Toxic L-tryptophan: Shedding Light on a Mysterious Epidemic -Was Genetic Enginee

[Guest guest]

There are 8 chapters altogether, plus a section on Acknowledgments and

another on Comments by Scientists and Others. On the left hand side of the

webpages are links to all the chapters/sections. There are aslo several links

in

each chapter - I have not put in the urls so please go to the websites.

I have wondered for years what happened to L-tryptophan. It seems it would

be very helpful to many of those with Fibromyalgia, sleep difficulties, pain,

and many other conditions. Well, I guess now I know.........except why is it

still not available? or was this just an excuse for the FDA and Health Canada

as they collaborate with Codex.......is this what the future holds for

Canada and the USA? best wishes, Shan

Toxic L-tryptophan: Shedding Light on a Mysterious Epidemic

_http://www.seedsofdeception.com/Public/L-tryptophan/index.cfm_

(http://www.seedsofdeception.com/Public/L-tryptophan/index.cfm)

by William E.Crist

Preface by Jeffrey Smith

In chapter 4 of my book Seeds of Deception, I present the story of a deadly

epidemic in 1989 that was linked to an L-tryptophan food supplement produced

by genetically engineered bacterium. Much of the material was based on the

research of William Crist, whose work is reproduced here.

I want to acknowledge William for his years of work, which has uncovered

important information about this tragic event.

The material is presented in sequence.

_http://www.seedsofdeception.com/Public/L-tryptophan/1Introduction/index.cfm_

(http://www.seedsofdeception.com/Public/L-tryptophan/1Introduction/index.cfm)

 

Introduction:

Was Genetic Engineering the Cause of

Contaminated L-tryptophan and the EMS Epidemic?

Knowing my long-standing interest in food purity issues, a friend in

California suggested in 1996 that I look into the cause of the tragic

eosinophilia-myalgia syndrome (EMS) epidemic that had struck seven years

earlier. The

outbreak was traced to consumption of L-tryptophan food supplements produced by

a

Japanese company using genetically engineered (GE) bacteria. Exactly what

role biotechnology played in the contamination of the supplement was hotly

debated. After thousands of hours of self-sponsored research, I believe I can

shed

some light on that issue.

I began by interviewing representatives of all the interested parties,

including many EMS patients, scientists, attorneys involved in the litigations,

the Food and Drug Administration (FDA) and the Centers for Disease Control

(CDC). I was generally met with courtesy and cooperation. I discovered sincere

differences of opinion among scientists about the possible causes of the

contamination that led to the EMS epidemic, and some interesting facts about

biotechnology. My follow-up questions — particularly to government agencies

about

its own findings and about the " problem with the purification process " —

generated obvious discomfort. Suddenly, agency personnel to whom I had spoken

were

" out of the country, " would not reply to my emails and/or letters, and in

one instance I was told that a CDC spokesperson refused to talk to me and had

" referred the matter to their attorney. "

Similarly, my several Freedom of Information Act requests were essentially

denied, sending me irrelevant information or saying the information requested

was proprietary, i.e., trade secret of the manufacturer. Later, I learned

from an attorney that the proprietary period had expired, so again I made an

FOIA request for specific agency reports. Again I came to a stonewall. An FOIA

spokesperson told me that the authors of the reports were no longer with the

agency. I listed at least four scientists who were still there, but the FOIA

representative simply repeated they were " no longer there " — as if unable to

deviate from following a script. This left me with the distinct feeling that

FOIA requests are a waste of time on closely guarded issues unless you are a

major media journalist or otherwise have the clout to back them up. No matter,

in this case, that scores had died from EMS and thousands had been left in

various degrees of debility.

I quickly realized that the government agencies were caught between two

conflicting mandates: both regulating and promoting biotechnology — and that

the

latter seemed to have the higher priority.

This review covers both the results of my research and the process of

getting to those results. The FDA's and CDC's sudden stonewalling made my work

more

difficult than it needed to be — and made it clear that they preferred to

leave all this " a mystery " rather than to investigate seriously all of the

factors that led to the epidemic. I invite a careful reading of the facts that

I

was able to extract from the parts of the L-tryptophan mess that they weren't

able to completely sweep under the carpet.

Background Information:

The EMS Epidemic, Initial Research Studies and News of Biotech Link

_http://www.seedsofdeception.com/Public/L-tryptophan/2BackgroundInformation/in

dex.cfm_ (http://www

..seedsofdeception.com/Public/L-tryptophan/2BackgroundInformation/index.cfm)

During the summer and autumn of 1989, an outbreak of a tragic and mysterious

disease swept across the U.S. First scores, then hundreds of people fell

seriously ill with a rare blood and muscle disorder. Doctors and hospital

staffs

were baffled by the unusual cluster of symptoms and were largely ineffective

in treating the victims.

The disease was characterized by an overproduction of white blood cells

called eosinophils (1000 or more cells per mm_3_

(http://www.seedsofdeception.com/Public/L-tryptophan/2BackgroundInformation/inde\

x.cfm#ref) ) and severe and

often debilitating myalgia (muscle pain). Hence, it was called

eosinophilia-myalgia syndrome or EMS._1_

(http://www.seedsofdeception.com/Public/L-tryptophan/2BackgroundInformation/inde\

x.cfm#ref) In addition to elevated eosinophils and

severe muscle pain, EMS patients exhibited a variety of other symptoms

including physical weakness, leg and arm swelling (edema), fever, breathing

difficulties, skin rashes, arthralgia, and pneumonia. In some instances,

patients

displayed signs of congestive heart failure and complete paralysis. An initial

report showed that as many as half of EMS patients were hospitalized._1-3_

(http://www.seedsofdeception.com/Public/L-tryptophan/2BackgroundInformation/inde\

x

..cfm#ref) (Refer also to _comments by patients and physicians_

(http://www.seedsofdeception.com/utility/showArticle?objectID=271) .)

By late October of 1989, three physicians in New Mexico discovered a

connection between the disorder and patients who had been taking food

supplements of

L-tryptophan._1_

(http://www.seedsofdeception.com/Public/L-tryptophan/2BackgroundInformation/inde\

x.cfm#ref)

On November 11, the U.S. Food and Drug Administration (FDA) issued a

nationwide warning that advised consumers to discontinue use of L-tryptophan

after

30 potential cases of EMS had been identified in New Mexico. Within a few days

of this first publicity alert, the Center for Disease Control (CDC) in

Atlanta received reports of 154 potential cases of a similar illness from

public

health agencies, physicians and the general public in 17 states and the

District of Columbia._1_

(http://www.seedsofdeception.com/Public/L-tryptophan/2BackgroundInformation/inde\

x.cfm#ref) The FDA quickly issued a recall of all

dietary supplements of L-tryptophan sold in doses of 100 mg. or more.

By early December of that same year, reports of EMS cases had soared to 707

in 48 states. One death had been reported and several others were under

investigation. Expanding its constraints, FDA imposed an automatic detention at

U.S. ports on all L-tryptophan products coming into the country._2_

(http://www.seedsofdeception.com/Public/L-tryptophan/2BackgroundInformation/inde\

x.cfm#ref)

 

For the next few months researchers and epidemiologists traced the multiple

retail brands of L-tryptophan involved with the disease back through hundreds

of wholesalers, distributors, tablet makers, encapsulators and importers.

On March 22, 1990, the FDA expanded its recall of L-tryptophan to include

any dosages of the dietary supplement after it was discovered that one person

with EMS had taken less than 100 milligrams a day. Meanwhile, the number of

EMS cases linked to use of L-tryptophan had risen to 1,411, including 19

deaths._3_

(http://www.seedsofdeception.com/Public/L-tryptophan/2BackgroundInformation/inde\

x.cfm#ref)

In late April, the first preliminary scientific data on the outbreak was

reported in meetings at the CDC that linked victims of EMS to L-tryptophan

produced by a single Japanese company, Showa Denko K.K. In two separate studies

virtually all EMS-case patients whose L-tryptophan consumption could be

reliably traced had taken product that came wholly or in part from this one

manufacturer._4_

(http://www.seedsofdeception.com/Public/L-tryptophan/2BackgroundInformation/inde\

x.cfm#ref)

L-Tryptophan: A Key Amino Acid

L-tryptophan is known to play a key role in the functioning of the human

body as a constituent of protein. It is important for the production of

serotonin, a neurotransmitter in the brain that helps regulate sleep, mood, and

perception of pain._5_

(http://www.seedsofdeception.com/Public/L-tryptophan/2BackgroundInformation/inde\

x.cfm#ref) For years, it was used by many Americans to

treat insomnia, depression, premenstrual syndrome and other disorders. The FDA

classified it as a food nutrient rather than a drug. Most people obtain

sufficient amounts of natural tryptophan in dietary sources contained in high

protein foods such as mother's milk, cow's milk, cheese, soybeans, fish, poultry

and meat._1,2,6,7_

(http://www.seedsofdeception.com/Public/L-tryptophan/2BackgroundInformation/inde\

x.cfm#ref)

All L-tryptophan marketed in the U.S. was manufactured by six companies in

Japan. It was available by mail order and through a wide variety of retail

outlets including pharmacies, grocery stores and health food stores. It was

sold

in tablet, capsule, powder and liquid form as a dietary supplement._8-10_

(http://www.seedsofdeception.com/Public/L-tryptophan/2BackgroundInformation/inde\

x

..cfm#ref)

Showa Denko Altered Manufacturing Process

On July 11, 1990, the Journal of the American Medical Association (JAMA)_7_

(http://www.seedsofdeception.com/Public/L-tryptophan/2BackgroundInformation/ind

ex.cfm#ref) published a study showing that 98 percent, and possibly 100

percent, of the EMS cases in Oregon had taken L-tryptophan product made by one

manufacturer, Showa Denko, and that there was a significant correlation

between these case patients and product manufactured by the company between

January

and June 1989. The JAMA study also noted that Showa Denko produced

L-tryptophan by bacterial fermentation using a genetically engineered Bacillus

species

that had been introduced in its manufacturing process in December 1988.

The following month, a study published in The New England Journal of

Medicine (NEJM),_10_

(http://www.seedsofdeception.com/Public/L-tryptophan/2BackgroundInformation/inde\

x.cfm#ref) reported that shortly after Showa Denko

introduced the newest strain of Bacillus (Strain V) in its manufacturing

process, it

reduced the amount of carbon powder used in the filtration of L-tryptophan

from 20kg. to 10kg. in most batches.

