Dr. John Martin

Statement from W. John Martin, M.D., Ph.D.

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I have generally disregarded the criticisms that have been leveled against the existence of stealth-adapted viruses. The individuals making or repeating the criticisms should never have been taken seriously. Unfortunately, negative comments are accessible on the internet and undoubtedly have caused considerable confusion and concern. In the interests of people reading such comments, I have decided to briefly discuss i) evidence for the existence of stealth-adapted viruses and ii) suspension of the clinical diagnostic testing for these viruses

The concept of stealth viruses was formulated in 1990 when I had the opportunity to examine a brain biopsy from a ?xml:namespace prefix = st1 ns = "urn:schemas-microsoft-com:office:smarttags" />Palm Springs’ school teacher with an unexplained severe neurological illness. In spite of a positive signal using a newly developed molecular probe assay (polymerase chain reaction or PCR), there was no inflammatory reaction (the accepted hallmark of an infectious process in an immunologically competent patient). By 1991, I had managed to culture a cell damaging (cytopathic) virus from a PCR positive patient with chronic fatigue syndrome (CFS). The virus caused cells to cluster, fuse and become foamy from lipid (fat) accumulation. A similar cytopathic virus was cultured from the cerebrospinal fluid (CSF) of a comatose patient with a 4 year history of a bi-polar manic-depressive illness. The second virus isolate was sent to the Los Angeles County Public Health Laboratory which subsequently forwarded the virus to the Californian Department of Health Laboratory.

DNA sequencing of two PCR amplified regions of the virus repeatedly isolated from the CFS patient was performed and the data published in 1994. One region showed partial, but highly significant, matching to a known sequence of human cytomegalovirus (HCMV). The other region could not be matched to HCMV. Cytopathic (cell damaging) viruses were being cultured from both blood and CSF of various other patients with neurological, psychiatric and auto-immune diseases, including autism, amyotrophic lateral sclerosis (ALS), schizophrenia and systemic lupus erythematosus. While some of these viruses showed molecular cross-reactivity with the virus isolated from the original CFS patient, other viruses were seemingly structurally unrelated. This led to the conclusion that stealth viruses comprise a broad grouping of viruses whose characteristic feature is the failure to provoke an inflammatory response in spite of being actively cytopathic in test tube cultures.

This conclusion did not sit well with traditional virologists who were accustomed to dealing with precisely definable entities with known and consistent DNA or RNA composition. Moreover, by focusing on the patient with CFS, I evoked the ire of several prominent investigators who had tried without success to culture viruses from CFS patients. Others researchers were more successful but thought that their findings simply reflected the reactivation of a conventional virus, such as human herpesvirus-6 (HHV-6), rather than revealing the primary cause of the patients’ illnesses. Nor could I satisfy those who wanted a laboratory test that would be specifically diagnostic for CFS and that could readily distinguished fatigued from non-fatigued patients; or at least would not be positive in some seemingly healthy individuals.

I was far more impressed with the clearly abnormal findings, especially when observed in cultures from patients with otherwise unexplained complex illnesses. Potential infectiousness was also suggested in patients in whom family members or even household pets had developed equally complex, yet clinically diverse diseases. Any doubt about the relevance of these findings was removed when cultured infected cells were shown to cause severe non-inflammatory illness in animals. In collaboration with Dr. Tom Glass of the University of Oklahoma, we observed the virus-induced transformation of healthy cats to reclusive, pained, irritable creatures creating bald spots on their heads from constant rubbing. Again, there was no inflammation in the brains or other tissues of these animals, only the foamy, lipid-filled cells that were being seen in the viral cultures.

Extended DNA sequencing of the original virus isolate revealed a region for which there were sequence data of the cytomegaloviruses from various other species, including African green monkeys. The DNA matching to the sequence of African green monkey simian cytomegalovirus (SCMV) was extremely close, indicating an unequivocal origin of the patient’s virus from SCMV. Partial DNA sequencing of the virus from the still comatose patient with a history of a bi-polar illness showed a similar origin from SCMV.

Having worked with the Food and Drug Administration (FDA), I immediately realized that the SCMV-derived stealth viruses could have entered into the human population from monkeys used to produce live polio virus vaccines. I called the FDA with my results and learned that they were still using African green monkeys to produce polio vaccines. I spoke of my work at an FDA meeting in June 1995 attended by vaccine manufacturers. I also wrote to the President of Wyeth about my research. I petitioned both FDA and CDC with unsolicited proposals to test polio vaccine lots and to request funding to continue DNA sequencing. Both proposals were dismissed. I presented my findings again to an Institute of Medicine meeting in November 1995. A lawyer immediately filed a legal suit against Wyeth for me to receive the polio vaccine lots administered to some of his clients with neurological diseases. He knew of an ongoing challenge by the mother of a stricken child who had surprisingly won a court order (resisted by both Wyeth and FDA) to have the vaccine she had received tested for human immunodeficiency virus (HIV). The court order specifically excluded any additional testing, including testing for monkey viruses. The lawyer involved in this case had recently arranged for me to receive a blood sample from the child for stealth virus testing. The lawyer could use a positive test result to seek an expansion of the scope of the court mandated testing on the polio vaccine lot administered to the child.

