**************************************************************

http://groups.yahoo.com/group/aspartameNM/message/1108
faults in 1999 July EPA 468-page formaldehyde profile:
Elzbieta Skrzydlewska PhD, Assc. Prof., Medical U. of Bialystok, Poland,
abstracts -- ethanol, methanol, formaldehyde, formic acid, acetaldehyde,
lipid peroxidation, green tea, aging, Lyme disease: Murray 2004.08.08
rmforall

Rich Murray, MA Room For All rmforall@...
1943 Otowi Road, Santa Fe, New Mexico 87505 USA 505-501-2298

Herein I offer abstracts and three full texts of dozens of studies by a
world-class biochemist and her associates, mostly experiments with rats, on
ethanol toxicity since 1984 and methanol toxicity since 1993. Enough
details are provided to show the competency and credibility of E.
Skrzydlewska and her colleagues over two decades, and to make access to
their literature more convenient for professionals.

For instance, anyone can click on this post at the above URL, and in Outlook
Express, use Control F to search the text for any word. Yahoo Groups also
includes a fine search function.

A conscientious, responsible review of any reseach that affects the
interests of vast commercial vested interests has to provide justified
criticism of dubious studies, reviews, and conclusions, that are
characteristically the main sources for private and professional
information. My experience since I first started investigating toxicity
issues in 1999 is, count on it, wolves guard the sheep.

To be effective, this criticism has to be calm, civil, detailed, specific,
reasonable, founded on evidence, focused on issues and not on persons, and
based on easily accessed public sources, so that anyone interested in basing
their conclusions on facts can start the laborous process of deciding for
themselves.

http://groups.yahoo.com/group/aspartameNM/message/667
25 rules of disinformation , Sweeney 1997: Murray 2001.07.04 rmforall

As a medical layman, age 62, earning my living the last 16 years as a home
health care giver, my volunteer public service for toxicity issues on the
Net since January 1999 depends entirely on earning credibility by deserving
it.

http://groups.yahoo.com/group/aspartameNM/messages
129 members, 1,108 posts in a public searchable archive

I regret that I almost never receive negative feedback based on any
specifics of this work, for this would enable me, in the finest tradition
of science, to either reverse, amend, or clarify my positions, to the
benefit of humanity, as well as deepening my own satisfaction and confidence
in a long-term effort.

However, it is indubitable that the best disinformation strategy is to
simply ignor any inconvenient researchers and their work, or, if that
becomes problematic due to the quality of the worker and his work, to firmly
repeat the exact opposite of truth, for example, "Aspartame is the most
tested food additive in history," while using ad hominen statements to
dismiss, ridicule, and marginalize the opposition. Fanning a "flame war" of
escalating incivility is a virtually infallible strategy for muddying the
waters, preventing any real examination of actual facts.

I should also here assert that, so far, I have not ever received a
penny of support for my toxicity service, save for about $ 300 of free
books, from both sides of the debate. Furthermore, when the happy day
ensues when I receive payment of any sort, I will immediately and forever
make this clearly and completely known on the Net, along with full details
for contacting the sources, who must agree to repond responsibly,
immediately, and publicly to all inquiries. All my financial information
about this work will be immediately, clearly, and forever public. I cannot
be bought, bent, or borrowed. I don't deal in secret.

A persistent, consistent campaign that provides facts about an actual toxin
can only succeed.

Fact is, the 11% methanol component of aspartame, readily released into the
G.I. tract, is within hours converted into potent amounts of formaldehyde
and formic acid, in amounts scores of times higher than that allowed by the
EPA for daily drinking water.

I have recently summarized mainstream evidence that the similar amounts of
methanol impurity, about one part in ten thousand, in dark wines and
liquors, are largely responsible for the famed "morning after" hangover:

http://groups.yahoo.com/group/aspartameNM/message/1106
hangover research relevant to toxicity of 11% methanol in aspartame
(formaldehyde, formic acid): Calder I (full text): Jones AW:
Murray 2004.08.06 rmforall

Similar potent levels of methanol, and its inevitable products in the human
body, formaldehyde and formic acid, can also ensue from fermentation of
fruits by certain yeast and bacteria in the colon:

http://groups.yahoo.com/group/aspartameNM/message/1110
methanol (formaldehyde and formic acid) from fermentation of fruit in the
colon: Lindinger W, 1997 Aug: Murray 2004.08.10

Alcohol Clin Exp Res. 1997 Aug; 21(5): 939-43.
Endogenous production of methanol after the consumption of fruit.
Lindinger W, Taucher J, Jordan A, Hansel A, Vogel W.
Institut fur Ionenphysik, Leopold Franzens Universitat Innsbruck, Austria.

After the consumption of fruit, the concentration of methanol in the human
body increases by as much as an order of magnitude.
This is due to the degradation of natural pectin (which is esterified with
methyl alcohol) in the human colon.
In vivo tests performed by means of proton-transfer-reaction mass
spectrometry show that consumed pectin in either a pure form (10 to 15 g)
or a natural form (in 1 kg of apples) induces a significant increase of
methanol in the breath (and by inference in the blood) of humans.
The amount generated from pectin (0.4 to 1.4 mg)
is approximately equivalent to the total daily endogenous production
(measured to be 0.3 to 0.6 mg/day)
or that obtained from 0.3 liters of 80-proof brandy
(calculated to be 0.5 mg).
[ typos corrected, g actually is mg for ethanol, methanol ]
This dietary pectin may contribute to the development
of nonalcoholic cirrhosis of the liver. PMID: 9267548

Alcohol Clin Exp Res. 1995 Oct; 19(5): 1147-50.
Methanol in human breath.
Taucher J, Lagg A, Hansel A, Vogel W, Lindinger W.
Institut fur Ionenphysik, Universitat Innsbruck, Austria.

Using proton transfer reaction-mass spectrometry for trace gas analysis of
the human breath, the concentrations of methanol and ethanol have been
measured for various test persons consuming alcoholic beverages and various
amounts of fruits, respectively.
The methanol concentrations increased from a natural (physiological) level
of approximately 0.4 ppm up to approximately 2 ppm a few hours after eating
about 1/2 kg of fruits,
and about the same concentration was reached after drinking of 100 ml brandy
containing 24% volume of ethanol and 0.19% volume of methanol.
[ 24 ml = 64 mg ethanol and 0.19 ml = 0.33 mg methanol ] PMID: 8561283


These three potent dietary sources of methanol, formaldehyde, and formic
acid, which impact many people, and cause the same symptoms in vulnerable
and sensitized people, are ignored in the following prestigious, official
source:

http://groups.yahoo.com/group/aspartameNM/message/1109
faults in 1999 July EPA 468-page formaldehyde profile, extracts & full
references: Murray 2004.08.09 rmforall

[ Extracts ] [ My comments are in square brackets. ]

http://www.atsdr.cdc.gov/toxprofiles/tp111.pdf

TOXICOLOGICAL PROFILE FOR FORMALDEHYDE
U.S. DEPARTMENT OF HEALTH AND HUMAN SERVICES
Public Health Service Agency for Toxic Substances and Disease Registry
July 1999

[ http://www.atsdr.cdc.gov atsdric@... 888-42-ATSDR
404-498-0110 fax 404-498-0093

Agency for Toxic Substances and Disease Registry Division of Toxicology
1600 Clifton Road NE, Mailstop E-29 Atlanta, GA 30333
* Information line and technical assistance Phone: (800) 447-1544 Fax:
(404) 639-6359

* To order toxicological profiles, contact
National Technical Information Service 5285 Port Royal Road Springfield,
VA 22161 Phone: (800) 553-6847 or (703) 487-4650

The National Center for Environmental Health (NCEH) focuses on preventing or
controlling disease, injury, and disability related to the interactions
between people and their environment outside the workplace.
http://www.cdc.gov/nceh/ 888-232-6789
http://www2.cdc.gov/nceh/contactnceh/frmSubmit.asp email contact
Contact: NCEH, Mailstop F-29, 4770 Buford Highway, NE, Atlanta, GA
30341-3724 . Phone: 770-488-7000 . FAX: 770-488-7015.

The National Institute for Occupational Safety and Health (NIOSH) conducts
research on occupational diseases and injuries, responds to requests for
assistance by investigating problems of health and safety in the workplace,
recommends standards to the Occupational Safety and Health Administration
(OSHA) and the Mine Safety and Health Administration (MSHA), and trains
professionals in occupational safety and health.
http://www.cdc.gov/niosh/homepage.html eidtechinfo@...
Contact: NIOSH, 200 Independence Avenue, SW, Washington, DC 20201 . Phone:
513-533-8328 800-356-4674 or NIOSH Technical Information Branch, Robert A.
Taft Laboratory, Mailstop C-19, 4676 Columbia Parkway, Cincinnati, OH
45226-1998 Phone: 800-35-NIOSH fax 513-533-8573

The National Institute of Environmental Health Sciences (NIEHS) is the
principal federal agency for biomedical research on the effects of chemical,
physical, and biologic environmental agents on human health and well-being.
http://www.niehs.nih.gov/
Contact: NIEHS, PO Box 12233, 104 T.W. Alexander Drive,
Research Triangle Park, NC 27709 . Phone: 919-541-3212.
Office of Communications 919-541-3345 TTY 919-541-0731


Referrals
The Association of Occupational and Environmental Clinics (AOEC) has
developed a network of clinics in the United States to provide expertise in
occupational and environmental issues. Contact:
AOEC, 1010 Vermont Avenue, NW, #513, Washington, DC 20005
Phone: 202-347-4976 FAX: 202-347-4950
Web Page: http://www.aoec.org/ e-mail: AOEC@...

AOEC Clinic Director: http://occ-envmed.mc.duke.edu/oem/aoec.htm.

The American College of Occupational and Environmental Medicine (ACOEM) is
an association of physicians and other health care providers specializing in
the field of occupational and environmental medicine.
http://www.acoem.org/ http://www.acoem.org/feedback/ email contact
Contact: ACOEM, 55 West Seegers Road, Arlington Heights, IL 60005
. Phone: 847-818-1800 . FAX: 847-818-9266. ]

FORMALDEHYDE page iii

UPDATE STATEMENT
Toxicological profiles are revised and republished as necessary, but no less
than once every three years. [ I could not locate any more recent updates
than July 1999 via Google. ]

FORMALDEHYDE ix

CONTRIBUTORS
CHEMICAL MANAGER(S)/AUTHORS(S):
Sharon Wilbur, M.A. [ Not a PhD level degree ]
[ Environmental Health Scientist ]
ATSDR, Division of Toxicology, Atlanta, GA

M. Olivia Harris, M.A. [ Not a PhD level degree ]
ATSDR, Division of Toxicology, Atlanta, GA
[ Environmental Health Scientist
1600 Clifton Road NE, E29 Atlanta, GA 30333
P: 404-639-5091 F: 404-639-6315 oxh0@... ]

Peter R. McClure, Ph.D., DABT [ Veterinarian ]
Syracuse Research Corporation, North Syracuse, NY
[ Syracuse Research Corporation Environmental Science Center
301 Plainfield Road Suite 350 Syracuse, New York 13212 (315) 452 8420
mcclure@... ]

Wayne Spoo, DVM, DABT, DABVT [ Veterinarian ]
Research Triangle Institute, Research Triangle Park, NC
[ Jerry Wayne Spoo Operations Director, Life Sciences and Toxicology
919-541-6000 jwspoo@... http://www.rti.org http://www.abvt.org/

CPT Spoo, HHC, USACAPOC, AOCP-MS, 910-432-2209.
jerry.w.spoo@... ]

FORMALDEHYDE xi

PEER REVIEW
A peer review panel was assembled for formaldehyde. The panel consisted of
the following members:
1. Carson Conaway, Research Scientist, American Health Foundation, Valhalla,
New York 10595;
[ http://www.ahf.org/contact/ 914-789-7210 914-789-7243
1 Dana Road Valhalla, NY 10595
300 E. 42nd. Street New York, NY 10017

http://www.ifcp.us/Scientists-Scientists-Carson_Conaway.cfm
Carson Clifford Conaway, Ph. D., DABT [ Veterinarian ]
Research Scientist phone: (914) 789-7210 email: cconaway@...
Institute for Cancer Prevention
In addition to his research work, Dr. Conaway is an Adjunct Associate
Professor in the Department of Pharmacology, New York Medical College. In
that capacity, he is called upon to present lectures in toxicology to
graduate students in the College of Basic Medical Sciences and in the School
of Public Health.

2. John Egle, Jr., Professor, Department of Pharmacology and Toxicology,
Medical College of Virginia, Smith Bldg., Room 656, Richmond, VA 23219; and
[ http://www.medschool.vcu.edu/ John L. Egle, Jr no longer listed. Last
PubMed study in 1995 . Studies on formaldehyde, 2 in 1974, 1 in 1972, no
PubMed abstracts for these. ]

3. Vincent Garry, Director, Environmental Medicine, University of Minnesota,
421 29th Ave., SE Minneapolis, MN 55414.

[
http://www.iatp.org/foodandhealth/library/admin/uploadedfiles/Vincent_Garry_Bio.\
pdf
Vincent F Garry Title: Professor
Department: Lab Medicine/Pathology (office: Lab Med/Pathology Department)
Dept Campus: UMN Twin Cities
E-mail Address: garry001@...
Office Address: Lab Med/Pathology Department
225 Mayo 8609 420 Delaware St SE Minneapolis, MN 55455
Campus Mail: Lab Medicine and Pathology
MMC 609 Mayo 8609 420 Delaware St SE Minneapolis, MN 55455
Office Phone:+1 612-626-3354 Fax:+1 612-626-3380
Address: 4829 Girard Ave So Minneapolis, MN 55409
Phone:+1 612-827-7316

Toxicol Appl Pharmacol. 2004 Jul 15; 198(2): 152-63.
Pesticides and children.
Garry VF.
Department of Laboratory Medicine and Pathology and Program in Toxicology,
University of Minnesota School of Medicine, Minneapolis, MN 55455, USA.

