BIOMEDICAL

The Utilization of Laboratory

Biomarkers to Predict and Prevent
Neuroimmune Disorders caused by
Environmental Stressors
By Kendal Stewart, MD and Lisa Hunter Ryden, MT (ASCP), MBA

Kendal Stewart, MD, is the chief
medical officer of the NeuroSensory
Center of Austin. He is board certified in
otolaryngology with fellowship training
in neurotology/skull base surgery. His
experience includes advanced training
in neurosurgical, neurological and
immunological disorders. Dr. Stewart
has extensive experience with advanced
neurological and audio-vestibular
techniques and has authored two
medically related patents in this area. He
has had specific research interests in vestibular disorders, athletic
injuries of the nervous system and processing/sensory integration
disorders. Dr. Stewart has developed innovative and highly effective
treatment protocols for neurological diseases including Meniere’s
disease, imbalance, vertigo, autism spectrum disorders, and “post-
concussion” syndrome. He has authored papers and is extensively
involved in lectures and instructional courses for physicians, therapists
and other health care professionals.
Lisa Hunter Ryden, MBA, also has a
degree in medical technology and post
graduate coursework in microbiology,
molecular genetics and immunology.
She began her career as a clinical medical
technologist and has spent the past
20 years in the medical diagnostics
industry. She communicates her extensive
knowledge of biomedical treatment to
the autism community as a means to help
physicians and parents develop a best
practices model and develop a partnership
to recover their children. Lisa hosts a program with Dr. Kendal Stewart
on Autism One Radio, has given numerous public presentations on
autism biomedical treatment, written several articles, and served as
a parent advocate in political and legislative autism issues. Lisa and
her husband have two boys. Their oldest son, Jake, suffered autistic
regression at age 12 months, and was nonverbal until age 5. Today
at age 8, he is recovering from his symptoms, speaking in sentences,
writing, and learning to read. 

A new case of autism is diagnosed every 20 minutes in the United States.

