Coca-Cola carcinogenicity in rats, Ramazzini Foundation, F Belpoggi, M
Soffritti, Annals NY Academy Sciences 2006 Sept, parts of 17 pages:
Murray 2006.12.02
http://groups.yahoo.com/group/aspartameNM/message/1385

[ For related information and links:

Fiorella Belpoggi & Morando Soffritti of Ramazzini Foundation prove
lifetime carcinogenicity of Coca-Cola, aspartame, and arsenic, Annals
of the NY Academy of Sciences: Murray 2006.11.28
http://groups.yahoo.com/group/aspartameNM/message/1382

http://groups.yahoo.com/group/aspartameNM/message/1383 aspartame

http://groups.yahoo.com/group/aspartameNM/message/1384 arsenic ]


http://www.ramazzini.it/fondazione/eng/eventidettagli.asp?id=33

NEWS AND EVENTS 01 November 2006
Results of 3 long term ERF carcinogenesis bioassays published in the
Annals of the New York Academy of Sciences

Aspartame, Sodium Arsenite, Coca-Cola

The proceedings of the Collegium Ramazzini's 2005 conference "Framing
the Future in Light of the Past: Living in a Chemical World" have been
published in the Annals of the New York Academy of Sciences.
The volume includes the results of three long term carcinogenesis
bioassays conducted by the European Ramazzini Foundation:


http://www.ramazzini.it/fondazione/docs/NYAS_Aspartame_Ramazzini.pdf
Ann. N.Y. Acad. Sci. 1076: 559-577 (2006)
Results of Long-Term Carcinogenicity Bioassay on Sprague-Dawley Rats
Exposed to Aspartame Administered in Feed
FIORELLA BELPOGGI, MORANDO SOFFRITTI, MICHELA PADOVANI, DAVIDE DEGLI
ESPOSTI, MICHELINA LAURIOLA, AND FRANCO MINARDI.

http://www.ramazzini.it/fondazione/docs/NYAS_Sodium_Arsenite_Ramazzini.pdf
Ann. N.Y. Acad. Sci. 1076: 578-591 (2006)
Results of a Long-Term Carcinogenicity Bioassay on Sprague-Dawley Rats
Exposed to Sodium Arsenite Administered in Drinking Water
MORANDO SOFFRITTI, FIORELLA BELPOGGI, DAVIDE DEGLI ESPOSTI, AND LUCA
LAMBERTINI.

http://www.ramazzini.it/fondazione/docs/NYAS_Coca-Cola_Ramazzini.pdf
Ann. N.Y. Acad. Sci. 1076: 736-752 (2006)
Results of Long-Term Carcinogenicity Bioassays on Coca-Cola
Administered to Sprague-Dawley Rats
Fiorella Belpoggi,
Morando Soffritti,
Eva Tibaldi,
Laura Falcioni,
Luciano Bua,
Francesca Trabucco.
Cesare Maltoni Cancer Research Center, European Foundation for Oncology
and Environmental Sciences "B. Ramazzini"�, European Ramazzini
Foundation, Castello di Bentivoglio, 40010 Bentivoglio, Bologna, Italy
crcfr@...


ABSTRACT:

Coca-Cola was invented in May 1886 in Atlanta, Georgia by a pharmacist
who, by accident or design, mixed carbonated water with the syrup of
sugar, phosphoric acid, caffeine, and other natural flavors to create
what is known as "the world's favorite soft drink."

Coca-Cola is currently sold in more than 200 countries
and in early 2000, the company sold its 10 billionth unit case of
Coca-Cola branded products.

Given the worldwide consumption of Coca-Cola,
a project of experimental bioassays to study its long-term effects when
administered as substitute for drinking water on male and female
Sprague-Dawley rats was planned and executed.

The objective of the project was to study whether and how long-term
consumption of Coca-Cola affects the basic tumorigram of test animals.

The bioassays were performed on rats beginning at different ages,
namely:
(a) on males and females exposed since embryonic life or from 7 weeks
of age; and
(b) on males and females exposed from 30, 39, or 55 weeks of age.

