of cer-
tain diphenylketone compounds show appreciable absorption of wavelengths
250 Chapter Eleven

longer than 320 nm. Alcohol solubility39 permits delivery of effective screening
amounts to the skin. Chemicals such as 2-hydroxy-4-methoxybenzophenone and
other benzophenones that are known to absorb UV -A radiation are often incorpo-
rated into an appropriate base and claimed to be effective sunscreens against
UV-A as well as UV-B. Most of these agents are effective absorbers up to 340
nm to 350 nm. Beyond this range, the absorption decreases, and hence their
photoprotective potency in the 360-400 nm spectrum becomes significantly less.
These agents are not particularly substantive to the skin under stress of perspira-
tion or immersion of skin in water, as they are easily washed off.
To provide protection against a wider spectrum of solar ultraviolet radiation,
further research is being done to combine benzophenones and carotenoids to
UV-B blockers and to find vehicles that provide substantivity and cosmetic ac-
ceptability. Naphthalene-l,5-bisureas can be modified to absorb in the UV_A40
and UV-B regIOns and therefore are being evaluated as sunscreens. Dihydroxy-
acetone and naphthoquinone have been used in an attempt to alter the stratum
corneum chemically to achieve decreased ultraviolet transmission of UV-B and
UV -A. 41 While clinical studies show light-sensitive patients to be improved,42
protection factors of normal skin in sunlight have not been impressive. 43
Evaluation of sunscreens designed to diminish UV-A radiant exposure can
be accomplished by the use of appropriately filtered sun or solar simulators, but
exposure times may be long. High-intensity UV-A sources that can irradiate
large areas of skin are not easily available. One testing approach has been to
evaluate the ability of sunscreens to prevent the erythema response of photosen-
sitizers whose action spectrum is known to be within the UV-A. After known
amounts of psoralens are given by mouth or applied topically, the exposure dose
of UV-A required to produce certain grades of erythema can be measured.
Sunscreens can be tested for their ability to protect against this delayed erythema.
One such study showed that topically applied 10% benzophenones prevented de-
layed erythema for UV-A exposures of more than 10 times that needed to cause
erythema in the absence of the sunscreen. 34
Many investigators are working to develop an oral photoprotectant, but no
such agents are yet practical. 44,45 Oral psoralens provide some protection, which
is best demonstrated after induction of melanogenesis. 46.47 Antimalarials are not
effective in altering the grossly visible responses of normal skin to sun. Vitamin
A and triprolidine are not effective in diminishing sunburn response. 44 The effec-
tiveness of oral beta carotene in treating erythropoietic proto porphyria, a light-
sensitivity disease in which patients are sensitive to 380-420 nm radiation, 48 en-
courages those seeking oral ultraviolet photoprotectants, but beta carotene does
not protect normal skin from sunburn. 45 There is some evidence, however, that
beta carotene protects against UV-induced carcinogenesis in experimental ani-
mals. 49 No practical and effective systemic UV-A photoprotectant is known to
date. Table 11-4 reviews photoprotectants and their uses over the optical spec-
trum of interest.

Radiation
Germicidal ultraviolet:
(less than 290 nm);
UV.c
Sunburn ultraviolet:
(290-320 nm); UV-B
Longwave ultraviolet:
(320-400 nm); UV-A
Visible light:
(400-700 nm)
a Modified from Fitzpatrick et al.• p. 752.'9
Table 11-4. Photoprotectants (1972-1973)a
Situations for which
photoprotectant may be required
Prevention against radiation from
artificial light in operating rooms and
industrial facilities.
Prevention of sunburn; polymorphous
photodermatitis; prevention of skin
cancer; possibly prevention of actinic
"aging."
Melasma; photosensitivity to drugs;
polymorphous photodermatitis; pre-
vention of sunburn (additive effects);
? prevention of wrinkling, aging, skin
cancer.
Melasma; erythropoietic protoporphyria.
Photoprotectant indicated
Topical Oral
Benzophenones. None.
5% para-aminobenzoic acid in Pigmentation from psoralens,
50-70% ethanol. 4% ethyl-hexyl- 8-methoxypsoralen, 4,5' ,8-tri-
paramethoxycinnamate + 3% methylpsoralen plus UV-A
2-hydroxy4' methoxycinnamate + exposure.
5.5% 2-phenylbenzimidazole
sulfonic acid (triethanol amine salt)
in cream base.
Benzophenones and benzotriazole; None.
titanium dioxide in colored, opaque
sunscreens.
Titanium dioxide in colored, opaque Beta carotene (60-180 mg
sunscreens. daily).
252 Chapter Eleven