I
n 2007, the Centers for Disease
Control and Prevention (CDC) Autism
and Developmental Disabilities
Monitoring (ADDM) Network released
data indicating that in many parts of
the United States, about one in 150
children who are 8 years old had an
autism spectrum disorder (ASD)1. The
autism community, including educators
and physicians who treat these children,
believes that prevalence is much
higher. While the cause for the autism
epidemic remains the subject of much
controversy, many parents report having
babies who met every milestone until
12-18 months of age, after which they
manifested regressive tendencies and
34 THE AUTISM FILE | www.autismfile.com | info@autismfile.com REPRINTED WITH PERMISSION © THE AUTISM FILE
loss of speech or eye contact, and
acquired peculiar behaviors, seizures
and/or gastrointestinal illness. These
same parents report that these occurred
following a round of childhood vaccines.
Especially in the case of a first or only
child, some parents do not realize that
their child is not meeting the standard
developmental milestones until the child
is 3 years old or older. Parents may notice
a speech delay and/or poor socialization
followed by strange behaviors such
as hand flapping, lack of eye contact,
toe walking, and a desire for repetitive
stimulation.
The National Vaccine Advisory
Committee 2 needs to fund independent
research toward the safety of vaccines
so public trust in these vaccines can be
restored and parents will have confidence
that the current vaccine schedule
is safe for all children of all genetic
predispositions. The recent court cases
of Poling and Banks have favored the
plaintiffs as the rulings declared that
vaccines contributed to both Hannah
Poling’s and Bailey Banks’ autism. In the
Poling case, Hannah was diagnosed with
a mitochondrial disorder which allegedly
caused her to have an adverse reaction to
childhood vaccines3. Despite thousands
of similar cases pending in the vaccine
injury court, a single judge without
a jury recently concluded the first of
ISSUE 33 2009
three cases lacked sufficient evidence to
prove that vaccines contributed to the
respective child’s autism4.
As a result of such recent court cases
and conflicting information, many
new parents have a hard time making
educated decisions about how to
safely vaccinate their children. In fact,
pediatricians are reporting a growing
public concern over vaccines, and many
parents are asking about vaccine safety.
To date, no adjustments have been made
to the childhood vaccine schedule by
the American Academy of Pediatrics or
the CDC. As a safeguard, however, many
parents have begun to request staggered
or delayed vaccination schedules, much
to the consternation of their pediatricians
and contrary to the CDC warning that
this is a dangerous practice that has not
been proven to be effective or a safe
community practice. Parents argue that
they have not seen sufficient safety data
to show that the current vaccine schedule
has been proven to be safe with regard
to all combinations and intervals of
vaccines5. In fact, the majority of safety
studies have been performed on one
vaccine at a time, not multiple vaccine
combinations. New vaccines are quickly
added to the CDC childhood schedule,
and more than 100 new vaccines are in
clinical trial or in the development phase.
Although there are specific genetic
tests and protein biomarkers that can
provide insight into the status of a
child’s neuroimmune system, they are
not used as an assessment tool for
vaccine safety. Geneticists continue to
look for a common gene or set of genes
implicated in autism; however, they
have found nothing conclusive after
10 years and millions of dollars spent
in genetic research. While autism has
grown to epidemic proportions, there
is a general consensus among genetic
researchers that there is no such thing
as a genetic epidemic. But it is widely
accepted by these same researchers
that there, most likely, is a genetic
predisposition increasing the potential for
specific children to undergo a regression
in development following one or more
vaccines, an environmental insult, or
another oxidative stressor.
Dr. Sandra Jill James, a research
professor in the college of medicine
department of pediatrics at the Arkansas
Children’s Hospital, mapped the
methylation pathway and demonstrated
how impairments in this pathway can
cause abnormal levels of specific amino
acid biomarkers that directly reflect
the potential for oxidative stress to
impact children with a diagnosis of
autism 6. Genetic polymorphisms in
the methylation and transsulfuration
pathways can lead to a significant
decrease in glutathione production,
thereby inhibiting the body’s ability
to clear heavy metals and other fat-
soluble toxins. In children with impaired
pathways who are subsequently impacted
by oxidative stressors, classic clinical
presentations include reduced levels
of methionine, cystathionine, cysteine,
glutathione, methyl-B12, methyl-folate,
and B6. Other notable biomarkers in
children with autism include reduced
levels of transferrin, cerumoplasmin,
and L-carnitine, and increased levels
of neopterin, biopterin, isoprostane,
8 OHG, ammonia, and lactic acid.
Children with these findings typically
present with many microbiological
pathogens, including persistent viral,
fungal, bacterial, and even parasitic
infections due to poor modulation of the
lymphocytic and, possibly, innate immune
system. They can present with intestinal
dysbiosis as indicated by an overgrowth
of yeast and bacterial pathogens, which
suppress the healthy balance of gut flora
and impact the body’s inflammatory
status, nutrient absorption, and intestinal
transition times. This abnormal state
creates the “leaky gut” syndrome,
which was first described by Dr. Andrew
Wakefield7,8 and later was confirmed
by multiple gastroenterologists. The
consequence of leaky gut is that patients
develop severe food intolerances and
food sensitivities due to microscopic
proteins leaking out of the gut and
causing immunomodulation. Therefore,
general clinical logic would link a group
of genetic polymorphisms, which should
produce specific identifiable protein or
vitamin biomarkers, to abnormalities of
immune function or modulation in these
children that directly impact their ability
to handle the oxidative stress of the
normal vaccination schedule or chronic
infectious state.
At this time, no research studies
have been performed on newborns
to determine if there are associated
abnormal biomarkers present at birth
for children who eventually develop
ASD. Therefore, as of this writing, no
specific biomarker has been identified in
infants who eventually develop ASD that
could be used as a newborn screening
tool, the usefulness of which would be

Geneticists continue to look for a common gene or set of genes implicated in autism; however, they have
found nothing conclusive after 10 years and millions of dollars spent in genetic research. While autism
has grown to epidemic proportions, there is a general consensus among genetic researchers that there is
no such thing as a genetic epidemic. But it is widely accepted by these same researchers that there, most
likely, is a genetic predisposition increasing the potential for specific children to undergo a regression in
development following one or more vaccines, an environmental insult, or another oxidative stressor.