Overall, the project included 1999 rats.

During the biophase, data were collected on fluid and feed consumption,
body weight, and survival.

Animals were kept under observation until spontaneous death and
underwent complete necropsy.

The results indicate:

(a) an increase in body weight in all treated animals;

(b) a statistically significant increase of the incidence in females,
both breeders and offspring, bearing malignant mammary tumors;

(c) a statistically significant increase in the incidence of exocrine
ademonas of the pancreas in both male and female breeders and
offspring; and

(d) an increased incidence, albeit not statistically
significant, of pancreatic islet cell carcinomas in females,
a malignant tumor which occurs very rarely in our historical controls.

On the basis of the results of this study, excessive consumption of
regular soft-drinks should be generally discouraged, in particular for
children and adolescents.
PMID: 17119251

KEYWORDS: Coca-Cola; carcinogenicity; long-term bioassay; rat

Address for correspondence: Morando Soffritti, M.D., Cesare Maltoni
Cancer Research Center, European Ramazzini Foundation, Castello di
Bentivoglio, 40010 Bentivoglio (BO), Italy.
Voice: +39-051-6640460; fax: +39-051-6640223.
e-mail: crcfr@... www.ramazzini.it

Funding for this research was provided entirely by the European
Foundation on Oncology and Environmental Sciences "B. Ramazzini".

c 2006 New York Academy of Sciences.
doi: 10.1196/annals.1371.078

INTRODUCTION

Cancer is one the principal public health problems faced by
industrialized countries, both because of its epidemiological dimension
and because of the growing environmental influences behind its increase
to today's epidemic proportions.

Diet is often cited as an important factor in the occurrence of cancer;
however, influence of diet on health cannot be easily isolated or
quantified given the varied composition of the average diet over a
lifetime and the complex mixture of substances present in food.

While the overall diet is difficult to analyze, specific nutrients and
food components have been implicated both in causing cancer and as
protective factors in the carcinogenic process.

There are several sources of food components believed to induce cancer.

One is the presence of naturally occurring substances, such as
aflatoxin found in ground nuts and corn.

Another potential source of risk are substances which are added
directly to food in order to improve its quality, taste, or stability.

It has long been suggested that total calorie intake may have an
important effect on the incidence of cancer.

Data by Tannenbaum in the 1940s and 1950s demonstrated that underfed
mice developed fewer spontaneous mammary tumors, primary lung adenomas,
and chemically induced tumors.1,2

In addition, human obesity has been identified as a risk factor for
some types of cancer.3,4

Long-term carcinogenicity bioassays on rodents, which reproduce
exposure situations experienced by humans as much as possible, is in
our opinion, a good approach to study the influence of dietary products
on the incidence of spontaneous tumors in a controlled environment.5,6

In an attempt to evaluate the interaction between one such product and
tumor incidence in rodents, an experimental carcinogenicity bioassay
was performed at the Cesare Maltoni Cancer Research Center (CMCRC) of
the European Ramazzini Foundation (ERF).

The product tested is a beverage mixture which is both caloric and
widely consumed in most of the world -- Coca-Cola.

Coca-Cola was invented in Atlanta, Georgia, on May 8, 1886 by
pharmacist John Pemberton.

The regular cola beverage contains water, sugar (about 10% by weight),
carbon dioxide, caramel coloring, acidulates, flavoring substances, and
caffeine, all of which are approved for use around the world.

Today Coca-Cola, recognized as the world's number one soft drink brand,
is currently sold in more than 200 countries.

In 2001, the company sold its 10 billionth unit case of Coca-Cola
branded products.7

The experimental project described in this article was designed to
evaluate the long-term effects of Coca-Cola on the spontaneous
development of tumors when administered as a substitute for drinking
water to Sprague-Dawley rats for the life span.

Since it is known that the age of the animals at the start of the
experiment may affect the possible modulating effects of the test
compounds, the project includes experiments performed on groups of male
and female breeder rats (30, 39, and 55 weeks old at the start of the
experiment), all the offspring of each litter descending from the
aforementioned breeders (exposed since intrauterine life), and one
group exposed beginning at 7 weeks of age.

