Environmental and Molecular Mutagenesis 59:79^90 (2018)

Research Article

Sesamol Ameliorates Radiation Induced DNA

Damage in Hematopoietic System of Whole
Body c-Irradiated Mice
Arun Kumar, Sandeep Choudhary, Jawahar S. Adhikari, and
Nabo K. Chaudhury*
Division of Radiation Biodosimetry, Institute of Nuclear Medicine and Allied
Sciences, Brig. SK Mazumdar Marg, Timarpur, Delhi 110054, India

Ionizing radiation exposure is harmful and at high
doses can lead to acute hematopoietic radiation syn-
drome. Therefore, agents that can protect hematopoi-
etic system are important for development of
radioprotector. Sesamol is a potential molecule for
development of radioprotector due to its strong free
radical scavenging and antioxidant properties. In the
present study, sesamol was evaluated for its role in
DNA damage and repair in hematopoietic system of
g-irradiated CB57BL/6 mice and compared with ami-
fostine. C57BL/6 male mice were administered with
sesamol 20 mg/kg (i.p.) followed by 2 Gy whole
body irradiation (WBI) at 30 min. Mice were sacri-
ficed at 0.5, 3, 24 h postirradiation; bone marrow,
splenocytes, and peripheral blood lymphocytes were
isolated to measure DNA damages and repair using
alkaline comet,g-H2AXand micronucleus assays. An
increase in % of tail DNA was observed in all organs
Key words: sesamol; amifostine; ionizing radiation; comet assay; c-H2AX assay; DNA damage
of WBI mice. Whereas in pre-administered sesamol
reduced %DNA in tail (P  0.05). Sesamol has also
reduced formation of radiation induced g-H2AX foci
after 0.5 h in these organs and further lowered to
respective control values at 24 h of WBI. Similar
reduction of % DNA in tail and g-H2AX foci were
observed with amifostine (P  0.05). Analysis of
mnPCE frequency at 24 h has revealed similar extent
of protection by sesamol and amifostine. Interestingly,
both sesamol and amifostine, alone and with radia-
tion, also increased the granulocytes count signifi-
cantly compared to the control (P  0.05). These
findings suggest that sesamol has strong potential to
protect hematopoietic system by lowering radiation
induced DNA damages and can prevent acute
hematopoietic syndrome in mice. Environ. Mol. Muta-
gen. 59:79–90, 2018. C 2017 Wiley Periodicals, Inc.
V

INTRODUCTION Since no radioprotector and mitigator is available for

Radiation exposure can cause irreversible damage in all
radiosensitive organs. The extent of damage depends on
the radiation dose absorbed, radiation quality, and organ
sensitivity. Development of radioprotector as a prophylac-
tic agent to prevent such toxicity is an unmet challenge
for radiation countermeasures. The hematopoietic system
is most sensitive to radiation and therefore protection of
this system is important for survival and quality of life
after exposure. Radioprotectors may help to prevent dele-
terious effects of radiation when administered prophylac-
tically. The requirement for such agents for radiation
emergency responders have been felt after nuclear power
plant accidents in the past, including the recent Fukush-
ima incident in Japan [Tokyo Electric Power Company
(TEPCO), 2011; Ten Hoeve and Jacobson, 2012]. In addi-
tion, combat soldiers in radiation environment, radiation
workers, and medical professionals also require radiopro-
tectors for working in planned radiation environments.
such exigency, extensive research has been carried out in
the last six decades. Though only amifostine has showed
promising results, the serious toxicities like nausea, vomit-
ing, and hypotension [Turrisi et al., 1986] associated with it
restricts its use in field conditions [Demiral et al., 2002;
Kuna et al., 2004]. At present, amifostine has US-FDA
approval for its use as an adjuvant (cytoprotectant) for
Additional Supporting Information may be found in the online version
of this article.
Grant sponsor: Defence Research and Development Organization, Govt.
of India; Grant number: NBC 1.29.
*Correspondence to: N. K. Chaudhury; E-mail: nkcinmas@rediffmail.com
Received 25 January 2017; provisionally accepted 24 June 2017; and in
final form 29 June 2017
DOI 10.1002/em.22118
Published online 2 August 2017 in
Wiley Online Library (wileyonlinelibrary.com).

C 2017 Wiley Periodicals, Inc.