Prevention and control of damage to health, crops, and property by insects,
fungi, and noxious weeds are the major goals of pesticide applications.
As with use of any biologically active agent, pesticides have unwanted
side-effects. In this review, we will examine the thesis that adverse
pesticide effects are more likely to occur in children who are at special
developmental and behavioral risk. Children's exposures to pesticides in the
rural and urban settings and differences in their exposure patterns are
discussed.
The relative frequency of pesticide poisoning in children is examined.
In this connection, most reported acute pesticide poisonings occur in
children younger than age 5.
The possible epidemiological relationships between parental pesticide use or
exposure and the risk of adverse reproductive outcomes and childhood cancer
are discussed.
The level of consensus among these studies is examined.
Current concerns regarding neurobehavioral toxicity and endocrine disruption
in juxtaposition to the relative paucity of toxicant mechanism-based studies
of children are explored. PMID: 15236951 ]

These experts collectively have knowledge of formaldehyde's physical and
chemical properties, toxicokinetics, key health end points, mechanisms of
action, human and animal exposure, and quantification of risk to humans.

All reviewers were selected in conformity with the conditions for peer
review specified in Section 104(I)(13) of the Comprehensive Environmental
Response, Compensation, and Liability Act, as amended.

Scientists from the Agency for Toxic Substances and Disease Registry (ATSDR)
have reviewed the peer reviewers' comments and determined which comments
will be included in the profile.

A listing of the peer reviewers' comments not incorporated in the profile,
with a brief explanation of the rationale for their exclusion, exists as
part of the administrative record for this compound. [ Not easily accessible
by public ]

A list of databases reviewed and a list of unpublished documents cited are
also included in the administrative record. [ Not easily accessible by
public ]

The citation of the peer review panel should not be understood to imply its
approval of the profile's final content. The responsibility for the content
of this profile lies with the ATSDR.... [ Apparently, the peer review
panel's opinions carry little weight. ]

1. PUBLIC HEALTH STATEMENT
This public health statement tells you about formaldehyde and the effects of
exposure....

1.2 WHAT HAPPENS TO FORMALDEHYDE WHEN IT ENTERS THE
ENVIRONMENT?

Most of the formaldehyde you are exposed to in the environment is in the
air.... [ A very misleading statement, as already pointed out above ]

There is usually more formaldehyde present indoors than outdoors.
[ Ignors the issue of dietary sources ]

Formaldehyde is released to the air from many home products and you may
breath in formaldehyde while using these products.

Latex paint, fingernail hardener, and fingernail polish release a large
amount of formaldehyde to the air. [ Bound to be much less than the potent
amounts in dietary sources, which have immediate strong effects on sensitive
and sensitivized persons and other vulnerable groups ]

Plywood and particle board, as well as furniture and cabinets made from
them, fiberglass products, new carpets, decorative laminates, and some
permanent press fabrics give off a moderate amount of formaldehyde.
[ This shows why new buildings, and especially mobile homes and RVs are
toxic for many people. ]

Some paper products, such as grocery bags and paper towels, give off small
amounts of formaldehyde.

Because these products contain formaldehyde, you may also be exposed on the
skin by touching or coming in direct contact with them.

You may also be exposed to small amounts of formaldehyde in the food you
eat. [ The amounts in dietary sources are the most potent sources for most
people. ]

You are not likely to be exposed to formaldehyde in the water you drink
because it does not last a long time in water. [ This ignores the fact that
methanol, always turned into formaldehyde and formic acid in humans, does
indeed
last a very long time in water and dietary sources. ]

Many other home products contain and give off formaldehyde although the
amount has not been carefully measured. [ Potent dietary sources have been
systematically ignored for decades. ]

These products include household cleaners, carpet cleaners, disinfectants,
cosmetics, medicines, fabric softeners, glues, lacquers, and antiseptics.
[ Notice "cosmetics", "medicines", "disinfectants", "antiseptics" -- to this
list of direct skin contact items, we can add hair care products and shoe
leather. ]

You may also breath formaldehyde if you use unvented gas or kerosene heaters
indoors or if you or someone else smokes a cigar, cigarette, or pipe
indoors.

The amount of formaldehyde in mobile homes is usually higher than it is in
conventional homes because of their lower air turnover. [ Evades the issue
that particleboard and other materials common in mobile homes are strong
formaldehyde sources, which this study showed caused symptoms and immune
system signs:

http://www.drthrasher.org/formaldehyde_1990.html full text Jack Dwayne
Thrasher, Alan Broughton, Roberta Madison. Immune activation and
autoantibodies in humans with long-term inhalation exposure to formaldehyde.
Archives of Environmental Health. 1990; 45: 217-223. "Immune activation,
autoantibodies, and anti-HCHO-HSA antibodies are associated with long-term
formaldehyde inhalation." PMID: 2400243 toxicology@... ]

People who work at or near chemical plants that make or use formaldehyde can
be exposed to higher than normal amounts of formaldehyde.

Doctors, nurses, dentists, veterinarians, pathologists, embalmers, workers
in the clothing industry or in furniture factories, and teachers and
students who handle preserved specimens in laboratories also might be
exposed to higher amounts of formaldehyde. The National

FORMALDEHYDE 4 1. PUBLIC HEALTH STATEMENT

Institute for Occupational Safety and Health (NIOSH) estimates that
1,329,332 individuals in the United States have had the potential for
occupational exposure to formaldehyde. [ Common sense suggests that health
professionals who have been exposed to formaldehyde and have become
sensitized and symptomatic naturally will have a prejudice against
discovering the real extent of the danger. ]

1.4 HOW CAN FORMALDEHYDE ENTER AND LEAVE MY BODY?

Formaldehyde can enter your body after you breath it in, drink or eat it, or
when it comes in contact with your skin....

Once absorbed, formaldehyde is very quickly broken down. [ Notice the phrase
"very quickly broken down", which skirts the issue that potent levels of
formaldehyde and formic acid are inevitably produced in humans, and retained
in complex, unresearch amounts. ]

Almost every tissue in the body has the ability to break down formaldehyde.
[ Again the dangers are waved away by definition. The correct way to say
this is that formaldehyde and formic acid toxicity affects every tissue in
the body. ]

It is usually converted to a non-toxic chemical called formate, which is
excreted in the urine. [ This is an astonishing, brazen deceit, defining
formic acid as "non-toxic". Notice the qualification "usually". ]

Formaldehyde can also be converted to carbon dioxide and breathed out of the
body. [ Notice the qualification "can" . ]

It can also be broken down so the body can use it to make larger molecules
needed in your tissues, or it can attach to deoxyribonucleic acid (DNA) or
to protein in your body.... [ In one sentence, formaldehyde is portrayed as
a useful food, while the very serious and complex issue of formaldehyde
adducts to DNA and proteins in all tissues and cells is minimalized:

C. Trocho (1998 July 26): [ Not cited in this lengthy tome. ]
"In all, the rats retained, 6 hours after administration, about 5% of the
label, half of it in the liver."

They used a very low level of aspartame ingestion, 10 mg/kg, for rats, which
have a much greater tolerance for aspartame than humans.
So, the corresponding level for humans would be about 1 or 2 mg/kg.
[ 60 to 120 mg aspartame for a 60-kg person, of which 11% is methanol,
6.6 to 13.2 mg ]
Many headache studies in humans used doses of about 30 mg/kg daily.
[ 1800 mg aspartame for a 60-kg person, of which 11% is methanol, 198 mg ]

http://groups.yahoo.com/group/aspartameNM/message/925
aspartame puts formaldehyde adducts into tissues, Part 1/2
full text, Trocho & Alemany 1998.06.26: Murray 2002.12.22 rmforall

http://ww.presidiotex.com/barcelona/index.html full text
Formaldehyde derived from dietary aspartame binds to tissue components in
vivo.
Life Sci June 26 1998; 63(5): 337-49.
Departament de Bioquimica i Biologia Molecular,
Facultat de Biologia, Universitat de Barcelona, Spain.
http://www.bq.ub.es/cindex.html Línies de Recerca: Toxicitat de
l'aspartame http://www.bq.ub.es/grupno/grup-no.html
Sra. Carme Trocho, Sra. Rosario Pardo, Dra. Immaculada Rafecas,
Sr. Jordi Virgili, Dr. Xavier Remesar, Dr. Jose Antonio
Fernandez-Lopez, Dr. Marià Alemany [male]
Fac. Biologia Tel.: (93)4021521, FAX: (93)4021559
Sra. Carme Trocho "Trok-ho" Fac. Biologia Tel.: (93)4021544,
FAX: (93)4021559 alemany@... ; bioq@...

Abstract:
Adult male rats were given an oral dose of 10 mg/kg aspartame,
14C-labeled in the methanol carbon.
At timed intervals of up to 6 hours, the radioactivity in plasma and several
organs was investigated.
Most of the radioactivity found (>98% in plasma, >75% in liver) was bound to
protein.
Label present in liver, plasma and kidney was in the range of 1-2% of total
radioactivity administered per g or mL, changing little with time.
Other organs (brown and white adipose tissues, muscle, brain, cornea and
retina) contained levels of label in the range of 1/12th to 1/10th of that
of liver.
In all, the rats retained, 6 hours after administration, about 5% of the
label, half of it in the liver.

The specific radioactivity of tissue protein, RNA and DNA was quite uniform.
The protein label was concentrated in amino acids, different from
methionine, and largely coincident with the result of protein exposure to
labeled formaldehyde.
DNA radioactivity was essentially in a single different adduct base,
different from the normal bases present in DNA.
The nature of the tissue label accumulated was, thus, a direct consequence
of formaldehyde binding to tissue structures.

The administration of labeled aspartame to a group of cirrhotic rats
resulted in comparable label retention by tissue components, which suggests
that liver function (or its defect) has little effect on formaldehyde
formation from aspartame and binding to biological components.

The chronic treatment of a series of rats with 200 mg/kg of non-labeled
aspartame during 10 days results in the accumulation of even more label when
given the radioactive bolus, suggesting that the amount of formaldehyde
adducts coming from aspartame in tissue proteins and nucleic acids may be
cumulative.

It is concluded that aspartame consumption may constitute a hazard because
of its contribution to the formation of formaldehyde adducts. PMID: 9714421

[ Extracts ]
"The high label presence in plasma and liver is in agreement with the
carriage of the label from the intestine to the liver via the portal vein.
The high label levels in kidney and, to a minor extent, in brown adipose
tissue and brain are probably a consequence of their high blood flows (45).
Even in white adipose tissue, the levels of radioactivity found 6 hours
after oral administration were 1/25th those of liver.
Cornea and retina, both tissues known to metabolize actively methanol
(21,28) showed low levels of retained label.
In any case, the binding of methanol-derived carbon to tissue proteins was
widespread, affecting all systems, fully reaching even sensitive targets
such as the brain and retina....

The amount of label recovered in tissue components was quite high in all the
groups, but especially in the NA rats.
In them, the liver alone retained, for a long time, more than 2 % of the
methanol carbon given in a single oral dose of aspartame, and the rest of
the body stored an additional 2 % or more.
These are indeed extremely high levels for adducts of formaldehyde, a
substance responsible of chronic deleterious effects (33), that has also
been considered carcinogenic (34,47).
The repeated occurrence of claims that aspartame produces headache and other
neurological and psychological secondary effects-- more often than not
challenged by careful analysis-- (5, 9, 10, 15, 48) may eventually find at
least a partial explanation in the permanence of the formaldehyde label,
since formaldehyde intoxication can induce similar effects (49).

The cumulative effects derived from the incorporation of label in the
chronic administration model suggests that regular intake of aspartame may
result in the progressive accumulation of formaldehyde adducts.

It may be further speculated that the formation of adducts can help to
explain the chronic effects aspartame consumption may induce on sensitive
tissues such as brain (6, 9, 19, 50).

In any case, the possible negative effects that the accumulation of
formaldehyde adducts can induce is, obviously, long-term.

The alteration of protein integrity and function may needs some time to
induce substantial effects.

The damage to nucleic acids, mainly to DNA, may eventually induce cell death
and/or mutations.

The results presented suggest that the conversion of aspartame methanol into
formaldehyde adducts in significant amounts in vivo should to be taken into
account because of the widespread utilization of this sweetener.

Further epidemiological and long-term studies are needed to determine the
extent of the hazard that aspartame consumption poses for humans." ]

Some people are more sensitive to the effects of formaldehyde than
others....
[ Again a very significant, complex, and problematic issue is mentioned and
minimalized in one sentence-- notice the phrase "some people". ]

The Department of Health and Human Services (DHHS) has determined that
formaldehyde may reasonably be anticipated to be a human carcinogen (NTP).

The International Agency for Research on Cancer (IARC) has determined that
formaldehyde is probably carcinogenic to humans.

This determination was based on specific judgements that there is limited
evidence in humans and sufficient evidence in laboratory animals that
formaldehyde can cause cancer.

The Environmental Protection Agency (EPA) has determined that formaldehyde
is a probable human carcinogen based on limited evidence in humans and
sufficient evidence in laboratory animals.... [ These extremely alarming
admissions ought to be emphasized and used to support calls for urgent
research, action, and public warning. ]

The most common way for children to be exposed to formaldehyde is by
breathing it. [ Again, kids are kept at risk with this policy of denial of
the potent role of dietary sources. ]

Children may also be exposed by wearing some types of new clothes or
cosmetics.