These events were too much for the Establishment. On November 15th, I was told by my University Department Chairman that he had closed my laboratory, dismissed all of the volunteers, and taken the more than $22,000 from my research account. The money he said had gone for back pay and pay-in-lieu of notice for the licensed medical technologist. The funds had been in a USC Gift Account specifically earmarked to trace the possible SCMV origin of cultured stealth viruses. I strongly protested the misappropriation of the patient donated funds and refused to sign a waiver of the University’s action. I knew the Department Chairman had received major funding from a subsidiary of Wyeth and felt suspicious that his abrupt actions were essentially in response to industry pressure. The blood sample from the sick child arrived on November 17th. I managed to store it and showed it was culture positive by temporarily spending time in the Los Angeles County Public Health Laboratory. I subsequently learned that the child had a brain biopsy. Although a diagnostic quandary to his treating physicians, it showed the vacuolated, foamy cells that I had come to recognize from earlier patients and from the virus inoculated cats. The child further deteriorated following the surgery and died.

I took leave from USC in January 1996 when confronted with demands for an additional $15,000 before my laboratory would be reopened. This amount was said to reflect the University’s overhead on the money disbursed from the Gift Account. It was also made clear that I could not resume any patient related testing. Under considerable duress, I eventually located a laboratory in a closed, bankrupt hospital and moved what equipment I could claim as my own into this facility. USC held firm to their insistence that any equipment I had purchased from prior grants belonged to the University.

Assisted by Zaki Saluhuddin, the discoverer of the HHV-6 virus, and by other volunteers, underpaid technicians, and a medical technologist, I was able to pursue the task of understanding the nature of stealth-adaptation and focus on how the diseases might best be treated. The laboratory was designated the Center for Complex Infectious Diseases or CCID. It was licensed under the Clinical Laboratory Improvement Act (CLIA) without any deficiencies cited in inspections held in 1996, 1998 and 2000.

Zaki Saluhuddin left in 1998 but not before he had independently validated the stealth virus testing in double blinded studies. Similar double-blinded validations were independently performed by Dr. Robert Gan, Ph.D., Russell Collins, CLS, and on various occasions by myself (with the help of an assistant to code the samples). As we all knew, positive tests were neither disease specific nor confined to patients with severe illnesses. Still a positive test was clearly and unmistakably abnormal. As an example, I recall in 1996 showing a positive culture to the technologist in the Virology Section of the Los Angeles County Public Health Laboratory. She immediately recalled seeing a similar cytopathic effect from a much earlier culture. As she retrieved her records it turned out to be the sample sent to her in 1991 from the comatose patient with the history of a bi-polar manic depressive illness.

DNA sequencing data continued to substantiate the model of stealth-adaptation as the loss or mutation of the relatively few genes that code for components that are recognized by the cellular immune system. Most virologists have yet to appreciate that since individual lymphocytes recognize distinct antigens, effective immune recognition of virally infected cells requires multiple copies of a few viral antigens at the cell surface rather than a plethora of many antigens that would impede effective lymphocyte binding. For example, approximately 60% of the anti-human cytomegalovirus cytotoxic T lymphocytes recognize the protein coded by the Unique Long segment gene number 83 (referred to as UL83). Twenty-five percent recognize UL55 and another 10% the UL123 protein. Deletion or mutation of these three genes and one can arrive at a virus that will not be effectively recognized by the cellular immune system. Stealth-adapted viruses show additional properties including the apparent capacity to assimilate other genes that can include genes of bacterial origins and cellular genes that can potentially cause cancer.

It is difficult to exaggerate the potential Public Health significance of the stealth-adaptation process to the microbial threat posed to mankind. The production of live vaccines, especially in tissues from foreign species, such as the use of African green monkeys to produce live polio vaccines, could clearly facilitate stealth-adaptation. Armed with an insider’s knowledge of the vaccine industry and the influence it can have on public policy, I continue to believe the message will eventually get through. On each trip to Washington or Atlanta, I make a point of visiting FDA or CDC, respectively. The following unanswered e-mail sent August 2000 to the then CDC Head of Herpesvirus Research (Dr. Phil Pellett) following a herpesvirus conference is typical:

"Dear Phil,

Thank you for the discussion during the last evening of the International Herpesvirus Workshop. You were willing to talk "bluntly," yet in a constructive manner, regarding CDC shunning of my research. As you said, CDC administrators look to you for scientific judgment on matters relating to herpesviruses. Without your support, there is little chance of any response to my requests that CDC pursue what I perceive to be a major Public Health problem. As I recall, the major points of our discussion were as follows:

You spent approximately 45 minutes at my poster and came away with the impression that some of the sequence data must be incorrect. You were concerned that sequence homology matching should be more uniform and not differ along a stretch of nucleotides. You asked if all of the sequencing had been fully double-stranded and whether I had reviewed all of the primary data. I indicated that most of the extended sequencing had been performed by Lark Technologies, at Houston Texas. While there was some internal overlapping, the sequences were primarily derived from one-way reactions. While, there is the possibility of an occasional nucleotide error, this would have had no effect on the conclusions that I was drawing from the data. I understand that you hold your own sequence-related studies up to a particularly rigorous standard, but this has more to do with the types of conclusions that you are trying to draw, rather than justifying dismissing any sequences that are not verified by double-stranded confirmation. Most of the sequences have been on GenBank for a long time and can be reviewed directly by anyone who's interested. The irregular matching that you noted is indeed interesting and goes along with an earlier publication on the genetic instability of the virus.