V
Environmental and Molecular Mutagenesis. DOI 10.1002/em
80 Kumar et al.
radiotherapy of head and neck cancer patients [McDonald
et al., 1994; Brizel et al., 2000; Koukourakis et al., 2000;
Anne, 2002; Anne et al., 2007]. Amifostine is a familiar
prodrug that dephosphorylate to active metabolite
WR-1065 by membranous alkaline phosphatase [Calabro-
Jones et al., 1985]. Its effectiveness on radiation induced
delayed genomic instability shows its antioxidant potential
[Dziegielewski et al., 2008; Murley et al., 2011]. It is known
to prevent radiation induced chromosomal aberrations in
bone marrow cells of mice [Uma Devi and Prasanna, 1990;
Hooker et al., 2009] by eliminating the aberrant cells via
apoptosis [Ormsby et al., 2014].
Radiation mediated free radicals attack DNA and cause a
variety of damages [Spotheim-Maurizot et al., 1991], including
single and double strand breaks (DSBs), and alkali-labile sites.
These damages, if not repaired, can be manifested as short- and
long term effects on health [Sankaranarayanan, 2006].
Although cells have mechanisms to repair such damages, if the
damage is nonrepairable then it may induce apoptosis [Branzei
and Foiani, 2008; Lobo Borges et al., 2008]. Therefore, an
investigating molecule having multiple roles like free radicals
scavenging, DNA repair, and anti-apoptosis would serve as
better candidates for developing radioprotectors.
Sesamol, (3, 4-methylenedioxyphenol) a natural phenolic
antioxidant [Joshi et al., 2005] found in sesame seeds, has a
lot of medicinal value [Mohamed and Awatif, 1998; Kuhad
et al., 2009]. Earlier studies have revealed that sesamol has
antimutagenic activity against reactive oxygen species
[Hiramoto et al., 1996; Kaur and Saini, 2000]. Sesamol is
able to provide protection against radiation-induced DNA
damage in human lymphocytes in vitro [Prasad et al.,
2005], and radiation induced histopathological and bio-
chemical changes in Swiss albino mice [Parihar et al.,
2006]. Apart from radioprotection, sesamol is being studied
for various pathologies such as neurodegenerative disorder,
cardiotoxicity, nephrotoxicity, etc. [Chandrasekaran et al.,
2009; Sonia Angeline et al., 2013; Geetha et al., 2015].
Strong antioxidant activity of sesamol has been reported
in comparison to standard antioxidants like vitamin C, cur-
cumin, melatonin, etc. [Mishra et al., 2012]; additionally, in
V79 cells it has shown higher radioprotection as compared
to melatonin [Mishra et al., 2011]. Radiation induced DNA
damage and its repair can be determined using comet assay
[Singh et al., 1988; Dhawan and Anderson, 2009] and g-
H2AX assay at cellular level [Rube et al., 2008; Paris et al.,
2011]. Comet assay is now incorporated in OECD guideline
to study the genotoxicity of chemical agents [OECD, 2014].
The g-H2AX foci formation is one of the earliest events at
the site of DSBs after radiation exposure. The DNA damage
induction and repair kinetics in different organs have been
reported using g-H2AX and were found to have tissue
specific wide variation [Rube et al., 2008; Paris et al.,
2011]. g-H2AX foci and its disappearance is dependent on
the radiation dose and time that can be monitored to assess
extent of damage and repair of DSBs. Cytogenetic damages
micronuclei, chromosomal aberrations includes genomic
instability are the manifestation of early DNA damage and
its aberrant repair [Jasin and Richardson, 2000].
In the present study, we have investigated the radiopro-
tective efficacy of sesamol against g-radiation-induced
DNA damage and repair in hematopoietic cells and com-
pared its efficacy with amifostine. The cytogenetic meas-
urements viz. micronuclei along with DNA damage assay
(alkaline comet assay and gH2AX assay) demonstrated
radioprotective efficacy of sesamol comparable to amifos-
tine. These findings strongly suggest sesamol as a strong
radioprotector which can be considered for further valida-
tion studies in NHP models.
MATERIALS AND METHODS
Chemicals and Reagents
Amifostine, Geimsa powder, soybean oil, DPX solution, normal-
melting-point agarose, low-melting-point agarose, bovine serum albumin
(BSA), and propidium iodide were procured from Sigma Aldrich chemi-
cals, USA. Sesamol was purchased from Acros organics, USA. Phosphate
buffer saline (PBS), May-Grunewald stain, EDTA disodium, sodium chlo-
ride, sodium hydroxide pellets, and N-sodium lauryl sarcosinate were
obtained from Hi-media, India. Fetal bovine serum (FBS) of Invitrogen,
USA was used. Sodium phosphate mono basic and dibasic buffers were
obtained from Calbiochem, India. Methanol, acetic acid, and dmethyl sulf-
oxide (DMSO) of analytical grade were purchased from Spectrochem,
India. Para formaldehyde, triton X-100, Tris base, and ethanol were used
from Merck, India. Anti-phospho H2AX mouse monoclonal antibody con-
jugate with FITC was purchased from Merck-Millipore, India.
Animals
Male C57BL/6 mice of age 8–10 weeks old were issued by experimen-
tal animal facility of Institute of Nuclear Medicine and Allied Sciences
(INMAS), Delhi, India. Mice were kept in cages and acclimatized under
optimum conditions of temperature (25 6 28C), humidity (50–60%), light
(14 and 10 h of light and dark), with standard food, and water ad libitum.
After acclimatization (5–6 days) mice were weighed and the average body
weight (bd wt) was 25.0 6 2 g. All study protocols were approved by insti-
tutional animal ethics committee (INM/IEAC/2012/06).
Experimental Design
The experiments were performed once with three mice per group
(n 5 3 mice/group). Mice were segregated into control, drug, radiation,
and drug plus radiation groups as follows:
Group I (C): Control
Group II (S20): Sesamol 20 mg/kg bd wt (intraperitoneally (i.p).
Group III (A20): Amifostine 20 mg/kg bd wt (i.p).
Group IV (R): Radiation (2 Gy WBI).
Group V (S20 1 2Gy): Sesamol 20 mg/kg bd wt (i.p), 30 min prior
to irradiation.
Group VI (A20 1 2Gy): Amifostine 20 mg/kg bd wt (i.p), 30 min
prior to irradiation.
Preparation and Administration of Antioxidants
Sesamol was dissolved in 2% (v/v) DMSO, and final volume was
made up with soybean oil to obtain dose of 20 mg/kg bd wt (200 mL)