Eye Protection against Ultraviolet Radiation

Because of the many applications of UV radiation in medicine, industry,

and the military, the need exists for suitable eye protection. Laser radiation in the
UV can reach many megawatts of peak power for short durations and can cause
irreversible eye damage (see Chapter 9). Brief exposures to germicidal lamps,
sunlamps, and welding arcs are also capable of damaging the eye. Photo-
chemotherapy, employing UV radiation in conjunction with photosensitizing
drugs, theoretically presents a hazard to the unprotected. eye. Solar UV may also
cause photokeratitis and is implicated in cataract formation (see Chapter 9).
The eye is incapable of detecting ultraviolet radiation. The natural protec-
tive reactions of contraction of the iris and the aversion response follow the eye's
perception of visible light. We must therefore rely on ultraviolet radiometry and
protective devices to ensure ocular safety from UV exposure.
The maximum protection is opaque, fully occlusive goggles, which are usu-
ally impractical and disorienting. Goggles or glasses that transmit the required
amount of visible light for comfortable sight but essentially eliminate other
wavelengths will generally offer acceptable protection, depending upon the use
and the user.
Use of glasses that absorb visible light much more than they ab&orb UV
reaching the eye may be harmful because the aversion response is diminished and
the pupil dilates in response to the decreased visible light levels, allowing greater
amounts of UV radiation to enter the pupil than would occur without the sunglas-
ses. While the total radiance of UV-A passing through the pupil may be in-
creased by use of these glasses, the UV-A irradiance at the pupil is in general less
than that without the glasses. Such potentially harmful glasses have been recently
reported and are commercially available. so Anderson and Gebepo calculated that
the worst of the sunglasses tested would approximately double the amount of
solar UV-A passing through the aperture of the human iris (sample #1, Fig.
11-2). Clearly, one cannot assume that "dark" sunglasses offer any ocular pro-
tection against ultraviolet radiation, unless manufacturers or governmental reg-
ulatory agencies impose ultraviolet-blocking standards for all commercial
sunglasses.
Patients receiving UV-A or UV-B" radiation as part of a phototherapy regi-
men must protect their eyes during treatment. Photochemotherapy patients or pa-
tients taking known photosensitizing drugs should additionally be protected for
the period after treatment during which the photosensitizing drug is still active.
Patients given oral psoralens require ocular protection against solar UV -A during
the daylight hours. Ideally, protective glasses for such patients should be com-
pletely opaque to wavelengths shorter than 400 nm, inexpensive, and cosmeti-
cally acceptable and should cover all directions of incoming radiation, be able to
be worn over prescription glasses, and transmit sufficient visible light in order for
Safety M~asures and Protection against Ultraviolet Exposure 253

100
RELATIVE EYE
SENSITIVITY

80

'\. SAMPLE #1
z
2
VI 60 I r#1 IV
VI
IjJ r'"'\\
I
r \\

1i /\\,
~
I, "
'IV1\
VI
z #2
«
a: 40
~

~ J!" SAMPLE #;',

I \'
20
/. #3 \\

If \ 'I'
.-
1-"1""
SAMPLE #3
/', I/1
I
0
400 500 600 700 800
WAVELENGTH (NM)