ISSUE 33 2009 REPRINTED WITH PERMISSION © THE AUTISM FILE info@autismfile.com | www.autismfile.com | THE AUTISM FILE 35
 BIOMEDICAL
to assess the potential adverse effects
of the oxidative stressors resulting
from vaccination or exposure to other
environmental toxins or chemicals.
Physicians who specialize in ASD and
other neurodevelopmental delays use
biomarkers in older children to assess
immune dysfunction, potential for toxic
burden, presence of pathogens, and
status of gastrointestinal function. Doing
research with younger children who
develop autistic tendencies can be quite
difficult and is impacted by the referral
pattern of pediatricians who typically
are first consulted when a child begins
showing early signs of abnormalities.
These children are usually referred to
a pediatric neurologist or psychiatrist
for developmental or behavioral issues.
Next, they may be referred to an
allergist for their food allergies or to a
gastroenterologist for gastrointestinal
issues. The abnormal laboratory findings
are typically only found once the child
is seen by a physician specializing in
autism, and this is usually later in the
child’s development. Unfortunately, the
American Medical Association and the
American Academy of Pediatrics have
not set up a best practices road map for
treatment of ASD. Autism “centers of
excellence” are emerging; but there are
too few specialists available to treat the
growing number of ASD patients.
The National Newborn Screening and
Genetics Resource Center (NNSGRC)
is a cooperative agreement between
the Maternal and Child Health Bureau’s
genetic services branch and the
University of Texas at San Antonio’s
Health Science Center’s department of
pediatrics. Its Web site provides a wealth
of information about newborn screening,
mainly in the United States, and lists the
congenital disorders that can be detected
in a newborn 9. While there is a set of
national recommendations for which
disorders are screened for in infants, each
state determines its own panel, based
The holy grail for medical care on the issue of vaccination safety
is identifying a consistent biomarker or group of biomarkers that
could indicate a compromised immune status or a potentially poor
response to oxidative stress.
36 THE AUTISM FILE | www.autismfile.com | info@autismfile.com REPRINTED WITH PERMISSION © THE AUTISM FILE
on cost to the state health department
or other factors such as demographics.
Newborn screening is performed within
the first few hours of birth via a heel-
stick sample spotted on filter paper in
the hospital. The dried blood samples are
sent to the state health department and
recorded for each newborn via a number
system. Standard screening for genetic or
protein abnormalities is then performed,
and results are usually available to
parents within two to four weeks.
Abnormal results are communicated to
physicians who refer parents to genetic
counselors to advise them about the
detected disorder. The filter papers for
all newborn screenings are saved for a
period of five to 10 years depending on
the state requirement. There are very
informative documents and transcripts
of past meetings on the bioethics of
newborn screening on the Web site for
the President’s Council of Bioethics10.
If we carefully examine the newborn
screening profile performed today,
we can find a long list of recognized
congenital disorders of amino acid
metabolism that potentially indicate
a poor response to oxidative stress.
Many of the abnormalities in amino acid
metabolism being screened for in the
program represent the same markers that
are recognized as abnormal in children
who ultimately develop ASD. What we
can’t determine, due to lack of scientific
research, is if the abnormal levels of
specific biomarkers found in children
who currently have autism were present
at the time of birth or in infancy. Based
on current research at our institution
and in other centers specializing in
children with autism, we have reason
to believe that the recognized amino
acid concentration abnormalities in
children with ASDs may have been
present at birth and represent the
genetic predisposition that defines the
potential for autism development when
environmental stressors are added to
the equation. This will be the topic for a
follow-up article.
The state-mandated newborn
screening program for metabolic core
disorders has established reference
ranges for only specific disease states;
thus, reports are made only if the
reference level is out of the defined
range. For example, a disorder known
as hypermethioninemia is only reported
if the methionine levels are too high.
It is widely known that many children
with regressive autism present with
hypomethioninemia (low levels of
ISSUE 33 2009
 Our hope is that autism will be redefined as a neuroimmune or
epigenetic disorder with autistic regression as a symptom.
methionine). In addition, according to
the Screening, Technology and Research
in Genetics (STAR-G) project’s own
literature, the amino acid disorder,
hyperhomocysteinemia (too much
homocysteine) is a disorder that can
cause mental retardation and/or
speech delays if left untreated. The
recommended treatment is folic acid,