A small number of studies have looked at the health effects of formaldehyde
in children. [ Notice the term "small number", which serves to both mention
and minimalize the problem of a dire shortage of adaquate research. ]

It is very likely that breathing formaldehyde will result in nose and eye
irritation (burning feeling, itchy, tearing, and sore throat). [ The focus
is placed on the most unimportant symptoms. ]

We do not know if the irritation would occur at lower concentrations in
children than in adults. [ Research that could threaten vested interests
somehow just doesn't get funded. ]

Studies in animals suggest that formaldehyde will not cause birth defects in
humans. [ Notice the qualification "suggest". ]

Inhaled formaldehyde or formaldehyde applied to the skin is not likely to be
transferred from mother to child in breast milk or to reach the developing
fetus.... [ Notice the qualification "not likely".

ttp://groups.yahoo.com/group/aspartameNM/message/915
formaldehyde toxicity: Thrasher & Kilburn: Shaham: EPA: Gold:
Wilson: CIIN: Murray 2002.12.12 rmforall

Thrasher (2001): "The major difference is that the Japanese demonstrated
the incorporation of FA and its metabolites into the placenta and fetus.
The quantity of radioactivity remaining in maternal and fetal tissues
at 48 hours was 26.9% of the administered dose." [ Ref. 14-16 ]

Arch Environ Health 2001 Jul-Aug; 56(4): 300-11.
Embryo toxicity and teratogenicity of formaldehyde. [100 references]
Thrasher JD, Kilburn KH. toxicology@...
Sam-1 Trust, Alto, New Mexico, USA.
http://www.drthrasher.org/formaldehyde_embryo_toxicity.html full text ]

Formaldehyde is usually found in the air.

Formaldehyde levels are also higher indoors than outdoors.

Opening windows or using a fan to bring in fresh air is the easiest way to
lower formaldehyde levels in the home and reduce the risk of exposure to
your family. [ This reassuring, simple advice is dangerous, since the potent
dietary sources are ignored. ]

Removing formaldehyde sources from the house will also reduce the risk of
exposure.

Since formaldehyde is found in tobacco smoke, not smoking or smoking outside
will reduce exposure to formaldehyde.

Unvented heaters, such as portable kerosene heaters, also produce
formaldehyde. If you do not use these heaters in your home or shop, you help
to prevent
the build up of formaldehyde indoors.

Formaldehyde is found in small amounts in many consumer products including
antiseptics, medicines, dish-washing liquids, fabric softeners, shoe-care
agents, carpet cleaners, glues, adhesives, and lacquers.

If you or a member of your family uses these products, providing fresh
outdoor air when you use them, this will reduce your exposure to
formaldehyde.

Some cosmetics, such as nail hardeners, have very high levels of
formaldehyde.

If you do not use these products in a small room, or if you have plenty of
ventilation when you use them, you will reduce your exposure to
formaldehyde.

If your children are not in the room when you use these products, you will
also reduce their exposure to formaldehyde.

Formaldehyde is emitted from some wood products such as plywood and particle
board, especially when they are new.

The amount of formaldehyde released from them decreases slowly over a few
months. [ Notice "slowly over a few months". ]

If you put these materials in your house, or buy furniture or cabinets made
from them, opening a window will lower formaldehyde in the house. [ What
happens in winter? ]

The amount of formaldehyde emitted to the house will be less if the wood
product is covered with plastic laminate or coated on all sides.

If it is not, sealing the unfinished sides will help to lower the amount of
formaldehyde that is given off.

Some permanent press fabrics emit formaldehyde. [ Why aren't there warning
labels? ]

Washing these new clothes before use will usually lower the amount of
formaldehyde and reduce your family's risk of exposure.

FORMALDEHYDE 7 1. PUBLIC HEALTH STATEMENT

1.8 IS THERE A MEDICAL TEST TO DETERMINE WHETHER I HAVE BEEN EXPOSED TO
FORMALDEHYDE?

We have no reliable test to determine how much formaldehyde you have been
exposed to or whether you will experience any harmful health effects....
[ Another casual mention of an alarming reality ]

1.9 WHAT RECOMMENDATIONS HAS THE FEDERAL GOVERNMENT MADE TO PROTECT HUMAN
HEALTH?

The federal government develops regulations and recommendations to protect
public health.

Regulations can be enforced by law.

Federal agencies that develop regulations for toxic substances include the
EPA, the Occupational Safety and Health Administration (OSHA), and the Food
and Drug Administration (FDA).

Recommendations provide valuable guidelines to protect public health but
cannot be enforced by law. [ Not reassuring... ]

Federal organizations that develop recommendations for toxic substances
include the Agency for Toxic Substances and Disease Registry (ATSDR) and the
NIOSH. [ ATSDR and NIOSH cannot enforce their recommendations. ]

Regulations and recommendations can be expressed in not-to-exceed levels in
air, water, soil, or food that are usually based on levels that affect
animals,
then they are adjusted to help protect people. [ This bypasses the
issue that the reseach on humans is very inadequate to determine the actual,
complex toxicity of methanol, formaldehyde, and formic acid. ]

Sometimes these not-to-exceed levels differ among federal organizations
because of different exposure times (an 8-hour workday or a 24-hour day),
the use of different animal studies, or other factors....
[ An outstanding example of disharmony in the EPA is the fact that the
1998.05.05 EPA IRIS level for oral methanol in humans (Oral Rfd) is 0.5
mg/kg/day, or 30 mg oral methanol daily for a 60 kg human. The animal
study used was:

U.S. EPA. 1986. Rat oral subchronic toxicity study with methanol. Office of
Solid Waste, Washington, DC.

I have not found on the Net any information as to the authors, institution,
abstract, or full text of this study.

But the EPA ATSDR limit for formaldehyde in drinking water is
1 ppm, or 2 mg daily for a typical daily consumption of 2 L of water:

http://groups.yahoo.com/group/aspartameNM/message/835
ATSDR: EPA limit 1 ppm formaldehyde in drinking water July 1999:
Murray 2002.05.30 rmforall

http://www.atsdr.cdc.gov/tfacts111.html
[excerpts]

Agency for Toxic Substances and Disease Registry Division of Toxicology
1600 Clifton Road NE, Mailstop E-29Atlanta, GA 30333 888-422-8737 FAX:
(404)498-0057 ATSDRIC@... http://www.atsdr.cdc.gov/contacts.html

"The EPA recommends that an adult should not drink water containing more
than 1 milligram of formaldehyde per liter of water (1 mg/L) for a
lifetime exposure, and a child should not drink water
containing more than 10 mg/L for 1 day or 5 mg/L for 10 days."

This stringent limit means that if over 13% of the oral methanol limit
results in production of formaldehyde in the human body by the liver, then
the formaldehyde limit would be exceeded. This is cutting things pretty
close.

http://www.epa.gov/iris/subst/0305.htm
also http://www.china-pops.net/enwww/IRIS-Mirror/subst/0305.htm 1998.05.05

USA Environmental Protection Agency EPA
Integrated Risk Information System IRIS

This site explains that the harmful rat dose of 500 mg/kg body weight per
day was
divided by 10 for "interspecies extrapolation" (the higher vulnerability of
humans than rats),
by 10 for "range of sensitivity" (the variation of individual human
vulnerability), and
by 10 for "subchronic to chronic exposure" (the increased danger from
lifetime as compared to the 3 month exposure in the rat test),
giving a total reduction of 10x10x10 = 1000 for the UF = Uncertainty Factor.

The human Oral RfD is the rat Oral RfD divided by 1000, so
500 mg/kg/day is reduced to 0.5 mg/kg/day , so that the allowed dose for a
60 kg human is 30 mg oral methanol daily.


Moreover, a recent study found that after 4 months of moderate oral
aspartame, rats took four times longer to finish a simple, one-turn maze--
an alarming level of neurotoxicity:

http://groups.yahoo.com/group/aspartameNM/message/1088
Murray, full plain text & critique:
chronic aspartame in rats affects memory, brain cholinergic receptors, and
brain chemistry, Christian B, McConnaughey M et al, 2004 May:
2004.06.05 rmforall

"Control and treated rats were trained in a T-maze to a particular side and
then periodically tested to see how well they retained the learned response.

Rats that had received aspartame (250 mg/kg/day) in the drinking water
for 3 or 4 months showed a significant increase in time to reach the reward
in the T-maze, suggesting a possible effect on memory due to the artificial
sweetener."

The 11% methanol component of aspartame is immediately released in the GI
tract, so these rats were being exposed to only 27.5 mg/kg/day methanol.

The EPA IRIS on 1998.05.05 used a 1986 90 day rat study to find a
No-Observed-Effect Level (NOEL) value of 500 mg/kg/day, which, divided by
1000, became their human long-term safe methanol level of 0.5 mg per kg body
weight per day, which for a 60 kg average person is 30 mg methanol daily,
for oral exposure.

However, the rat level is 18 times greater than that for the level of
dramatic memory loss and clear-cut brain changes found by McConnaughey M,
May 2004.

This suggests reducing the human long-term safe level twenty times to
.025 mg/kg/day = 25 micrograms per kg body weight per day,
which for a 60 kg average person is 1.5 mg oral methanol per day.


It is certain that high levels of aspartame use, above 2 liters daily for
months and years, must lead to chronic formaldehyde-formic acid toxicity.

Fully 11% of aspartame is methanol-- 1,120 mg aspartame in 2 L diet soda,
almost six 12-oz cans, gives 123 mg methanol (wood alcohol), about 22 mg
methanol per can.

If only 10% of the methanol accumulates daily as formaldehyde, that would
give 12 mg daily formaldehyde accumulation-- about 60 times more than the
0.2 mg from 10% retention of the 2 mg EPA daily limit for formaldehyde in
drinking water.

If about 30% of oral methanol is retained as formaldehyde and formic acid,
then this EPA ATSDR formaldehyde limit of 2 mg daily for 2 L drinking water
suggests a corresponding methanol limit of 6.7 mg daily, about 4.5 times the
safe limit based on the McConnaughey data. This is much closer than the
1998 EPA IRIS limit of 30 mg daily oral methanol, which is 20 times the
McConnaughey data
limit. ]
**************************************************************



Returning to the voluminous work of Elzbieta Skrzydlewska, it is important
that many of her studies suggest that many safe substances may prevent or
treat toxicity from methanol and its inevitable toxic human body products,
formaldehyde and formic acid:

N-acetylcysteine (2000); U-83836E containing a trolox ring (1997);
green tea (2004); vitamins E, C, A, and beta-carotene (2004);
glutathione (2001); N-Acetylcysteine (NAC) (2001); melatonin (2001);
low and medium levels of cysteine (1990).



"Methanol, when introduced into all mammals, is oxidized into formaldehyde
and then into formate, mainly in the liver.

Such metabolism is accompanied by the formation of free radicals....

The consequences of methanol metabolism and toxicity distinguish the human
and monkey from lower animals.

Formic acid is likely to be the cause of the metabolic acidosis and ocular
toxicity in humans and monkeys,
which is not observed in most lower animals.

Nevertheless, chemically reactive formaldehyde and free radicals may damage
most of the components of the cells of all animal species, mainly proteins
and lipids...."

http://taylorandfrancis.metapress.com/openurl.asp?genre=article&eissn=1537-6524&\
volume=13&issue=4&spage=277

Toxicology Mechanisms and Methods
Publisher: Taylor & Francis Health Sciences, part of the Taylor & Francis
Group Issue: Volume 13, Number 4 / Oct-Dec 2003 Pages: 277 - 293

Toxicological and Metabolic Consequences of Methanol Poisoning
Elzbieta Skrzydlewska, Assoc. Professor, MSc, PhD, Deputy Dean of Faculty of
Pharmacy, Head of Department of Analytical Chemistry, Medical University of
Bialystok, Mickiewicza 2A 15-230 Bialystok 8, P.O. Box 14, Poland
skrzydle@...
http://www.amb.edu.pl/en/sites/university.html dzss@...
Kilinskiego 1 15-089 Bialystok, Poland fax (48 85)7485408

Abstract:
Methanol, when introduced into all mammals, is oxidized into formaldehyde
and then into formate, mainly in the liver.

Such metabolism is accompanied by the formation of free radicals.

In all animals, methanol oxidation, which is relatively slow, proceeds via
the same intermediary stages, usually in the liver,
and various metabolic systems are involved in the process, depending on the
animal species.

In nonprimates, methanol is oxidized by the catalase-peroxidase system,
whereas in primates, the alcohol dehydrogenase system takes the main role in
methanol oxidation.

The first metabolite (formaldehyde is rapidly oxidized by formaldehyde
dehydrogenase) is the reduced glutathione (GSH)-dependent enzyme.

Generated formic acid is metabolized into carbon dioxide with the
participation of H4folate and two enzymes, 10-formyl H4folate synthetase and
dehydrogenase,
whereas nonprimates oxidize formate efficiently.

Humans and monkeys possess low hepatic H4folate and 10-formyl H4folate
dehydrogenase levels
and are characterized by the accumulation of formate after methanol
intoxication.

The consequences of methanol metabolism and toxicity distinguish the human
and monkey from lower animals.

Formic acid is likely to be the cause of the metabolic acidosis and ocular
toxicity in humans and monkeys,
which is not observed in most lower animals.

Nevertheless, chemically reactive formaldehyde and free radicals may damage
most of the components of the cells of all animal species, mainly proteins
and lipids.

The modification of cell components results in changes in their functions.

Methanol intoxication provokes a decrease in the activity and concentration
of antioxidant enzymatic as well as nonenzymatic parameters,
causing enhanced membrane peroxidation of phospholipids.

The modification of protein structure by formaldehyde as well as by free
radicals results changes in their functions,
especially in the activity of proteolytic enzymes and their inhibitors,
which causes disturbances in the proteolytic-antiproteolytic balance toward
the proteolytics and
enhances the generation of free radicals.

Such a situation can lead to destructive processes because components of the
proteolytic-antiproteolytic system during enhanced membrane lipid
peroxidation may penetrate from blood into extracellular space, and an
uncontrolled proteolysis can occur.

This applies particularly to extracellular matrix proteins.

Keywords:
Free Radicals, Methanol Metabolism, Methanol Poisoning, Proteases, Protease
Inhibitors

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J Pharm Pharmacol. 2000 May; 52(5): 547-52.
Protective effect of N-acetylcysteine on rat liver cell membrane during
methanol intoxication. kasacka@...
Dobrzynska I, Skrzydlewska E, Kasacka I, Figaszewski Z.
Institute of Chemistry, University in Bialystok, Poland.