Fig. 11-2. The relative eye response and spectral transmission curves for some commercially availa-
ble sunglasses. (From Anderson, W. J., and Gebel, K. H. Ultraviolet windows in commercial
sunglasses. Appl. Optics /6:515-517, 1977. Copyright, Optical Society of America, Washington,
D.C., 1977. Used with pennission.)

the patient to perform tasks and perceive colors. Although there are numerous
types of ultraviolet-blocking eyewear available, many of these are impractical or
too costly. However, some commercial sunglasses, prescription glasses, and
specially designed ultraviolet-blocking glasses meet these criteria to varying ex-
tents.
If UV radiation entering the central portion of the pupil is attenuated by 10- 3
(0.1%), glasses can be considered essentially UV-opaque and protective against
solar UV. When considering the total UV received by the anterior surface of the
eye, transmission characteristics of the glasses lens is not the only consideration
because with most glasses some leakage around the side of the frames is practi-
cally unavoidable.
Solar spectroradiometric measurements taken by a small cosine-corrected
UV fiber optic detector at the position of the cornea in a mannequin with both
commercial and special UV-blocking sunglasses revealed that the best-fitting
glasses, short of occlusive goggles, allow a few percent of incident solar UV-A
to "leak" around the edge of the glasses and reach the eye. The worst-fitting
254 Chapter Eleven

glasses may allow greater than 25% leakage. This variation, plus variations of
the UV and visible spectral transmissions of the lenses, yield a very wide range
for solar-UV protection provided by sunglasses and protective eyewear. It is im-
possible to judge the degree of solar-UV protection provided by colored glasses
on the basis of their appearance. Interestingly, with the head in an upright posi-
tion, the shape of the eye socket alone (no glasses) reduces solar UV-A incident
on the eye to 10% of the maximum value obtained by staring into the sun at noon.
In the case of psoralen photochemotherapy patients, exposed skin areas de-
velop great phototoxicity to solar UV-A. Because ofthis transient skin photosen-
sitivity caused by 8-methoxypsoralen, patients are instructed to avoid sun expo-
sure. This instruction greatly reduces the theoretical ocular hazards associated
with psoralen photochemotherapy. Moreover, the approach of using essentially
UV-opaque, visible transmissive, widely occlusive, and cosmetically acceptable
glasses that fit over prescription glasses is sound practice for essentially any ul-
traviolet phototherapy regimen.

References
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2. Webb, R. B., and Lorenz, J. R. Oxygen dependence and repair of lethal effects of near ul-
traviolet and visible light. Photochem. Photobiol. 12:283-289, 1970.
3. Webb, R. B., and Malina, M. M. Mutagenic effects of near ultraviolet and visible radiant energy
on continuous cultures of Escherichia coli. Photochem. Photobiol. 12:457-468, 1970.
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14. Mills, J. N. Human circadian rhythms. Physiol. Rev. 46:128-171,1966.
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Safety Measures and Protection against Ultraviolet Exposure 255

Wurtman, R. Stimulation by artificial lighting of calcium absorption in elderly human subjects.