B12, and B6 supplementation and a low
methionine diet (no dairy or wheat)11
which is a recognized amino acid
metabolic deficiency in some children
with autism. In a study by Sarah Aldred,
measured plasma amino acid levels in
autistic patients, their siblings, and
parents showed that children with ASD
come from a family background of
dysregulated amino acid metabolism,
providing further evidence for an
underlying biochemical basis for their
condition12. In summary, it should be
possible to retrospectively examine the
amino acid concentrations of blood
samples collected at birth in any state for
abnormalities in amino acid biomarkers in
children who were later diagnosed with
regressive autism.
The holy grail for medical care on
the issue of vaccination safety is
identifying a consistent biomarker or
group of biomarkers that could indicate
a compromised immune status or a
potentially poor response to oxidative
stress. Once recognized, a rapid, bedside
test cartridge for these biomarkers could
be developed for use prior to vaccinating
the child. Additionally, a biomarker
panel determination that a child could
not safely be vaccinated according to
the recommended vaccine schedule
could reduce, if not eliminate, the need
for parents to be questioned about
medically-based vaccine exemptions.
This biomarker panel would be valuable
in states that do not allow religious or
philosophical exemption from vaccines.
We feel quite confident the identification
of these biomarkers is close at hand.
Pending the advancements in medical
technology, parents are expected to
vaccinate their children without question.
ISSUE 33 2009 REPRINTED WITH PERMISSION © THE AUTISM FILE info@autismfile.com | www.autismfile.com | THE AUTISM FILE
The best advice that we can give them is
to educate themselves before vaccinating
their children and to consider the legal
and ethical options available in their
state.
Newborn screening presents
significant bioethical challenges. There
is increased liability in the event of a
false-positive or false-negative test,
which can dramatically alter a course of
treatment in congenital disorders. Most
likely, many parents would not want their
child to have a label of “autistic” at birth.
In the age of personalized medicine,
based on individual genomic differences,
it will eventually be possible for every
single individual to have a genetic profile
performed at birth to assess genetic
risk factors for certain diseases. For
insurance reporting, the implications
of genomic profiling are likely to cause
many to fear discrimination by disease
state, and this information is likely
to be used to determine the cost of
insurance coverage. We will probably
need legislation to protect individuals
from discrimination based on genomic
profiling. We know certain individuals
can smoke without developing lung
cancer or emphysema, yet others,
non-smokers even, develop smoking-
related diseases based on their genetic
predisposition. There is growing research
that we can change the way our genes
are expressed with biomedical or dietary
intervention, even if we cannot change
our genes. In short, we should be
discussing issues now. As there continues
to be a surge in epigenetics (the co-
dependent role of environment and
genes) we will need to be proactive with
the ethical issues surrounding our future
generations. Our hope is that autism
will be redefined as a neuroimmune
or epigenetic disorder with autistic
regression as a symptom. For parents of
a child with autism, these ethical issues
are of little concern when we discuss the
possibility of recognizing risk factors at
birth and preventing even one more case
of autism … perhaps even staunching
the epidemic.
References
1
CDC Web site, http://www.cdc.gov/
ncbddd/autism/overview.htm
2
NVAC Web site, http://www.hhs.gov/
nvpo/nvac/index.html
3
Generation Rescue Web site, http://
www.generationrescue.org/cases/
index.htm
4
Autism Omnibus Web site, http://
www.uscfc.uscourts.gov/omnibus-
autism-proceeding
5
Fourteen Studies Web site, http://
www.fourteenstudies.org/studies.html
6
James SJ et al., Metabolic biomarkers
of increased oxidative stress and
impaired methyllation capacity in
children with autism. Am J Clin Nutr.
2004 Dec; 80(6):1611-7. http://www.
ajcn.org/cgi/reprint/80/6/1611
7
Presentation on Autistic Enterocolitis,
http://www.thoughtfulhouse.
org/0405-conf-awakefield.htm
8
Wakefield AJ, Anthony A, Murch SH,
Thomson M, Montgomery SM, Davies
S, Walker-Smith JA. Enterocolitis in
children with developmental disorder.
American Journal of Gastroenterology
2000;95:2285-2295
9
National Newborn Screening and
Genetic Resource Center (NNSGRC)
Web site: http://genes-r-us.uthscsa.
edu/
10
The President’s Council on Bioethics
Web site documents: http://www.
bioethics.gov/reports/newborn_
screening/chapter3.html
http://www.bioethics.gov/reports/
newborn_screening/Newborn%20
Screening%20for%20the%20web.pdf
11
The Screening, Technology and
Research in Genetics (STAR-G) Project
http://www.newbornscreening.info/
index.html
12
Aldred S. Plasma Amino Acid Levels in
Children with Autism and Their Families,
J Autism Dev Disord. 2003 Feb; 33(1):
93-7
37