Sesamol Ameliorates Radiation Induced DNA Damage in Hematopoietic System
Fig. 1. Differential staining pattern of May–Grunewald Giemsa in bone
marrow smear under 1003 objective (oil) of light microscope. A. PCE
(polychromatic erythrocytes) and NCE (normochromatic erythrocytes), B.
and C. mnPCE (micronucleated polychromatic erythrocytes), D. fre-
quency of mnPCE is shown for control (C), 2Gy (R), sesamol 20 mg/kg
(S20), amifostine 20 mg/kg (A20), sesamol 20 mg/kg 1 2Gy (S20 1 R),
amifostine 20 mg/kg 1 2Gy (A20 1 R) groups, radiation induced mnPCE
for each animal. While same dose of amifostine was prepared in distilled
water. Mice were administered (i.p.) sesamol and amifostine in their
respective groups.
c-Irradiation and Tissue Harvesting
Animals were irradiated to whole body g-rays (60Co) with 2Gy radia-
tion dose using Cobalt Tele-therapy Unit, Bhabhatron-II (Panacea Bio-
tech, India) at a dose rate of 1 Gy/min and field size 35 3 35 cm2. The
system is routinely calibrated as a part of quality assurance by Radiation
Safety Officer of the Institute. Animals were scarified by cervical dislo-
cation after 0.5, 3, and 24 h of WBI. Both the femurs and spleen of ani-
mal were dissected and immediately the bone marrow was flushed out
with chilled PBS. Spleen was minced with glass slides on an ice bed to
prepare single cell suspension and filtered through 100 micron mesh.
Peripheral blood lymphocytes were isolated using Hi-Sep LSM 1084
(Hi-media, India), as per manufacturer’s protocol. Cells were counted
using haemocytometer and diluted the cell suspension to maintain 1 3
106 cells/mL.
Bone Marrow Micronucleus (MN) Assay
For MN assay bone marrow cell suspension was processed according
to the method of Schmid with minor modifications [Schmid, 1975].
Briefly, after centrifugation at1000 rpm for 10 min, the supernatant was
discarded and cell pellet was resuspended in 2–3 drops of fetal bovine
Environmental and Molecular Mutagenesis. DOI 10.1002/em
81
frequency after 24 h of whole body irradiation (WBI) compared with con-
trol (#P < 0.001); whereas sesamol and amifostine significantly reduced
radiation induced mnPCE frequency,**(P < 0.001), E. WBI decreases the
PCE/NCE ratio in bone marrow after 24 h. Preadministered sesamol and
amifostine recovered radiation decreased ratio significantly (*P < 0.05).
Error bars on data points represent the SD.
serum (FBS). The cell suspension was smeared on clean dry slides, air
dried, and fixed with methanol. The slides were stained with cocktail of
May Grunewald and Giemsa (1:1) diluted in Sorensen’s phosphate
buffer (pH 6.8) at 1:4 ratio for 20–25 min followed by washing with
same buffer. All slides were blinded and analyzed under 1003 objec-
tives with light microscope (Axio Imager M2, Carl Zeiss) . A minimum
of 1000 polychromatic erythrocytes (PCE) were scored along with nor-
mochromatic erythrocytes (NCE) for each mouse. The frequency of
micronucleated polychromatic erythrocytes (mnPCE) and normochro-
matic erythrocytes (mnNCE) were calculated with respect to total PCE
and NCE scored. The ratio of PCE and NCE (P/N) was also calculated.
All slides were decoded after completion of scoring and analysis.
Alkaline Comet Assay
DNA damage was studied using alkaline comet assay by the proce-
dure adopted earlier [Kumar et al., 2015]. Briefly, 40,000 cells were sus-
pended in low-melting-point agarose (0.7%) and layered on precoated
frosted slides with 1% normal-melting-point agarose. The slides were
immersed in lysis solution (2.5 M NaCl, 1% N-sodium lauryl sarcosi-
nate, 30 mM Na2EDTA, 10 mM Tris, 1% Triton X-100, and 10%
DMSO, pH 5 10.0) for 1 h at 48C. After lysis, slides were transferred in
alkaline buffer (200 mM NaOH, 100 mM Na2EDTA, pH 13.0) for 20
min and proceeded for alkaline electrophoresis (300 mM NaOH, 1 mM
Na2EDTA, pH  13.0) at 0.7 V/cm, 300 mA at 48C for 20 min. After
electrophoresis, the slides were washed gently with 0.4M Tris base
Environmental and Molecular Mutagenesis. DOI 10.1002/em
82 Kumar et al.
Fig. 2. Effect of sesamol and amifostine on radiation induced DNA
damage and its repair in hematopoietic system of C57BL/6 mice at 0.5,
3, and 24 h post-WBI using alkaline comet assay; A–D. Representative
images of comet observed under 203 objective of fluorescent micro-
scope; E–F. Comet images analyzed through MetaCyte comet scan
buffer (pH 7.4), fixed in 70% ethanol for 15 min followed by dehydra-
tion on a hot plate and stored in a dry place at room temperature. Prior
to analysis, the slides were rehydrated in distilled water, stained with
propidium iodide (25 mg/mL) for 10 min and visualized at 203 magnifi-
cation using automated scanning fluoresence microscope Metafer 4
(Meta-system, Germany) to determine the migration of damaged DNA.
Automated analysis of the comets was performed by using MetaCyte
Comet Scan module. At least 1000 nuclei were scanned to calculate the
relevant parameters; tail length, tail moment, olive tail moment, and
%DNA in tail to measure migrated nuclear DNA of cells. Another
important parameter, viz., % of apoptotic comets, comets having more
than 60% DNA in tail were classified as apoptotic comet (fragment
nuclei), counted separately in each group and expressed as % of apopto-
tic comets in total number of cells counted.
c-H2AX Assay
About 5 3 105 cells were spread on charged slides and allowed for
adhering at 48C for 20 min according to the method of Venkateswarlu
et al. [2015] with minor modification. Briefly, after adhesion, remaining
cells were decanted and fixed with 2% paraformaldehyde for 15 min.
The slides were washed with chilled PBS and kept in 70% ethanol.
Again washed with 0.5% Triton-X 100 for 15 min and blocked with 3%
BSA for 0.5 h. After blocking, BSA solution was decanted and added
50 mL of FITC conjugated antiphospho H2AX mouse monoclonal
software; G. Comet report after analysis; H., I., J. Percentage of tail
DNA in bone marrow, splenocytes, and peripheral blood lymphocytes,
respectively at different time points. Error bars on data points represent
the SD (*P < 0.05), (**P < 0.001), (#P < 0.001) (n 5 3 mice/group).
antibody (1:20,000 dilution in PBST) on slides for 1 h at room tempera-
ture as per manufacturer’s instruction. After antibody incubation, slides
were rinsed twice with PBST. Finally, slides were mounted using vecta-
shield mounting medium with DAPI (Vector Laboratory, CA, USA).
Each slide was observed under 103 objective for pre scan and 633
magnification for g-H2AX foci counting using MetaCyte g-H2AXfoci
scan software of automated Metafer microscope (Metasystem, Germany).
A minimum 300 cells were scanned and each analyzed manually to cal-
culate the frequency of g-H2AX foci per cell. The % of cells containing
g-H2AX foci was also calculated to check foci distribution in hemato-
poietic cells. During microscopic observation of slides an interesting
observation was recorded as FITC1 cellular bodies with no uptake of
DAPI stain (appeared to be RBC population) along with FITC1/DAPI1
or FITC–/DAPI1 nucleated cells. These cells (FITC1/DAPI–) were sep-
arately counted in minimum five randomly captured fields per slide by
Image J software. The data obtained was further normalized with respect
to nucleated cells. The % of FITC positive cells of this population was
also plotted to understand any difference between all groups.
Hematology
Blood samples were collected in K2EDTA coated microvette tubes
via cardiac puncture immediately after cervical dislocation of mice.
Complete blood cell counts were measured using automated hematology
analyzer MEK-6450 (Nihon Kohden, Japan). The cell counts included