Methanol is oxidized in vivo to formaldehyde and then to formate, and these
processes are accompanied by the generation of free radicals.
We have studied the effect of N-acetylcysteine on liver cell membrane from
rats intoxicated with methanol (3.0 g kg(-1)).
Evaluation of the effect was achieved by several methods.
Lipid peroxidation and surface charge density were measured.
An ultrastructural study of the liver cells was undertaken.
The concentration of marker enzymes of liver damage (alanine
aminotransferase and aspartate aminotransferase) in blood serum was
measured.
Methanol administration caused an increase in lipid peroxidation products
(approximately 30%) as well as in surface charge density (approximately
60%).
This might have resulted in the membrane liver cell damage visible under
electron microscopy and a leak of alanine aminotransferase and aspartate
aminotransferase into the blood (increase of approximately 70 and 50%,
respectively).
Ingestion of N-acetylcysteine with methanol partially prevented these
methanol-induced changes.
Compared with the control group, lipid peroxidation was increased by
approximately 3% and surface charge density by approximately 30%.
Alanine aminotransferase and aspartate aminotransferase activity increased
by 9 and 8%, respectively, compared with the control group.
The results suggested that N-acetylcysteine was an effective antioxidant in
methanol intoxication.
It may have efficacy in protecting free radical damage to liver cells
following methanol intoxication. PMID: 10864143



"Changes in protein structure resulted both from free radical action and
formaldehyde generation during methanol intoxication."

J Appl Toxicol. 2000 May-Jun; 20(3): 239-43.
Effect of methanol intoxication on free-radical induced protein oxidation.
Skrzydlewska E, Elas M, Farbiszewski R, Roszkowska A.
Department of Analytical Chemistry, Medical University, 15-230 Bialystok 8,
Poland.

Oxygen free radicals are generated during methanol-induced liver injury, as
was shown for ethanol.
The effect of methanol intoxication (6 g kg(-1) body wt.) on protein
modification in the liver of rats was investigated.
Electron spin resonance determination indicated an increase in the free
radical signal 6 and 12 h after intoxication.
After 7 days of treatment, the contents of malondialdehyde and carbonyl
groups in proteins were significantly increased.
The level of amino groups and sulphydryl groups and the amount of tryptophan
in proteins were decreased,
whereas the amount of bi-tyrosine was increased significantly.
Changes in protein structure resulted both from free radical action and
formaldehyde generation during methanol intoxication.
Copyright 2000 John Wiley & Sons, Ltd. PMID: 10797478



Toxicology. 2000 Dec 7; 156(1): 47-55.
N-acetylcysteine or trolox derivative mitigate the toxic effects of methanol
on the antioxidant system of rat brain.
Farbiszewski R, Witek A, Skrzydlewska E.
Department of Analytical Chemistry, Bialystok Medical Academy, Mickiewicza
Str 2, P.O. Box 14, 15-230 Bialystok 8, Poland.

The effect of two compounds: N-acetylcysteine (NAC) and trolox derivative
(U-83836E) on the methanol induced impairment of the antioxidant system of
the rat brain was studied in male Wistar rats (approx. 250 g body weight).
The animals were divided into six main groups:
control group (0.5 ml of physiological saline intragastrically),
NAC group (150 mg/kg intraperitoneally-i.p),
U-83836E group (10 mg/kg i.p.),
methanol group (3 g/kg intragastrically),
NAC+methanol and U-83836E+methanol groups.
In these particular groups the changes in antioxidant parameters were
observed for 6,12,24,48 h and 5 and 7 days.
The results proved that the use of methanol and N-acetylcysteine increased
the activities of Cu,Zn-superoxide dismutase, glutathione peroxidase and
glutathione reductase by about 15,15 and 41%, respectively, in comparison to
the group of rats receiving methanol alone.
Similarly, the level of GSH increased by about 17%, the concentration of
ascorbate by 20%, while the thiobarbituric acid-reactive substances (TBA-rs)
diminished to the values as in control group.
The use of new antioxidant U8383E and methanol showed less beneficial effect
in the measured parameters however,
it serves as a better protector for the methanol induced decrease in
GSH-content. These data suggest that NAC and U-83836E mitigate the toxic
effects of
methanol on the antioxidant system of the rat brain. PMID: 11162875




Rocz Akad Med Bialymst. 1999; 44: 89-101.
Morphological changes in the liver of rats intoxicated with methanol.
Kasacka I, Skrzydlewska E.
Department of Histology and Embryology, Medical Academy of Bialystok.

On the basis of morphological examinations in light and electron microscope,
the evaluation of methanol influence on the liver of rats was conducted.
The examination was carried out in the group of 36 rats that were given a
single dose of methanol (1.5 g/kg b.w.) into the stomach through a gastric
tube.
The liver was taken from rats under the ether anaesthesia after 6, 12, and
24 hours as well as after 2, 5, and 7 days of methanol administration.
Results showed that methanol intoxication caused visible changes in the
examined organ.
Only 6 h after intoxication, lobular peripheral hepatocytes presented
characteristic features of vacuolar degradation persisting up to 48 h.
Since the second day of intoxication, many cells with double nuclei were
found more frequently than in controls.
Single hepatocytes or small hepatocytic clusters with the features of
deliquescent necrosis could be seen after 5 and 7 days of examination.
All animals intoxicated with methanol showed distinct weakness of glycogen
reaction.
The loss of glycogen resources was highest at 24 h after methanol
administration.
The results indicate, that methanol causes morphological changes in the rat
liver and that intensification of these changes depends on the time after
intoxication. PMID: 10697423



Rocz Akad Med Bialymst. 1999; 44: 76-88.
Activity of lysosomal proteases in the liver and in the plasma from rats
intoxicated with methanol.
Skrzydlewska E.
Department of Analytical Chemistry, Medical Academy of Bialystok.

The activity of lysosomal proteolytic enzymes (cathepsin A, B, C, D and E)
in cytosol and in the whole homogenate of the liver and in the blood plasma
from rats intoxicated with 1.5, 3.0 and 6.0 g methanol/kg b.w. was measured
6, 12 and 24 h and 2, 5 and 7 days after the intoxication.
The activity of all proteases was increased in the cytosol from 12 h to 5
days of intoxication, whereas the activity of these enzymes was decreased in
the whole liver homogenate during the same time.
The magnitude of the decrease in proteolytic activity in the whole
homogenate of the liver depended on the amino acid active center of the
enzyme.
The greatest decrease was observed for sulfhydryl and hydroxyl proteases and
smaller one for carboxyl proteases.
The proteases activity in the plasma was increased from 12 h to 5 days after
methanol intoxication.
These results suggest that during methanol intoxication the cellular and
lysosomal membranes are impaired and proteases are translocated into the
blood. However, changes in proteases activities and proteases distribution
within
the hepatocytes may lead to disturbances in the catabolism of cell proteins
and to destruction of liver cells. PMID: 10697422



"The primary metabolic appropriation of methanol is oxidation to
formaldehyde and then to formate.
These processes are accompanied by formation of superoxide anion and
hydrogen peroxide....
Methanol administration,[ by ] increasing concentration of the lipid
peroxidation products, decreased the liver glutathione-peroxidase and
glutathione reductase (GSSG-R) activities, GSH concentration and total
antioxidant status (TAS)."

Drug Alcohol Depend. 1999 Nov 1; 57(1): 61-7.
Protective effect of N-acetylcysteine on reduced glutathione, reduced
glutathione-related enzymes and lipid peroxidation in methanol intoxication.
Skrzydlewska E, Farbiszewski R. skrzydle@...
Department of Analytical Chemistry, Bialystok Medical University, Poland.

The primary metabolic appropriation of methanol is oxidation to formaldehyde
and then to formate.
These processes are accompanied by formation of superoxide anion and
hydrogen peroxide.
This paper reports data on the effect of N-acetylcysteine (NAC) on reduced
glutathione (GSH) and on activity of some GSH-metabolising enzymes in the
liver, erythrocytes and serum of rats intoxicated with methanol (3 g/kg
b.w.) during 7 days after intoxication.
Methanol administration,[ by ] increasing concentration of the lipid
peroxidation products, decreased the liver glutathione-peroxidase and
glutathione reductase (GSSG-R) activities, GSH concentration and total
antioxidant status (TAS).
The use of NAC after methanol ingestion apparently diminished lipid
peroxidation, elevated the GSH level in the liver and erythrocytes, and
increased activity of GSH-related enzymes in the serum, erythrocytes and in
the liver.
These results suggest that NAC exerts its protective effect by acting as a
precursor for glutathione, the main low molecular antioxidant and as a free
radical scavenger. PMID: 10617314



"Methanol ingestion in humans caused changes in activities of proteases and
their inhibitors with similar direction as in rats.
These changes in activity of proteases and their inhibitors produce signific
ant disturbances in proteolytic-antiproteolytic balance after methanol
administration."

J Toxicol Environ Health A. 1999 Jul 23; 57(6): 431-42.
Activity of cathepsin G, elastase, and their inhibitors in plasma during
methanol intoxication.
Skrzydlewska E, Szmitkowski M, Farbiszewski R.
Department of Analytical Chemistry, Medical University, Bialystok, Poland.
skrzydle@...

Methanol oxidation in the liver is accompanied by formation of formaldehyde
and free radicals.
These compounds can react with biologically active proteins, including
proteolytic enzymes and their inhibitors.
The activity of cathepsin G and elastase and their inhibitors such as
alpha-1-antitrypsin and alpha-2-macroglobulin in plasma of rats given
methanol orally in doses of 1.5, 3, and 6 g/kg was investigated for 7 days.
The activity of cathepsin G and elastase was increased from 12 h to 5 d,
proportionally to methanol dose.
At the same time, activity of their inhibitors was reduced.
Methanol ingestion in humans caused changes in activities of proteases and
their inhibitors with similar direction as in rats.
These changes in activity of proteases and their inhibitors produce signific
ant disturbances in proteolytic-antiproteolytic balance after methanol
administration. PMID: 10478824


Folia Histochem Cytobiol. 1999; 37(2): 111-2.
Parenchymal cell mitochondria in the liver of rats after methanol
intoxication.
Kasacka I, Skrzydlewska E.
Department of Histology, Medical University, Bialystok, Poland.
PMID: 10352983


"Our findings indicate decreased antioxidative potential both in the brain
and in the liver of rats after methanol ingestion."

Comp Biochem Physiol C Pharmacol Toxicol Endocrinol. 1998 Aug; 120(2):
289-94.
The comparison of the antioxidant defense potential of brain to liver of
rats after methanol ingestion.
Skrzydlewska E, Witek A, Farbiszewski R.
Department of Instrumental Analysis, Bialystok Medical University, Poland.

The antioxidant enzymatic and nonenzymatic potential in the brain of rats
given methanol orally was investigated for 7 days consecutively and compared
to that one in the liver.
Glutathione (GSH) and the activities of superoxide dismutase (Cu, Zn-SOD),
glutathione peroxidase (GSH-Px) and glutathione reductase (GSSG-R) were
reduced in the brain after the first 24 h, whereas
in the liver these parameters were diminished after 6 h.
The brain catalase (CAT) activity was very low and constant in contrast to
high and changeable CAT in the liver.
At the beginning of intoxication, the activities of Cu, Zn-SOD and CAT in
the liver were increased;
after 5 days they were restored to normal values
while Cu, Zn-SOD diminished gradually in the brain.
An early change that occurred 6 h after intoxication was a decrease of
ascorbate in the brain and in the liver.
The increase in thiobarbituric acid-reactive substances (TBA-rs) in the
liver was preceded by their increase in the brain.
Our findings indicate decreased antioxidative potential both in the brain
and in the liver of rats after methanol ingestion.
The regulatory mechanisms of the antioxidant enzymes in the brain of
intoxicated rats differ from those ones in the liver. PMID: 9827043



Acta Biol Hung. 1998; 49(2-4): 345-52.
Formaldehyde-induced modification of hemoglobin in vitro.
Farbiszewski R, Skrzydlewska E, Roszkowska A.
Department of Analytical Chemistry, Medical University, Bialystok, Poland.

Formaldehyde is known to react with proteins.
The purpose of our experiments was to analyse in vitro the effect of
formaldehyde on the physicochemical and biological properties of hemoglobin
molecules.
The effect of formaldehyde concentration, reaction time, pH and temperature
on hemoglobin free amino groups was estimated.
The modified hemoglobin was analysed using electrophoretic, potentiometric
and spectrophotometric techniques.
Reaction between formaldehyde and hemoglobin was accelerated by increasing
concentration of formaldehyde and higher temperature.
This reaction was most intensive during the first few hours at pH 7.4 so the
amount of free amino groups of hemoglobin was significantly diminished by
directly mixing formaldehyde with hemoglobin.
The modified protein was characterized by the increase in electrophoretic
mobility and the decrease in maximum absorption derived from porphyrin
rings. Formaldehyde modified hemoglobin was less susceptible to the action
of
cathepsin D. PMID: 10526979



"These results indicate that methanol intoxication in rats leads to an
increase in the lipid peroxidation and impairment in the antioxidant
mechanisms in liver, erythrocytes, and blood serum."

J Toxicol Environ Health A. 1998 Apr 24; 53(8): 637-49.
Lipid peroxidation and antioxidant status in the liver, erythrocytes, and
serum of rats after methanol intoxication.
Skrzydlewska E, Farbiszewski R.
Department of Instrumental Analysis, Medical Academy, Bialystok, Poland.

Lipid peroxidation products measured as a malondialdehyde and activities of
superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), glutathione
reductase (GSSG-R), and concentrations of ascorbic acid, alpha-tocopherol,
and glutathione (GSH) were measured in the liver, erythrocytes, and serum of
rats 6, 14, and 24 h and 2, 5, and 7 d after treatment with 3 g methanol/kg.
GSH-Px and GSSG-R activities, GSH level, and ascorbate concentration in the
liver, erythrocytes, and blood serum were significantly decreased.
In addition, SOD and alpha-tocopherol in erythrocytes were diminished, while
malondialdehyde (MDA) in liver, erythrocytes, and serum were elevated.
Further, erythrocyte counts, hemoglobin levels, hematocrit, and mean
corpuscular volume (MCV) were reduced.
These results indicate that methanol intoxication in rats leads to an
increase in the lipid peroxidation and impairment in the antioxidant
mechanisms in liver, erythrocytes, and blood serum. PMID: 9572161



Rocz Akad Med Bialymst. 1997; 42 Suppl 2: 47-55.
Ultrastructural evaluation of lysosomes and biochemical changes in cathepsin
D distribution in hepatocytes in methanol intoxication.
Skrzydlewska E, Szynaka B.
Department of Instrumental Analysis, Medical Academy of Bialystok.