The final product still exceeded the standards specified by the United

States Pharmacopoeia, of 98.5 percent purity. The NEJM study's coauthors noted,

" Although the powdered carbon may have contributed to the removal of the

etiologic agent, it does not explain how the agent was introduced into the

product. " They said that the newly introduced Strain V bacterium " may have

produced

larger quantities of the etiologic agent than earlier strains. "

High performance liquid chromatography (HPLC) analysis of Strain V samples

revealed a number of peak trace contaminants, but only one, Peak E, was

initially found to be significantly associated with the EMS epidemic. The

researchers observed that because the change in carbon used during manufacturing

happened at about the same time as the introduction of the new bacterial strain,

it was difficult to assess the contribution of the bacterial strain to the

onset of EMS.

Hence, the authors concluded, " The outbreak of eosinophilia-myalgia syndrome

in 1989 resulted from the ingestion of a chemical constituent that was

associated with specific tryptophan-manufacturing conditions at one company.

The

chemical constituent represented by Peak E may contribute to the pathogenesis

of eosinophilia-myalgia syndrome or it may be a surrogate to another chemical

that induces the syndrome. "

News of Biotech Link to Epidemic

A few days later, on August 14, 1990, Newsday_11_

(http://www.seedsofdeception.com/Public/L-tryptophan/2BackgroundInformation/inde\

x.cfm#ref) ran a story

that linked genetic engineering to the EMS epidemic. In the article, Dr.

Michael Osterholm, coauthor of the NEJM study on EMS and epidemiologist at the

Minnesota Health Department, said, " Strain V was cranked up to make more

L-tryptophan and something went wrong. This obviously leads to that whole

debate

about genetic engineering. "

A flurry of newspaper headlines ensued on genetic engineering gone awry.

In late August, Science magazine published an article_12_

(http://www.seedsofdeception.com/Public/L-tryptophan/2BackgroundInformation/inde\

x.cfm#ref) in

which Sam Page, chief of natural products and instrumentation branch at FDA,

" blasted Osterholm for 'propagating hysteria.' The whole question: Is there

any relation to genetic engineering? is premature-especially given the impact

on the industry. "

According to the article, Osterholm seemed somewhat bewildered by Page's

response. " Anyone who looks at the data comes to the same conclusion, " he said,

namely, that genetic engineering could be involved. " I think FDA doesn't want

it to be so because of the implications for the agency. "

The article said that the FDA knew for months that the batches of

L-tryptophan implicated in EMS were produced by a genetically engineered

organism, but

government officials had withheld the information from the public " apparently

hoping to keep the recombinant link quiet until they could determine whether

it in fact did play a role in the outbreak. "

The article also reported that researchers had identified the chemical

structure of the contaminant Peak E as a " dimer " — essentially, two

tryptophan

molecules linked together.

FDA Knew of the Danger

The FDA knew that genetic engineering could potentially promote toxicity in

a plant or organism. According to its 1992 Statement of Policy: Foods

Derived From New Plant Varieties:

Plants are known to produce naturally a number of toxicants and

anti-nutritional factors… which often serve the plant as natural defense

compounds

against pests and pathogens… Many of these toxicants are present in today's

foods

at levels that do not cause acute toxicity… Plants, like other organisms,

have

metabolic pathways that no longer function due to mutations that occurred

during evolution. Products or intermediates of some such pathways may include

toxicants. In rare cases, such silent pathways may be activated by mutations,

chromosomal rearrangements, or new regulatory regions introduced during

breeding, and toxicants hitherto not associated with a plant species may

thereby

be produced. Similarly, toxicants ordinarily produced at low levels in a plant

may be produced at high levels in a new variety as a result of such

occurrences [e.g., using recombinant DNA techniques, also known as genetic

engineering]._13_

(http://www.seedsofdeception.com/Public/L-tryptophan/2BackgroundInformation/inde\

x.cfm#ref)

Four years later in a telephone interview, James Maryanski, FDA biotech

policy coordinator, had a different view. I asked him whether the genetically

engineered bacteria used by Showa Denko during fermentation could have created

toxic impurities in their L-tryptophan product.

“We have no evidence of that, none whatsoever that the technique causes

those kind of effects, differently from other methods of modifying

organisms,†he

said._14_

(http://www.seedsofdeception.com/Public/L-tryptophan/2BackgroundInformation/inde\

x.cfm#ref)

Apparently Maryanski was unaware that a year earlier, in 1995, a study by

scientists at Japan’s Research Institute for Food Science, reported that

genetically engineered yeast produced an accumulation of a highly toxic

compound in

yeast cells, compared to non-transformed control cells. The authors

concluded, “These results illustrate that careful thought should be given to

the

potential metabolic products and their safety when a genetically engineered

yeast

is applied to food-related fermentation processes._15_

(http://www.seedsofdeception.com/Public/L-tryptophan/2BackgroundInformation/inde\

x.cfm#ref)

Michael Antoniou, PhD., Reader in Molecular Genetics in the U.K., commented,

“[This example] illustrates that a product derived from a GE organism can be

devoid of genetic material, but can still unexpectedly contain potentially

harmful alterations to a GE product, a novel toxin or elevated levels of a

known hazardous substance._16_

(http://www.seedsofdeception.com/Public/L-tryptophan/2BackgroundInformation/inde\

x.cfm#ref)

Later in my interview with Maryanski, he said that while theoretically

genetic engineering could have played a role in causing EMS, it was not likely

the

cause:

We can not rule it (genetic engineering) out. However, close to two dozen

cases of L-tryptophan linked EMS (cases) occurred before Showa Denko began

using their engineered strain. So, there would have to be a cause other than

just

the mere engineering of the strains. I can't say that definitively because

we don't have a lot of information on these earlier cases.

We can not say definitely that the engineering strains were not a

contributing factor. However, we do have enough information that suggests that

there

are probably other factors involved, that either L-tryptophan itself, or

L-tryptophan in combination with something that was the result of the

purification

process, was the more likely cause. Until the science advances far enough, we

really can't say._14,17_

(http://www.seedsofdeception.com/Public/L-tryptophan/2BackgroundInformation/inde\

x.cfm#ref)

That was before new information surfaced regarding _the pre-epidemic cases

(next section).>>_

(http://www.seedsofdeception.com/utility/showPage/index.cfm?objectID=public,4974\

)

See also:

* _The Severity of the Disease—Comments by Victims and Physicians_

(http://www.seedsofdeception.com/utility/showArticle?objectID=271)

_http://www.seedsofdeception.com/utility/showArticle/?objectID=271_

(http://www.seedsofdeception.com/utility/showArticle/?objectID=271)

 

References

1. Centers for Disease Control, " Eosinophilia-Myalgia Syndrome-New

Mexico, " Morbidity and Mortality Weekly Report (MMWR) (November 17, 1989), Vol.

38, No. 45, pp. 765-767.

2. Food & Drug Administration, " Update on L-Tryptophan " (December 5,

1989), _http://www.fda.gov/bbs/topics/ANSWERS/ANS00114.html_

(http://www.fda.gov/bbs/topics/ANSWERS/ANS00114.html) .

3. Food & Drug Administration, " Recall of L- 6 Tryptophan, " P90-21,

(March 22, 1990), _http://www.fda.gov/bbs/topics/NEWS/NEW0004.html_

(http://www.fda.gov/bbs/topics/NEWS/NEW0004.html) .

4. Food & Drug Administration, " L-Tryptophan Update: Company Link

Suggested " (April 26, 1990),

_http://www.fda.gov/bbs/topics/ANSWERS/ANS00089.html_

(http://www.fda.gov/bbs/topics/ANSWERS/ANS00089.html) .

5. Daily Appellate Report (April 22, 1996), " L-tryptophan Manufacturer

is Per Se Negligent Under Food, Drug And Cosmetic Act in Products Liability

Action, " pp. 3534.

6. David Swinbanks & Christopher Anderson " Search for contaminant in

EMS outbreak goes slowly, " Nature (July 9, 1992), Vol. 358, pp. 96.

7. L Slutsker, et al., Journal of the American Medical Association

(July 11, 1990), Vol. 264, No. 2, pp. 213-217.

8. Food & Drug Administration, " L-Tryptophan Recall, " P89-49 (November

17, 1989), _http://www.fda.gov/bbs/topics/NEWS/NEW00091.html_

(http://www.fda.gov/bbs/topics/NEWS/NEW00091.html) .

9. Food & Drug Administration, " FDA's Regulatory Response to

L-Tryptophan Situation, " (July 18, 1991).

10. Belongia, EA, et al., New England Journal of Medicine (August 9,

1990), Vol. 323, No. 6, pp. 357-365.

11. Laurie Garrett, Newsday, " Genetic engineering flaw blamed for toxic

deaths, " The Ann Arbor News (August 14, 1990), pp C-1.

12. Leslie Roberts, " L-tryptophan puzzle takes new twist, " Science

(August 31, 1990).

13. Federal Register, Notices (May 29, 1992), Vol. 57, No. 104, pp.

22987.

14. Jim Maryanski, biotech coordinator at FDA, personal interview July

5, 1996

15. Tomoko Inose & Kousaku Murata, Department of Applied Microbiology,

Research Institute for Food Sciences, Kyoto University, Japan, “Enhanced

accumulation of toxic compound in yeast cells having high glycolytic activity:

a

case study on the safety of genetically engineered yeast,†International

Journal of Food Sciences and Technology (1995), Vol. 30, pp. 141-146.

16. Michael Antoniou, PhD, Reader in Molecular Genetics, London, UK, “Is

GM Food Devoid of DNA Safe?†Nutritional Therapy Today (1996).

17. See also James Maryanski’s comments in “Cornucopia of Biotech Food

Awaits Labeling,“ by Paul Jacobs, The Los Angeles Times, January 31, 2000.

The Pre-epidemic Cases: A Key to the EMS Puzzle?

_http://www.seedsofdeception.com/Public/L-tryptophan/3Pre-epidemicCases-KeytoE

MSPuzzle/index.cfm_

(http://www.seedsofdeception.com/Public/L-tryptophan/3Pre-epidemicCases-KeytoEMS\

Puzzle/index.cfm)

 

" The existence of cases prior to those that constituted the great bulk of

the epidemic is one of the most peculiar and interesting aspects of the EMS

outbreak. "

— Edwin M. Kilbourne,_1_

(http://www.seedsofdeception.com/Public/L-tryptophan/3Pre-epidemicCases-KeytoEMS\

Puzzle/index.cfm#ref) epidemiologist, Center for

Disease Control

 

For more than a decade now, researchers have been puzzled by cases of EMS

that occurred for several years prior to the epidemic.