Sesamol Ameliorates Radiation Induced DNA Damage in Hematopoietic System
Fig. 3. Effect of sesamol and amifostine on radiation induced apoptotic
comet in hematopoietic system of C57BL/6 mice at 0.5, 3, and 24 h post-
WBI using alkaline comet assay; A.,B. Representative images of normal
cell and apoptotic comet observed under 203 objective of fluorescent
microscope. C., D. Comet images analyzed through MetaCyte comet scan
white blood cells (WBCs), red blood cells (RBCs), hemoglobin, lympho-
cytes, and granulocytes.
Total Antioxidant Capacity Using 2,20 -Azinobis -(3-
Ethylbenzothiazoline-6-Sulfonic Acid) (ABTS1) Assay
Plasma was isolated from peripheral blood of each mouse after 0.5 h of ses-
amol and amifostine administration, and stored at 2808C till use. The stock of
preformed radical monocation of 2, 20 -azinobis-(3-ethylbenzothiazoline-6-
sulfonic acid) (ABTS•1) was generated by dissolving 7 mM ABTS with
2.25 mM potassium persulfate in MilliQ water [Re et al., 1999]. ABTS•1stock
was diluted to attain a working solution of absorbance 0.7 6 0.02 at wave-
length 732 nm. Presence of antioxidants decreased the concentration of
ABTS•1. Plasma was diluted five times with PBS and 10 mL was used for 300
mL reaction mixture of ABTS•1. The absorbance was recorded at 732 nm for
60 min using kinetic scan mode in UV-multiplate reader (Varioskan, Thermo-
fisher, USA). The percentage of total antioxidant capacity in plasma was cal-
culated and plotted against each group.
Statistical Analysis
Data presented are mean 6 SD of all samples in the experiment.
The comet parameters, g-H2AX foci per cells, and MN frequencies of
different groups were compared using Student’s t test and P < 0.05 was
considered as significant.
Environmental and Molecular Mutagenesis. DOI 10.1002/em
83
software shown more than 60% DNA in tail; E., F., G. Percentage of
apoptotic comets in bone marrow, splenocytes, and peripheral blood lym-
phocytes of different grouped mice, respectively. Error bars on data points
represent the SD (**P < 0.001), (#P < 0.001), (*P < 0.05) (n 5 3 mice/
group).
RESULTS
Sesamol Ameliorates Radiation Induced Micronuclei
in Bone Marrow Cells
PCE and NCE were observed under microscope and
scored for micronuclei frequency and ratio of PCE and NCE
in each group was determined. The results showed that sesa-
mol 20mg/kg administration caused no alteration of mnPCE/
1000PCE (6.33 6 1.17) compared to control (7.48 6 1.26).
In contrast, 2 Gy WBI significantly induced mnPCE/
1000PCE (43.05 6 1.57), which was significantly lowered
by sesamol pre-administration (20.65 6 0.72). Similarly,
amifostine alone did not induce mnPCE/1000PCE
(6.30 6 0.51), while reduced the radiation induced mnPCE/
1000PCE (20.97 6 1.23). Sesamol and Amifostine both sig-
nificantly reduced the mnPCE frequency per1000PCE in
radiation group (P < 0.001). The plot of mnPCE per 1000
PCE for different groups has shown protective effect of sesa-
mol and amifostine (Fig. 1). Many studies have reported
reduction of P/N ratio in bone marrow of rodents due to radi-
ation exposure [Badr et al., 1999; Rastogi et al., 2010]. In
Environmental and Molecular Mutagenesis. DOI 10.1002/em
84 Kumar et al.
Fig. 4. Effect of sesamol and amifostine on radiation induced double
stand break and its repair in hematopoietic system of C57BL/6 mice at
0.5, 3, and 24 h post-WBI using g-H2AX assay; representative images of
foci distribution are shown in cellular nuclei. A. DAPI stained nuclei, B.
FITC stained g-H2AX foci, C. merge g-H2AX foci in nuclei, D., E. foci
this study, WBI significantly (P < 0.05) reduced P/N ratio
from 1.17 6 0.07 to 0.59 6 0.02, while pre-administration of
sesamol and amifostine restored the P/N ratio to 0.72 6 0.02
and 0.73 6 0.03, respectively. Therefore, the results suggest
that this dose of sesamol or amifostine provided protection in
bone marrow cells.
Sesamol Reduces DNA Damage in Hematopoietic
Organs of Irradiated Mice
Alkaline Comet Assay
g-radiation induced DNA damages include DSBs, SSBs
(single strand breaks), and fragments in individual cells. The
comet head and tail contains intact DNA, and fragmented
DNA, respectively [Singh et al., 1988; Olive and Banath,
2006]. The results of comet assay showed significant increase
in % DNA in tail 33.5 6 3.4, 27.8 6 6.6, and 40.5 6 4.7
in bone marrow cells after 0.5, 3, and 24 h of WBI, respec-
tively. The corresponding value of %DNA in tail in control
was 2.5 6 0.9. Whereas, pre-administration of sesamol low-
ered the % DNA in tail to 16.0 6 1.0,7.2 6 1.6, and 8.1 6 1.3
counted by MetaCyte g-H2AX foci scan software of automated Metafer
microscope, F., G., H., frequency of g-H2AX foci/cell in bone marrow,
splenocytes, and peripheral blood lymphocytes of different grouped mice,
respectively. Error bars on data points represent the SD (#P < 0.001),
(*P < 0.05) (n 5 3 mice/group).
after 0.5, 3, and 24 h of WBI, respectively. Similar protective
effect of amifostine was observed in bone marrow cells except
at 3h post WBI time point. The % DNA in tail was 7.0 6 2.0,
15.6 6 2.7, and 3.1 6 1.7 after 0.5, 3, and 24 h of WBI, respec-
tively (Fig. 2). In spleenocytes, sesamol pre-treatment reduced
the radiation induced % DNA in tail from 30.1 6 5.1,
26.3 6 5.0, and 31.1 6 5.6 to 15.9 6 3.5, 15.3 6 1.2, and
12.3 6 3.3 after 0.5, 3, and 24 h WBI, respectively. Similarly,
Amifostine reduced % DNA in tail from 9.7 6 1.2,
19.1 6 1.1, and 11.1 6 2.1 (Fig. 2). Whereas in lymphocytes,
sesamol tend to protect in terms of % DNA in tail (11.6 6 1.7,
13.6 6 3.8, and 8.4 6 1.0) compared to radiation group
(16.6 6 2.2, 20.4 6 6.3, and 17.3 6 4.3) after 0.5, 3, and 24 h
WBI, respectively. Amifostine reduced % DNA in tail
5.1 6 3.2, 15.2 6 5.1, 2.6 6 0.5 after 0.5, 3, and 24 h WBI in
lymphocytes, respectively (Fig. 2J). The values of other
parameters viz.; tail length, tail moment, and Olive tail
moment also showed similar patterns (Supporting Information
Table I). Irradiation subsequently increased % of apoptotic
comets up to 24 h of WBI. Pre-administration of sesamol
reduced the radiation induced % of apoptotic comets (Fig. 3).