Nature 229:255-257, 1971.
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cinogenesis in man. Fed. Proc. 30:1766-1771, 1971.
18. Blum, H. F. Carcinogenesis by Ultraviolet Light. Princeton University Press, Princeton, New
Jersey, 1959.
19. Michaelson, S. M. Human exposure to nonionizing radiant energy-Potential hazards and
safety standards. Proc. IEEE 60:389-421, 1972.
20. Matelsky, I. The non-ionizing radiations. In Industrial Hygiene Highlights, Vol. 1 (L. V. Cral-
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21. Leach, W. M. Biological aspects of ultraviolet radiation: A review of hazards. Division of
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23. Biologic Effects and Measurements of Light Sources. Bureau of Radiological Health, FDA Doc.
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24. Occupational Exposure to Ultraviolet Radiation. USDHEW, National Institute for Occupational
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26. Sliney, D. H. The merits of an envelope action spectrum for ultraviolet radiation exposure
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27. Pathak, M. A., Fitzpatrick, T. B., and Frenk, E. Evaluation of topical agents that prevent
sunburn-Superiority of para-aminobenzoic acid and its ester in ethyl alcohol. N. Engl. J. Med.
280:1459-1463, 1969.
28. Willis, I., and Kligman, A. M. Aminobenzoic acid and its esters. Arch. Dermatol. 102:405-
417, 1970.
29. Fitzpatrick, T. B., Pathak, M. A., and Parrish, J. A. Protection of the human skin against the
effects of the sunburn ultraviolet (290-320 nm). In Sunlight and Man: Normal and Abnormal
Photobiologic Responses (M. A. Pathak, L. C. Harber, M. Seiji, and A. Kukita, Eds.; T. B.
Fitzpatrick, Consulting Ed.). University of Tokyo Press, Tokyo, 1974, pp. 751-765.
30. Finnerty, E. F. The results of exposure to solar radiation. Southwestern Med. 43:112-120,
1962.
31. Kahn, G., and Wilcox, G. Comparison of in vitro and in vivo sunscreen testing methods. J. Soc.
Cosmet. Chem. 20:807-824, 1969.
32. Parrish, J. A., Pathak, M. A., and Fitzpatrick, T. B. Protection of skin from germicidal ul-
traviolet radiation in the operating room by topical chemicals. N. Engl. J. Med. 284:1257-1258,
1971.
33. Dahlen, R. F., Shapiro, S. I., Berry, C. Z., and Schreiber, M. M. A method of evaluating
sunscreen protection from longwave ultraviolet. J. Invest. Dermatol. 55:164-169, 1970.
34. Parrish, J. A., Pathak, M. A., and Fitzpatrick, T. B. Prevention of unintentional overexposure
in topical psoralen treatment of vitiligo. Arch. Dermatol. 104:281-283, 1971.
35. Pathak, M. A., Parrish, J. A., and Fitzpatrick, T. B. New effective sunscreens for protection
against erythemogenic and carcinogenic spectrum of solar radiation. Program and Abstracts,
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36. Van Allen, J., and Tinker, J. F. Derivatives of benzoylresorcinol. J. Org. Chem. 19:1243-
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37. Purvis, J. E., and McCleland, N. P. The absorption spectra of some substances containing two
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256 Chapter Eleven