For more than a decade now, researchers have been puzzled by cases of EMS

that occurred for several years prior to the epidemic.

The Center for Disease Control (CDC) surveillance data revealed nearly 100

cases with onset of illness prior to May 1989, the beginning of the epidemic

period._2_

(http://www.seedsofdeception.com/Public/L-tryptophan/3Pre-epidemicCases-KeytoEMS\

Puzzle/index.cfm#ref) This represented about 7% of the patients

for whom complete information was available._3_

(http://www.seedsofdeception.com/Public/L-tryptophan/3Pre-epidemicCases-KeytoEMS\

Puzzle/index.cfm#ref) CDC

estimated that the number of actual EMS cases could be about four times what

was reported. This is because of the agency's passive surveillance system and

its surveillance case definition, which stressed specificity over

sensitivity. The definition was not sensitive enough to recognize the broad

spectrum of

EMS symptoms and excluded milder cases._3, 4_

(http://www.seedsofdeception.com/Public/L-tryptophan/3Pre-epidemicCases-KeytoEMS\

Puzzle/index.cfm#ref)

CDC researchers calculated that with a more sensitive case definition there

could be as many as 5,000 to 10,000 EMS cases in the U.S._4_

(http://www.seedsofdeception.com/Public/L-tryptophan/3Pre-epidemicCases-KeytoEMS\

Puzzle/index.cfm

#ref) Using the 7% figure from CDC surveillance data, this would mean that

there could be as many as 350 to 700 pre-epidemic cases of EMS. These early

EMS victims would have consumed L-tryptophan (LT) pills that were contaminated

prior to the manufacturing changes attributed to the etiology of the

epidemic. In that scenario, Showa Denko's introduction of the Strain V

bacterium and

their reduction in activated carbon powder in the filtering process could

not have been solely responsible.

Some researchers suggested that a low level of contaminants may have always

been present in L-tryptophan-containing products,_5_

(http://www.seedsofdeception.com/Public/L-tryptophan/3Pre-epidemicCases-KeytoEMS\

Puzzle/index.cfm#ref)

while others suggested that Showa Denko L-tryptophan might have contained the

etiologic agent in lower concentrations or in fewer lots before the

manufacturing changes were implemented._6,7_

(http://www.seedsofdeception.com/Public/L-tryptophan/3Pre-epidemicCases-KeytoEMS\

Puzzle/index.cfm#ref)

Evidence Points to Showa Denko

While limited data has been available on pre-epidemic EMS, findings from

product trace-back studies on epidemic cases have been unambiguously clear.

In four epidemiological studies (Oregon, Minnesota, New York and South

Carolina) 96-100 percent of epidemic cases traced to Showa Denko._6_

(http://www.seedsofdeception.com/Public/L-tryptophan/3Pre-epidemicCases-KeytoEMS\

Puzzle/index.

cfm#ref) In the two cases that didn't initially appear to trace to Showa

Denko, tablets from one (in Minnesota) were analyzed and found to have a

chromatographic pattern that was characteristic of Showa Denko L-tryptophan.

The

other victim (in Oregon) had consumed two different brands, one of which was

untraceable and therefore could have been from Showa Denko._1_

(http://www.seedsofdeception.com/Public/L-tryptophan/3Pre-epidemicCases-KeytoEMS\

Puzzle/index.cf

m#ref) CDC researchers concluded, " Of 6 manufacturers of LT, only LT

manufactured by Showa Denko KK was clearly associated with illness. " _8_

(http://www.seedsofdeception.com/Public/L-tryptophan/3Pre-epidemicCases-KeytoEMS\

Puzzle/ind

ex.cfm#ref)

Surprisingly little trace-back data is available on the pre-epidemic EMS

cases. In the Minnesota study on the epidemic, four 1988 cases (September to

November) were excluded from the analysis because they were pre-epidemic. Two of

these cases were traced to Showa Denko and two were untraceable._6_

(http://www.seedsofdeception.com/Public/L-tryptophan/3Pre-epidemicCases-KeytoEMS\

Puzzle/i

ndex.cfm#ref) In a CDC study, two case patients were found to have consumed

pills manufactured by Showa Denko in April 1988, but they also had consumed

pills from 1989 lots. Again, only Showa Denko product was found to be

case-associated; no other manufacturer's product was linked to EMS._9_

(http://www.seedsofdeception.com/Public/L-tryptophan/3Pre-epidemicCases-KeytoEMS\

Puzzle/index

..cfm#ref)

Curious if pre-epidemic or epidemic EMS cases were linked to other

manufacturers' L-tryptophan, I faxed and called about a dozen law firms who had

handled Showa Denko cases. None had handled or knew of any definite case

associated

with another manufacturer.

Stephen Sheller, a Philadelphia attorney whose firm handled over 100 EMS

cases, including about 10 pre-epidemic, commented,

" We have always been suspicious that there were EMS cases caused by other

L-tryptophan... However, we have never had a case that we could confirm that

with. All cases that we've had (have) been traced to Showa Denko.

" I thought I had a case with Ajinomoto.... We went through hoops in certain

tracing. We had affidavits from companies who insisted they had no Showa Denko

product. A diet food, for example, put out by some companies, we traced....

product to a time when Hormel had a strike and they sub-contracted out the

work to another company that used Showa Denko L-tryptophan. We were able to

confirm direct shipments of Showa Denko product to that company in 1986. " _10_

(http://www.seedsofdeception.com/Public/L-tryptophan/3Pre-epidemicCases-KeytoEMS

Puzzle/index.cfm#ref)

In response to how many EMS victims claimed early onset of illness, Don

Morgan, an attorney representing Showa Denko in the litigations, replied in a

phone interview, " I don't have a good idea of that... See, there have been

thousands of cases... My general impression is that there have been a good

number

of those cases of pre-epidemic onset of symptoms... more and more cases came

up that involved onset before 1989. " He said that SDKK has settled some EMS

claims related to such tryptophan, and many epidemic-period claims, without

admitting causation or legal liability._11_

(http://www.seedsofdeception.com/Public/L-tryptophan/3Pre-epidemicCases-KeytoEMS\

Puzzle/index.cfm#ref)

Gerald Gleich, M.D., a leading researcher on L-tryptophan/EMS at the Mayo

Clinic, recently summed up the matter, saying, " Tryptophan itself clearly is not

the cause of EMS in that individuals who consumed products from other

companies, other than Showa Denko, did not develop EMS. The evidence points to

Showa Denko product as the culprit and to the contaminants as the cause. " _12_

(http://www.seedsofdeception.com/Public/L-tryptophan/3Pre-epidemicCases-KeytoEMS\

Pu

zzle/index.cfm#ref)

Intrigued by the elusive pre-epidemic cases, I contacted a woman with the

National EMS Network, a nonprofit organization composed of EMS victims and their

families. She put me in touch with several pre-epidemic EMS victims, some

with onset of illness dating back to 1984-85. I also found two early cases who

had given testimony at the Congressional Committee Hearings on EMS._13_

(http://www.seedsofdeception.com/Public/L-tryptophan/3Pre-epidemicCases-KeytoEMS\

Puzz

le/index.cfm#ref) All of these had received settlements from Showa Denko

(except for two who were offered settlements but declined them and then later

lost their cases on statutes of limitation).

One case was particularly striking, because the man had onset of illness in

November 1987 and stopped using L-tryptophan in February 1988. In his legal

proceedings, his pills were tested and identified as from Showa Denko._14_

(http://www.seedsofdeception.com/Public/L-tryptophan/3Pre-epidemicCases-KeytoEMS\

Puz

zle/index.cfm#ref) His tablets had to have been from lots manufactured

about a year and a half before October 1988, — the alleged date when

L-tryptophan

product became contaminated._15_

(http://www.seedsofdeception.com/Public/L-tryptophan/3Pre-epidemicCases-KeytoEMS\

Puzzle/index.cfm#ref) (It took several

months for product to be packaged in Japan, shipped to the U.S., pass through

the distribution chain, and then be consumed for a few weeks before onset of

symptoms.)

Birth of a New Disease: A Problem with Diagnosis

In late October 1989, when three physicians in New Mexico first linked

L-tryptophan ingestion to patients with debilitating myalgia (muscle pain) and

elevated eosinophil levels, the EMS epidemic had already been six months in

progress. Prior to that time, no medical classification existed to accommodate

the cluster of symptoms associated with EMS._1_

(http://www.seedsofdeception.com/Public/L-tryptophan/3Pre-epidemicCases-KeytoEMS\

Puzzle/index.cfm#ref)

Consequently, physicians were either baffled by this new disease and gave no

clear

diagnosis, or they gave a diagnosis that fit only one of the patient's

predominant symptoms.

In addition to severe myalgia and elevated eosinophils, EMS patients often

exhibited fasciitis, scleroderma, rashes, perimyositis, peripheral edema

(swelling), and/or other symptoms._14_

(http://www.seedsofdeception.com/Public/L-tryptophan/3Pre-epidemicCases-KeytoEMS\

Puzzle/index.cfm#ref) Before news of the

epidemic emerged in November 1989, patients seeking medical treatment were

sometimes diagnosed with one of these illnesses. Of the eleven pre-epidemic

cases on whom I collected information, two were initially diagnosed with

eosinophilic fasciitis (EF), one with fibromyalgia, one with scleroderma, and

one

with neuritis and fibromyalgia. Six had no clear diagnosis, though their

physicians had considered some of the above diseases. All were re-diagnosed

with

EMS after news of epidemic emerged.

One woman who had onset of illness in October 1988 told me she was initially

diagnosed as having eosinophilic fasciitis " even though some of the symptoms

didn't quite match. " A man with onset in March 1989 said that he was not

given a diagnosis though " fibromyalgia was tossed around. " And a woman who

testified at the Congressional Committee Hearings on L-tryptophan stated that

her

doctor noted elevated eosinophils, but gave no diagnosis because " he knew of

no illness which had this cluster of symptoms. "

Other Diseases Linked to L-Tryptophan Ingestion

I started searching the medical literature for L-tryptophan-related illnesses

other than EMS, and found several studies and a few isolated case reports.