Methanol oxidation is accompanied by free radicals and formaldehyde
formation.
It is likely to cause damage of lysosomal membranes.
Lysosomal ultrastructure under transmission electron microscope and
biochemical
localization of cathepsin D were estimated after rats intoxication with
methanol.
The examination was carried out 6, 12 and 24 h and 2.5 and 7 days after
intoxication.
Ultrastructural examination showed that methanol causes
extension of Golgi apparatus cisterns and an increase in a number of
lysosomes.
From 12 h to 2 days lysosomes were characterized by damage of structure of
membrane enclosing lysosomes.
During the first days of intoxication activity of cathepsin D decreased in
lysosomes and increased in cytosol.
These changes may lead to uncontrolled extralysosomal proteolysis in the
liver cells and to the onset of liver tissue destruction. PMID: 9646682



Rocz Akad Med Bialymst. 1997; 42 Suppl 2: 39-46.
Ultrastructural evaluation of hepatocytes membranes and changes in cytosolic
enzymes distribution in methanol intoxication.
Skrzydlewska E, Szynaka B.
Department of Instrumental Analysis, Medical Academy of Bialystok.

Acute methanol intoxication causes metabolic and structural disturbances of
liver cells.
The aim of this paper, therefore, was to evaluate the
ultrastructure of liver cells membrane and the amount of lipid peroxidation
products, as well as the concentration of marker enzymes of liver damage
(ALT and AST) in blood serum.
The experiment was done on Wistar rats which once received intragastrically
6.0 g methanol/kg b.w. as a 50% solution.
The animals were decapitated 6, 12 and 24 h and 2, 5 and 7 days after the
methanol administration.
The liver was evaluated under transmission electron microscope and lipid
peroxidation products were determined in the liver homogenate.
The serum ALT and AST activity were also assayed.
The biochemical results indicate the increase in lipid peroxidation
products.
The consequence of this is probably the membrane liver cell damage visible
in the electron microscope. PMID: 9646681




"The primary metabolic fate of methanol is oxidation to formaldehyde and
then to formate by enzymes of the liver....

Changes due to methanol ingestion may indicate impaired antioxidant defense
mechanisms in the liver tissue. "

Free Radic Res. 1997 Oct; 27(4): 369-75.
Decreased antioxidant defense mechanisms in rat liver after methanol
intoxication.
Skrzydlewska E, Farbiszewski R.
Department of Instrumental Analysis, Medical Academy, Poland.

The primary metabolic fate of methanol is oxidation to formaldehyde and then
to formate by enzymes of the liver.
Cytochrome P-450 and a role for the hydroxyl radical have been implicated in
this process.
The aim of the paper was to study the liver antioxidant defense system in
methanol intoxication, in doses of 1.5, 3.0 and 6.0 g/kg b.w., after
methanol administration to rats.
In liver homogenates, the activities of Cu,Zn-superoxide dismutase and
catalase were significantly increased after 6 h following methanol ingestion
in doses of 3.0 and 6.0 g/kg b.w. and persisted up to 2-5 days,
accompanied by significant decrease of glutathione reductase and glutathione
peroxidase activities.
The content of GSH was significantly decreased during 6 hours to 5 days.
The liver ascorbate level was significantly diminished, too, while MDA
levels were considerably increased after 1.5, 3.0 and 6.0 g/kg b.w. methanol
intoxication.
Changes due to methanol ingestion may indicate impaired antioxidant defense
mechanisms in the liver tissue. PMID: 9416465



Arch Toxicol. 1997; 71(12): 741-5.
Glutathione consumption and inactivation of glutathione-related enzymes in
liver, erythrocytes and serum of rats after methanol intoxication.
Skrzydlewska E, Farbiszewski R.
Department of Instrumental Analysis, Medical Academy, Bialystok, Poland.

The primary metabolic fate of methanol is oxidation to formaldehyde and then
to formate.
These processes are accompanied by formation of superoxide anion
and further hydrogen peroxide.
Glutathione plays a unique role in the cellular defense system against
xenobiotics.
The glutathione (GSH) content and glutathione peroxidase (GSH-Px) and
glutathione reductase (GSSG-R) activities were measured in liver,
erythrocytes and serum of rats.
Rats were intoxicated with 3.0 and 6.0 g methanol/kg body wt. and
measurements taken after 6, 12 and 24 h and 2, 5 and 7 days of intoxication.
The decrease in GSH content and in GSH-related enzyme activity was observed
during the whole time-course of the intoxication.
The most significant changes were observed in the erythrocytes.
The results obtained show that the protection against oxidative damage due
to methanol intoxication in rats seems to be less efficient than in control
rats. PMID: 9388006



Acta Biochim Pol. 1997; 44(2): 339-42.
Activity of liver proteases in experimental methanol intoxication.
Skrzydlewska E, Skrzydlewski Z, Worowski K.
Department of Instrumental Analysis, Medical Academy, Bialystok, Poland.

Intoxication of rats with methanol (1.5 and 3.0 g/kg body weight) led to a
significant, time- and dose-dependent decrease in the activities of
cathepsins A, B and C, while the activity of cathepsin D was unaffected.
The decrease was associated with a different partial release of individual
cathepsins to the post-lysosomal fraction. PMID: 9360724



Fundam Clin Pharmacol. 1997; 11(5): 460-5.
Trolox-derivative antioxidant protects against methanol-induced damage.
Skrzydlewska E, Farbiszewski R.
Department of Instrumental Analysis, Medical Academy, Bialystok, Poland.

This paper reports data on the effect of a new antioxidant, U-83836E, on the
lipid peroxidation and antioxidant status of liver, red blood cells (RBCs)
and blood serum of rats intoxicated with methanol (3.0 g/kg body weight).
Methanol administration slightly increased the levels of peroxidation
products in the liver, and markedly increased them in RBCs and serum.
In contrast, glutathione-peroxidase, glutathione-reductase activity, reduced
glutathione concentration and total antioxidant status were decreased.
The use of U-83836E, containing a trolox ring, appeared to be beneficial in
reducing lipid peroxidation products and in partially in preventing the
decrease in glutathione and antioxidant enzymes induced by methanol in liver
and serum.
These results show that antioxidant U-83836E may partially prevent methanol
toxicity. PMID: 9342600

"These results indicate that methanol in rats leads to the impairment of
antioxidant mechanisms in the liver, erythrocytes, and blood serum. "

Alcohol. 1997 Sep-Oct; 14(5): 431-7.
Antioxidant status of liver, erythrocytes, and blood serum of rats in acute
methanol intoxication.
Skrzydlewska E, Farbiszewski R.
Department of Analytical Chemistry, Medical Academy, Poland.

SOD, CAT, GSH-Px, GSSG-R, ascorbic acid, alpha-tocopherol, nonprotein- and
protein-bound sulfhydryl compounds, and TBA-rs content
in the liver, erythrocytes, and blood serum of rats treated with methanol
after 6, 12, and 24 h and 2, 5, and 7 days were investigated.
Furthermore, hematological parameters of erythrocytes were analysed.
GSH-Px, GSSG-R, sulfhydryl compounds, and ascorbic acid in the liver,
erythrocytes, and in blood serum were significantly decreased.
In addition, Cu,Zn-SOD and tocopherol in erythrocytes were diminished,
whereas TBA-rs in the three biological materials was enhanced.
Simultaneously, erythrocytes amount, hemoglobin level, hematocrit, and MCV
were reduced.
These results indicate that methanol in rats leads to the impairment of
antioxidant mechanisms in the liver, erythrocytes, and blood serum. PMID:
9305457



Alcohol. 1997 May-Jun; 14(3): 295-9.
Influence of methanol and its metabolites on the activity of
alpha1-antitrypsin.
Skrzydlewska E, Mielczarska J.
Department of Instrumental Analysis, Medical Academy, Bialystok, Poland.

Among methanol and its metabolites, formaldehyde was found to have the
strongest inactivating effect on the activity of alpha1-antitrypsin
preparation and inhibitor existing in blood serum.
The influence of formaldehyde on the activity of serum alpha1-antitrypsin is
lower in comparison with purified inhibitor.
alpha1-Antitrypsin modified by formaldehyde inactivates the trypsin in its
action on the BAPA to a smaller degree than on the hemoglobin.
The effective formaldehyde concentration in the case of the BAPA is about 64
mM and in the case of the hemoglobin is about 256 mM.
The significant inhibitory effect of methanol on alpha1-antitrypsin appears
only at a high concentration of this compound.
Formate does not decrease alpha1-antitrypsin activity.
In people intoxicated with methanol, alpha1-antitrypsin activity decreases,
whereas the content of this inhibitor does not change. PMID: 9160807



Acta Biochim Pol. 1997; 44(1): 139-45.
Liver and serum antioxidant status after methanol intoxication in rats.
Skrzydlewska E, Farbiszewski R.
Department of Instrumental Analysis, Medical Academy, Bialystok, Poland.

Activities of superoxide dismutase (SOD), catalase, glutathione peroxidase
(GSH-Px) and glutathione reductase (GSSG-R) and concentration of ascorbate,
alpha-tocopherol, non-protein and protein-bound sulfhydryl compounds and
thiobarbituric acid-reactive substances (TBA-rs) were measured in liver and
serum of rats 6, 12 and 24 h and 2, 5 and 7 days after intoxication with
1.5 g or 3.0 g methanol/kg b.w.
Liver GSH-Px and GSSG-R activities and SH-groups and ascorbate content were
significantly diminished at 6 and 24 h,
while TBA-rs were increased.
Serum SOD, GSH-Px and GSSG-R activities and SH-groups concentration were
reduced, while TBA-rs were elevated.
The changes were more intensive after application of the higher dose of
methanol.
It is concluded that methanol impairs the liver and blood serum antioxidant
mechanisms in rats. PMID: 9241366




Vet Hum Toxicol. 1996 Dec; 38(6): 429-33.
Diminished antioxidant defense potential of liver, erythrocytes and serum
from rats with subacute methanol intoxication.
Skrzydlewska E, Farbiszewski R.
Department of Analytical Chemistry, Medical Academy, Bialystok, Poland.

The activities of superoxide dismutase (SOD), catalase glutathione
peroxidase (GSH-Px) and glutathione reductase (GSSG-R) and the concentration
of ascorbate, alpha-tocopherol, non-protein and protein-bound sulfhydryl
compounds, and thiobarbituric acid-reactive substances (TBA-rs)
in liver, erythrocytes and serum of rats dosed with 1.5 g methanol/kg bw
were measured after 6, 12 and 24 h and 2, 5 and 7 d.
Hematological erythrocyte parameters were also determined.
Liver GSH-Px and GSSG-R activities, SH-groups and ascorbate were
significantly diminished at 12 and 24 h, while TBA-rs increased.
Blood SOD, GSH-Px and GSSG-R activities and sulfhydryl-group concentrations
were reduced while TBA-rs were elevated.
Methanol given to rats impaired liver, erythrocyte and blood serum
antioxidant mechanisms. PMID: 8948074



Rocz Akad Med Bialymst. 1996; 41(2): 397-404.
Decreased antioxidant status and increased lipid peroxidation in rats after
methanol intoxication.
Skrzydlewska E.
Department of Instrumental Analysis, Medical Academy of Bialystok.

The liver is the main metabolic place where the methanol is oxidized to
formaldehyde and to formate.
The aim of this paper was to study the liver antioxidant system in acute
methanol intoxication, after 6, 12, 24 hours and 2, 5 and 7 days of alcohol
administration into rats.
In liver homogenates the superoxide dismutase, catalase, peroxidase and
reductase glutathione activity and content of malondialydehyde (MDA),
SH-compounds in protein and non-protein fraction and ascorbate were
estimated.
Activity of superoxide dismutase and catalase was significantly increased
after 6 hours following methanol ingestion and persisted up to 2-5 days of
intoxication.
It was accompanied by significant decreased of reductase and peroxidase
glutathione activities.
The protein and non-protein SH-groups were significantly decreased during 6
hours to 5 days following methanol ingestion.
The liver MDA content was considerably increased.
After 2 days since methanol intoxication the liver vitamin C content was
significantly decreased in comparison with the control group.
The obtain results demonstrated that during methanol induced liver injury
there are increase of lipid peroxidation and impairment of proantioxidant
equilibrium in favour to prooxidant. PMID: 9020552



Pol Tyg Lek. 1993 May 3-10; 48(18-19): 433-6.
[Metabolism and toxic effects of methanol]
[Article in Polish]
Skrzydlewska E.
Zakladu Analizy Instrumentalnej AM, Bialymstoku.
Publication Types: Review Review, Tutorial PMID: 8309827



Cell Mol Biol Lett. 2003; 8(2): 391-413.
DNA damage caused by lipid peroxidation products.
Luczaj W, Skrzydlewska E.
Department of Analytical Chemistry, Medical Academy of Bialystok,
Mickiewicza 2A, P.O. Box 14, 15-230 Bialystok 8, Poland.