38. Tronnier, H., and Hoppe-Seyler, G. Praktisch-dermatologische Gesichtspunkte fiir den

Lichtschutz und die Depigmentierung. Anaesthet. Med. (Berlin) 17:221-228, 1968.
39. Hodgman, C. D. (Ed.). Handbook of Chemistry and Physics (38th ed.). Cleveland, Chemical
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40. Hoppe, U. Photostabilitat und Hautaffinitat-zwei Kriterien fiir kosmetische
Lichtschutzsubstanzen am Beispiel der Naphthalin-l,5-bis-hamstoffe. J. Soc. Cosmet. Chem.
25:667-680, 1974.
41. Fusaro, R. M., Runge, W. J., Lynch, F. W., and Watson, C. J. Sunlight protection in normal
skin-By absorptive filter chemically induced in stratum corneum. Arch. Dermatol. 93:106-
111,1966.
42. Fusaro, R. M., and Johnson, J. A. Photoprotection of patients sensitive to short and/or long
ultraviolet light with dehydroxyacetone/naphthoquinone. Dermatologica 148:224-227, 1974.
43. Macleod, T. M., and Frain-Bell, W. The study of the efficacy of some agents used forthe pro-
tection of the skin from exposure to light. Br. J. Dermatol. 84:266-281, 1974.
44. Findlay, G. H. Oral interceptives that do not work. In The Biologic Effects of Ultraviolet Radia-
tion (with r.mphasis on the Skin) (F. Urbach, Ed.). Pergamon Press, Oxford, 1969, pp. 693-
695.
45. Mathews-Roth, M. M., Pathak, M. A., Parrish, J. A., Fitzpatrick, T. B., Kass, E. H., Toda,
K., and Clemens, W. A clinical trial of the effects of oral beta-carotene on the responses of
human skin to solar radiation. J. Invest. Dermatol. 59:349-353, 1972.
46. Imbrie, J. D., Daniels, F., Jr., Bergeron, L., Hopkins, C. E., and Fitzpatrick, T. B. Increased
erythema threshold six weeks after a single exposure to sunlight plus oral methoxsalen. J. Invest.
Dermatol. 32:331-337, 1959.
47. Imbrie, J. D., Bergeron, L., and Fitzpatrick, T. B. Follow-up stUdy of effect of oral methoxsa-
len (8-methoxypsoralen) in sunburn and suntan. Arch. Dermatol. 82:617-620, 1960.
48. Mathews-Roth, M. M., Pathak, M. A., Fitzpatrick, T. B., Harber, L. C., and Kass, E. H.
Beta-carotene as a photoprotective agent in erythropoietic protoporphyria. N. Engl. J. Med.
282:1231-1234, 1970.
49. Epstein, J. Effects of J3-carotene on ultraviolet-induced cancer formation in the hairless mouse.
Photochem. Photobiol. 25:211-213, 1977.
50. Anderson, W. J., and Gebel, K. H. Ultraviolet windows in commercial sunglass. Appl. Optics
16:515-517, 1977.
Index
Absorption, of photons, 3
by excised skin, 71
in photochemical reactions, 87
of radiation, by molecules, 2
by skin, 65
Actinic reticuloid, 149-150
Action spectrum, of photocarcinogenesis,
166-168
for erythema, 115-125
of ocular effects, 188-193
for UV-induced cataracts, 208
Aging, cross-linkage hypothesis, 172
lens protein in, 206
and UV radiation, 171-172
Amino acids, in cataract formation, 206,
209
Animal cells, effect of UV-A, 98-99
Anterior epithelium of lens, ·183-184
Ap~,80,179,203
Aqueous humor, radiation effects, 188
Arc lamps, 15-16
Bacteria, effects ofUV-A, 96-98
Bactericidal action and absorption spectra,
88
Blackbody incandescent source, 22
Biologic interaction phenomena in UV-A
radiation, 100
Blood, radiation absorption, 76
Bowman's layer of cornea, 181
Broadband mters, 24-25
Capsule of crystalline lens, 183
Carcinogenesis, UV, 157-175
action spectrum, 166-168
animal studies, 163-165
chemicalintluences, 169
dose-dependency, 165
mechanisms, 165-166
Carcinoma of skin; basal cell, 157, 159-
161
incidence, 158-159
role of sunlight, 157-158
squamous cell, 157, 161
sun exposure in, 159, 161
Cataracts. See also Permanent lenticular
opacities
animal studies, 208-211