A study at the University of Miami School of Medicine reported (1991) that

65% of EF cases (11 of 17) and 20% of scleroderma patients (2 of 10) had

ingested L-tryptophan prior to onset of disease. Eight of the eleven EF cases

and

both scleroderma patients had pre-epidemic onset, with one scleroderma case

dating back to 1985._15_

(http://www.seedsofdeception.com/Public/L-tryptophan/3Pre-epidemicCases-KeytoEMS\

Puzzle/index.cfm#ref)

A study at the University of Pennsylvania reported (May 1990) that all eight

patients with EF had taken L-tryptophan before the onset of their disease.

All had myalgias and high peripheral eosinophil counts. Only one of 40 patients

with scleroderma had used L-tryptophan preceding illness._16_

(http://www.seedsofdeception.com/Public/L-tryptophan/3Pre-epidemicCases-KeytoEMS\

Puzzle/index.

cfm#ref)

An article on the Los Alamos Conference on EMS (1990) stated, " It is of

interest that since 1986, one-half of the patients diagnosed with EF at the Mayo

Clinic had been exposed to L-tryptophan; and 9 of 45 patients in the ARAMIS

database with EF (1985-1986) remembered using L-tryptophan before onset of

EF. " _4_

(http://www.seedsofdeception.com/Public/L-tryptophan/3Pre-epidemicCases-KeytoEMS\

Puzzle/index.cfm#ref)

EF is a rare disease with unknown cause, characterized by scleroderma-like

skin changes and peripheral eosinophilia, two common symptoms of EMS. It was

first diagnosed in 1974 by Shulman and by 1988 some 200-plus cases had been

reported worldwide._17_

(http://www.seedsofdeception.com/Public/L-tryptophan/3Pre-epidemicCases-KeytoEMS\

Puzzle/index.cfm#ref)

Other studies on L-tryptophan-related EF reported similar findings._18,19_

(http://www.seedsofdeception.com/Public/L-tryptophan/3Pre-epidemicCases-KeytoEMS\

P

uzzle/index.cfm#ref) Some commentators have concluded that L-tryptophan

products are the first agents to be implicated as a cause of EF in some

patients._5_

(http://www.seedsofdeception.com/Public/L-tryptophan/3Pre-epidemicCases-KeytoEMS\

Puzzle/index.cfm#ref) Others have suggested that L-tryptophan-related

EF and EMS are the same disease._15_

(http://www.seedsofdeception.com/Public/L-tryptophan/3Pre-epidemicCases-KeytoEMS\

Puzzle/index.cfm#ref) One 1988 case

of EF was contacted after news of the EMS epidemic and it was found that she

had used L-tryptophan before onset of her illness; she was re-diagnosed as

having EMS._1_

(http://www.seedsofdeception.com/Public/L-tryptophan/3Pre-epidemicCases-KeytoEMS\

Puzzle/index.cfm#ref)

Isolated cases of other diseases have also been linked to L-tryptophan

ingestion, including eosinophilic perimyositis_20_

(http://www.seedsofdeception.com/Public/L-tryptophan/3Pre-epidemicCases-KeytoEMS\

Puzzle/index.cfm#ref) and

lichen sclerosus et atrophicus and acanthosis nigricans._15_

(http://www.seedsofdeception.com/Public/L-tryptophan/3Pre-epidemicCases-KeytoEMS\

Puzzle/index.cfm#r

ef)

Although L-tryptophan usage is also associated with cases of these other

diseases, researchers never conducted trace back studies to determine whose

product was causing all this damage. Commentators assumed that

L-tryptophan-containing products in general were probably responsible, but no

one did follow-up

research to verify this.

To date, the available evidence, from both L-tryptophan trace back studies

and EMS legal proceedings, appears to point to Showa Denko product as the

source of the problem.

In retrospect, these pre-epidemic L-tryptophan-related illnesses may provide

vivid demonstration of the mechanics of the birth of a disease, and how

medical science grapples with diagnosing and accommodating a new entity into

its

medical repertoire. For several years, cases of the new disease were occurring

unbeknownst to the manufacturer, government regulators and medical

authorities. It took a tragic epidemic in 1989 to bring EMS into focus. For

more than

a decade since then, scientists have been trying to fit the pieces of the EMS

puzzle together. These puzzle pieces fit together only one way, namely, when

the scientific model and understanding are correct. The pre-epidemic EMS and

other L-tryptophan-related diseases appear to be a key piece in solving the

puzzle.

According to attorney Sheller, there is a huge lesson in all this. Because

EMS cases went unrecognized for so many years, he said, " One has to wonder

about a lot of different things that are in the food supply, because no one has

really tested it to find out. The biggest problem is that most of the testing

is done for short periods of time. " _10_

(http://www.seedsofdeception.com/Public/L-tryptophan/3Pre-epidemicCases-KeytoEMS\

Puzzle/index.cfm#ref)

For more information on the pre-epidemic cases of EMS, see:

* _1987 EMS Victim - Initially Diagnosed with Fibromyalgia_

(http://www.seedsofdeception.com/utility/showArticle?objectID=233)

* _Summary Profiles of 11 Pre-epidemic EMS Cases_

(http://www.seedsofdeception.com/utility/showArticle?objectID=230)

* _Excerpt from Attorneys Frank Silvestri and John Massicot article on

'EMS Lawsuits'_

(http://www.seedsofdeception.com/utility/showArticle?objectID=272)

With respect to Showa Denko's L-tryptophan, the next piece in the puzzle is

to address the question, _Where did the contaminants come from?_

(http://www.seedsofdeception.com/utility/showPage/index.cfm?objectID=public,4975\

)

References

1. Edwin M Kilbourne, " Eosinophilia-Myalgia Syndrome: Coming to Grips

with a New Illness, " Epidemiological Reviews (1992), Vol. 4, pp. 16-36.

2. Leslie A. Swygert, et al., " Eosinophilia-Myalgia Syndrome: Results

of National Surveillance, " Journal of the American Medical Association

(October 3, 1990), Vol. 264, No. 13, pp. 1698-1703.

3. Lee D. Kaufman and Rossanne M. Philen, " Tryptophan: Current Status

and Future Trends for Oral Administration, " Drug Safety (1993), Vol. 8, No. 2,

pp. 89-98.

4. Phillip A. Hertzman, et al., " The Eosinophilia-Myalgia Syndrome: The

Los Alamos Conference, " Journal of Rheumatology (1991), Vol. 18, No. 6, pp.

867-873.

5. Richard W. Martin and Joseph Duffy, " Eosinophilic Fasciitis

Associated With Use of L-Tryptophan: A Case- Control Study and Comparison of

Clinical

and Histopathologic Features, " Mayo Clinic Proceedings (September 1991),

Vol. 66, pp. 892-898.

6. Edward A. Belongia and Gerald J. Gleich, Editorial, " The

Eosinophilia-Myalgia Syndrome Revisited, " Journal of Rheumatology (1996), Vol.

23, No.

10, pp. 1682-1685.

7. Arthur N. Mayeno and Gerald J. Gleich, " Eosinophilia-myalgia

syndrome and tryptophan production: a cautionary tale, " Trends in Biotechnology

(TIBTECH) (September 1994), Vol. 12, pp. 346-352.

8. Edwin M. Kilbourne, et al., " Tryptophan Produced by Showa Denko and

Epidemic Eosinophilia-Myalgia Syndrome, " Journal of Rheumatology Supplement

(October 1996), Vol. 23, No. 46, pp. 81-92.

9. Samuel P. Caudill, et al., " Important Issues Affecting

Eosinophilia-Myalgia Syndrome Investigations Based on Analyses of L-Tryptophan

Samples, "

Journal of Occupational Medicine and Toxicology (1993), Vol. 2, No. 1, pp.

41-52.

10. Stephen Sheller, attorney, personal interviews, March 25 and May 1,

1998.

11. Don Morgan, attorney, Cleary, Gottlieb, Steen & Hamilton,

Washington, D.C., personal interview, May 21, 1998.

12. National Eosinophilia-Myalgia Syndrome Network, position statement,

approved quote by Gerald J. Gleich, M.D., Mayo Clinic and Foundation, May 25,

2000.

13. Dorothy C. Wilson and Paul L. Hauts, statements, Subcommittee on

Human Resources and Intergovernmental Relations, House Committee on Government

Operations, July 18, 1991.

14. Anonymous pre-epidemic case, personal correspondences, January 10,

2000 to April 13, 2001.

15. Andrew Blauvelt and Vincent Falanga, " Idiopathic and

L-Tryptophan-Associated Eosinophilic Fasciitis Before and After L-Tryptophan

Contamination, "

Archives of Dermatology (August 1991), Vol. 127, pp. 1159-1166.

16. Frendlich B, et al., " L-tryptophan ingestion associated with

eosinophilic fasciitis but not progressive systemic sclerosis, " Annuals of

Internal

Medicine (May 15, 1990), Vol. 112, No. 10, pp. 758-762.

17. Lakhanpal S, et al., " Eosinophilic Fasciitis: Clinical Spectrum and

Therapeutic Response in 52 Cases, " Seminars in Arthritis and Rheumatism

(1988), Vol. 17, No. 4, pp. 221-231.

18. Hibbs JR, et al., " L-tryptophan-associated eosinophilic fasciitis

prior to the 1989 eosinophilia-myalgia syndrome outbreak, " Arthritis

Rheumatology (1992), Vol. 35, No. 3, pp. 299-303.

19. Richard M. Silver, et al., " Scleroderma, Fasciitis, and Eosinophilia

Associated with the Ingestion of Tryptophan, " The New England Journal of

Medicine (March 29, 1990), Vol. 322, No. 13, pp. 874-881.

20. Richard M. Silver, editorial, " Unraveling the Eosinophilia-Myalgia

Syndrome, " Archives of Dermatology (August 1991), Vol. 127, pp. 1214-1216.

 

Where Did the Contaminants Come From?

_http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsC

omeFrom/index.cfm_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm)

 

“Although the [20kg/batch] powdered carbon may have contributed to the

removal of the etiological [causal] agent, it does not explain how the agent

was

introduced into the product.â€

—Edward Belongia, M.D., et al., The New England Journal of Medicine _1_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eF

rom/index.cfm#_edn1)

It’s been more than a decade since Belongia and his colleagues raised this

salient point about what created the contaminants that caused EMS.

Surprisingly, few researchers have addressed it.