You also suggested that real science ought to be obvious to the intelligent scientist and that it was my responsibility to present the work so that it would be more widely accepted. Again I disagree. Most scientists are pretty fixed in their belief system, and historically any shift in a prevailing paradigm is met with resistance. The average scientists can not be expected to plow through loads of someone else's raw data, or as you said "interpret my data for me." The CDC is something of an exception, however, since its mission is to be vigilant for possible threats of emerging infections. The poster provided a good opportunity for a scientific discussion, but you chose to view it in my absence. I, therefore, do not know if you fully understood and appreciated the significance of what was being presented.

I was surprised by your suggestion that I sought a chance to discuss my work at CDC as a "cheap" way of claiming CDC recognition. I view my challenge as primarily to get CDC to listen and to take some action. I hope you will continue to provide some assistance by engaging in more meaningful discussions of actual sequence data and patients' histories. In particular, I would like to know your responses to the following issues that I have raised.

1. Do you doubt that the virus for which I have extensive sequence data, was derived from an African green monkey simian cytomegalovirus.

2. Do you doubt that the virus came from a patient, or that it can induce severe illness when inoculated into cats.

3. Are you convinced that the virus has some unusual sequences that at least qualifies it as being an atypically structured virus.

4. Do you feel the apparent absence of UL83 and UL55 related genes could provide the virus a way of evading recognition by the cellular immune system.

5. Are you willing to accept that the virus has recombined with cellular sequences, including the CXC chemokine coding gene, melanoma growth stimulatory activity, a potential oncogene.

6. Do you see any significance in the apparent amplification of the US28 chemokine receptor coding gene.

7. Do you accept the presence of bacteria-derived sequences within the viral genome.

8. Are you aware of the high proportion of patients with unexplained encephalitis-like illnesses, including cases in which brain biopsies have been submitted to CDC for review.

9. Given our positive tissue culture findings in such patients, can you dismiss the possibility that these patients are infected with atypically structured viruses.

10. Don't you think we owe it to those responsible for the Nation's Public Health to have some of these topics more openly discussed?

Enough questions for now. Most of the papers dealing with stealth-adapted viruses are on the web site www.ccid.org I hope we will continue to dialogue and thanks again for the time provided in Portland.

Kind regards, W. John Martin, M.D., Ph.D."

The stakes with CDC got higher when I identified a potential linkage between the testing of cytomegalovirus contaminated polio vaccines in Africa and the emergence of HIV from simian immunodeficiency virus (SIV). This connection was based on the increased number of US28 genes in both SCMV and rhesus CMV that provide a receptor for the cellular entry of HIV and SIV. CDC had been storing sera from early polio vaccine studies in the Congo. Analysis of these stored sera could show that the vaccine used in Africa was indeed contaminated. Complacent bureaucrats are reluctant to bring skeletons out from the closets, such as the 1972 study showing that all 11 African green monkeys tested jointly by Wyeth and FDA were infected with SCMV, or the CDC employee who told me of CDC culturing SCMV from a recipient of an experimental rubella vaccine.

Doing scientific battle to learn some of Nature’s secrets is fun and uplifting. Interacting with Government officials can be frustrating, but at least the Government is in the position of achieving major Public Health changes. Assisting patients and especially children with baffling illnesses is a rewarding privilege especially when the promise of research justifies continuing optimism. On the other hand listening to patient support group advocates wanting to promote their own financial needs, and having to deal with a few dishonest, vindictive patients seeking some perceived entitlement for their sorry state, are decidedly negative experiences. I refused to perform a stealth virus culture on a patient who had been previously tested at USC. The patient was intent on using a positive result to justify receiving ganciclovir, a potentially toxic anti-viral drug, and yet was not being seen by an infectious disease specialist. There were also the confounding issues of possible Lyme disease, babesia infection, multiple previous diagnoses and anger at the medical system’s failure to provide effective therapies.

Seemingly based on prior CDC communications and triggered by this disgruntled patient, the California Department of Health conducted a CLIA inspection of the laboratory in 2002. My detailed responses to a series of stated deficiencies were deemed inadequate and the matter was referred to the Center for Medicare and Medicaid Services (CMS), the Federal body that issues CLIA certificates. They suspended the CLIA License. I did not appeal the finding mainly because a failed appeal leads to a two year disqualification rather than a suspension. In addition, I could not get a straight answer whether a cytopathic effect seen in culture would ever be accepted as adequate evidence for the presence of a reportable abnormality. As indicated in the following series of letters sent to CMS, I wholeheartedly tried to use the inspection process to raise the Federal Government’s Public Health concern that our Nation is in the midst of a serious epidemic.