Lipid peroxidation is a process involving the oxidation of polyunsaturated
fatty acids (PUFAs), which are basic components of biological membranes.
Reactive electrophilic compounds are formed during lipid peroxidation,
mainly alpha, beta-unsaturated aldehydes.
These compounds yield a number of adducts with DNA.
Among them, propeno and substituted propano adducts of deoxyguanosine with
malondialdehyde (MDA), acrolein, crotonaldehyde and etheno adducts,
resulting from the reactions of DNA bases with epoxy aldehydes, are a very
important group of adducts.
The epoxy aldehydes are more reactive towards DNA than the parent
unsaturated aldehydes.
The compounds resulting from lipid peroxidation mostly react with DNA
showing both genotoxic and mutagenic action;
among them, 4-hydroxynonenal is the most genotoxic, while MDA is the most
mutagenic.
DNA damage caused by the adducts of lipid peroxidation products
with DNA can be removed by the repairing action of glycosylases.
The formed adducts have been hitherto analyzed using the IPPA
(Imunopurification-(32)P-postlabelling assay) method and via gas
chromatography/electron capture negtive chemical ionization/mass
spectrometry (GC/EC NCI/MS).
A combination of liquid chromatography with electrospray tandem mass
spectrometry (LC/ES-MSMS) with labelled inner standard has mainly been used
in recent years. PMID: 12813574



Hepatogastroenterology. 2003 Jan-Feb; 50(49): 126-31.
Antioxidant potential in esophageal, stomach and colorectal cancers.
Skrzydlewska E, Kozuszko B, Sulkowska M, Bogdan Z, Kozlowski M,
Snarska J, Puchalski Z, Sulkowski S, Skrzydlewski Z.
skrzydle@...
Department of Analytical Chemistry Medical Academy, 15-230 Bialystok 8, P.O.
Box 14, Poland.

BACKGROUND/AIMS: The gastrointestinal tract is particularly susceptible to
reactive oxygen species attack which lead to carcinogenesis.
An important role in defense strategy against reactive oxygen species is
played by antioxidants.
The present study aims at examining antioxidant parameters and
malondialdehyde-- the product of lipid peroxidation as well as the marker of
cancer progression-- and cancer procoagulant in esophageal, gastric and
colorectal cancer patients.
METHODOLOGY: The activity of superoxide dismutase, catalase, glutathione
peroxidase and reductase and the level of glutathione, vitamin C,
malondialdehyde and cancer procoagulant were determined in tumors and normal
mucous from 18 patients with esophageal cancer, 18 patients with stomach
tumor and 62 patients with colorectal cancer.
RESULTS: In esophageal tumor the activity of all enzymes has been increased
compared with normal mucous.
Stomach tumor has been also characterized by an increase in antioxidant
enzymes activity except glutathione peroxidase and reductase whose
activities have been decreased.
However in colorectal tumor the activity of enzymes has been increased apart
from catalase.
In all cases the glutathione level has been increased while the vitamin C
content has been significantly decreased.
Tumor malondialdehyde level was significantly increased, too.
The level of cancer procoagulant also increased in cancer tissues as well as
in the serum.
CONCLUSIONS: Antioxidant potential in all cases of
gastrointestinal tract cancer has been unbalanced which has lead to increase
in reactive oxygen species action and enhancement of lipid peroxidation and
cancer procoagulant generation. PMID: 12630007



J Toxicol Environ Health A. 2004 Apr 9; 67(7): 595-606.
Green tea protection against age-dependent ethanol-induced oxidative stress.
Luczaj W, Waszkiewicz E, Skrzydlewska E, Roszkowska-Jakimiec W.
Department of Analytical Chemistry, Medical University of Bialystok,
Bialystok, Poland.

Ethanol intoxication leads to oxidative stress, which may be additionally
enhanced by aging.
The aim of this study was to investigate the influence of green tea as a
source of water-soluble antioxidants on the ability to prevent oxidative
stress in aged rats sub-chronically intoxicated with ethanol.
Two-, 12-, and 24-mo-old male Wistar rats were divided into 4 experimental
groups: (1) control, (2) green tea, (3) ethanol, and (4) ethanol and green
tea. Ethanol intoxication produced age-dependent decrease in the activity of
serum superoxide dismutase, glutathione peroxidase, and reductase and in
levels of glutathione (GSH), vitamins C, E, and A, and beta-carotene.
Changes in the serum antioxidative ability were accompanied by enhanced
oxidative modification of lipid (increase in lipid hydroperoxides,
malondiadehyde, and 4-hydroxynonenal levels) and protein (rise in carbonyl
group levels).
Green tea partially protected against changes in antioxidant enzymatic as
well as nonenzymatic parameters produced by ethanol and enhanced by aging.
Administration of green tea significantly protects cellular components such
as lipids and proteins against oxidative modification.
Results indicate that green tea effectively protects blood serum against
oxidative stress produced by ethanol as well as aging. PMID: 15129554



Postepy Hig Med Dosw (Online). 2004 Mar 30; 58: 194-201.
[Antioxidative abilities during aging] [Article in Polish]
Augustyniak A, Skrzydlewska E.
Zaklad Chemii Nieorganicznej i Analitycznej Akademii Medycznej w
Bialymstoku.

Biological aging is associated with increased cellular levels of reactive
oxygen species (ROS) as well as the formation and accumulation of oxidized
biomolecules.
During evolution, organisms developed a highly-efficient and adaptive
antioxidant defense system.
Antioxidants can generally be divided into two categories: enzymatic and
non-enzymatic. During the aging process the activity of antioxidant enzymes,
e.g. SOD, CAT, GSH-Px, and GSSG-R, depends on factors such as race, gender,
tissue and subcellular localization of enzymes.
The age-dependent decrease in antioxidant enzyme activity may be attributed
to oxidative modifications of enzymes.
During the aging process, ROS may also lead to the induction of some enzyme
activity which is explained as an adaptive phenomenon.
The decrease in GSH concentration with age can be explained by decreased GSH
synthesis and/or increased GSH consumption in the removal of peroxides and
xenobiotics.
In plasma albumin, ferritin, transferrin, and caeruloplasmin exert
protective action.
Plasma proteins can inhibit ROS generation and lipid peroxidation by
chelating free transition metals.
Plasma protein concentrations changes with age.
The major exogenous antioxidants, mostly derived from the diet, are vitamin
E, C, A, and beta-carotene.
During the aging process the level of vitamins may decrease or increase,
depending on such factors as diet, and diseases. PMID: 15077054



Folia Morphol (Warsz). 2004 Feb; 63(1): 123-6.
Green tea as an antioxidant which protects against alcohol induced injury in
rats -- a histopathological examination.
Baltaziak M, Skrzydlewska E, Sulik A, Famulski W, Koda M.
Department of General Pathology, Medical University, Bialystok, Poland.
drbal@...

Our study with animal models was designed to test the hypothesis that green
tea protects against chronic (over 4 weeks) alcohol induced liver injury in
rats.
The research was conducted on Wistar male rats divided into 4 research
groups:
I - received the Libera-De Carli control diet (L-DC),
II - received (L-DC) and green tea,
III - received (L-DC) and ethanol and
IV - received (L-DC), green tea and ethanol.
When comparing groups I and II we saw less intensive steatosis in group II
than in group I, which can suggest that green tea may affect the
accumulation of fat in the hepatocytes and protect them against steatosis
and disruption.
In III, the ethanol group, the steatosis of the liver increased considerably
and
the green tea which was given with ethanol in group IV did not halt this,
as in group IV we also observed intensive steatosis in the liver.
From this data we conclude that green tea has an important, although not
fully understood role in preventing liver injury. PMID: 15039917



Alcohol. 2004 Jan; 32(1): 25-32.
Green tea protects against ethanol-induced lipid peroxidation in rat organs.
Ostrowska J, Luczaj W, Kasacka I, Rozanski A, Skrzydlewska E.
Department of Analytical Chemistry, Medical Academy of Bialystok,
PO Box 14, 15-230 Bialystok, Poland.

Ethanol metabolism is accompanied by generation of free radicals, which
stimulates lipid peroxidation.
Natural antioxidants are particularly useful in such a situation.
The current study was designed to investigate the efficacy of green tea, as
a source of water-soluble antioxidants (catechins), on lipid peroxidation in
liver, brain, and blood induced by chronic (4 weeks) ethanol intoxication in
rats. Feeding of ethanol led to a significant increase in lipid
peroxidation, as
measured by increased concentrations of lipid hydroperoxides,
4-hydroxynonenal, and malondialdehyde.
Feeding of ethanol also changed the glutathione-dependent lipid
hydroperoxide decomposition system, resulting in a decrease in both reduced
glutathione concentration and activity of glutathione peroxidase.
Observed changes were statistically significant in all examined tissues.
Enhancement in lipid peroxidation was associated with disruption of
hepatocyte cell membranes, as observed through electron microscopic
evaluation.
Green tea protects phospholipids from enhanced peroxidation and prevents
changes in biochemical parameters and morphologic changes observed after
ethanol consumption.
These results support the suggestion that green tea protects membranes from
peroxidation of lipids associated with ethanol consumption. PMID: 15066700




Journal of Toxicology and Environmental Health Part A
Publisher: Taylor & Francis Health Sciences, part of the Taylor & Francis
Group Issue: Volume 67, Number 7 / April 9, 2003 Pages: 595 - 606

Green Tea Protection Against Age-Dependent Ethanol-Induced Oxidative Stress
Wojciech Luczaj A1, Ewa Waszkiewicz A1, Elzbieta Skrzydlewska A1,
Wiesawa Roszkowska-Jakimiec A2

A1 Department of Analytical Chemistry, Medical University of Bialystok,
Bialystok, Poland
A2 Department of Instrumental Analysis, Medical University of Bialystok,
Bialystok, Poland

Abstract:
Ethanol intoxication leads to oxidative stress, which may be additionally
enhanced by aging.
The aim of this study was to investigate the influence of green tea as a
source of water-soluble antioxidants on the ability to prevent oxidative
stress in aged rats sub-chronically intoxicated with ethanol.
Two-, 12-, and 24-mo-old male Wistar rats were divided into 4 experimental
groups: (1) control, (2) green tea, (3) ethanol, and (4) ethanol and green
tea. Ethanol intoxication produced age-dependent decrease in the activity of
serum superoxide dismutase, glutathione peroxidase, and reductase and in
levels of glutathione (GSH), vitamins C, E, and A, and g-carotene.
Changes in the serum antioxidative ability were accompanied by enhanced
oxidative modification of lipid (increase in lipid hydroperoxides,
malondiadehyde, and 4-hydroxynonenal levels) and protein (rise in carbonyl
group levels).
Green tea partially protected against changes in antioxidant enzymatic as
well as nonenzymatic parameters produced by ethanol and enhanced by aging.
Administration of green tea significantly protects cellular components such
as lipids and proteins against oxidative modification.
Results indicate that green tea effectively protects blood serum against
oxidative stress produced by ethanol as well as aging.




Addict Biol. 2002 Jul; 7(3): 307-14.
Green tea as a potent antioxidant in alcohol intoxication.
Skrzydlewska E, Ostrowska J, Stankiewicz A, Farbiszewski R.
Department of Analytical Chemistry, Medical Academy of Bialystok, Bialystok,
Poland. skrzydle@...

Ethanol oxidation to acetaldehyde and next to acetate is accompanied by free
radical generation.
Free radicals can affect cell integrity when antioxidant mechanisms are no
longer able to cope with the free radical generation observed in ethanol
intoxication.
Natural antioxidants are particularly useful in such a situation.
The present study was designed to investigate the efficacy of green tea as a
source of water-soluble antioxidants (catechins) on the liver and blood
serum antioxidative potential of rats chronically (28 days) intoxicated with
ethanol.
Alcohol caused a decrease in liver superoxide dismutase, glutathione
peroxidase and catalase activities and an increase in activity of
glutathione reductase.
Moreover, a decrease in the level of reduced glutathione, ascorbic acid,
vitamins A and E and beta-carotene were observed.
The activity of serum glutathione peroxidase decreased while glutathione
reductase activity increased.
The level of serum non-enzymatic antioxidants was also decreased in the
liver.
Alcohol administration caused an increase in the liver and serum lipid
peroxidation products, measured as thiobarbituric acid-reactive substances.
However, green tea prevents the changes observed after ethanol intoxication.
Green tea also protects membrane phospholipids from enhanced peroxidation.
These results indicate a beneficial effect of green tea in alcohol
intoxication. PMID: 12126490



Phytomedicine. 2002 Apr; 9(3): 232-8.
Protective effect of green tea against lipid peroxidation in the rat liver,
blood serum and the brain.
Skrzydlewska E, Ostrowska J, Farbiszewski R, Michalak K.
Department of Analytical Chemistry, Medical Academy of Bialystok, Poland.
skrzydle@... michalak@...

This paper reports data on the effect of green tea on the lipid peroxidation
products formation and parameters of antioxidative system of the liver,
blood serum and central nervous tissue of healthy young rats drinking green
tea for five weeks.
The rats were permitted free access to solubilized extract of green tea.
Bioactive ingredients of green tea extract caused in the liver an increase
in the activity of glutathione peroxidase and glutathione reductase and in
the content of reduced glutathione as well as marked decrease in lipid
hydroperoxides (LOOH), 4-hydroksynonenal (4-HNE) and malondialdehyde (MDA).
The concentration of vitamin A increased by about 40%.
Minor changes in the measured parameters were observed in the blood serum.
GSH content increased slightly, whereas the index of the total antioxidant
status increased significantly.
In contrast, the lipid peroxidation products, particularly MDA was
significantly diminished.
In the central nervous tissue the activity of superoxide dismutase and
glutathione peroxidase decreased while the
activity of glutathione reductase and catalase increased after drinking
green tea.
Moreover the level of LOOH, 4-HNE and MDA significantly decreased.
The use of green tea extract appeared to be beneficial to rats in reducing
lipid peroxidation products.
These results support and substantiate traditional consumption of green tea
as protection against lipid peroxidation in the liver, blood serum, and
central nervous tissue. PMID: 12046864



Rocz Akad Med Bialymst. 2001; 46: 240-50.
The influence of green tea on the activity of proteases and their inhibitors
in plasma of rats after ethanol treatment.
Skrzydlewska E, Roszkowska A, Makiela M, Skrzydlewski Z.
Department of Analytical Chemistry, Medical Academy of Bialystok, Bialystok,
Poland.