Instead, scientists have largely focused on identifying the contaminants in

Showa Denko L-tryptophan and trying to replicate EMS in bioassay (test tube)

and animal feeding experiments. Unfortunately, their experiments have not

yielded satisfactory results._2_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_edn2) While solid

epidemiological evidence supports the conclusion that Showa Denko L-tryptophan

caused

the EMS epidemic, it is unclear precisely which compound(s) in the implicated

product caused the illness._3_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_edn3)

A leading EMS researcher, Gerald Gleich at the Mayo Clinic, said in an

interview, “It doesn’t really make a difference where they [the

contaminants]

came from. The fact of the matter is the thing is contaminated.â€_4_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index

..cfm#_edn4)

This has been the prevailing view of many scientists and regulatory

officials, but it misses the big picture.

Baseball analogy: The pitcher’s or catcher’s fault?

 

An analogy from baseball may help clarify this crucial issue:

If a new pitcher enters a baseball game in the late innings and throws a

pitch that gets by the catcher, causing the winning run to score, who’s

responsible: the pitcher or the catcher?

If the pitch was what the catcher had called for and/or was thrown within

the batting zone or a reasonable distance outside of it, an umpire would

probably call it a “passed ball†and it would go against the catcher’s

record. The

catcher should have caught the pitch because it was within a normal range of

performance.

On the other hand, if the pitcher throws a ball that was not anticipated by

the catcher and/or was significantly outside his normal range, perhaps a

knuckleball that bounces in the dirt, then even a good catcher may not have

adequate time to react and catch the ball. Thus, the umpire would rule it a

“wild

pitch†and it would go against the pitcher’s record.

Of course, this is an oversimplification of Showa Denko’s manufacturing

process. Nevertheless, it illustrates the key roles of both bacterial

fermentation, which produced the L-tryptophan, and purification, which was

supposed to “

catch†and remove the impurities.

Anyone familiar with baseball knows that it would be foolish for someone to

say that it doesn’t matter who the pitcher is, the catcher should be able to

catch anything that he throws.

With respect to Showa Denko’s manufacturing process, scientists have largely

avoided this critical issue involving the role that the genetically

engineered (GE) bacteria played in creating impurities in the product. The

whole

question surrounding Showa Denko’s use of GE bacteria appears to have been

downplayed or dismissed outright by researchers and government agencies,

especially

here in the United States—where the biotech industry would stand to lose the

most if GE was implicated in, or linked to, a major epidemic like EMS.

Showa Denko’s genetically modified bacteria

Showa Denko K.K. is the fourth largest petrochemical manufacturer in Japan.

In December 1981, at its Oita complex, the company began construction of a

new plant to manufacture L-tryptophan food supplement.

On December 14, 1982, Showa Denko received a U.S. patent (#4,363,875) for

its L-tryptophan production process using a novel mutant microorganism,

Bacillus amyloliquefaciens strain IAM 1521, obtained from the Institute of

Applied

Microbiology, University of Tokyo._5_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_edn5) A mutant of

this parent strain was developed by Showa Denko as Strain I, and was used from

the start of production on December 16, 1982 to October 22, 1984, when Strain

II was introduced._5_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_edn5)

Strain II was the first of several genetically engineered—recombinant DNA—

strains of the parent bacterium used by the company to bolster L-tryptophan

yields (_see table of genetic modifications_

(http://www.seedsofdeception.com/utility/showArticle?objectID=247) )._6_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_edn6) Strain III

was used in commercial production from February 23, 1986 to November 21, 1988;

Strain IV from November 22 to December 25, 1988 and Strain V from December

26, 1988 until November 21, 1989,_7_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_edn7) when production

ceased following news of the L-tryptophan-linked epidemic in America.

Showa Denko, like most Japanese manufacturers of L-tryptophan at that time,

used a fermentation process, where a selected bacterial strain was grown from

specific precursors under specific conditions._8_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_edn8)

Showa Denko’s bacteria were fed intermittently with sterilized glucose

(sugar) and anthranilic acid to produce a broth containing L-tryptophan and

impurities. The liquid broth was then heat treated and sent to a cell

separator.

From there it was sent to the purification process, including ion exchange

resin

columns, a membrane for removal of high molecular weight substances, and

towers with activated and granulated carbon powder to remove trace

impurities._9_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_edn9)

Don Morgan, an attorney representing Showa Denko in the legal proceedings in

the U.S., commented about the purity of the company’s product:

“Showa Denko assured that its LT [L-tryptophan] was at least 98.5% pure, and

generally the purity was around 99.5%, and purchasers were advised of the

purity.â€_5_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_edn5)

In the summer of 1988, a German company, A.S. Biologische, tested Showa

Denko L-tryptophan and found impurities, one of which was called Peak D.

According to internal Showa Denko documents, when Showa Denko was questioned

about

the Peak D impurity, they admitted that they couldn’t determine a lack of

toxicity of the impurity because they couldn’t figure out what the impurity

was._10_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_edn10) ,_11_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_edn11)

John Baker, a Denver-based attorney who represented many EMS victims and was

a member of the National Steering Committee for litigation against Showa

Denko, commented:

“After reviewing the company documents and the depositions of company

employees, expert scientists retained by Plaintiffs in the EMS litigation in

the

United States have opined that Showa Denko appears to have destroyed some of

the serial chromatograms that showed contaminants in their L-tryptophan product

in 1988.â€_12_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_edn12)

1988 was nearly a year before the U.S. epidemic began.

However, all L-tryptophan preparations contain various minor impurities,

which vary with different manufacturing processes. The high performance liquid

chromatography (HPLC) “fingerprint†profiles of impurities_13_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.c

fm#_edn13) are relatively consistent from lot to lot for each manufacturer.

The chromatograms from EMS patient-related L-tryptophan (i.e., Showa Denko’s

product) showed many more small peaks of impurities and much higher levels

of several impurities than other manufacturers’ products._14_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm

#_edn14)

The search to identify contaminants

Using HPLC, researchers have found as many as 60-69 trace contaminants in

Showa Denko L-tryptophan. _Six of these were associated with EMS cases_

(http://www.seedsofdeception.com/utility/showArticle?objectID=234) ._15_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index

..cfm#_edn15) The chemical structures of five of these case-associated

contaminants have been identified, and the sixth, Peak AAA, still has not been

identified. Four of the case-associated contaminants were tryptophan

derivatives

and one was an aniline derivative._16_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_edn16) ,_17_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom

/index.cfm#_edn17)

According to Showa Denko attorney Don Morgan,

“There is no evidence to suspect that any external materials got into the

production process and ‘contaminated’ the product. The manufacturing

process

was carefully controlled to assure, among other things, that contaminants did

not get into the process. All fermentation products contain a number of

impurities. For example, beer contains numerous impurities, most of which I

believe have never been identified or isolated.â€_5_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_edn5)

If the contaminants did not get into the product externally, where did they

come from?

One of the world’s leading authorities on the biosynthesis of L-tryptophan,

Charles Yanofsky, PhD, with the Department of Biological Sciences at Stanford

University, said that impurities could be created in several ways:

“If you significantly overproduce a natural substance, such as tryptophan,

it is likely that one or more enzymes of the bacterium will modify tryptophan

and produce an unnatural product or products [during fermentation].

Furthermore, tryptophan is unstable at extreme pH’s and therefore during

purification

it is possible that under the conditions used some other compounds produced

by the bacterium, or that are used during purification, modify the tryptophan

at some step in purification.

“Thus depending upon the organism used for overproduction, the level of

expression, and the conditions of growth, some fraction of the synthesized

tryptophan could be modified. In addition, during purification modification is

also

possible. In fact there is also a third possibility, namely that a modified

form of tryptophan produced during bacterial growth is further altered during

purification, i.e., toxic forms of tryptophan could be generated by a

two-stage process. It should be possible to determine exactly what happened

from

analysis of a typical bacterial preparation of tryptophan, before purification,

and what contaminants are present before and at different stages of

purification.â€_18_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_edn18)

Yanofsky raises a key point, which suggests that if contaminant(s) are

formed at some stage of purification, it doesn’t necessarily eliminate what

happened during bacterial fermentation as a possible cause or contributing

factor.

In an _article in the Medical Post_

(http://www.seedsofdeception.com/utility/showArticle?objectID=245) in 1990,

Yanofsky explained that the more

L-tryptophan that is produced in the fermentation cell, the greater the chance

that

some side reaction will occur at a greater rate, producing more of some

contaminant: “It’s possible that one purification scheme may be quite

adequate

when producing low levels of tryptophan, but at higher levels, it might not be

good enough.â€_19_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_edn19)

Showa Denko’s genetically modified strains (II-V) were used to increase the

biosynthesis of L-tryptophan through bacterial fermentation. Interestingly,

the introduction of these higher yielding GE strains over several years prior

to the EMS epidemic appears, on the surface, to correspond directly to the

gradual increase in the incidence of EMScases reported by researchers._20_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFr

om/index.cfm#_edn20)

Regarding overproduction of L-tryptophan through bacterial fermentation,

Yanofsky said (emphasis added):

“If Showa Denko engineered the bacterium to overproduce tryptophan [which

Showa Denko did], then there are many unknowns that would be associated with

its overproduction. They probably engineered the strain to overproduce

chorismate [which they did], the common aromatic precursor of tryptophan, as

well as

overproduce all the enzymes of the tryptophan biosynthetic pathway. Overall

this would mean that the bacterium is producing large amounts of about 10-15

metabolites that are not normally produced in excess. The accumulation of

these metabolites would, in some cases, lead to the modification by other

enzymes, to give products that normally are never produced by the bacterium.

One or

more of these products could be a compound toxic to man.

“Similarly the production of enzymes of the aromatic and tryptophan

biosynthetic pathways could lead to the synthesis of unnatural products by side

reactions that normally do not occur. Again, toxic products could be

produced….â€

_21_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_edn21)

In fact, in an unpublished study_7_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_edn7) Showa Denko

scientists reported that in Strain V, genetic engineering was used to increase

and amplify genes for two enzymes used in the biosynthesis of L-tryptophan:

“The principal difference between strains III and V is that the prs and serA

genes for two enzymes required for the biosynthesis of tryptophan PRPP

[5-phosphoribosyl-1-pyrophosphate] and serine were increased and their

expression

amplified in Strain V by standard genetic engineering techniques.â€

Dr. Yanofsky continued:

“Genetic engineering results in the formation of higher than normal

concentrations of certain enzymes and products; these could provide the basis

for the

synthesis of higher levels of toxic substances.â€_21_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_edn21

)

Thus, from a theoretical perspective, genetic engineering could have played

a role in creating metabolites, enzymes and other compounds during

fermentation that directly or indirectly caused increased toxins in Showa

Denko’s

product.