Copies of all letters (sent on CCID letterhead) to CMS regarding CLIA inspection

July 24, 2002

Ms. Karen Fuller,

CMS, DSO/CLIA Division

75 Hawthorne Street, Suite 408

San Francisco, CA 94105

Fax 415-744-2692

Dear Karen,

In good faith, I provided the California Department of Health and your Office a comprehensive response to a Statement of Deficiencies. My response was received by the State on Monday afternoon. Yesterday, I was notified by Fax that my response was unsatisfactory and a recommendation for sanctions has been forwarded to your Office. I trust your Office will not be so quick to judgment.

I would like to work within the system to "establish the existence of atypically structured cytopathic viruses and to demonstrate a culture method for their detection." In this regard, I have reviewed the Inspector General Report on CLIA Regulation of Unestablished Laboratory Test. I realize that CMS does not validate tests and I am not asking you to do so. What I would like, however, is to have your support and guidance in my interactions with Los Angeles County Public Health Laboratory, CDC and CMS.

The Nation's health is our common goal. Before you take any action, I would appreciate an opportunity for an introductory phone conversation. I would also like to contact the author of the Inspector General's report. Some years ago it was suggested that I apply for a "Demonstration Project" with HCFA. I will also see if I can present the work at CDC and NIH. It would be encouraging if some of these efforts were to become part of your determinative process. I look forward to contacting you later today.

Sincerely,

W. John Martin, M.D., Ph.D.

Laboratory Director

July 27, 2002

Ms. Karen Fuller,

CMS, DSO/CLIA Division

75 Hawthorne Street, Suite 408

San Francisco, CA 94105

Dear Karen,

I had a pleasant and constructive telephone conversation with Ms. Karen Nickel, Head of the Laboratory Field Services of the California Department of Health. She kindly explained that repeated complaints had been received by the Health Department concerning CCID and in particular the testing for stealth viruses. The complaints came from CDC, certain State Health Departments and individual physicians. The complaints were essentially that patients were being misled into believing they were infected with viruses that did not exist. The California Department of Health was in no position to question the opinion of CDC or to assess how the testing was being used, or possibly abused. Staff shortage prevented action being taken last year, but with continuing complaints, closure of the laboratory was given a higher priority this year. Consequently, the California Department of Health undertook a Laboratory Field Service inspection with a predetermined outcome.

I was not surprised by this forthright explanation. Clearly I need to address the concerns of CDC. For several years, I have been trying to engage CDC in a scientific evaluation of my work. I believe the data are irrefutable, but they obviously can be ignored. In March of this year, I presented a Poster at a CDC sponsored International Workshop on Emerging Infectious Diseases. Disappointedly, it did not evoke further inquiry. I had prepared more data for an International Herpesvirus Workshop that I was unable to attend because of the limited time allowed to respond to the inspection report. At this stage, I am seeking the opportunity for an informal seminar at CDC at which I could present the data and answer questions. I would also like to demonstrate the culture techniques to laboratory technologists.

CMS is in a justified position of helping me by asking for CDC input as to whether my data "establish the existence of atypically structured cytopathic viruses." I really want to get CDC, FDA, NIH and others involved so that I can pursue hopeful leads regarding potential therapies. I am including a copy of the poster that I had presented last March.

Sincerely,

W. John Martin, M.D., Ph.D.

cc. Karen Nickel

August 7, 2002

Ms. Karen Fuller,

CMS, DSO/CLIA Division

75 Hawthorne Street, Suite 408

San Francisco, CA 94105

Dear Karen,

A serious criticism expressed by the State Examiner related to her interpretation that what she observed in some culture tubes was overgrowth of the cell sheet, or inoculum or reagent toxicity rather than viral cytopathic effect (Tag D6115). This deficiency was highlighted in another citation referring to the lack of confirmation of a positive result with acridine orange or secondary and tertiary culture or other method (Tag D6087). It also related to the issue of verification in Tag D6086, in which the examiner distinguishes between a result being reproducible and being accurate. She cited the laboratory "for not ensuring that testing methods (serology, PCR, electron microscopy, etc), that were described in the patient report, were used to differentiate 'Stealth Viruses' from conventional viruses and to individually characterize different 'Stealth Virus' isolates. Also, that those tests were (not also) performed by an outside laboratory to verify the patient 'Stealth Virus' result."

I fully appreciate the distinction between claiming isolation and characterization of a specific virus, as opposed to describing the development of a cytopathic effect, presumably caused by a virus. I have shown the characteristic cytopathic effect that I routinely observe to various expert virologists, such as Zaki Salahuddin (the discoverer of HHV-6), Luc Montagnier (the discoverer of HIV) and others. They agreed with my own opinion that the cytopathic effect was presumably viral. I have included photographs of positive cultures in several peer reviewed publications and meeting presentations without pre or post publication objections. I have submitted several isolates to the American Type Culture Collection (ATCC). They confirm viral growth prior to placing material in long term storage. I have retrieved several aliquots from ATCC and have experienced no difficulties in re-establishing positive long term cultures.