MATERIALS AND METHODS

The Coca-Cola used was supplied by an Italian reseller, in 1-L glass
bottles delivered every 2-3 weeks.
During the experiments, the bottles were stored at a room temperature
of 22 +- 3 deg C.
Before administering the Coca-Cola to the rodents, the CO2 was first
eliminated by mechanical shaking for 60 minutes.

The male (M) and female (F) Sprague-Dawley rats used in this experiment
were bred from the colony used at the CMCRC/ERF laboratories for nearly
30 years.

Extensive historical data on more than 15,000 rodents are available on
the tumor incidence among untreated rats.

At 4 weeks of age, the breeders were identified by ear punch, separated
by sex, and assigned to experimental groups so as to have no more than
one male and one female from each litter in the same group.
Rats were housed 5 per cage until the time of mating, at which time
parents were placed in breeding cages (one male with one female).
After 7 days, males were re-housed 5 per cage and females were housed
individually.
After 8 weeks, the females were also re-housed 5 per cage.
All available pups of all litters from treated and untreated dams were
included in the various groups of offspring.
After weaning (at 4-5 weeks), offspring were identified by ear punch,
weighed, separated by sex, and assigned sequentially, litter by litter,
to the exposed and control group, respectively.
The two groups of 7-week-old rats were randomized in order to have no
more than one male and one female from each litter in the same group.
Animals were housed in makrolon cages (41 cm X 25 cm X 15 cm) with
stainless-steel wire tops and a shallow layer of white wood shavings as
bedding.
Cages were kept in rooms used exclusively for this experiment at a
temperature of 21 +- 2 deg C and relative humidity of 50-60%.
A light-dark cycle of 12 hours was maintained using both natural and
artificial light sources.
Animals were given the standard Corticella pellet diet (Corticella
S.p.A., Bologna, Italy), analyzed for nutritional components and
possible contaminants.
Every 24 hours, Coca-Cola or drinking water (control) were disposed of
and the bottles were cleaned and refilled.
The animals were checked three times a day from Monday to Saturday, and
twice on Sundays and holidays for clinical symptoms and behavior.
Coca-Cola was administered ad libitum instead of drinking water to
groups of breeders (55-110 per sex per group) 30, 39, or 55 weeks old
at the start of the experiment, to their offspring exposed since
intrauterine life (24-110 per sex per group), and to a group of 80
males and 80 females, 7 weeks old at the start of the experiment.
The administration of Coca-Cola lasted until spontaneous death.
Equivalent groups of breeders and offspring administered drinking water
served as controls.
The entire project involved 1999 rodents.
Mean daily drinking water and feed consumption were measured on the day
prior to the start of the breeders' treatment, after 7 days of
treatment before mating and then, post-weaning, once weekly for the
first 13 weeks from the start of the experiment, then every 2 weeks
until 104 weeks.
Individual animal weight of breeders and of offspring after weaning was
measured once weekly for the first 13 weeks, every 2 weeks until 104
weeks, and then every 8 weeks until the end of the experiments.
In order to detect and register all gross lesions, the animals were
examined every week for the first 13 weeks, and then every 2 weeks
until the end of the experiment.

Upon death, all animals underwent complete necropsy.
Histopathology was routinely performedon all macroscopically observed
pathological lesions (with a margin of surrounding normal tissue) and
on skin and subcutaneous tissue, the brain, pituitary gland, Zymbal
glands, salivary glands, Harderian glands, cranium (with oral and nasal
cavities and external and internal ear ducts, 5 levels), tongue,
thyroid and parathyroid, pharynx, larynx, thymus and mediastinal lymph
nodes, trachea, lung and mainstem bronchi, heart, diaphragm, liver,
spleen, pancreas, kidneys and adrenal glands, esophagus, stomach (fore
and glandular), intestine (4 levels), bladder, prostate, uterus,
gonads, interscapular fat pad, subcutaneous and mesenteric lymph nodes,
and any other organ or tissue with pathological lesions.