Is there any evidence to support this view? Unfortunately, there’s very

little, because researchers have not been able to study samples of the

genetically engineered (GE) bacteria used in Showa Denko’s fermentation.

FDA officials say that Showa Denko never gave them samples of their GE

bacterial cultures._22_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_edn22) ,_23_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_edn23)

but an attorney representing the manufacturer claims that FDA never followed

up on the manufacturer’s offer to supply the GE bacteria to FDA via a trained

courier rather than by shipping. (Shipping could cause mutations, creating

impurities not present in Showa Denko’s cultures—_see Discrepancy Over

Genetically Engineered Cultures_

(http://www.seedsofdeception.com/utility/showArticle?objectID=249) )._24_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_edn24) The manufacturer

eventually destroyed the cultures in 1996.

In the absence of the GE cultures, researchers have studied how the

case-associated contaminants could have been formed during the purification

process.

One case-associated contaminant that has received significant attention is

3-phenylamino-L-alanine (3-PAA, _25_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_edn25) also referred

to as Peak I or UV-5 _26_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_edn26) ,_15_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_ed

n15) ). 3-PAA is remarkably similar to 3-PAP (3-phenylamino-1,

2-propanediol), an impurity implicated in the 1981 toxic oil syndrome (TOS)

that seriously

injured 20,000 people (causing 839 deaths) in Spain with an EMS-like

disease._17_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_edn17) ,_6_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_edn6) Both 3-PAA and

3-PAP are aniline derivatives._6_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_edn6)

Showa Denko used anthranilic acid in the biosynthetic pathway during

fermentation to create L-tryptophan:_9_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_edn9) Anthranilic acid +

5-phosphoribosyl-1-pyrophosphate (PRPP) + serine = LT.

According to an _Asahi News Service report_

(http://www.seedsofdeception.com/utility/showArticle?objectID=246) (1992),

Showa Denko said that it “did not

use any aniline compounds anywhere in the manufacturing process.â€_27_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/

index.cfm#_edn27) But an article in the Journal of the American Medical

Association (JAMA) reported that the chemical structures of anthranilic acid

and

aniline are similar._20_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_edn20) Could anthranilic acid,

which has a similar structure to aniline, get modified to form an

aniline-derived compound, 3-PAA?

The Asahi News Service article states, “Showa Denko genetically altered the

Bacilli to increase the bacteria’s production of the serine used to

manufacturer L-tryptophan.â€_27_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_edn27) Serine is a non-toxic,

natural amino acid, but its overproduction via the genetically engineered

bacteria could have created the contaminant 3-PAA, according to Yanofsky:

“It is conceivable that by overproducing serine the manufacturer [showa

Denko] caused, or increased, the production of [3-] PAA.â€_28_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_e

dn28)

So, theoretically, Showa Denko’s genetically engineered bacteria, which were

used to overproduce serine in the biosynthesis of L-tryptophan, could quite

feasibly have created the toxic aniline derivative 3-PAA by modifying

anthranilic acid during fermentation.

However, a study by Toyoda, et al., found that 3-PAA could be formed under

the purification conditions used by Showa Denko from anthranilic acid and

serine through a 2-step process: Aniline was made first by the heat degradation

of anthranilic acid under acidic conditions, and this aniline subsequently

reacted with L-serine under basic conditions to produce 3-PAA. The authors

stated that formation of 3-PAA from anthranilic acid and serine had not yet

been

investigated (i.e., during fermentation) and cautioned, “more precise

experiments are needed before any quantitative statements can be made regarding

the

formation of PAA under fermentation and purification conditions.â€_29_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/i

ndex.cfm#_edn29)

Interestingly, in the paper’s abstract, it states, “These results suggest

that PAA could be formed under the fermentation and purification conditions

used to produce L-tryptophan on an industrial scale.â€(emphasis added). Why

did

the authors include “fermentation†in the abstract, if the conditions of

their experiment pertained to Showa Denko’s purification procedures?

This appears confusing, but a description of Showa Denko’s manufacturing

process may offer clarification. It states that the fermentation broth—

containing anthranilic acid, serine, L-TRP, glucose, anti-foaming agent and

impurities—

underwent “heat treatment†immediately following fermentation, while the

broth was still in the vat, and before it went to the cell separator and onto

the filtration system._9_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_edn9) Technically, the results of

the study would suggest that aniline and case-associated contaminant 3-PAP

were formed between fermentation and filtration.

Is there evidence of other steps in the process where the case-associated

contaminants may have been formed?

A study by Thomas Simat and colleagues at the University of Hamburg,

Germany, reported that two case-associated contaminants, IMT and HIT, were

2-tryptophan derivatives formed by the reaction of excess L-tryptophan during

purification._30_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_edn30)

Showa Denko used GE bacteria specifically to produce excess L-tryptophan,

i.e., to amplify the biosynthetic pathway of tryptophan to increase yields.

This suggests that the GE bacteria did, in fact, play a role in creating

case-associated contaminants IMT and HIT, because if excess L-tryptophan had

not

been produced during fermentation in the first place, then IMT and HIT would

not

have been formed, or at least not to the same extent, in the downstream

processing (filtration).

From the foregoing discussion it is apparent that the formation of these and

other contaminants associated with EMS (i.e., EBT and PIC) were the result

of specific features of Showa Denko’s manufacturing process—both during

fermentation and purification.

Simat and his colleagues reported, “All results indicate that the

purification process, not the fermentation, governs the pattern of contaminants

in

biotechnologically derived Trp.â€_16_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_edn16) But this claim is

contrary to findings of a CDC priority case lot study, which showed “peaks

predictive of case status reflected principally the differences in trace

components between the bacterial strain V (and IV2) fermentation processes and

bacterial strain III fermentation processes.â€_15_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_edn15)

This contradiction in reports is clarified by Yanofsky’s earlier point, that

contaminants in SDK product could have formed in two-step processes, where

the overproduction or excess of L-tryptophan produced from the GE bacteria

during fermentation subsequently reacted to some condition during purification.

At first glance, some researchers may fault Showa Denko’s purification

process. But the two aspects of the manufacturing process—fermentation and

purification—are not as separate as some have suggested. Contaminants formed

during

purification could have been directly influenced by what happened during the

GE-enhanced fermentation, which created the initial conditions for the

reactions.

Coming back to the baseball analogy, the unbiased umpire must decide whether

or not the pitcher threw the ball in the normal range of the catcher, such

that he should have caught it. Similarly, unbiased observers must query, Did

Showa Denko’s GE bacteria create an unexpected situation during fermentation

that made proper filtration more difficult and/or out of the normal range of

catching impurities?

The view of many scientists has been that it doesn’t matter where the

contaminants came from—the manufacturer should be able to clean up its

product.

From that perspective, the problem was due to defective filtration._30_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/i

ndex.cfm#_edn30) This is similar to saying that it doesn’t matter what the

pitcher did, a good catcher should be able to catch any ball thrown to him—

which anyone familiar with baseball knows is ludicrous.

Nevertheless, let’s assume for the time being that their logic is correct.

This raises several important questions: Why didn’t Showa Denko have proper

filtration in place to purify its L-tryptophan? Why were these contaminants so

toxic at such low concentrations—well within the U.S. Pharmacopoeia limit—

that they caused EMS, a seriously debilitating disease? And, why didn’t FDA

have more strict regulatory standards in place to prevent such a public health

tragedy?

These questions and related issues are addressed in the subsequent sections.

For more on this subject, see also:

* _Discrepancy Over Genetically Engineered Cultures_

(http://www.seedsofdeception.com/utility/showArticle?objectID=249)

* _Table of Genetic Modifications of the different strains of Bacillus

amyloliquefaciens used [by Showa Denko] to manufacture L-tryptophan,

TIBTECH, Sept. 1994 (Vol. 12), p. 348_

(http://www.seedsofdeception.com/utility/showArticle?objectID=247)

* _Six EMS case-associated L-tryptophan contaminants_

(http://www.seedsofdeception.com/utility/showArticle?objectID=234)

* _Comments by Charles Yanofsky, PhD, Stanford University_

(http://www.seedsofdeception.com/utility/showArticle?objectID=235)

* _“EMS deaths: Is recombinant DNA technology involved?†The Medical

Post, Nov. 6, 1990_

(http://www.seedsofdeception.com/utility/showArticle?objectID=245)

* _“Japanese Identify Second Impurity in L-Trypt.,†Asahi News

Service, June 23, 1992_

(http://www.seedsofdeception.com/utility/showArticle?objectID=246)

_Next section: Problems with Identification and Testing for Trace

Contaminants>>_

(http://www.seedsofdeception.com/utility/showPage/index.cfm?objectID=public,5003\

)

_http://www.seedsofdeception.com/Public/L-tryptophan/5ProblemswithIdentificati

onTesting/index.cfm_

(http://www.seedsofdeception.com/Public/L-tryptophan/5ProblemswithIdentification\

Testing/index.cfm)

References

 

 

_1_ (http

://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsComeFrom\

/index.cfm#_ednref1) Belongia EA, et al., “An Investigation of

the Cause of the Eosinophilia-Myalgia Syndrome Associated with Tryptophan

Use,â€

The New England Journal of Medicine (August 9, 1990), Vol. 323, No. 6, pp.

357-365.

_2_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_ednref2) Article by Gerald Gleich, MD, National EMS

Network Newsletter, February 1998.

_3_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_ednref3) Kilbourne EM, et al., “Tryptophan Produced

by Showa Denko and Epidemic Eosinophilia-Myalgia Syndrome,†The Journal of

Rheumatology (1996), Vol. 23, Supplement 46, pp. 81-88.

_4_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_ednref4) Gerald Gleich, MD, Mayo Clinic, personal

interview, April 13, 1998.

_5_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_ednref5) Don Morgan, attorney, Cleary, Gottlieb,

Steen & Hamilton, Washington, DC, personal correspondence (email), April 19,

2001.

_6_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_ednref6) Mayeno AN and Gleich GJ, “

Eosinophilia-myalgia syndrome and tryptophan production: a cautionary tale,â€

TIBTECH

(September 1994), Vol. 12, pp. 346-352.

