Guest guest Posted September 23, 2003 Report Share Posted September 23, 2003 What can jump massive cell junctions in a single bound?Dodge " bullets " faster than the speed of light?Is it a bird?Is it a plane?Is it Superman?no!!!It's HIV!!!! http://www.virusmyth.net/aids/data/ehremarks.htm REMARKS ON METHODS FOR RETROVIRAL ISOLATION By Etienne de Harven Continuum Spring 1998 Dr. Etienne de Harven is emeritus Professor of Pathology, University of Toronto. He worked in electron microscopy (EM) primarily on the ultrastructure of retroviruses throughout his professional career of 25 years at the Sloan Kettering Institute in New York and 13 years at the University of Toronto. In 1956 he was the first to report on the EM of the Friend virus in murine (mouse) leukemia, and in 1960, to coin the word " budding " to describe steps of virus assembly on cell surfaces. He will deliver a speech at the 12th World AIDS Conference in Geneva (June 28-July 3) at the session " HIV-testing: Open Questions about Specificity " . The most impressive developments of molecular genetics over the past 20 years do not make Robert Koch's postulates obsolete. The first of these postulates indicates that to be considered as pathogenic, a microorganism should be isolated in every single case of the disease. Still, according to E. Papadopulos et all and S. Lanka (2) isolation of HIV from fresh plasma of AIDS patients has never been achieved under any circumstances. Moreover, and most surprisingly, the " efficiency " of current antiviral therapeutic protocols (AZT tri-therapy) is being measured by determining " viral load " in the plasma of treated patients. " Viral load " implies viremia I.e. the presence of circulating viral particles in the peripheral blood. The virus incriminated being allegedly a retrovirus, this would have been the time to remember that the morphology of such viruses in several animal experimental tumors and leukemias had been extensively characterised by transmission electron microscopy (EM) over the past 40 years, the viral particles having a characteristic ultrastructure and a diameter ranging between 100 and 120 um. Some of them had been studied by methods of high resolution transmission electron microscopy.(3) In the 1960s, transmission electron microscopy was by far the best available method to identify viruses within or around diseased cells. Consequently, many cancer research centers all around the world, started to compete for the best equipment and training in EM, aiming at the demonstration in human malignancies of viruses similar to those which had just been recognized as significantly associated with tumors and leukemias of several laboratory animals. This approach to cancer research appeared highly justified when Lwoff, Horne and Tournier proposed to classify all viruses primarily on the basis of their morphological features demonstrated by electron microscopy.(4) Identification of viruses by EM in leukemic animal tissues became unambiguous when steps in virus assembly, i.e. the 'budding' of complete virions from the surface of the infected cells, were described.(5) In spite of considerable efforts, the search for similar, typical viruses in human malignancies remained entirely negative. Pleomorphic membranous microvesicles, approaching viral size, and frequently described in the literature on human malignancies as " virus-like particles " were without any pathogenic significance. As stated in 1965, typical RNA tumor viruses have never been observed in association with human neoplasia.(6) Concentrations of retroviruses from murine and avian leukemic tissue homogenates were reproducibly achieved permitting titration of infectivity into receptive laboratory animals. This was not, however, an easy approach to the problem of virus purification, large amounts of microvesicles and cell debris being usually present. As far as virus purification was concerned, it soon became evident that when viremia is present, blood plasma was far better than tissue homogenates for efficient virus isolation and purification. In the case of RNA tumor viruses, now called retroviruses, the demonstration of viremia in the blood plasma of experimental leukemic animals (chickens and mice) was published more than 35 years ago. A most efficient purification method including ultrafiltration and ultracentrifugation of a 1/1 dilution of plasma in heparinized Ringer's solution, allowed me to demonstrate packed retroviruses by transmission electron microscopy (7) in thin sections of pellets obtained by high speed centrifugation of the purified virus, quite clearly establishing that the amount of contaminating cell debris was remarkably small, a conclusion which could never have been reached by using the negative staining EM method. Using this simple ultrafiltration procedure, virions were never exposed to hypertonic shock. However, sedimentation in sucrose density gradients, at the density of 1.16 gm/ml, soon became the most popular method for retrovirus purification.