causes, 178, 185
and solar radiation, 205
thermal mechanism, 205
Cell(s)
animal, effects of UV, 85-101
optical properties, 76-77
protein, radiation effects, 88-89
Cement substance oflens, 184-185
Chemicals, environmental, in skin tumors,
169
Chromophores, in UV-A induced reactions,
95-96
Cold quartz lamps, 16
Collagen decrease in premature aging, 177
Conjunctivitis, 177-17 8
Conversion of irradiance units, 9
Cornea, action spectrum exposure thresh-
old, 189-191, 195-198
criteria for damage, 195
257
258
Cornea, action spectrum exposure thresh-
old (cont.)
effect of high exposure dose, 197-198
effect oflongwave UV radiation, 193-
202
histologic radiation changes, 187-188
mechanisms ofUV damage, 191-193
morphology and histology, 179-182
radiation transmittance, 79
ultraviolet absorption, 179
Cosine-weighted response, 52-53
Deexcitation, molecular, mechanisms of,
86-87
Dermatoses,
management of, 151-153
Dermis, histologic radiation effects, 124,
125
optical properties, 72-73
structure, 62
Descemet's membrane of cornea, 181-182
Detectors for UV energy measurement,
42-45
Diagnoses, use of UV-A, 234-235
Dichroic mirror, as broadband filter, 25
DNA, adducts, in aging, 172
cross-linkage, UV radiation in, 172
damage and UV carcinogenesis, 165-166
in lens damage, 210
radiation effects, 88-89
repair, 89-95
repair error and UV carcinogenesis, 166
synthesis, biphasic response, 126-128
in UV-A lethality, 96-97
Eczema, PUVA treatment, 231
Electromagnetic energy, classiflcation, 2
Electromagnetic radiation, properties, 1
Electromagnetic spectrum, 2-6
Electrophoretic changes in corneal
epithelium protein, 192
Energy of radiation wavelengths, 85-86
Epidermal cell cycle, 126
Epidermis, histological radiation effects,
124,125-126
optical properties, 65-71
structure, 61-62
Epithelium ofrabbit cornea, 180-181
Erythema, 109-124
action spectrum, 114-115
Index
Erythema (cont.)
angle of incidence, 113-114
delayed phase, 109
exposure dose, 112-113
mediators, 109-112
prolonged, in skin cancer, 168-169
radiation wavelengths, 116-120
reciprocity law, 112-113
to UV-A and UV-B, 122-124
variables in, 115-116
vascular response, 109
Excision repair of DNA, 90-91
Excitation, triplet state, 87
Exposure dose, 39
in erythema, 112
Exposure area, biologic response, 113
Eye,
damage, UV sources of, 185-186
at UV wavebands, 199-201
effect ofpsoralens, 212-214
gross structure, 78, 179
histologic effects of UV radiation, 185-
188
protection, ideal glasses, 252-253
use of sun glasses, 252
for UV radiation, 252-254
radiation absorption, 78
ultraviolet optics, 77-81
ultraviolet radiation effects, 177-219
ultraviolet reflection, 80
Filters, glass absorption, 48-49
interference, 49-50
neutral density,' SO-52
terminology, 49-50
Fluorescence of singlet state molecule, 86
Fluorescent lamps, 18-20
Gas discharge sources, 15-20
Glass, transmission and reflection, 28-30
Goniometric measurements, in skin
radiation, 66
Heat and tumor production, 169
Hematoporphyrin, in malignant photo-
sensitization, 225
Histamine, release in erythema, 109
Index
Histology, changes with UV-A, UV-B,
UV-C, 124-126
electron microscope studies, 126
of radiation response, 124-126
History, in dermatoses, 151-152
Immune response to skin tumors, 166
Incandescent sources, temperature effects,
21
ofUV-A,21-22
Indomethacin, in erythema, 110
Inflammation, components of, 108
skin response, 108
Infrared radiation, 3
Input optics, 52-54
Ionization, and photon energy, 3-4
Iris, radiation effects, 188
Keratin absorption in spectral transmit-
tance,68
Keratinocytes, of skin, 60, 61
Laboratory fmdings in dermatoses, 152