_7_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_ednref7) Sakimoto K and Torigoe Y, Showa Denko K.K.,

Tokyo, Japan, “Study of Mechanism of Peak E Substance Formation in Process

of Manufacture of L-tryptophan,†unpublished study, April 1, 1992.

_8_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_ednref8) Steven B. Auerbach and Henry Falk, “

Eosinophilia-myalgia syndrome: CDC update,†Cleveland Clinical Journal of

Medicine

(May-June 1991), Vol. 58, No. 3, pp. 215-217.

_9_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_ednref9) Fax from Food and Drug Administration,

Washington, DC, “Showa Denko’s Manufacturing Process,†September 18,

1990.

_10_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_ednref10) Dateline NBC, NBC News, “Bitter Pill,â€

August 22, 1995.

_11_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_ednref11) NHK Special in Japan, “Product Liability

Litigation in America,†August 5, 1995. _

12_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_ednref12) John Baker, attorney, Denver, Colorado,

personal correspondence (email), April 30, 2001.

_13_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_ednref13) Trucksess MW, “Separation and isolation

of trace impurities in L-tryptophan by high performance liquid

chromatography,â€

Journal of Chromatology (1993), Vol. 630, pp. 147-150.

_14_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_ednref14) Caudill S, et al., “Important Issues

Affecting Eosinophilia-Myalgia Syndrome Investigations Based on Analysis of

L-tryptophan Samples,†Journal of Occupational Medicine and Toxicology

(1993),

Vol. 2, No. 1, pp. 41-52.

_15_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_ednref15) Hill R, et al., “Contaminants in

L-tryptophan Associated with Eosinophilia-Myalgia Syndrome,†Archives of

Environmental Contamination and Toxicology (1993), Vol. 25, pp. 134-142.

_16_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_ednref16) Thomas Simat, et al., “Synthesis,

formation, and occurrence of contaminants in biotechnologically manufactured

L-tryptophan,†Adv Exp Med Biol (1999) Vol. 467, pp. 469-480.

_17_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_ednref17) Arthur N. Mayeno, Ph.D., et al., “

3-(Phenylamino)alanine, a Novel Aniline-Derived Amino Acid Associated With the

Eosinophilia-Myaliga Syndrome: A Link to Toxic Oil Syndrome?†Mayo Clinic

Proceedings (1992), Vol. 67: pp. 1134-1139.

_18_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_ednref18) Charles Yanofsky, Ph.D., Dept. of

Biological Sciences, Stanford University, personal correspondence (email), June

2,

1998.

_19_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_ednref19) Philip Raphals, “EMS deaths: Is

recombinant DNA technology involved?â€, The Medical Post, November 6, 1990.

_20_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_ednref20) Slutsker L, et al., “Eosinophilia-Myalgia

Syndrome Assoicated with Exposure to Tryptophan from a Single Manufacturer,â€

Journal of the American Medical Association (July 11, 1990), Vol. 264, No.

2, pp. 213-217.

_21_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_ednref21) Charles Yanofsky, personal correspondence

(email), February 21, 2001.

_22_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_ednref22) James Maryanski, Ph.D., biotechnology

coordinator, FDA, phone interview, July 5, 1986.

_23_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_ednref23) Sam Page, M.D., Chief, Natural Products,

Center for Food Safety and Applied Nutrition, FDA, Congressional Hearing,

Subcommittee, July 18, 1991.

_24_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_ednref24) Don Morgan, attorney, personal

correspondences (emails), April 9 and March 5, 2001.

_25_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_ednref25) Goda Y, et al., “3-aniline-L-alanine,

structural determination of UV-5, a contaminant in EMS-associated L-tryptophan

samples†Chem Pharm Bull (Tokyo) (August 1992), Vol. 40 (8): pp. 2236-2238.

_26_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_ednref26) Toyooka T, et al., “Characterization of

contaminants in EMS-associated L-tryptophan samples by high-performance liquid

chromatography,†Chem Pharm Bull (1991)(Japan), Vol. 39, pp. 820-822.

_27_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_ednref27) Asahi News Service, “Japanese Identify

Second Impurity in L-trypt,†June 23, 1992.

_28_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_ednref28) Yanofsky, personal correspondence

(email), March 22, 2001.

_29_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_ednref29) Toyoda M, et al., Bioscience,

Biotechnology, Biochemistry (1894) Vol. 58, pp. 1318.

_30_

(http://www.seedsofdeception.com/Public/L-tryptophan/4WhereDidtheContaminantsCom\

eFrom/index.cfm#_ednref30) Thomas Simat, et al., “Contaminants in

biotechnologically manufactured L-tryptophan,†Journal of Chromatology B

(1996), Vol. 685, pp. 41-51.

Problems with Identifying and Testing for Trace Contaminants

_http://www.seedsofdeception.com/Public/L-tryptophan/5ProblemswithIdentificati

onTesting/index.cfm_

(http://www.seedsofdeception.com/Public/L-tryptophan/5ProblemswithIdentification\

Testing/index.cfm)

“I must emphasize that the presence of the contaminants in the [showa Denko]

L-tryptophan is astonishingly small and so you require very sophisticated

instrumentation and a lot of hard work to even come close to determining the

structures.â€

—Stephen Naylor, Mayo Clinic_1_

(http://www.seedsofdeception.com/Public/L-tryptophan/5ProblemswithIdentification\

Testing/index.cfm#ref)

Researchers have had enormous difficulty identifying the specific micro

contaminant(s) in Showa Denko L-tryptophan that caused EMS.

In June 1990, the Los Alamos National Laboratory hosted a research

conference on EMS in cooperation with the CDC, FDA, NIH and the New Mexico

Department

of Health and Environment. Phillip Hertzman, one of three New Mexico

physicians who discovered the link between EMS and L-tryptophan consumption,

summarized the conference proceedings in a paper published in the Journal of

Rheumatology. Regarding toxicity tests on Showa Denko L-tryptophan, he and his

colleagues stated:

“Radio-chemical studies have found no contamination. Analyses for 37

inorganic elements and for gross chemical contamination have been negative.

Microbiological analyses have detected no significant contamination. While

endotoxin

has been detected in some lots, this contamination has not been associated

with the illness. Several toxicological studies have been unable to produce the

syndrome [EMS] in an animal model. Although the L-tryptophan was generally

greater than 98.5% pure, high performance liquid chromatography analysis of

material from the implicated manufacturer indicates the presence of more than

30 trace contaminants._2_

(http://www.seedsofdeception.com/Public/L-tryptophan/5ProblemswithIdentification\

Testing/index.cfm#ref)

Other analyses such as ion chromatography and gel-permeation chromatography

identified no case associated contaminants in Showa Denko product from the

comparison of batches of control and case-associated L-tryptophan._3_

(http://www.seedsofdeception.com/Public/L-tryptophan/5ProblemswithIdentification\

Testing/

index.cfm#ref)

Using high performance liquid chromatography (HPLC), researchers have

identified six case-associated contaminants in Showa Denko L-tryptophan._4_

(http://www.seedsofdeception.com/Public/L-tryptophan/5ProblemswithIdentification\

Testin

g/index.cfm#ref) Two of these were novel amino acids: EBT_5_

(http://www.seedsofdeception.com/Public/L-tryptophan/5ProblemswithIdentification\

Testing/index

..cfm#ref) (also called Peak 97 and Peak E) and 3-PAA (also called Peak I

and UV-5). EBT and 3-PAA were significantly correlated to each other and

patients with EMS ingested significantly greater amounts of both (10 and 15

times

respectively) than did control L-tryptophan users.6 Others reported, “The

quantities of the known EMS case-associated contaminants, EBT and 3-PAA, were

remarkably small, of the order of .01%, and could easily escape detection._7_

(http://www.seedsofdeception.com/Public/L-tryptophan/5ProblemswithIdentification\

T

esting/index.cfm#ref) ; EBT and 3-PAA levels were 105 and 220 parts per

million (ppm) respectively in one Showa Denko (SDK) L-tryptophan lot._8_

(http://www.seedsofdeception.com/Public/L-tryptophan/5ProblemswithIdentification\

Testing

/index.cfm#ref) Others reported 3-PAA levels at 89 ppm_5_

(http://www.seedsofdeception.com/Public/L-tryptophan/5ProblemswithIdentification\

Testing/index.cfm

#ref) and 100 ppm,_9_

(http://www.seedsofdeception.com/Public/L-tryptophan/5ProblemswithIdentification\

Testing/index.cfm#ref) while EBT levels varied

widely from zero to nearly 300 ppm._10_

(http://www.seedsofdeception.com/Public/L-tryptophan/5ProblemswithIdentification\

Testing/index.cfm#ref)

In one study, Center for Disease Control (CDC) researchers found contaminant

EBT in case-associated L-tryptophan lots dating back to August 19, 1986,_10_

(http://www.seedsofdeception.com/Public/L-tryptophan/5ProblemswithIdentificati

onTesting/index.cfm#ref) more than two years before the alleged date of

contamination._11_

(http://www.seedsofdeception.com/Public/L-tryptophan/5ProblemswithIdentification\

Testing/index.cfm#ref) High levels of EBT were positively

associated with EMS, but the association lacked statistical significance. The

CDC researchers wrote: “While these findings do not rule out the possibility

that EBT is the etiologic agent in EMS, they raise the possibility that other

chemical contaminants in manufactured tryptophan modify the effects of EBT

or that the causal agent of EMS is an entirely distinct compound._10_

(http://www.seedsofdeception.com/Public/L-tryptophan/5ProblemswithIdentification\

Testing

/index.cfm#ref)

Still, epidemiological data showed that EBT was the impurity most strongly

associated with EMS_12_

(http://www.seedsofdeception.com/Public/L-tryptophan/5ProblemswithIdentification\

Testing/index.cfm#ref) and the Wilcoxon rank-sum

test showed Peak 97 (EBT) was the single most predictive peak of case

associated

L-tryptophan lots._13_

(http://www.seedsofdeception.com/Public/L-tryptophan/5ProblemswithIdentification\

Testing/index.cfm#ref)

Researchers said, “It is also possible that the causative agent (1) may not

absorb in the UV [ultraviolet] range [used in chromatography] or (2) may be

present in a peak that is hidden beneath another peak (e.g., the large

L-tryptophan peak) on HPLC._2_

(http://www.seedsofdeception.com/Public/L-tryptophan/5ProblemswithIdentification\