Sesamol Ameliorates Radiation Induced DNA Damage in Hematopoietic System
Fig. 5. Effect of Sesamol and amifostine on % of FITC positive cells in hematopoietic system of C57BL/6 mice at
0.5 h post-WBI. (**P < 0.001).
c-H2AX Assay
Phosphorylation of histone H2AX is one of the initial
events in DNA damage signaling and is an important bio-
marker for DSBs. The kinetic of this is characterized by the
induction phase, usually after few minutes postexposure
and then the removal phase, which occurs up to 24 h of
postexposure and beyond. In an early time point (0.5 h post-
WBI), substantial increase of g-H2AX foci in all organs of
mice was observed. The g-H2AX foci/cell in different
hematopoietic organs of irradiated mice ranges from
10.4 6 1.5 to 12.5 6 1.5 compared 0.9 6 0.4 to 1.0 6 0.4 in
control mice significantly indicated the induction of DSBs
after 0.5h of WBI (Fig. 4). Pre-administration of sesamol
decreased radiation induced g-H2AX foci/cell to 8.5 6 0.4,
6.3 6 0.8, and 8.2 6 1.4 in bone marrow, splenocytes, and
lymphocytes, respectively. Similar extent of reduction in g-
H2AX foci/cell was observed with amifostine followed by
WBI (Fig. 4). The kinetics of g-H2AX foci/cell decreased
with time up to minimal level after 24 h of WBI in radiation
group, whereas, pre-administration of sesamol or amifostine
lowered the initial values foci/cell in irradiated groups
(Fig. 4).
Environmental and Molecular Mutagenesis. DOI 10.1002/em
85
Radiation induced % of g-H2AX cells were 94.1 6 1.7,
95.0 6 1.1, and 97.3 6 1.7 after 0.5 h of WBI in bone
marrow, splenocytes and lymphocytes respectively, At the
same time interval, sesamol tend to reduce the radiation
induced % of g-H2AX cells to 88.5 6 2.8, 85.9 6 4.4, and
86.1 6 23.2 in bone marrow, splenocytes, and lympho-
cytes, respectively. Amifostine also showed similar reduc-
tion in radiation induced % of g-H2AX cells (Supporting
Information Data Table II). The % of FITC positive cells
was also increased by sesamol and amifostine at 0.5 h
while by radiation at 24 h (Figs. 5 and 6).
Hematology
The alteration of total WBC count was observed in all
groups, radiation alone slightly increased WBC count at 3 h
and then significantly decreased at 24 h of post WBI
(P  0.001) (Fig. 7). Sesamol alone did not alter the WBC
count and the difference was nonsignificant when compared
with control (P  0.05). The analysis of differential leucocytes
count revealed an interesting observation; both sesamol and
amifostine increased the granulocytes count significantly com-
pared to control after 0.5 h of administration (P  0.05) (Fig.
Environmental and Molecular Mutagenesis. DOI 10.1002/em
86 Kumar et al.
Fig. 6. Effect of Sesamol and amifostine on % of FITC positive cells in hematopoietic system of C57BL/6 mice at 3
and 24 h post-WBI (**P < 0.001).
7). This elevated granulocytes count was observed up to 24 h
after sesamol administration, whereas radiation showed this
elevation in granulocytes count after 3 h and then decreased at
24 h WBI (Fig. 7). Radiation decreased the lymphocytes count
after 24 h of WBI, while both pre-administrated sesamol and
amifostine tend to protect radiation induced depletion in
lymphocytes. The ratio of granulocytes to non-granulocytes
(lymphocytes) counts was significantly high in sesamol and
amifostine alone and followed by WBI groups compared to
control (P  0.05) shown in Figure 7.
Total Antioxidant Capacity
The total antioxidant capacity in plasma after 0.5 h of
drug administration was determined by decolorization of
ABTS_1 by measuring reduction of absorbance as percent-
age inhibition of absorbance; there was no significant dif-
ference observed among the control, radiation alone,
sesamol; and amifostine treated groups (P  0.05) (Fig. 8).
DISCUSSION
Protection of hematopoietic system against radiation
exposure is the key strategy for development of radiation
countermeasures. Protection of bone marrow, the produc-
tion center of blood lineages, during radiation exposure is
necessary for survival and quality of life after exposure.
Most of the studies on radioprotectors have focused on effi-
cacy and related mechanisms with high radiation dose even
up to supra lethal [Parihar et al., 2006; Shirazi et al., 2013;
Khan et al., 2015; Singh et al., 2016]. However, the proba-
bility of such high radiation dose exposure is unlikely dur-
ing planned scenarios. Therefore, the present study was
designed for low dose radiation environment with an
attempt to compare the efficacy of sesamol and amifostine
in hematopoietic system of mice. The results have shown
similar extent of reduction (P  0.001) in radiation induced
micronuclei frequency in bone marrow of mice pre-
administered with sesamol or amifostine (Fig. 1). Further,
observation of similar level of protection after 24 h of WBI
and no genotoxicity by sesamol at this drug dose is promis-
ing for radioprotector development. Radiation induced
DNA damage either single or double stand breaks, are man-
ifested as cytogenetic damages, and can be assessed using
alkaline comet assay. The eventual decrease in % DNA in
tail post irradiation from 0.5 to 3 h has shown the extent of
DNA damage and its repair. Furthermore, the elevated %
 Environmental and Molecular Mutagenesis. DOI 10.1002/em
Sesamol Ameliorates Radiation Induced DNA Damage in Hematopoietic System 87

Fig. 7. Influence of sesamol/amifostine in hematological alterations of mice at 0.5, 3, and 24 h post-WBI. Error bars on
data points represent the SD (*P < 0.05) (#P < 0.001), (n 5 6 mice/group).