There are important Public Health implications if a viral cytopathic activity can be summarily dismissed by qualified medical technologists, such as the Examiner. In response to the Inspection Report, I proposed having at least six positive cultures periodically reviewed by an experienced outside virologist. I am also seeking the opportunity to demonstrate the culture techniques to CDC and other Public Health officials. I was hopeful that CMS would retain an open opinion on this critical issue until I could provide input from such authorities.

For some cultures, I do have extensive DNA sequence data and numerous electron micrographs. I did feel that these data justified the statement that techniques such as serology, PCR, electron microscopy, etc can (or could) be used to differentiate among different stealth virus isolates. I have never inferred however that these sophisticated techniques were anything other than research methods. I have never issued a clinical report with the results of serology, PCR, or electron microscopy. Nevertheless, I am more than prepared to delete all reference to such techniques in subsequent reports.

The extent of cellular abnormalities seen in positive cultures is far beyond any of the changes ever seen in an uninoculated culture. I have taken a series of illustrative photographs as part of an ongoing validation study. I will send you and the State copies by regular mail for review. As stated in my previous correspondence, uninoculated cultures were included in each experiment with written documentation of a lack of cytopathic activity included in the sterility quality control that was performed on each batch of MRC-5 cells received.

Sincerely,

W. John Martin, M.D., Ph.D.

Laboratory Director

cc California Department of Health

August 11, 2002

Ms. Karen Fuller,

CMS, DSO/CLIA Division

75 Hawthorne Street, Suite 408

San Francisco, CA 94105

Dear Karen,

The argument for Immediate Jeopardy was the conviction that patients were being harmed. This conviction was prompted in part by complains received by the California Department of Health from CDC and other Public Health authorities. These authorities doubted the existence of atypical viral infections that I have been describing over the last several years. The immediate threat to Public Health was explained to me as resulting from possible toxicity of medicines prescribed on the basis of a positive test result.

I was observing and reporting on a cytopathic effect with the qualification that it was presumably viral. I had been assured during prior CLIA inspections that this was a reasonable and legitimate statement. Nevertheless, in the most recent CLIA inspection, the point was made by the examiner that the "findings were consistent with overgrowth of the cell sheet, reagent or inoculum toxicity, rather than viral cytopathic effect indicating that test procedures were not properly verified" (Tag D6115). This opinion naturally led to concerns that I had not documented a lack of toxicity in the uninoculated tubes on each occasion that I had read a positive culture. It also raised a question regarding the possible toxicity of the Ficoll-Paque reagent. Finally, details of the double blind assays certified by Robert Gan and Russ Colins were brought into question, as was the subsequently provided affidavit of Zaki Salahuddin. To establish her case, a wide range of other issues were mentioned in the Examiner's report.

I have responded by restating the argument that I was indeed reporting on a cytopathic effect that, based on an extensive amount of data and on the opinion of acknowledged experts, can reasonably be ascribed as presumably being viral. I have re-examined a number of specimens to demonstrate the reliability and reproducibility of the assay. I have shown features in positive cultures, such as the formation of lipid crystals, ribbon-like structures and pigmented inclusions, that can not be dismissed as artifacts of simple toxicity. I have also sought the opportunity to share samples and to demonstrate culture techniques with State and Federal Public Health laboratories.

I know of no patient who has received therapy solely on the basis of a culture result. The medications that are being prescribed are based primarily on the patients' symptoms and the experience of the prescribing physicians. None of the various medications being used is considered particularly toxic. Moreover, patients will continue to be treated whether or not pre and post therapy cultures are performed. There is really no immediate jeopardy to the nation's health.

I trust you will allow me the opportunity to continue efforts to engage CDC in a scientific dialogue regarding the existence of atypically structured cytopathic viruses. As requested in my previous communications, I would appreciate if either CMS or the State could second my request for an expedited meeting to discuss the scientific data that I have compiled and to demonstrate the cytopathic effect. While I cannot force the federal government to join me in the pursuit of a viral cause of many complex neurological and neuropsychiatric illnesses, I can expect protection from a capricious and unjustified stifling of the work. I firmly believe that imposing sanctions that will prevent the continued examination of patient samples is an inappropriate exercise of the CLIA authority entrusted to CMS. I hope that I can continue to work co-operatively with you and your staff.

Sincerely,

W. John Martin, M.D., Ph.D.

Laboratory Director

cc California Department of Health

August 11, 2002

Ms. Karen Fuller,

CMS, DSO/CLIA Division

75 Hawthorne Street, Suite 408

San Francisco, CA 94105

Dear Karen,

I am pleased to enclosed both the protocol and the results of a study that validates the existing cytopathic assay conducted in my laboratory. I have maintained that processed blood from certain patients is able to produce a characteristic cytopathic effect (CPE) in MRC-5 cells. The CPE is characterized by the normal spindle shaped, translucent, closely packed MRC-5 cells becoming enlarged, rounded and fusing into small, and later into larger, three dimensional cell syncytia and clusters. The cellular cytoplasm displays a vacuolated, lipid filled appearance, often accompanied by fine, darkly pigmented inclusions. In many stealth virus cultures the excess production of lipid is so marked that it deposits as extra-cellular crystals. The pigmented material can also take on the form of ribbons and large aggregates. Neither the formation of extra-cellular lipids or other structures can be easily explained on the basis of a simple toxic reaction of the cells.