Testing/index.cfm#ref)

Rossanne Philen and Robert Hill at CDC commented on the biological potency

of Showa Denko L-tryptophan trace contaminants:

“The L-tryptophan associated with EMS met the standards for purity

established by the United States Pharmacopoeia (USP). Nevertheless, more than

60 micro

contaminants have been found in L-tryptophan in amounts of 10 or fewer parts

per million. This finding implies that if one of these 60 or more

contaminants is the etiologic agent for EMS, the contaminant must be a

biologically

potent compound to cause such a serious disease at such a low dosage (emphasis

added)._14_

(http://www.seedsofdeception.com/Public/L-tryptophan/5ProblemswithIdentification\

Testing/index.cfm#ref)

Animal Studies Fail to Reproduce Full EMS Pathology

An early study of implicated L-tryptophan (L-TRP) in Lewis rats produced

encouraging results, replicating many of the pathological features of human

EMS._15_

(http://www.seedsofdeception.com/Public/L-tryptophan/5ProblemswithIdentification\

Testing/index.cfm#ref)

In late 1990, FDA and NIH researchers reported that Lewis rats treated with

implicated L-TRP, but not USP grade L-TRP or vehicle control, developed many

of the specific features of L-TRP EMS, including perimyositis, fasciitis, and

perivascular inflammation. However, peripheral blood eosinophilia, a common

feature of human EMS, was not observed. The large dosage of L-TRP given to

the animals was equivalent to human consumption of about 5 grams/day of L-TRP

(a recommended human dosage was 1-2 grams/day_16_

(http://www.seedsofdeception.com/Public/L-tryptophan/5ProblemswithIdentification\

Testing/index.cfm#ref) )

and treatment was for 38 days._15_

(http://www.seedsofdeception.com/Public/L-tryptophan/5ProblemswithIdentification\

Testing/index.cfm#ref)

“Our study provides evidence that the female Lewis rat is a suitable animal

in which to study the effects of implicated L-tryptophan as well as the

complex factors involved in the pathogenesis of inflammatory, fibrosing

syndromes

of muscle and fascia,†the researchers concluded.

Douglas L. Archer, Ph.D., the deputy director of FDA’s Center for Food

Safety and Applied Nutrition, testified before a subcommittee hearing of the

House

of Representatives on July 18, 1991: “One of the most significant scientific

breakthroughs was the development of an animal model for EMS…. After suspect

LT [L-tryptophan] was fed to a special stain of rats, several pathological

changes that were comparable to those in the EMS patients were observed.â€_17_

(http://www.seedsofdeception.com/Public/L-tryptophan/5ProblemswithIdentificatio

nTesting/index.cfm#ref)

But FDA’s initial enthusiasm for an animal model of EMS was short lived.

In March 1993, government researchers led by Laurie Love, M.D., Ph.D.,

reported that while all animals (Lewis rats) treated with case-associated L-TRP

or

EBT developed significant myofascial thickening, compared with two control

groups, even those animals receiving the control L-TRP showed a mild but

significant increase in the thickness of the myofascia, compared with

vehicle-treated control animals. The basic difference between this study and

the previous

one was that it was with a higher dosage (approx. 5-6 grams/day) of

implicated L-TRP for a slightly longer period (42 days)._12_

(http://www.seedsofdeception.com/Public/L-tryptophan/5ProblemswithIdentification\

Testing/index.cfm#ref)

 

The study demonstrated for the first time the pathological effects of the

EBT contaminant, but the results did not rule out the possibility that other

impurities in the EMS-case-associated L-TRP might also contribute to some of

the features of EMS. “Our study implicates EBT as one compound found in

case-associated L-TRP that can cause some pathological changes in Lewis rats

that

are similar to some pathological features of L-TRP-associated EMS,†according

to the researchers._12_

(http://www.seedsofdeception.com/Public/L-tryptophan/5ProblemswithIdentification\

Testing/index.cfm#ref)

Myofascial thickening and immune cell changes were most prominent in test

animals after administration of case-associated L-TRP, but EBT when combined

with control L-TRP was also found to cause immune cell activation in the

peripheral blood.

The researchers stated, “This study also strongly suggests that control

L-TRP alone [in high doses] plays an important role in this [EMS] and possibly

other fibrosing illnesses, because it is associated with mild but significant

myofascial thickening and alterations in peripheral mononuclear cell

phenotypes, as well as with significant pancreatic pathology.â€_12_

(http://www.seedsofdeception.com/Public/L-tryptophan/5ProblemswithIdentification\

Testing/index.cfm#

ref)

Edwin Kilbourne, M.D., a CDC epidemiologist, commented, “The meaning of the

experimental findings in Lewis rats is not yet clear. The histopathologic

changes produced by implicated tryptophan and by EBT mimic a part of the

pathology of human EMS but fall short of reproducing the illness.â€_18_

(http://www.seedsofdeception.com/Public/L-tryptophan/5ProblemswithIdentification\

Testing/inde

x.cfm#ref)

In 1998, a leading EMS researcher at the Mayo Clinic, Gerald Gleich, MD,

updated the _status of EMS research_

(http://www.nemsn.org/Articles/Gleich%20summary%2098.htm) after scores of

studies:

“The failure of both the bioassay and the animal feeding experiments to

yield robust and reproducible results has been a major disappointment…. As a

consequence of the investigations by public health authorities, L-tryptophan

produced by Showa Denko KK has been implicated as a cause of EMS. Certain

contaminants present in the L-tryptophan have been implicated as candidates for

causation. However, we do not know the exact structures and without this

knowledge one fears that another epidemic will occur at some time.â€_19_

(http://www.seedsofdeception.com/Public/L-tryptophan/5ProblemswithIdentification\

Testing/inde

x.cfm#ref)

For more information on this subject, see also:

* _“Current Status of Research on EMS†by Gerald J. Gleich, M.D.,

Mayo Clinic_ (http://www.nemsn.org/Articles/Gleich%20summary%2098.htm)

_Next section: Government Agencies Disagree on Cause of EMS>>_

(http://www.seedsofdeception.com/utility/showPage/index.cfm?objectID=public,5004\

)

References:

1. National EMS Network Newsletter, winter 2000, interview with Stephen

Naylor, researcher, Mayo Clinic.

2. Herzman PA, et al., “The Eosinophilia-Myalgia Syndrome: The Los

Alamos Conference,†Journal of Rheumatology (1991), Vol. 18, No. 6, pp.

867-873.

3. Toyo’oka T, et al., “Characterization of Contaminants in

EMS-Associated L-tryptophan Samples by High-Performance Liquid

Chromatography,†Chem.

Pharm. Bull. (1991), Vol. 39(3): pp. 820-822.

4. Hill R, Caudill S, el al, “Contaminants in L-Tryptophan Assoicated

with Eosinophilia Myalgia Syndrome,â€Archives of Environmental Contamination

and Toxicology (1993), Vol. 25, pp. 134-142.

5. Arthur N. Mayeno, et al., “Characterization of ‘Peak E,†a Novel

Amino Acid Associated with Eosinophilia-Myalgia Syndrome,†Science (December

21, 1990), Vol. 25: pp. 1707-1708.

6. Mayeno AN, et al., “3-(Phenylamino)alanine, a novel aniline-derived

amino acid associated with the eosinophilia myalgia syndrome: a link to the

toxic oil syndrome?†Mayo Clinic Proceedings (1992), Vol. 67, pp. 1134-1139.

7. Mayeno AN, Gleich, JG, “Eosinophilia-Myalgia Syndrome and tryptophan

production: a cautionary tale,†TIBTECH (Trends of Biotechnology) September

1994, Vol. 12, pp. 346-352.

8. Simat T, et al., “Contaminants in biotechnologically manufactured

L-tryptophan,†Journal of Chromatography B (1996), Vol. 685, pp 41-51.

9. Goda Y, et al., “3-anilino-L-alanine, structural determination of

UV-5, a contaminant in EMS-associated L-tryptophan samples,†Chem. Pharm.

Bulletin (1992), Vol. 40(8), pp. 2236-2238.

10. Philen, RM, et al., “Tryptophan Contaminants Associated with

Eosinophilia-Myalgia Syndrome,†American Journal of Epidemiology (1993), Vol.

138,

No. 3, pp 154-159.

11. Blauvelt A and Falanga V, " Idiopathic and L-Tryptophan-Associated

Eosinophilic Fasciitis Before and After L-Tryptophan Contamination, " Archives

of Dermatology (August 1991), Vol. 127, pp. 1159-1166.

12. Love LA, et al., “Pathological and Immunological Effects of

Ingesting L-Tryptophan and 1,1’-Ethylidenebis (L-Tryptophan) in Lewis Rats,â€

Journal

of Clinical Investigation (March 1993), Vol. 91, pp. 804-811.

13. MMWR, “Analysis of L-tryptophan for the Etiology of

Eosinophilia-Myalgia Syndrome,†(August 24, 1990), Vol. 39(34): pp. 589.

14. Philen RM, et al., “3-(Phenylamino)alanine—a Link Between

Eosinophilia-Myalgia Syndrome and Toxic Oil Syndrome?†Mayo Clinic

Proceedings (1993),

Vol. 68, pp. 197-200.

15. Crofford L, et al., “L-Tryptophan Implicated in Human

Eosinophilia-Myalgia Syndrome Causes Fasciitis and Perimyositis in the Lewis

Rat,†Journal

of Clinical Investigation (November 1990), Vol. 86: pp. 1757-1763.

16. Author’s personal communications with EMS patients.

17. Douglas L. Archer, Ph.D., Deputy Director, Center for Food Safety

and Applied Nutrition, FDA, Hearing before the Subcommittee on Human Resources

and Intergovernmental Relations Subcommittee of the Committee on Government

Operations House of Representatives, First Session, July 18, 1991.

18. Edwin M. Kilbourne, “Eosinophilia-myalgia syndrome: Coming to Grips

with a New Illness,†Epidemilogical Reviews, (1992), Vol. 4, pp. 16-36.

19. Gerald J. Gleich, MD, “Current Status of Research on EMS,†National

EMS Network Newsletter, February (winter 1998).

 

 

(http://www.seedsofdeception.com/Public/L-tryptophan/5ProblemswithIdentification\

Testing/index.cfm)

 

 

© Copyright 2005 William E. Crist. All Rights Reserved