All organs and tissues were preserved in 70% ethyl alcohol, except for
the bones which were fixed in 10% formalin and then decalcified with
10% formaldehyde and 20% formic acid in water solution.
The normal specimens were trimmed, following Standard Operating
Procedures (SOP) at the CMCRC/ERF laboratories.
Trimmed specimens were processed as paraffin blocks, and 3-5 micrometer
sections of every specimen were obtained.
Sections were routinely stained with hematoxylin-eosin.
Specific stainings were performed when needed.
All slides were examined microscopically by the same group of
pathologists;
a senior pathologist reviewed all tumors and any other lesion of
oncological interest.
Statistical analysis was performed using the X2 test in order to
evaluate the significance in tumor incidence differences between
treated and control groups.

RESULTS

In this article, we present the results of data aggregated from all
breeders and all offspring.

Offspring include both rodents whose treatment began during embryonic
life and those rodents whose treatment began at 7 weeks of age.

A treatment-related difference in fluid consumption was observed, in
both sexes of breeders and offspring, with treated animals consuming
more than two times as much fluid compared to controls.

An opposite treatment-related difference in food consumption was
observed in both sexes of breeders and offspring during the experiment,
with treated animals consuming about 40% less feed compared to
controls.

An increase in body weight was observed, in both breeders and
offspring, among both sexes in treated groups compared to the controls.

No differences in survival were observed among treated males and
females breeders compared to the controls.

A slight decrease in survival was observed in female offspring compared
to the controls.

The occurrence of benign and malignant tumors among male and female
rats is shown in TABLE 1.

The differences observed among treated and control animals were as
follows:

1. a slight increase in the incidence of malignant tumor-bearing
animals was observed in female breeders compared to controls (TABLE 2);

2. a statistically significant increase in females bearing malignant
mammary tumors and total malignant mammary tumors, both among breeders
(P < 0.01) and offspring (P < 0.01), compared to the controls (TABLE
3);

3. a statistically significant increased incidence in animals bearing
adenomas of the exocrine component of the pancreas was observed among
male (P < 0.01) and female (P < 0.05) breeders and male (P < 0.01) and
female (P < 0.01) offspring compared to the controls.

No exocrine carcinomas were observed (TABLE 4).

Concerning the incidence of islet cell adenomas, no differences were
observed in treated breeders or offspring of either sex compared to the
controls.

An increased incidence of islet cell carcinomas was observed in treated
female breeders and treated female offspring compared to the controls.

Only one case of carcinoma was observed in treated male offspring.

It must be noted that in our historical controls, we observed only 1
(0.04%) islet cell carcinoma out of 2274 untreated females and 5 (0.2%)
islet cell carcinomas out of 2265 males.

CONCLUSIONS

Although Coca-Cola is one of the most consumed soft-drinks worldwide,
to our knowledge this study represents the first time that a long-term
carcinogenesis bioassay has been conducted to evaluate the beverage
mixture for its potential carcinogenic effects.

The results of the study conducted at the CMCRC/ERF have shown the
following:

1. an increase of fluid consumption in all animals, both breeders and
offspring, administered Coca-Cola as a substitute for drinking water
and a general decrease in food consumption;

2. an increase in body weight in all treated animals.

When compared to controls, animals treated with Coca-Cola also
demonstrated the following oncological effects:

1. a statistically significant increase of the incidence in females,
both breeders and offspring, bearing malignant mammary tumors.

In our opinion, this observation confirms the correlation between the
increase in body weight and an increased risk of mammary cancer;

2. a statistically significant increase in the incidence of exocrine
adenomas of the pancreas in both male and female breeders and
offspring;

3. an increased incidence, albeit not statistically significant, of
pancreatic islet cell carcinomas in females.

Because of the rarity of pancreatic islet cell carcinomas in our
historical controls, the biological significance of the increased
incidence of this malignant tumor cannot be underestimated.

Although humans do not consume this beverage under the same conditions
designed in our experiment, the results nevertheless confirm that an
exaggerated ingestion of high caloric beverages, such as regular
soft-drinks, can lead to a marked increase in body weight which in turn
presents an increased risk for developing cancer.