after 0.5 h WBI and the value attained near to control in

Fig. 8. Percentage of total antioxidant capacity in plasma after 30 min
of sesamol and amifostine administration to C57BL/6 mice using ABTS
assay. Error bars on data points represent the SD (n 5 6 mice/group).
DNA in tail in radiation after 24 h suggests apoptosis
of damaged cells [Woo et al., 2002; Milone et al., 2005]
as observed in comet images of bone marrow cells and
splenocytes. Pre-administration of sesamol significantly
(P  0.05) reduced the radiation induced % DNA in tail
these hematopoietic organs after 24 h of irradiation (Fig. 2).
The significant (P  0.001) reduction in % of apoptotic
comets in sesamol administered group reveals its antiapop-
totic activity (Fig. 3) [Prasad and Ramachandran, 2012;
Khan et al., 2015]. Whereas, amifostine showed higher abil-
ity to reduce the % DNA in tail at 0.5 h postirradiation com-
pared to sesamol, probably due to its strong antiradical
capacity. The increased % DNA in tail by amifostine at 3 h
of postirradiation has shown unanticipated DNA damage
(Fig. 2). However, no such induction was observed in ami-
fostine alone group. These results depict its differential
role, i.e., amifostine induces the apoptosis in cells with non-
repairable DNA damage after radiation exposure [Warters
et al., 1997; Lee et al., 2003; Mazur et al., 2003; Ormsby
et al., 2014] which is presently supported by relatively
higher number of apoptotic comets in bone marrow and
spleen of amifostine treated mice at 3 h postirradiation
(Fig. 3). Induction of DNA damage by radiation, viz., DSBs
is not easily repaired and causes more consequences [van
Gent et al., 2001; Kim et al., 2015] depending upon the radi-
ation dose and tissue sensitivity. A molecule with multiple
Environmental and Molecular Mutagenesis. DOI 10.1002/em
88 Kumar et al.
properties, including ability to mediate DSBs repair, is
expected to be a better radioprotector. The role of radiopro-
tector in tissue specific DNA damage and repair using
H2AX assay is not studied comprehensively. Therefore, an
attempt was made with sesamol to study its role in DSBs
repair and compared with amifostine using g-H2AX assay.
The analysis of observed frequency of g-H2AX foci/cell
has revealed that sesamol 20mg/kg dose significantly
(P  0.05) provide protection in the bone marrow and the
spleen by lowering radiation induced DSBs after 0.5 h WBI
and also facilitated its repair at subsequent post irradiation
time points. However, no significant reduction of radiation
induced g-H2AX foci/cell was observed in peripheral blood
lymphocytes of sesamol administered mice at 0.5 h WBI
compared to radiation group (P  0.05), but at a later time
at 24 h of WBI, sesamol decreased radiation induced g-
H2AX foci frequency. Thus, present observation has indi-
cated that sesamol facilitated slow repair of radiation
induced DSBs in lymphocytes as compared to amifostine.
Exposure to radiation leads to hemocytopenia. The analy-
sis of hematological parameters in post irradiated mice has
shown a drastic decrease of WBC count, whereas pre-
administration of sesamol tends to recover this radiation
induced devastation in WBC with nonsignificant difference
(P > 0.05). Pre-administration of sesamol or amifostine
alone had increased the granulocytes count at 30 min, while
in radiation group at 3 h. The increase in granulocyte count
may be induced through granulocytes colony stimulating
factor, that help in cell proliferation, differentiation, mobili-
zation of hematopoietic stem and progenitor cells
[Schneider, Kr€ uger, et al., 2005; Schneider, Kuhn, et al.,
2005; Publicover et al., 2013] and recovery of radiation
damaged cellularity [Kulkarni et al., 2013; Plett et al.,
2014]. G-CSF is also reported to prevent apoptosis [Herodin
et al., 2007; Jun et al., 2011; Kojima et al., 2011]. Thus,
increased granulocytes count by sesamol and amifostine
may be related to induction of G-CSF and radioprotection
in the form of reduction in cytogenetic damages. The ability
of sesamol mediated protection against DNA damage
immediately after radiation (0.5 h) may be due to its domi-
nant role as an antioxidant and free radical scavenger.
Whereas, better protection by amifostine at 0.5 h may be
due to its multiple roles as antioxidant, differences in phar-
macokinetic behavior, and possible molecular pathways as
compared to sesamol. The total antioxidant capacity of
plasma after 0.5 h of both sesamol and amifostine adminis-
tration (i.p) showed no significant difference between sesa-
mol, and amifostine as well as in control group. The
possible reasons could be due to low sensitivity of the assay
at this dose. In conclusion, sesamol appears substantially
good as amifostine and therefore may be embarked upon
for further drug development process as a promising radio-
protector. Further studies are being planned to elucidate the
role of this antioxidant molecule on gene expression of G-
CSF and its downstream pathways.
AUTHOR CONTRIBUTIONS
AK, JSA, and NKC designed the study and NKC
applied for institutional animal ethics committee approval
after finalization of the design. AK, SC performed the
experiments under supervision of NKC. AK analyzed the
data, and prepared the draft manuscript. All authors
reviewed the draft manuscript and NKC finalized manu-
script for submission.
ACKNOWLEDGMENTS
Arun Kumar is thankful to DRDO for fellowship, and
to Dr. AK Singh, Director INMAS for encouragement.
The authors are also thankful to Dr. B. G. Roy and Ms
Anjali Sharma for providing experimental animals and g-
irradiation facility.
CONFLICT OF INTEREST
None.
REFERENCES
Anne PR. 2002. Phase II trial of subcutaneous amifostine in patients
undergoing radiation therapy for head and neck cancer. Semin
Oncol 29:80–83.
Anne PR, Machtay M, Rosenthal DI, Brizel DM, Morrison WH, Irwin
DH, Chougule PB, Estopinal NC, Berson A, Curran WJ. 2007. A
Phase II trial of subcutaneous amifostine and radiation therapy in
patients with head-and-neck cancer. Int J Radiat Oncol 67:445–
452.
Badr FM, Habit OHM, El, Harraz MM. 1999. Radioprotective effect of
melatonin assessed by measuring chromosomal damage in mitotic
and meiotic cells. Mutat Res 444:367–372.
Branzei D, Foiani M. 2008. Regulation of DNA repair throughout the
cell cycle. Nat Rev Mol Cell Biol 9:297–308.
Brizel DM, Wasserman TH, Henke M, Strnad V, Rudat V, Monnier A,
Eschwege F, Zhang J, Russell L, Oster W, et al. 2000. Phase III
randomized trial of amifostine as a radioprotector in head and
neck cancer. J Clin Oncol 18:3339–3345.
Calabro-Jones PM, Fahey RC, Smoluk GD, Ward JF. 1985. Alkaline
phosphatase promotes radioprotection and accumulation of WR-
1065 in V79–171 cells incubated in medium containing WR-
2721. Int J Radiat Biol Relat Stud Phys Chem Med 47:23–27.
Chandrasekaran VRM, Hsu D-Z, Liu M-Y. 2009. The protective effect
of sesamol against mitochondrial oxidative stress and hepatic
injury in acetaminophen-overdosed rats. Shock 32:89–93.
Demiral AN, Yerebakan O, Simşir V, Alpsoy E. 2002. Amifostine-
induced toxic epidermal necrolysis during radiotherapy: A case
report. Jpn J Clin Oncol 32:477–479.
Dhawan A, Anderson D. 2009. Comet assay in toxicology. 2nd Edition.
Cambrige: Royal Society of Chemistry. 590 p.
Dziegielewski J, Baulch JE, Goetz W, Coleman MC, Spitz DR, Murley
JS, Grdina DJ, Morgan WF. 2008. WR-1065, the active metabo-
lite of amifostine, mitigates radiation-induced delayed genomic
instability. Free Radic Biol Med 45:1674–1681.
Geetha T, Kapila M, Prakash O, Deol PK, Kakkar V, Kaur IP. 2015.
Sesamol-loaded solid lipid nanoparticles for treatment of skin
cancer. J Drug Target 23:159–169.
van Gent DC, Hoeijmakers JH, Kanaar R. 2001. Chromosomal stability
and the DNA double-stranded break connection. Nat Rev Genet
2:196–206.