The validation test consisted of 41 culture tubes. Ten of the tubes were inoculated with stored aliquots of processed blood samples that were previously shown to give a positive CPE. Ten of the tubes were inoculated with stored aliquots of processed blood samples that did not previously cause a positive CPE. Ten samples came from previously untested blood samples provided by the University of California Irvine Blood Transfusion Service. Ten tubes were designated as no-sample or uninoculated controls. Finally, as in all routine studies, a uninoculated control, designated as a Blank tube was included. X Vivo-15 medium was used for all of the tubes. Incubation, feeding and viewing of tubes were performed throughout the experiment in an identical manner without regard to the original inoculum. The tubes were examined daily by myself and independently looked at by Russ Collins, CLS at day 5. I also took photographs of all of the cultures. The results can be summarized as follows:

There was no difficulty in distinguishing a positive from a negative culture

All of the samples that previously tested positive again showed the development of the characteristic CPE

None of the samples that previously tested negative showed the development of a CPE

Three of the blood donor samples did develop a positive CPE, one of which (tube #17) was quite striking

None of the uninoculated tubes (without an added processed blood sample) showed any CPE

The control blank tube also did not show a CPE.

There was a 100% concordance with my day 5 readings and those of Mr. Russ Collins, CLS

None of 20 previously left over tubes from earlier time points showed any signs of the characteristic CPE.

I have typically saved an aliquot from many of the samples that I have tested over the last several years. I can repeat the study at any time intervals that you feel is appropriate. I can also send both positive and negative aliquots of previously tested processed blood samples to an outside laboratory. Among the selected photographs of positive cultures, several show the types of lipid crystals commonly seen in positive cultures. Other photos show the ribbon-like structures and the pigmented aggregated inclusions. I hope this information will forestall the imposition of sanctions, so that I can learn to better understand the clinical importance of such striking findings.

Sincerely,

W. John Martin, M.D., Ph.D.

Laboratory Director

cc California Department of Health

August 14, 2002

Ms. Karen Fuller,

CMS, DSO/CLIA Division

75 Hawthorne Street, Suite 408

San Francisco, CA 94105

Dear Karen,

I would like to correct the impression given to Dan Hersh that no one had confirmed my findings. As with much of science, technologists can observe the same phenomena but interpret the findings differently. The Wisconsin Viral Research Group, Ltd., offers a Rapid Culture Assay for human herpesvirus-6 that is essentially similar to what I do. The difference is that they stain the cultured MRC-5 fibroblasts with polyclonal antibodies reactive with HHV-6. I have spoken with their virologist and explained that stealth virus cultures will typically stain with a variety of polyclonal anti-viral antibodies, including anti-HHV-6 and anti-CMV. They have their proprietary reasons for wanting to promote their assay as a test for HHV-6, especially since they argue that from 40-70% of chronic fatigue syndrome (CFS) patients are positive for HHV-6 by culture.

I have shown the CPE to several bench technologists. It is not that they have not seen the changes, it just that they did not know how to interpret the changes. The waning of the CPE in infrequently fed cultures has also encouraged a disregard for the changes. Some other patients have been told they have an atypical CMV infection.

Additional support for an involvement of atypical herpesviruses in CFS patients is provided by clinical trials that use anti-herpesvirus drugs. For example a patent was recently issued to Dr. Martin Lerner on valganciclovir therapy in CFS patient. A brief summary of the patent is as follows:

United States Patent 6,399,622; Lerner June 4, 2002.

Method for diagnosing and alleviating the symptoms of chronic fatigue syndrome

Abstract

A method for alleviating chronic fatigue syndrome with the administration of antiviral agents. Based on clinical tests, chronic fatigue syndrome is a persistent herpes virus infection including incomplete virus multiplication and thus administration of antiviral agents are shown to alleviate the symptoms associated with the disorder. Based on therapeutic trials, patients receiving the recommended antiviral treatment, have experienced significant reduction or elimination of the symptoms associated with chronic fatigue syndrome. A method of diagnosis of the chronic fatigue syndrome is further disclosed.

I also spoke today with Dr. John Stewart of CDC. He is circulating my poster among their virologists. Given the current concern about West Nile virus, I suspect there will be strong interest in my work. It is known for example that herpesviruses can be activated by West Nile virus and that dual infection is more serious than either infection alone.

I trust this information is helpful in your deliberations and that you will not abruptly close CCID clinical laboratory. I will work diligently with you and the State if their are remaining Condition-Level deficiencies or concerns.