(8) Interestingly, it was very well known by electron microscopists in the 1960s, that sharp bands sedimenting at the density of 1.16 frequently contained large amounts of microvesicles and cell debris of non-viral nature. These debris just happened to sediment in sucrose gradients at a density very similar to that of retroviruses clearly indicating that finding a " sharp band " at the density of 1.16 gm/ml was of little significance and was certainly far from any demonstration of retroviruses isolation. But this conclusion was based on EM findings, and around 1970 the faith in retroviral oncology was assuming quasi-religious proportions! If EM cannot demonstrate viruses in the 1.16 bands, let us forget about EM and rely on other " markers " ! When around 1980, R. Gallo and his followers attempted to demonstrate that certain retroviruses can be suspected of representing; human pathogens, to the best of my bibliographical recollection, electron microscopy was never used to demonstrate directly viremia in the studied patients. Why? Most probably, EM results were negative and swiftly ignored! But over-enthusiastic retrovirologists continued to rely on the identification of so-called " viral markers " , attempting to salvage their hypothesis. When retrovirus particles are legion, the study of molecular markers can be useful, and provide an approach to quantification probably better than direct particle counting under the EM (which I always found very difficult). But when, using EM, retrovirus particles are absent relying exclusively on 'markers' is a methodological nonsense. 'Markers' of what? Nevertheless, for the past ten years, HIV research and clinical therapeutic trials have been primarily based on the study of several HIV " markers " . First the antibody. Elisa, then Western Blot tests were hastily developed (at sizable financial profit eagerly split between the Pasteur Institute and the US). " Seropositivity " became synonymous with the disease itself, plunging an entire generation into behavioral panic, and exposing hundreds of thousands of people to 'preventive' antiviral AZT therapy which actually hastened the appearance of severe or lethal immunodeficiency syndrome. Appropriate controls were apparently never carried out or were never published. Still, back in 1993 it became clear that the so-called HIV antibody tests badly lacked specificity, (9) cross-reactivity being observed with patients suffering from a long list of pathological conditions including malaria, leprosy, auto-immune diseases and many more. Secondly, 'viral proteins'. Several proteins have been identified as 'HIV markers', most frequently because they were identified in a variety of 1.16 bands. The case of the p24 " viral " antigen is a significant example and its lack of viral specificity has been well documented.(10) Third, reverse transcription. If reverse transcriptase activity were a unique feature of retroviruses, it could have been an interesting molecular marker. Unfortunately, it has been shown that reverse transcriptase is found in the uninfected cells of yeasts, insects and mammals (11) and " has nothing to do with retroviruses as such " as well referenced in a recent report from S. Lanka. Moreover, K. Mullis himself does not support the use - to amplify and quantify the " HIV genome " - which is being made of the PCR methodology he developed, which is the current method of " measuring the viral load " in AIDS patients. More disturbing is the fact that some 'markers' are searched for in the 1.16 gradient sedimenting material which is the density where intact virions are expected to be found, but not their molecular fragments. If lysed retrovirus particles released molecular markers, the 1.16 samples should at least initially allow investigators to demonstrate virus particles by EM. They don't. however after 15 years of most intensive HIV research, two independent groups finally decided to explore by electron microscopy the ultrastructural features of the material sedimenting at the 1.16 density. Working on " HIV-1 infected T-cell " cultures supernatants, both groups found that it contains primarily cellular debris and cell membrane vesicles which could definitely not be identified with HIV particles and rare " virus-like " particles.