Ethanol oxidation in the liver is accompanied by formation of acetaldehyde
and free radicals.
These compounds can react with biologically active proteins, including
proteolytic enzymes and their inhibitors.
The aim of this paper was to determine the influence of green tea on the
activity of cathepsin G and elastase and their inhibitors such as
alpha-1-antitrypsin and alpha-2-macroglobulin, total antioxidant status and
lipid peroxidation in plasma of young rats chronically intoxication with
ethanol.
The activity of cathepsin G and elastase was increased, while the activity
of their inhibitors was reduced after ethanol treatment.
At the same time, the total antioxidant status was significantly decreased
while lipid peroxidation measured as malondialdehyde and 4-hydroxynonenal
was significantly increased.
Giving green tea to rats did not change the proteases and their inhibitors
activity, but significantly increased total antioxidant status and decreased
lipid peroxidation.
Drinking green tea with ethanol partially prevents the changes observed
after ethanol intoxication. PMID: 11780568



J Toxicol Environ Health A. 2001 Oct 12; 64(3): 213-22.
Antioxidant status and lipid peroxidation in colorectal cancer.
Skrzydlewska E, Stankiewicz A, Sulkowska M, Sulkowski S, Kasacka I.
Department of Analytical Chemistry, Medical Academy of Bialystok, Poland.
skrzydle@...

Colon carcinogenesis is a multistep process where oxygen radicals were found
to enhance carcinogenesis at all stages: initiation, promotion, and
progression.
Since insufficient capacity of protective antioxidant system can result in
cancer, the aim of this study was to examine the activity of antioxidant
enzymes (superoxide dismutase, catalase, glutathione peroxidase, and
glutathione reductase) and the levels of reduced glutathione, vitamin C, and
vitamin E.
The lipid peroxidation products were also determined by measuring
malondialdehyde and 4-hydroxynonenal levels in colorectal cancer tissue
collected from 55 patients.
In these cases the activity of superoxide dismutase, glutathione peroxidase,
and glutathione reductase was significantly increased
while the activity of catalase was significantly decreased in cancer tissue.
However, the level of nonenzymatic antioxidant parameters (glutathione,
vitamin C, and vitamin E) was significantly decreased in cancer tissue.
Further lipid peroxidation was enhanced during cancer development,
manifested by a significant increase in malondialdehyde and 4-hydroxynonenal
levels.
The obtained results indicate significant changes in antioxidant capacity of
colorectal cancer tissues, which lead to enhanced action of oxygen radicals,
resulting in lipid peroxidation. PMID: 11594700



Postepy Hig Med Dosw. 2001; 55(6): 871-89.
[Melatonin as an antioxidant] [Article in Polish]
Skrzydlewska E.
Zaklad Chemii Nieorganicznej i Analitycznej Akademii Medycznej w
Bialymstoku.

This review describes the structure and properties of melatonin.
The interaction of melatonin with reactive oxygen species and its protective
action in relation to DNA, lipids and proteins are presented.
The effect of melatonin on antioxidant and prooxidant enzymes is discussed,
too. Publication Types: Review Review, Tutorial PMID: 11875783



Rocz Akad Med Bialymst. 2001; 46: 133-44.
Ethanol and N-acetylcysteine influence on the development of liver changes
in experimental methanol intoxication.
Kasacka I, Skrzydlewska E.
Departments of Histology & Embriology, Medical Academy of Bialystok,
Bialystok, Poland.

The evaluation of ethanol and N-Acetylcysteine (NAC) influence on
histopathological changes in rat liver intoxicated with 3 g of methanol/kg
b.w. was conducted, based on morphological examinations in light and
electron microscope.
The rats received intragastrically 3.0 g of methanol/kg b.w. as a 50%
solution, 10% ethanol for 24 hours before methanol and next 48 hours after
methanol ingestion and NAC (150 mg/kg b.w.) after 15 min. methanol
administrated.
The results indicate that methanol intoxication causes pronounced
morphological changes in the examined organ.
Ethanol administered to methanol-intoxicated rats caused intensification of
certain parameters of hepatocytes morphological damage.
A simultaneous administration of methanol and NAC resulted in a lower degree
of parenchymal damage. PMID: 11780556




Psychiatr Pol. 1992 Sep-Oct; 26(5): 421-9.
[The influence of disulfiram on the metabolism of ethanol] [Article in
Polish]
Skrzydlewska E.
Zakladu Analizy Instrumentalnej AM, Bialymstoku.

The author discusses the metabolism of disulfiram and the enzymes which
metabolize ethanol.
The restraining of the activity of ALDH in the liver by disulfiram causes an
accumulation of acetaldehyde which in their turn cause a series of
psychophysical symptoms which are unpleasant and in some instances dangerous
for the patient.
Thus, it is important to monitor changes in the activity of ALDH after
administration of disulfiram.
Publication Types: Review Review, Tutorial PMID: 1302340



Postepy Hig Med Dosw. 1992; 46(1): 117-30.
[Metabolism of liver proteins in ethanol poisoning] [Article in Polish]
Skrzydlewska E, Worowski K, Roszkowska-Jakimiec W.
Zaklad Analizy Instrumentalnej AM, Bialymstoku.

The present paper reviews the literature on influence of ethanol and
acetaldehyde on synthesis, export and degradation of liver proteins.
Direction and intensification changes caused by ethanol and acetaldehyde
depend on concentration, time of activity and the way of administration of
these compounds, and the way of feeding.
Publication Types: Review Review Literature PMID: 1641374



Postepy Hig Med Dosw. 1992; 46(2): 159-72.
[Proteolytic enzymes of the digestive tract in ethanol poisoning] [Article
in Polish]
Skrzydlewska E, Worowski K, Roszkowska-Jakimiec W.
Zaklad Analizy Instrumentalnej Akademii Medycznej, Bialymstoku.

The present paper reviews the literature on changes of proteolytic enzymes
activity, disorders of protein digestion and absorption of protein
degradation products from digestive tract in ethanol intoxication.
Magnitude of the change depends on concentration, dose and time of ethanol
consumption.
Acute ethanol intoxication causes increase in gastric and pancreatic
proteolytic enzymes secretion and reduces amino acids and peptides
absorption.
Chronic ethanol consumption results in reduced synthesis and secretion of
gastric and pancreatic proteinases.
Publication Types: Review Review Literature PMID: 1470579



Postepy Hig Med Dosw. 1992; 46(4): 417-30.
[Interaction of acetaldehyde and proteins] [Article in Polish]
Skrzydlewska E, Roszkowska-Jakimiec W.
Zaklad Analizy Instrumentalnej Akademii Medycznej, Bialymstoku.

A review of literature dealing with acetaldehyde-proteins reactions in vitro
and in vivo was done.
The changes in proteins structure and functions resulting from acetaldehyde
binding were discussed.
Publication Types: Review Review, Academic PMID: 1293589


Rocz Akad Med Bialymst. 1990-91; 35-36: 119-27.
[Histopathological evaluation of protective effect of L-cysteine in
ethanol-induced liver damage in rats] [Article in Polish]
Worowski K, Chyczewski L, Skrzydlewska E.
Zakladu Analizy Instrumentalnej, Akademii Medycznej Bialymstoku.

Rats fed standard diet were intoxicated during 4 weeks with ethanol at the
dose of 0.6 g/100 g of the body weight.
This poisoning causes vacuolar degeneration, disappearance of glycogen
granules, steatosis of hepatocytes and focal necrosis changes in the liver.
The intake of food with cysteine at the dose of 0.012 and 0.024 g/100 g/24
hrs markedly prevents histopathological changes in the liver of rats
intoxicated with ethanol.
Larger amounts of cysteine (0.044 g/100 g/24 hrs) intensify
histopathological changes caused by ethanol in the liver of rats. PMID:
2136542



Rocz Akad Med Bialymst. 1990-91; 35-36: 129-41.
[Effect of cysteine on protein metabolism in the liver of rats with
ethanol-induced liver damage] [Article in Polish]
Skrzydlewska E, Worowski K, Chyczewski L.
Zakladu Analizy Instrumentalnej, Akademii Medycznej, Bialymstoku.

Rats intoxicated with ethanol at the dose of 0.6 g/100 g of the body weight
during 4 weeks were fed on standard diet and the one containing 0.125, 0.25
and 0.5% L-cysteine.
Intoxication of rats fed standard food causes an increase in the activity
of cathepsin D and gamma-glutamyl-transpeptidase in the liver and an
increase in the activity of alanine aminotransferase and
gamma-glutamyl-transpeptidase in the blood serum.
Consuming by rats food containing small and medium quantity of cysteine
causes normalization of the activity of all enzymes,
whereas consuming food containing large quantity of cysteine does not give
such effect. PMID: 1983788



Rocz Akad Med Bialymst. 1990-91; 35-36: 163-75.
[Effect of immunomodulating drugs on the release and activities of lysosomal
proteinases of the liver of rats with ethanol poisoning] [Article in
Polish]
Skrzydlewska E, Chyczewski L, Worowski K.
Zakladu Analizy Instrumentalnej, Akademii Medycznej, Bialymstoku.

Increased activity of cathepsin A and D in the cytosol fraction and
homogenate of the liver of rats intoxicated for 4 weeks with ethanol (0.6
g/100 g of the body weight) was found.
The cytosol cathepsin A and D activities were unaffected under the influence
of Levamisole and isoprinosine++.
Encorton reduced the activity of both cathepsins in the cytosol fraction
while it did not diminish their activities in the liver homogenates.
Encorton, and to a markedly lesser degree, Levamisole and isoprinosine++
caused a regression of vacuolar degeneration and of necrotic lesions and
an increase in the number of glycogen granules in the livers of
ethanol-intoxicated rats. PMID: 1726679



Mater Med Pol. 1989 Jul-Sep; 21(3): 225-7.
Inhibitory effect of ethanol and acetaldehyde on the amidolytic activity of
trypsin and chymotrypsin.
Skrzydlewska E, Worowski K, Zakrzewska I, Prokopowicz J, Puchalski Z,
Piotrowski Z.
Department of Instrumental Analysis, Bialystok, Poland.

Ethanol and in higher degree acetaldehyde displayed inhibitory effect
directed against amidolytic activity of trypsin and chymotrypsin.
The decrease of the activity of both enzymes is related to the concentration
of these compounds.
The rate of inhibition of amidolytic activity of chymotrypsin with both
reagents is more evident in comparison to trypsin. PMID: 2491274



Acta Med Pol. 1988; 29(1-2): 41-5.
Effect of ethanol and acetaldehyde on the enzymatic activity of human
pancreatic juice in vitro. I. Inhibition of alpha-amylase and lipase
activity. Zakrzewska I, Worowski K, Skrzydlewska E, Prokopowicz J, Puchalski
Z,
Piotrowski Z. PMID: 3267162



Acta Med Pol. 1988; 29(1-2): 47-52.
Effect of ethanol and acetaldehyde on the enzymatic activity of human
pancreatic juice in vitro. II. Inhibition of the activity of proteolytic
enzymes. Skrzydlewska E, Worowski K, Zakrzewska I, Prokopowicz J, Puchalski
Z,
Piotrowski Z. PMID: 3076740



Folia Med Cracov. 1987; 28(1-2): 205-14.
[Effect of ethanol and acetaldehyde on the release and activity of proteases
and protein degradation in the rat liver (in vitro and in vivo studies)]
[Article in Polish] Skrzydlewska E. PMID: 3623321



Rocz Akad Med Im Juliana Marchlewskiego Bialymst. 1986-87; 31-32: 3-18.
[Effects of ethanol and acetaldehyde on proteolytic enzyme activity of the
stomach] [Article in Polish] Skrzydlewska E. PMID: 3152147



Rocz Akad Med Im Juliana Marchlewskiego Bialymst. 1986-87; 31-32: 19-28.
[Effects of ethanol and acetaldehyde on proteolytic enzyme activity of the
small intestine and pancreas] [Article in Polish]
Skrzydlewska E. PMID: 3152145



Acta Biochim Pol. 1985; 32(3): 271-7.
Release of acid proteolytic activity from lysosomes and degradation of
protein in organs of rats intoxicated with ethanol and acetaldehyde.
Skrzydlewski Z, Worowski K, Skrzydlewska E.

Intoxication with ethanol and acetaldehyde resulted in a marked increase of
the acid proteolytic activity in the post-lysosomal supernatant of rat
kidney, lung, and liver, while the content of protein and acid-soluble
tyrosine remained practically unchanged.
Proteins of the post-lysosomal supernatant were degraded in vitro by the
endogenous proteinase(s) of lysosomal origin at pH 3.5 and 5.5 but not at pH
7.0. PMID: 3911694



Rocz Akad Med Im Juliana Marchlewskiego Bialymst. 1984-85; 29-30: 59-76.
[Effect of ethanol and acetaldehyde on the activity and release of
cathepsins from lysosomes of the canine liver (studies in vitro)]
[Article in Polish] Skrzydlewska E, Worowski K. PMID: 6443826



Rocz Akad Med Im Juliana Marchlewskiego Bialymst. 1984-85; 29-30: 153-62.
Influence of ethanol and acetaldehyde on blood coagulation (examinations in
vitro). Skrzydlewska E, Worowski K. PMID: 6443814



Rocz Akad Med Im Juliana Marchlewskiego Bialymst. 1984-85; 29-30: 163-73.
Influence of ethanol and acetaldehyde on fibrinolytic system (examinations
in vitro). Skrzydlewska E, Worowski K. PMID: 6242841
********************************************************************

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Postepy Higieny I Medycyny Doswiadczalnej http://www.phmd.pl

Postepy Hig Med Dosw (Online). 2004 Mar 30; 58: 194-201.
[Antioxidative abilities during aging]
[Article in Polish] [ Abstract and 80 References, in English, are given
here. ]
Augustyniak A, Skrzydlewska E.
Zaklad Chemii Nieorganicznej i Analitycznej Akademii Medycznej w
Bialymstoku.