Sesamol Ameliorates Radiation Induced DNA Damage in Hematopoietic System
Herodin F, Grenier N, Drouet M. 2007. Revisiting therapeutic strategies
in radiation casualties. Exp Hematol 35:28–33.
Hiramoto K, Ojima N, Sako K, Kikugawa K. 1996. Effect of plant phe-
nolics on the formation of the spin-adduct of hydroxyl radical
and the DNA strand breaking by hydroxyl radical. Biol Pharm
Bull 19:558–563.
Ten Hoeve JE, Jacobson MZ. 2012. Worldwide health effects of the
Fukushima Daiichi nuclear accident. Energy Environ Sci 5:8743.
Hooker AM, Grdina DJ, Murley JS, Blyth BJ, Ormsby RJ, Bezak E, Giam
KA, Sykes PJ. 2009. Low doses of amifostine protect from chromo-
somal inversions in spleen in vivo when administered after an occu-
pationally relevant X-radiation dose. Int J Low Radiat 6:43–55.
Jasin M, Richardson C. 2000. Frequent chromosomal translocations
induced by DNA double-strand breaks. Nature 405:697–700.
Joshi R, Kumar MS, Satyamoorthy K, Unnikrisnan MK, Mukherjee T.
2005. Free radical reactions and antioxidant activities of sesamol:
Pulse radiolytic and biochemical studies. J Agric Food Chem 53:
2696–2703.
Jun HS, Lee YM, Song KD, Mansfield BC, Chou JY. 2011. G-CSF
improves murine G6PC3-deficient neutrophil function by modu-
lating apoptosis and energy homeostasis. Blood 117:3881–3892.
Kaur IP, Saini A. 2000. Sesamol exhibits antimutagenic activity against
oxygen species mediated mutagenicity. Mutat Res Toxicol Envi-
ron Mutagen 470:71–76.
Khan S, Kumar A, Adhikari JS, Rizvi MA, Chaudhury NK. 2015. Pro-
tective effect of sesamol against Co gamma-ray-induced hemato-
poietic and gastrointestinal injury in C57BL/6 male mice. Free
Radic Res 49:1344–1361.
Kim J, Sturgill D, Tran AD, Sinclair DA, Oberdoerffer P. 2015. Con-
trolled DNA double-strand break induction in mice reveals post-
damage transcriptome stability. Nucleic Acids Res 44:1–10.
Kojima H, Otani A, Oishi A, Makiyama Y, Nakagawa S, Yoshimura N.
2011. Granulocyte colony-stimulating factor attenuates oxidative
stress-induced apoptosis in vascular endothelial cells and exhibits
functional and morphologic protective effect in oxygen-induced
retinopathy. Blood 117:1091–1100.
Koukourakis MI, Kyrias G, Kakolyris S, Kouroussis C, Frangiadaki C,
Giatromanolaki A, Retalis G, Georgoulias V. 2000. Subcutaneous
administration of amifostine during fractionated radiotherapy: A
randomized phase II study. J Clin Oncol 18:2226–2233.
Kuhad A, Sachdeva AK, Chopra K. 2009. Attenuation of renoinflamma-
tory cascade in experimental model of diabetic nephropathy by
sesamol. J Agric Food Chem 57:6123–6128.
Kulkarni S, Singh PK, Ghosh SP, Posarac A, Singh VK. 2013. Granulo-
cyte colony-stimulating factor antibody abrogates radioprotective
efficacy of gamma-tocotrienol, a promising radiation countermea-
sure. Cytokine 62:278–285.
Kumar A, Selvan TG, Tripathi AM, Choudhary S, Khan S, Adhikari JS,
Chaudhury NK. 2015. Sesamol attenuates genotoxicity in bone
marrow cells of whole-body g-irradiated mice. Mutagenesis 30:
651–661.
Kuna P, Dostal M, Neruda O, Knajfl J, Petyrek P, Podzimek F, Severa J,
Svoboda V, Simsa J, Spelda S, et al. 2004. Acute toxicity and radio-
protective effects of amifostine (WR-2721) or cystamine in single
whole body fission neutrons irradiated rats. J Appl Biomed 2:43–49.
Lee EJ, Gerhold M, Palmer MW, Christen RD. 2003. p53 protein regu-
lates the effects of amifostine on apoptosis, cell cycle progres-
sion, and cytoprotection. Br J Cancer 88:754–759.
Lobo Borges H, Linden R, Wang JY. 2008. DNA damage-induced cell
death: lessons from the central nervous system. Cell Res 181:17–
26.
Mazur L, Augustynek A, Halicka HD, Deptała A. 2003. Induction of
apoptosis in bone marrow cells after treatment of mice with WR-
2721 and gamma-rays: Relationship to the cell cycle. Cell Biol
Toxicol 19:13–27.
Environmental and Molecular Mutagenesis. DOI 10.1002/em
89
McDonald S, Meyerowitz C, Smudzin T, Rubin P. 1994. Preliminary
results of a pilot study using WR-2721 before fractionated irradi-
ation of the head and neck to reduce salivary gland dysfunction.
Int J Radiat Oncol Biol Phys 29:747–754.
Milone M, Tsai D, Hodinka R, Silverman L, Malbran A, Wasik M,
Nichols K. 2005. Treatment of primary ebb infection in patients
with X-linked lymphoproliferative disease using B-cell-directed
therapy. Blood 105:994–996.
Mishra K, Ojha H, Chaudhury NK. 2012. Estimation of antiradical prop-
erties of antioxidants using DPPH assay: A critical review and
results. Food Chem 130:1036–1043.
Mishra K, Srivastava PS, Chaudhury NK. 2011. Sesamol as a potential
radioprotective agent: In vitro studies. Radiat Res 176:613–623.
Mohamed HMA, Awatif II. 1998. The use of sesame oil unsaponifiable
matter as a natural antioxidant. Food Chem 62:269–276.
Murley JS, Baker KL, Miller RC, Darga TE, Weichselbaum RR, Grdina
DJ. 2011. SOD2-mediated adaptive responses induced by low-
dose ionizing radiation via TNF signaling and amifostine. Free
Radic Biol Med 51:1918–1925.
OECD. 2014. Test No. 489: In vivo Mammalian Alkaline Comet Assay,
OECD Guidelines for the Testing of Chemicals. OECD Publ,
section 4:1–21.
Olive PL, Banath JP. 2006. The comet assay: A method to measure
DNA damage in individual cells. Nat Protoc 1:23–29.
Ormsby RJ, Lawrence MD, Blyth BJ, Bexis K, Bezak E, Murley JS,
Grdina DJ, Sykes PJ. 2014. Protection from radiation-induced