ACKNOWLEDGMENTS

We thank the research staff of the CMCRC/ERF and Kathryn Knowles for
her support in the preparation of the manuscripts.

REFERENCES

1. TANNENBAUM, A. 1940.
The initiation and growth of tumors. I: effects of underfeeding.
Am. J. Cancer 38: 335-350.

2. TANNENBAUM, A. & H. SILVERSTONE. 1953.
Nutrition in relation to cancer.
Adv. Cancer Res. 1: 451-501.

3. CALLE, E. et al. 2003.
Overweight, obesity, and mortality from cancer in a prospectively
studied cohort of U.S. Adults.
N. Engl. J. Med 348: 1625-1638.

4. RAPP, K. et al. 2005.
Obesity and incidence of cancer: a large cohort study of over
145,000 adults in Austria.
Br. J. Cancer 93: 1062-1106.

5. SOFFRITTI, M. et al. 1999.
Mega-experiments to identify and assess diffuse carcinogenic risks.
Ann. N. Y. Acad. Sci. 895: 43-55.

6. SOFFRITTI,M. et al. 1996.
Results of experimental bioassays on the chemopreventive
effects of vitamin A and N-(4-hydroxyphenyl)-retinamide (HPR) on
mammary
cancer.
In The Scientific Bases of Cancer Chemoprevention.
C. Maltoni, M. Soffritti & W. Davis, Eds.:
International Congress Series 1120 : 241-248. Elsevier.
Amsterdam, The Netherlands.

7. THE COCA-COLA COMPANY ANNUAL REPORT. 2005.
http://www.cocacola.com [accessed 16 May 2006]
*******************************************************


short aspartame (methanol, formaldehyde) toxicity research summary:
Murray 2006.12.02
http://groups.yahoo.com/group/aspartameNM/message/1379

"Of course, everyone chooses, as a natural priority,
to actively find, quickly share, and positively act upon the facts
about healthy and safe food, drink, and environment."

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

http://groups.yahoo.com/group/aspartameNM/messages
group with 79 members, 1,385 posts in a public, searchable archive
http://RMForAll.blogspot.com

http://groups.yahoo.com/group/aspartameNM/message/1340
aspartame groups and books: updated research review of 2004.07.16:
Murray 2006.05.11

http://groups.yahoo.com/group/aspartameNM/message/1378
11 members of New Mexico legislature sign letter to ban aspartame as a
source of toxic methanol and formaldehyde, Stephen Fox, NM Senator
Gerald Ortiz y Pino: Murray 2006.10.22

http://groups.yahoo.com/group/aspartameNM/message/1374
47 UK Members of Parliament now support aspartame ban initiative of
Roger Williams, MP: Murray 2006.10.16

http://groups.yahoo.com/group/aspartameNM/message/1271
combining aspartame and quinoline yellow, or MSG and brilliant blue,
harms nerve cells, eminent C. Vyvyan Howard et al, 2005
education.guardian.co.uk, Felicity Lawrence: Murray 2005.12.21

http://groups.yahoo.com/group/aspartameNM/message/1277
50% UK baby food is now organic -- aspartame or MSG
with food dyes harm nerve cells, CV Howard 3 year study
funded by Lizzy Vann, CEO, Organix Brands,
Children's Food Advisory Service: Murray 2006.01.13

http://groups.yahoo.com/group/aspartameNM/message/1279
all three aspartame metabolites harm human erythrocyte [red blood cell]
membrane enzyme activity, KH Schulpis et al, two studies in 2005,
Athens, Greece, 2005.12.14: 2004 research review, RL Blaylock:
Murray 2006.01.14

http://groups.yahoo.com/group/aspartameNM/message/1366
toxicity in rat brains from aspartame, Vences-Mejia A, Espinosa-Aguirre
JJ et al 2006 Aug: Murray 2006.09.06