Laser, argon-ion, 203, 204
helium-cadmium, 203
nitrogen, 203-204
thermal lenticular damage, 204
tunable, 23
UV, effects on eye, 203-205
UV-A radiation emission, 22-24
Latitude, influence on skin cancer, 160,
162
Lens, action spectrum for exposure
threshold, 195-197
biochemical alterations, 206-211
biomicroscopic damage, 201
criteria of damage, 195
effects of longwave UV radiation, 193-
202
fibers, 185
morphology and histology, 182-185
pigment, 205
and UVexposure, 210
sutures, 185
UV-A absorption, 80
Lenticular opacity,
permanent,changesin,201-202
exposure in, 200-202
transient, wavelength and exposure, 201
259
Light reactivity, persistent, 149
Light source, research criteria, 23-24
Longwave UV radiation, experimental
procedures, 193-195
Lysosome, in erythema, 110-111
rupture, effects of, 111
Macromolecular synthesis,
bacterial, ultraviolet effects, 89
epidermal, ultraviolet effects, 126-128
Malignant melanoma, 157,161-163
sun exposure in, 162-163
Melanin, protective role, 62
radiation absorption, 133-134
in radiation transmission, 73
Melanocytes, 61-62
dendrites of, 132
response to radiation, 129-130, 131-133
Melanosomes, distribution pattern, 132-
133
pigment transfer, 62, 73
Melasma, hyperpigmentation in, 151
Mercury lamps, 16-17
Metal surfaces, reflectance of, 32-33
Methoxsalen, in radiation studies, 125-
126
ocular effects, 212-214
and photo carcinogenesis experiments,
170
Microorganisms, effects of UV radiation,
85-105
Minimal erythema dose (MED), 114-115
Minimum phototoxic dose (MPD), 147
Mitosis, effect of radiation, 126-127
Monochromatic radiation, 23
Mucopolysaccharides, of corneal epi-
thelium, 193
Mucous membrane, effects of UV-A, 128
Mutagenesis, of UV-A, 98-99
Mycosis fungoides, PUVA treatment, 230
Nonmetallic surfaces, reflectance of, 32
Nucleoproteins, in corneal UV absorption,
191-192
Optical radiation, skin measurements,
65-77
Organ systems, radiation effects, 99
260
Oxygen dependence, ofUV-A lethality, 96
Ozone layer, as UV shield, 163
Para-aminobenzoic acid, as sunscreen,
248-249
Phage multiplication, effect of UV, 98
Phosphorescence, of triplet state, 87
Photoaddition, in erythema, 121-122
Photoallergy, 147-149
agents, 148
clinical response, 147
mechanisms, 148
Photo carcinogenesis, effect ofUV-A, 168
experimental studies, 164-165
and phototoxic agents, 169-170
Photochemical lamps, 17
Photochemical reactions, and radiant
energy, 85-87
Photochemotherapy, 225
Photoconductive mode, 225
Photodiodes, solid state, 45-46
vacuum, 44-45
manufacturers, 55-57
Photokeratitis, clinical picture, 177-178
Photomultipliers, 42-44
amplification\ 43
disadvantages, 43-44
Photons, absorption of, 3
energy and frequency, 1
Photoprotectants, 251
Photoreactivating enzyme (PRE), 90
Photoreactivation, in DNA repair, 90
of bacteria, UV inhibition, 98
Photorecovery, 122, 124
Photosensitivity, chemical, 141-149
drug-induced, 143
in niacin deficiency syndrome, 151
Photo sensitizers, common chemical, 141
Phototesting, in dermatoses, 152-153
Phototherapy, 221-224
Phototoxicity,142-147
chlorpromazine, 146
mechanisms of, 145
other chemicals in, 146
phototoxic index (PI), 147
psoralens, 145
quantification of response, 146-147
tissue response, 144
variables in, 147
Index
Photovoltaic mode, 45-46
Pigments, skin, effects of, 73-77
radiation penetration, 65
Plastics, transmission, 30-32
Polymorphous light eruption, 150
Porphyrias photosensitization syndrome,
150-151
Postreplication, in DNA repair, 91-95
gap filling mechanisms, 94
Prostaglandins, in erythema, 110
Protein synthesis, biphasic response, 126-
128
Psoralen, DNA interaction, 229