apoptosis by the radioprotector amifostine (WR-2721) is radiation
dose dependent. Cell Biol Toxicol 30:55–66.
Parihar VK, Prabhakar KR, Veerapur VP, Kumar MS, Reddy YR, Joshi
R, Unnikrishnan MK, Rao CM. 2006. Effect of sesamol on
radiation-induced cytotoxicity in Swiss albino mice. Mutat Res
Genet Toxicol Environ Mutagen 611:9–16.
Paris L, Cordelli E, Eleuteri P, Grollino MG, Pasquali E, Ranaldi R,
Meschini R, Pacchierotti F. 2011. Kinetics of gamma-H2AX
induction and removal in bone marrow and testicular cells of
mice after X-ray irradiation. Mutagenesis 26:563–572.
Plett PA, Chua HL, Sampson CH, Katz BP, Fam CM, Anderson LJ,
Cox GN, Orschell CM. 2014. PEGylated G-CSF (BBT-015),
GM-CSF (BBT-007), and IL-11 (BBT-059) analogs enhance sur-
vival and hematopoietic cell recovery in a mouse model of the
hematopoietic syndrome of the acute radiation syndrome. Health
Phys 106:7–20.
Prasad N, Ramachandran S. 2012. Sesamol modulates ultraviolet-B-
induced apoptotic and inflammatory signaling in human skin der-
mal fibroblasts. Int J Nutr Pharmacol Neurol Dis 2:31–39.
Prasad NR, Menon VP, Vasudev V, Pugalendi KV. 2005. Radioprotec-
tive effect of sesamol on g-radiation induced DNA damage, lipid
peroxidation and antioxidants levels in cultured human lympho-
cytes. Toxicology 209:225–235.
Publicover A, Richardson DS, Davies A, Hill KS, Hurlock C, Hutchins
D, Jenner MW, Johnson PW, Lamb J, Launders H, et al. 2013.
Use of a biosimilar granulocyte colony-stimulating factor for
peripheral blood stem cell mobilization: An analysis of mobiliza-
tion and engraftment. Br J Haematol 162:107–111.
Rastogi L, Feroz S, Narain Pandey B, Jagtap A, Prasad Mishra K. 2010.
International Journal of Radiation Biology Protection against
radiation-induced oxidative damage by an ethanolic extract of
Nigella sativa L. Int J Radiat Biol 86:719–731.
Re R, Pellegrini N, Proteggente A, Pannala A, Yang M, Rice-Evans C.
1999. Antioxidant activity applying an improved ABTS radical
cation decolorization assay. Free Radic Biol Med 26:1231–1237.
Rube CE, Grudzenski S, K€ uhne M, Dong X, Rief N, L€obrich M, R€ube C.
2008. DNA double-strand break repair of blood lymphocytes and
normal tissues analysed in a preclinical mouse model: Implications
for radiosensitivity testing. Clin Cancer Res 14:6546–6555.
Environmental and Molecular Mutagenesis. DOI 10.1002/em
90 Kumar et al.
Sankaranarayanan K. 2006. Estimation of the genetic risks of exposure
to ionizing radiation in humans: Current status and emerging
perspectives. J Radiat Res 47: B57–B66.
Schmid W. 1975. The micronucleus test. Mutat Res Mutagen Relat
Subjects 31:9–15.
Schneider A, Kr€uger C, Steigleder T, Weber D, Pitzer C, Laage R,
Aronowski J, Maurer MH, Gassler N, Mier W, et al. 2005. The
hematopoietic factor G-CSF is a neuronal ligand that counteracts
programmed cell death and drives neurogenesis. J Clin Invest
115:2083–2098.
Schneider A, Kuhn HG, Sch€abitz WR. 2005. A role for G-CSF (granulo-
cyte-colony stimulating factor) in the central nervous system.
Cell Cycle 4:1753–1757.
Shirazi A, Mihandoost E, Ghobadi G, Mohseni M, Ghazi-Khansari M.
2013. Evaluation of radio-protective effect of melatonin on whole
body irradiation induced liver tissue damage. Cell J 14:292–297.
Singh NP, McCoy MT, Tice RR, Schneider EL. 1988. A simple tech-
nique for quantitation of low levels of DNA damage in individual
cells. Exp Cell Res 175:184–191.
Singh VK, Fatanmi OO, Wise SY, Newman VL, Romaine PLP, Seed TM.
2016. The potentiation of the radioprotective efficacy of two medi-
cal countermeasures, gamma-tocotrienol and amifostine, by a com-
bination prophylactic modality. Radiat Prot Dosim 172:302–310.
Sonia Angeline M, Sarkar A, Anand K, Ambasta RK, Kumar P. 2013.
Sesamol and naringenin reverse the effect of rotenone-induced
PD rat model. Neuroscience 254:379–394.
Spotheim-Maurizot M, Franchet J, Sabattier R, Charlier M. 1991. DNA
radiolysis by fast neutrons. II. Oxygen, thiols and ionic strength
effects. Int J Radiat Biol 59:1313–1324.
Tokyo Electric Power Company (TEPCO). TEPCO : Press Release |
Plant Status of Fukushima Daiichi Nuclear Power Station (as of
11PM March 12th): 2Press Release.
Turrisi AT, Glover DJ, Hurwitz S, Glick J, Norfleet AL, Weiler C,
Yuhas JM, Kligerman MM. 1986. Final report of the phase I trial
of single-dose WR-2721 [S-2-(3-aminopropylamino)ethylphos-
phorothioic acid]. Cancer Treat Rep 70:1389–1393.
Uma Devi P, Prasanna PG. 1990. Radioprotective effect of combinations
of WR-2721 and mercaptopropionylglycine on mouse bone mar-
row chromosomes. Radiat Res 124:165–170.
Venkateswarlu R, Tamizh SG, Bhavani M, Kumar A, Alok A,
Karthik K, Kalra N, Vijayalakshmi J, Paul SFD, Chaudhury
NK, et al. 2015. Mean frequency and relative fluorescence
intensity measurement of g-H2AX foci dose response in PBL
exposed to g-irradiation: An inter- and intra-laboratory com-
parison and its relevance for radiation triage. Cytom A 87:
1138–1146.
Warters RL, Roberts JC, Wilmore BH, Kelley LL. 1997. Modulation of
radiation-induced apoptosis by thiolamines. Int J Radiat Biol 72:
439–448.
Woo RA, Jack MT, Xu Y, Burma S, Chen DJ, Lee PWK. 2002. DNA
damage-induced apoptosis requires the DNA-dependent
protein kinase, and is mediated by the latent population of p53.
EMBO J 23:4877.
Accepted by—
S. D. Dertinger