http://groups.yahoo.com/group/aspartameNM/message/1373
aspartame rat brain toxicity re cytochrome P450 enzymes, expecially
CYP2E1, Vences-Mejia A, Espinosa-Aguirre JJ et al, 2006 Aug,
Hum Exp Toxicol: relevant abstracts re formaldehyde from methanol
in alcohol drinks: Murray 2006.09.29

http://groups.yahoo.com/group/aspartameNM/message/1369
Bristol, Connecticut, schools join state program to limit artificial
sweeteners, sugar, fats for 8800 students, Johnny J Burnham, The
Bristol Press: Murray 2006.09.22

http://groups.yahoo.com/group/aspartameNM/message/1341
Connecticut bans artificial sweeteners in schools, Nancy Barnes,
New Milford Times: Murray 2006.05.25

http://groups.yahoo.com/group/aspartameNM/message/1353
carcinogenic effect of inhaled formaldehyde, Federal Institute of Risk
Assessment, Germany -- same safe level as for Canada:
Murray 2006.06.02

http://groups.yahoo.com/group/aspartameNM/message/1352
Home sickness -- indoor air often worse, as our homes seal in
pollutants
[one is formaldehyde, also from the 11% methanol part of aspartame],
Megan Gillis, WinnipegSun.com: Murray 2006.06.01


http://groups.yahoo.com/group/aspartameNM/message/1143
methanol (formaldehyde, formic acid) disposition: Bouchard M
et al, full plain text, 2001: substantial sources are
degradation of fruit pectins, liquors, aspartame, smoke:
Murray 2005.04.02

http://groups.yahoo.com/group/aspartameNM/message/1349
NIH NLM ToxNet HSDB Hazardous Substances Data Bank
inadequate re aspartame (methanol, formaldehyde, formic acid):
Murray 2006.08.19

http://toxnet.nlm.nih.gov/cgi-bin/sis/search/f?./temp/~HwoSfJ:1
HSDB Hazardous Substances Data Bank: Aspartame

ASPARTAME CASRN: 22839-47-0
METHANOL CASRN: 67-56-1
FORMALDEHYDE CASRN: 50-00-0
FORMIC ACID CASRN: 64-18-6

http://groups.yahoo.com/group/aspartameNM/message/1307
formaldehyde from 11% methanol part of aspartame or from red wine
causes same toxicity (hangover) harm: Murray 2006.05.24

Dark wines and liquors, as well as aspartame, provide
similar levels of methanol, above 120 mg daily, for
long-term heavy users, 2 L daily, about 6 cans.

Within hours, methanol is inevitably largely turned into formaldehyde,
and thence largely into formic acid -- the major causes of the dreaded
symptoms of "next morning" hangover.

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). If 30% of the methanol is turned
into formaldehyde, the amount of formaldehyde, 37 mg,
is 18.5 times the USA EPA limit for daily formaldehyde in
drinking water, 2.0 mg in 2 L average daily drinking water.

Any unsuspected source of methanol, which the body always quickly
and largely turns into formaldehyde and then formic acid, must be
monitored, especially for high responsibility occupations, often with
night shifts, such as pilots and nuclear reactor operators.

http://groups.yahoo.com/group/aspartameNM/message/1052
DMDC: Dimethyl dicarbonate 200mg/L in drinks adds methanol 98 mg/L
( becomes formaldehyde in body ): EU Scientific Committee on Foods
2001.07.12: Murray 2004.01.22

http://www.HolisticMed.com/aspartame mgold@...
Aspartame Toxicity Information Center Mark D. Gold
12 East Side Drive #2-18 Concord, NH 03301 603-225-2100

http://www.holisticmed.com/aspartame/abuse/methanol.html
"Scientific Abuse in Aspartame Research"

http://groups.yahoo.com/group/aspartameNM/message/1371
Russell L. Blaylock, MD discusses MSG, aspartame, excitotoxins
with Mike Adams: Murray 2006.09.27

http://groups.yahoo.com/group/aspartameNM/message/1372
Mike Adams interviews Randall Fitzgerald on "The Hundred Year Lie:
How Food and Medicine are Destroying Your Health" 2006.06.21:
Murray 2006.09.28
*******************************************************