Sincerely,

W. John Martin, M.D., Ph.D.

Laboratory Director

cc California Department of Health

October 12, 2002

Ms. Karen Fuller

Acting Director, CLIA Operations

Center for Medicare and Medicaid

75 Hawthorne Street, Suite 408

San Francisco CA 94105

Dear Karen,

I am responding to your letter dated October 9, 2002. I did not pursue the option of appealing your determination primarily because of the punitive consequence of an unsuccessful appeal.

Culture results from CCID have only been recorded and reported from cultures that were completed prior to August 27, 2002. In anticipation of reestablishing a diagnostic service, that I truly believe will be of potential Public Health value, I would appreciate clarification on the following issue: The phase "all patient testing operations" appears to me to be broader than the stipulation of allowing to "test human specimens but do not report patient-specific results for the diagnosis, prevention, or treatment of any disease or impairment of, or the assessment of the health of, individual patients." Especially given a challenge of needing to submit "a credible allegation of compliance and … acceptable evidence of correction" the opportunity should exist for some additional testing. Specifically, could you please clarify for me which if any of the following are acceptable practices given the current suspension of the CLIA certificate.

1. Testing my own blood sample.

2. Testing stored frozen aliquots of previously collected samples.

3. Testing a fresh sample submitted by a physician, obtained from either him/herself or donated by a patient, in which a clinical diagnosis might be provided, but for which no verbal or written report would be issued.

4. Testing donated samples submitted by a physician for which a verbal report could be discussed with the physician, but with the physician’s written assertion that the information is not being used for any purpose related to the patient’s medical care, and with the added assurance that the information would not be communicated to the patient.

5. Testing unlabeled samples from blood banks given the appropriate Institutional Review Board (IRB) and donor approvals with, of course, no reporting of results to the blood bank.

6. Testing unlabeled samples from a hospital or clinical practice with no reporting of results.

7. Participation in a research program for which an IRB has approved testing of samples at CCID.

8. Testing samples requested by a patient or his/her attorney for forensic purposes.

9. Testing samples collected from animals.

The way the Attestation is written is somewhat confusing and possibly open to misunderstanding. I have noted the enormity of the penalty and have no interest in violating CLIA laws. May I ask, however, for you to incorporate your responses to the above questions into a more clearly defined use of the term "patient testing" in line 4 of the Attestation. I am not a lawyer and will still probably need to have some help in interpretation of the revised document. I will be in Washington D.C. for the week of October 20th and will be spending most of this week preparing for this trip. If possible, I would like to delay final signing of the attestation till the following week. No experiments will be performed in the interim period. The web site is managed by a volunteer. I have conveyed to her the need to make additional changes to the web site in accordance with your instruction. For various reasons, this too may take her several days.

Sincerely,

W. John Martin, M.D., Ph.D.

3-3-2003

 Ms Karen Fuller,

Manager

State Oversight and CLIA Branch

Division of Survey and Certification

Centers for Medicare and Medicaid Services

Department of Health Services

San Francisco Regional Office

75 Hawthore Street, 4th Floor

San Francisco, CA 94105

 Re: Your response dated February 20, 2003

Dear Karen,

I read your response on March 1, 2003, having been away from the laboratory during the week. I fully understand that "a laboratory that reports any patient-specific results, either in writing or verbally, must have a valid CLIA certificate in effect." I note CLIA’s unwillingness to accept statements that "no verbal or written report would be issued," or that test results "would not be communicated to the patient or their physician." There is apparently an underlying presumption that having access to a patient identity is essentially equivalent to relaying results of any research testing to the patient or to the patient’s physician. You have my assurance that I wish to remain in full compliance with all CLIA mandated laws. I have not reported any results to patients or physicians for any testing performed since suspension of the CLIA license. Nor have I maintained any patient identification records for the few samples that were occasionally delivered to the laboratory since the CLIA suspension.

Sincerely,

W. John Martin, M.D., Ph.D.

Declaration of Mr. Russ Collins, CLS

I, Russell Collins am a duly licensed clinical laboratory scientist. I have known Dr. W. John Martin for over 5 years and have been a staff member of the Center for Complex Infectious Diseases (CCID) since September 1999. I can attest to the following facts:

CCID has conscientiously and reliably performed clinical laboratory testing on human and on occasional animal derived blood samples for evidence consistent with the presence of a viral infection, of the type referred to as a stealth-adapted virus. The primary indication for the presence of such viruses is the ability of the sample to induce characteristic cellular damage in cultured fibroblast cells. The type of damage being sought was the development of foamy vacuolated cells that form clusters, and that commonly become pigmented. This type of change occurs with known stealth-adapted viruses, including the well characterized virus that Dr. Martin had previously isolated from the patient with initials D.W.

I have personally performed numerous stealth virus cultures that were requested by Dr. Martin and other physicians. Most of the testing was done on blood samples obtained from patients, but on occasions were done on control individuals, including blood donors. I have also tested several blood samples from patient D.W.

I have reported and personally signed the results of the cultures that I have performed to the requesting physicians.