Biological aging is associated with increased cellular levels of reactive
oxygen species (ROS) as well as the formation and accumulation of oxidized
biomolecules.
During evolution, organisms developed a highly-efficient and adaptive
antioxidant defense system.
Antioxidants can generally be divided into two categories: enzymatic and
non-enzymatic. During the aging process the activity of antioxidant enzymes,
e.g. SOD, CAT, GSH-Px, and GSSG-R, depends on factors such as race, gender,
tissue and subcellular localization of enzymes.
The age-dependent decrease in antioxidant enzyme activity may be attributed
to oxidative modifications of enzymes.
During the aging process, ROS may also lead to the induction of some enzyme
activity which is explained as an adaptive phenomenon.
The decrease in GSH concentration with age can be explained by decreased GSH
synthesis and/or increased GSH consumption in the removal of peroxides and
xenobiotics.
In plasma albumin, ferritin, transferrin, and caeruloplasmin exert
protective action. Plasma proteins can inhibit ROS generation and
lipid peroxidation by chelating free transition metals.
Plasma protein concentrations changes with age.
The major exogenous antioxidants, mostly derived from the diet, are vitamin
E, C, A, and beta-carotene.
During the aging process the level of vitamins may decrease or increase,
depending on such factors as diet, and diseases. PMID: 15077054

www.phmd.pl
Review
Postepy Hig Med Dosw (online), 2004; 58: 194-201

page 194

Zdolnosci antyoksydacyjne w starzejacym
sie organizmie
Antioxidative abilities during aging
Agnieszka Augustyniak, Elzbieta Skrzydlewska
Zaklad Chemii Nieorganicznej i Analitycznej Akademii Medycznej w Bialymstoku
Streszczenie
Received: 2003.04.22
Accepted: 2003.08.08
Published: 2004.03.30
Key words: aging . antioxidant enzymes . non-enzymatic antioxidants
Full-text PDF: http://www.phmd.pl/pub/phmd/vol_58/5343.pdf
Word count: 3953
Tables: -
Figures: 1
References: 80
Source of support: Praca finansowana z grantu KBN 6PO5F01720.
Adres autorki: dr hab. Elzbieta Skrzydlewsdka, Zaklad Chemii Nieorganicznej
i Analitycznej AM, ul. Mickiewicza 2a,
15-230 Bialystok, e-mail: skrzydle@...

page 196

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page 200

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page 201

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********************************************************************

"CONCLUSION
There is much strong evidence that the background of etheno-DNA and
propano-DNA detected in tissues from unexposed humans and rodents arises
from endogenous lipid peroxidation products, such as MDA.

With the advent of ultrasensitive, specific detection methods for this kind
of DNA damage in human tissues and cells, new insights can be gained into
the mechanisms involved in human cancers.

Moreover, etheno-DNA adducts can now be used as biomarkers to investigate
the potential role of lipid peroxidation in human cancers."

Cell Mol Biol Lett. 2003; 8(2): 391-413.
DNA damage caused by lipid peroxidation products.
Luczaj W, Skrzydlewska E.
Department of Analytical Chemistry, Medical Academy of Bialystok,
Mickiewicza 2A, P.O. Box 14, 15-230 Bialystok 8, Poland.

Lipid peroxidation is a process involving the oxidation of polyunsaturated
fatty acids (PUFAs), which are basic components of biological membranes.
Reactive electrophilic compounds are formed during lipid peroxidation,
mainly alpha, beta-unsaturated aldehydes.
These compounds yield a number of adducts with DNA.
Among them, propeno and substituted propano adducts of deoxyguanosine with
malondialdehyde (MDA), acrolein, crotonaldehyde and etheno adducts,
resulting from the reactions of DNA bases with epoxy aldehydes,
are a very important group of adducts.
The epoxy aldehydes are more reactive towards DNA than the parent
unsaturated aldehydes.
The compounds resulting from lipid peroxidation mostly react with DNA
showing both genotoxic and mutagenic action;
among them, 4-hydroxynonenal is the most genotoxic,
while MDA is the most mutagenic.
DNA damage caused by the adducts of lipid peroxidation products
with DNA can be removed by the repairing action of glycosylases.
The formed adducts have been hitherto analyzed using the IPPA
(Imunopurification-(32)P-postlabelling assay) method and via gas
chromatography/electron capture negtive chemical ionization/mass
spectrometry (GC/EC NCI/MS).
A combination of liquid chromatography with electrospray tandem mass
spectrometry (LC/ES-MSMS) with labelled inner standard has mainly been used
in recent years. PMID: 12813574

http://www.cmbl.org.pl/vol8/V8Page391.pdf

CELLULAR & MOLECULAR BIOLOGY LETTERS
Volume 8, (2003) pp 391 - 413
http://www.cmbl.org.pl
Received 6 January 2003
Accepted 16 April 2003

Abbreviations used: eA - 1,N6-ethenoadenine;
eC - 3,N4-ethenocytosine; edA - 1,N6-
ethenodeoxyadenosine; edC - 3,N4-ethenodeoxycytidine;
AdG - acrolein-derived 1,N2-propanodeoxyguanosine;
CdG - crotonaldehyde-derived 1,N2-propanodeoxyguanosine;
EH - 2,3-epoxy-4-hydroxynonanal;
GC/EC NCI/MS - gas chromatography with mass spectrometry with electron
capture negative chemical ionization detection;
GSH - gluthatione; HNE - trans-4-hydroxy-2-nonenal;
IPPA - Imunopurification-32Ppostlabelling assay;
LC/ES-MSMS - liquid chromatography with electrospray tandem mass
spectrometry;
M1 C - N4-(3-oxo-propenyl)deoxycytidine;
M1 G - pirymido[1,2a]purin-10(3H)-one); M1A -
N6-(3-oxo-propenyl)deoxyadenosine; MDA - malondialdehyde; PUFAs -
polyunsaturated fatty acids.

DNA DAMAGE CAUSED BY LIPID PEROXIDATION PRODUCTS

WOJCIECH LUCZAJ and ELZIBIETA SKRZYDLEWSKA
Department of Analytical Chemistry, Medical Academy of Biaystok,
Mickiewicza 2A, P.O. Box 14, 15-230 Biaystok 8, Poland

Abstract: Lipid peroxidation is a process involving the oxidation of
polyunsaturated fatty acids (PUFAs), which are basic components of
biological
membranes. Reactive electrophilic compounds are formed during lipid
peroxidation, mainly a,b-unsaturated aldehydes. These compounds yield a
number of adducts with DNA. Among them, propeno and substituted propano
adducts of deoxyguanosine with malondialdehyde (MDA), acrolein,
crotonaldehyde and etheno adducts, resulting from the reactions of DNA bases
with epoxy aldehydes, are a very important group of adducts. The epoxy
aldehydes are more reactive towards DNA than the parent unsaturated
aldehydes. The compounds resulting from lipid peroxidation mostly react with
DNA showing both genotoxic and mutagenic action; among them, 4-
hydroxynonenal is the most genotoxic, while MDA is the most mutagenic. DNA
damage caused by the adducts of lipid peroxidation products with DNA can be
removed by the repairing action of glycosylases. The formed adducts have
been hitherto analyzed using the IPPA (Imunopurification-32P-postlabelling
assay) method and
via gas chromatography/electron capture negtive chemical
ionization/mass spectrometry (GC/EC NCI/MS).
A combination of liquid
chromatography with electrospray tandem mass spectrometry (LC/ES-MSMS)
with labelled inner standard has mainly been used in recent years.

Key Words: Lipid Peroxidation, Etheno Adducts, Propano Adducts, á,â-
Unsaturated Aldehydes

CELL. MOL. BIOL. LETT. Vol. 8. No. 2. 2003

page 392

INTRODUCTION
Carcinogenesis was induced by chronic infections caused by viruses, bacteria
or parasites and the inflammation accompanying them. A hypothesis explaining
carcinogenesis mechanisms suggested that endogenic compounds causing DNA
damage are formed by the organism in the inflammation state [1]. The main
species/compounds involved in them are oxygen and nitrogen (species). The
production of these species markedly increases in such conditions, and they
can induce enhanced lipid peroxidation in direct reactions and/or cause DNA
damage [1].

Various reactive electrophilic compounds can be formed by lipid
peroxidation and some of them, showing mutagenic and genotoxic properties,
reaily react with proteins and DNA. This is particularly true for trans-4-
hydroxynonenal, malondialdehyde, and crotonaldehyde, which form adducts
with DNA [7]. The formation of these compounds was described as early as in
the eighties, but their identification and quantitative determination in
animal organisms under physiological conditions has only been possible in
recent
studies, in which ultrasensitive analytical methods such as IPPA
(Imunopurification-32P-postlabelling assay) and gas chromatography/electron
capture negtive chemical ionization/mass spectrometry (GC/EC NCI/MS) were
used [86-89]. This also created chances to obtain evidence of the toxic and
genetic consequences of such DNA damage [1].

LIPID PEROXIDATION
Lipid peroxidation occurs in physiological conditions. It involves the
oxidation of polyunsaturated fatty acids (PUFAs), which are basic biological
membrane components. Many unsaturated components, mainly aldehydes, are then
formed [2]. Fundamental reactions occurring during peroxidation are showing
in Fig. 1 [2].

Polyunsaturated fatty acids contain active methylene groups situated between
cis double bonds. Such groups readily react with oxidizing agents and their
hydrogen atoms are removed to form carbon-centered radicals (compound 1,
Fig. 1) [2]. These radicals react with molecular oxygen-yielding peroxyl
radicals, which are the initial products of polyunsaturated fatty acid
oxidation [3].

Further transformations of peroxyl radicals depend on their position in
the carbon chain of the fatty acids [3]. If the peroxyl radical exists at
the
end of a double bond system (compound 2, Fig. 1) then it can be reduced to a
hydroperoxide. Conjugated diene hydroperoxides formed in this way (compound
4, Fig. 1) are relatively stable lipid peroxidation products in the absence
of transition metal ions [2]. Peroxyl radicals can be also reduced to
hydroperoxides by other fatty acid molecules or by vitamin E [4].

The reduction of a peroxyl radical by another fatty acid molecule results in
the formation of a new carbon centered radical which propagates the fatty
acid oxidation. In this way, an oxidized molecule can induce the oxidation
of many other fatty acid molecules. Approximately 60 linoleic acid molecules
and 200 molecules of arachidonic acid

CELLULAR & MOLECULAR BIOLOGY LETTERS 393

are oxidized as the result of transformations initiated by one free-radical
reaction [4]. The length of the free-radical reaction chain depends on many
factors.

The main factor determining the free-radical reaction chain length in vivo
is
vitamin E concentration in the lipid bilayer [5]. This vitamin reduces
peroxyl
radicals to hydroperoxides, thereby breaking the reaction chain and slowing
the rate of lipid peroxidation [6].
However, vitamin E can initiate other free-radical chain reactions if
present in very small concentrations [6].

Fig. 1. The pathways of lipid peroxidation [3].

If the peroxyl radical is located within a fatty acid chain
(compound 3, Figure 1) then it can undergo cyclization owing to a
neighbouring double bond [7],
yielding a cyclic peroxide located in vicinity of a carbon-centered radical
(compound 5, Fig. 1). This radical can undergo further transformations. It
can bind an oxygen molecule, yielding a peroxyl radical which can be reduced
to
hydroperoxide (compound 6, Fig. 1), as described earlier, or it can undergo
a new cyclization, yielding a bicyclic peroxide. This can bind another
oxygen
molecule and be reduced to a compound (compound 7, Fig. 1) structurally
analogous to prostaglandin endoperoxide but lacking a stereochemical control
[7]. The chemical conversion of the bicyclic peroxide group of this compound
gives malondialdehyde (MDA) and isoprostanes [8] and 17-carbon fatty acids
are simultaneously formed as side products (compound 8, Fig. 1) [9].

There is a growing belief that the most valuable biomarkers of lipid
peroxidation in the human body are the isoprostanes [10-14]. Highly
sensitive and accurate
spectrophotometric methods have hitherto been used to determine isoprostane

CELL. MOL. BIOL. LETT. Vol. 8. No. 2. 2003 394

concentration in blood serum [13, 15, 16]. Isoprostates undergo relatively
rapid metabolic transformations. Therefore, lipid peroxydation in various
organs
can be readily monitored by determining the level of their decomposition
products in urine [13, 17].

It is known that transition metal ions initiate free-radical reactions
including lipid peroxidation because they participate in generating reactive
oxygen species
(O2-*, *OH). At the same time, they contribute to the propagation of the
process by reducing lipid hydroperoxides [18, 19]. Compounds of such ions as
Cd(II), Co(II), Cu(II), Hg(II), Ni(II), Pb(II), Sn(II), V(V), Fe(II), and
Fe(III)
provoke enhanced lipid peroxidation under both in vitro and in vivo
conditions
[20-25].

All the hydroperoxides presented in Fig. 1, as well as their regio- and
stereoisomers, can be reduced to their akoxyl radicals by transition metal
cations and then they can undergo b-cleavage yielding many products [26,
27].

This results in a number of epoxy compounds (e.g. 2,3-epoxybutanal or
2,3-epoxy-4-hydroxynonanal), hydroperoxides (e.g. compounds 4, 6, 7, Fig. 1)
and saturated and á,â-unsaturated aldehydes (e.g. acrolein, crotonaldehyde,
4-hydroxynonenal, 2,4-nonadienal, 2,4-decadienal) [28]. These compounds are
formed from fatty acids in various amounts depending on their stucture and
oxidation conditions.

Recently, a new product of lipid peroxidation, 4-oxo-2-nonenal, was reported
to be generated by the decomposition of linoleic acid hydroperoxide [13(S)-
hydroperoxy-(Z,E)-9, 11-octadecadienoic acid] [29].

All lipid peroxidation pathways leading to product formation are not known
in sufficient detail. Nevertheless, it has been found that crotonaldehyde is
mainly generated from a-linoliec acid and linoleate.

Crotonaldehyde is also formed
in small amounts by the peroxidation of arachidonate and such acids as cis-
5,8,11,14,17-eicosapentaenoic and cis-4,7,10,13,16,19-docosahexaenoic [30].
It is interesting that arachidonate and w-3-polyunsaturated fatty acids were
earlier stated to be primary compounds of acrolein generation [31].

It was also found that hexanal and 4-hydroxynonenal (HNE) were formed from
lipids containing
w-6-polyunsaturated fatty acids (18:2, 20:4), whereas 4-hydroxyh

(Message over 64k, truncated.)