Psoralen photochemistry, 225-233
Psoraien and UV-A (PUVA),
erythema reaction, 225-226
immunological effects, 231-232
in macromolecular synthesis, 127
pigmentation, 226-227
ocular effects, 212-214
therapy risks, 232-233
Psoriasis, Goeckerman treatment, 223-
224
PUVA treatment, 228-230
UV treatment, 222-224
Pyrimidine dimers, from DNA radiation
88 '
in postreplication repair, 91-94
in tumor development, 165
UV-A induced, 97
Pyroelectric detectors, 41-48
spectral response, 47
Radiometers, calibration, 38
Radiometric terminology, 7-9
Radiometry,37-57
considerations, 37-42
factors in, 41
geometric factors, 39,40
goal,38
Reciprocity, law, 112-113
in photocarcinogenesis, 164
and temperature, 113
Reflectance, of corneal surface, 80
direct, of skin, 63-64
total spectral, of skin, 71
Resinous dental restorati()ns, UV-A
polymerization, 233
Retina, effects ofUV-A, 211-212
histologic changes in radiation, 188
Index
RNA synthesis, effect of radiation, 126-
128
Safety standards, 242-248
NIOSH proposal, calculation, 243
complications, 245, 248
envelope action spectrum, 242
examples of application, 244-245
Scattering, in dermis, 72
relationships, 64-65
of solar radiation, 12-13
Silicon photodiodes, use of, 46
Singlet excited state, 86-87
Sister chromatid exchange (SCE), in
PUVA therapy, 233
Skin, adverse reactions to UV-A, 141-155
aging from radiation, 157-175
color, constitutive, 128
faculta~ve, 128, 131
factors in UV radiation, 62-65
immediate and short term radiation,
107-139
layers of, 63
optical properties, 59-77
optical radiation path, 62-64
radiation penetration, 73, 74-75
structure, 59-62
Solar elastosis, dermal changes, 171-172
Solarization of filters, 30
Solar radiation, 9-15
variations in, 10
Solar urticaria, 150
Spectral absorption, of skin, 71
Spectral irradiance, 7
curve, 9-10
measurement, 38-39
and wavelengths, 11-12
Spectral reflectance, of skin,,71-72
Spectral response, 37-38
Spectral transmission, of common mate-
rials, 26-33
in skin, 66-69
racial differences, 68
and wavelengths, 68
Spectroradiometers, use of, 52, 54
Spectrum of electromagnetic radiation,
1-6
Stratum corneum, protective role, 60, 73
Stroma, of cornea, 181
Sunburn cells, 111, 124, 126
261
Sunburn reac1J.on, 122
Sun exposure and carcinoma of skin, 157-
158,159,161-163
Sunscreen(s),248-252
evaluation, 250
ideal agent, 249
photoprotectants, 251
UV absorption, 249-250
Tanning, 128-134
delayed,132-134
action spectrum, 132, 134
mechanisms, 132
immediate, 131-132
mechanisms, 131
Tetrachlorosalicylanilide (TCSA), in
photoallergy studies, 148-149
Thermal effects of radiation, 86
Thermopiles, 48
Threshold, cataract, 210
cornea and lens exposure, 195-198
corneal damage, 192,202
photokeratitis, 190, 191
Tissue culture medium, irradiation of, 99-
100
photoproducts in, 100
Transforming DNA, effect of UV, 98
Treatment, for dermatoses, 153
use of UV radiation, 221-222
Tryptophan, in lens changes, 206-209
oxidation products, 207
role in UV-induced cataracts, 208-209
UV-A, artificial sources, 15-24
effects of, 95-101
experimental data, 95
lethal,96
sensitizer, action of, 87
significance of, 5-6
sources of, 7-35
comparison of, 22, 23
wavelength, 4
UV-B
defInition, 3-5
erythema, 109, 113, 117-126
reactions in eye, 177 -1 78
UV-C
defInition, 3-5
262 Index

UV-c (cont.) Vitiligo, treatment of, 227-228

erythema, 115-117
germicidal effects, 88
responses of eye, 177-17 8 Water cell fllter, 25
Uveitis, anterior, 198-199 Wood's lamp, in fluorescent diagnosis, 234

Visible light, 2 Xenon arc lamp, 17-18

Vitamin D, and UV radiation, 107 Xeroderma pigmentosum, 94, 161, 165