A positive report indicated that I observed the development of a very characteristic cell damaging (cytopathic) effect. This effect is consistent with the presence in the blood sample of a viral agent of the type defined as a stealth-adapted virus. I reported this interpretation to the requesting physician.

A negative report indicated that I did not observe the development of a cell damaging (cytopathic) effect. For these samples, therefore, I reported to the requesting physician that there was no evidence for the existence in the sample of a viral agent of the type defined as a stealth-adapted virus.

In doing these cultures, I strictly followed the detailed procedure manuals maintained at CCID, including any revisions approved by Dr. Martin in consultation with myself and other members of the laboratory. These revisions essentially involved the process of better documentation of the rapidity of onset and intensity of the cytopathic effect.

I have, to the extent required of providing a highly reliable efficient clinical laboratory, diligently followed all procedures and maintained all needed records, as well as followed other requirements expected of the laboratory.

On numerous occasions, I would review cultures with Dr. Martin and confirm that we were in full agreement, not only in distinguishing a positive from a negative culture, but also in grading a positive culture as having a mild, moderate or severe degree of cellular damage.

On regular occasions, Dr. Martin and I would separately record our findings and subsequently review and discuss the very occasional occurrence when we disagreed as to the severity of a cytopathic effect. I can recall no instance of our disagreeing as to whether a culture was positive or negative.

I also independently conducted double blind studies in which blood samples from a group of blood donors not known to be ill were randomly assigned numbers and mixed with blood samples of patients that were assigned the remaining numbers. Without knowing the source of the blood samples, I, and on occasions, Dr. Robert Gan and/or Dr. Martin, would examine the cultures and independently record our findings and interpretation. In each of several double blind studies, the occurrence of a positive culture among the blood donors occurred at a frequency far less than among the patients. Moreover, my results and interpretations were essentially identical to those of Dr. Gan and Dr. Martin. In my opinion, the assay was validated by these double-blind studies.

In performing patient related testing, I can confidently assert that all studies were performed with a very high degree of diligence and attention to detail. I take my responsibilities as a licensed medical technologist very seriously and recognize the importance of timely, accurate and reliable clinical results. I also recognize that stealth virus testing is breaking new grounds with regards to understanding the cause and the manifestations of chronic persisting fatiguing illnesses. I have spent many hours listening to patients describing their illnesses. I have also had regular telephone contact with the patient D.W. and have cultured blood samples from her on several occasions. In my opinion, the positive blood cultures seen in many of the patients can provide a likely explanation of their illness.

I have observed Dr. Martin performing cultures and discussing results with physicians and patients. He is a very dedicated, conscientious individual. His primary concern is to learn more about the viruses he has identified so that he might be of more assistance to virally infected patients. He has made numerous original observations on the cultures which he readily shared with me and asked for my input. Together, we have gained an in-depth understanding of the series of events leading to a positive culture and the role of inhibitory factors in suppressing the cytopathic effects seen in infrequently fed cultures. Dr. Martin has termed such factors "Epione." He has frequently expressed the belief that characterization of Epione will provide a means of potentially curing stealth virus infected patients.

I am also aware that running the laboratory has cost Dr. Martin considerable amounts of money. He feels compelled to pursue his passion.

I participated in a laboratory inspection conducted by the State of California inspectors in March, 2002. In my opinion, neither Dr. Martin nor I were provided the opportunity to explain or to fully demonstrate the culture methods. I reviewed the citations from this inspection and found many of them to be unreasonable and somewhat inappropriate. Nevertheless, along with Dr. Martin, I tried to answer each of the cited deficiencies and helped perform yet another double-blind study, the results of which were submitted to the State in August 2002.

As did Dr. Martin, I fully complied with the directive from CMS and the State that we cease to perform patient care (or physician reported) testing as of late August 2002.

In addition, to patient care related testing, I also participated with Dr. Martin in numerous research studies on positive stealth virus cultures. These studies involved identifying crystals and pigmented structures that would never be expected to occur in normal cultures.

Sworn to on this day of February, 2003 at Rosemead in the State of California, under perjury.

End of Letters to CMS

Upon reflection, I was spending so much time doing the cultures that I was not doing justice to the observations that had already been made. I set about writing two overdue manuscripts on alternative cellular energy pigments (ACE-pigments). I also set about learning enough physics to call myself a biophysicist. I began to meet a wonderful array of individuals, including the gentleman who kindly provided the MI-Hope non-profit foundation. It became the BioPhysics Institute and then S3Support. Other individuals have each contributed to an understanding of stealth viruses that I would probably not have gained from continuing with diagnostic cultures. More importantly, I now see a clear rationale for well designed "energy medicine" based clinical trials. Possibly only when a therapy is proven will our Public Health system be willing to address the likelihood of a widespread infectious component impacting on our Nation’s health. For now, I feel patients should at least expect the Government to be performing stealth virus cultures. The methods are fully described in various publications and the original stealth virus has been available from the American Type Culture Collection (ATCC) since 1992. There is a serious epidemic and it needs to be officially addressed.

W. John Martin, M.D., Ph.D.