(12, 13) Still, this is the type of sample in which " viral markers " are currently identified and used to measure the effects of anti-viral drugs in current clinical trials. In conclusion, and after extensive reviewing of the current AIDS research literature, the following statement appears inescapable: neither electron microscopy nor molecular markers have so far permitted a scientifically sound demonstration of retrovirus isolation directly from AIDS patients. This conclusion fully confiens the recent reports published in Continuum by E. Papadopulos and by S. Lanka. Harvey Bialy, editor of the journal Bio/Technology has stated that (14) " A powerful hypothesis has to explain and predict. What kind of scientist continues to support a hypothesis that fails to explain and fails to predict? " The HIV/AIDS hypothesis fails to explain the considerable drop of T4 circulating lymphocytes in AIDS patients. It predicted a dramatic AIDS epidemic which was never observed (unless we accent the CDC's most surprising redefinition of AIDS as including some 31 " AIDS defining illnesses " !). Obviously, the HIV/AIDS hypothesis has to be scientifically reappraised.(15) And, most urgently, the funding for AIDS research should no longer be restricted to laboratories working on an hypothesis which has never been proven. * References 1. Papadopulos-Eleopulos E, Turner VF, Papadimitriou JM, Causer D, Hedland-Thomas 13, Page B, 1994. A critical analysis of the HIV-T4-AIDS hypothesis. Genetica 95:5-24 2. Lanka, Stefan, 1994. Fehldiagnose AIDS? Wechselwirkung, Aachen, December, 48-53. 3 de Harven, E, 1974. remarks on the ultrastructure of type A, B and C virus particles. Advances in Virus Research 19: 221-264. Academic Press, Inc., publ., New York. 4. Lwoff A, Horne R, Tournier P, 1962. Cold Spring Harbour Symposium on Quantitative Biology 27:51. 5 de Harven E, and Friend C, 1960. Further electron microscope studies of a mouse leukemia induced by cell-free filtrates. J. Biophysic. and Biochem. Cytol., vol 7, 747-752. Rockefeller University Press, New York 6. de Harven E, 1965. Remarks on viruses, leukemia and electron microscopy. In Methodological approaches to the study of leukemias. V Defendi, edit., The Wistar Institute Press publ, Philadelphia, pp147-156 7. de Harven E, 1965. Viremia in Friend leukemia: the electron microscope approach to the problem Pathologie-Biologie,13:125-134 de Harven E, 1998. Pioneer deplores " HIV " . Continuum vol 5, page 24 8. Sinoussi F, Mendiola L, Chermann JC, 1973. Purification and partial differentiation of the particles of murine sarcoma virus (M.MSV) according to their sedimentation rates in sucrose density gradients. Spectra 4:237-24 9. Papadopulos-Eleopulos E, Turner VF and Papadimitnou JM, 1993. Is a positive Western Blot proof of HIV infection? Bio/Technology 11:696-707 10. Todak C, Klein E, Lange M et al., 1991. A clinical appraisal of the p24 antigen test. International Conference on AIDS, vol 1, Florence, Italy 11. Varmus H, 1987. Reverse transcription Sci. Am. 257:48-54 12. Gluschankof P. Mondor I, Gelderblom HR, and Sattentau QJ, 1997. Cell Membrane vesicles are a major contaminant of gradient-ennched human immunodeficiency virus type-l preparations. Virology 230:125-133 13. Bess JW Jr., Gorelick WJ, Bosche WJ, Henderson LE, and Arthur LO, 1997. Microvesicles are a source of contaminating cellular proteins found in purified HIV-I preparations. Virology 230:134-144 14. Farber, C, 1992. Fatal distraction, Spin Magazine, June 1992 15. Philpott P, 1997. The isolation question. Reappraising AIDS, vol 5 number 6, 1-12 Author's address: Dr Etienne de Harven le Mas Pitou, 2879 Route de Grasse 06530 Saint Cezaire sur Siagne, France VIRUSMYTH HOMEPAGE -Allen Graves IBWB ( U.S.A. ) " Do not let either the medical authorities nor the politicians mislead you. Find out what the facts are, and make your own decisions about how to live a happy life and how to work for a better world. " - Linus Pauling Sites that I've found to be helpful/informative ; aidscured A good forum for those who are seeking a non-pharmaceutical solution to illness(es). AIDSsoc This forum focuses on the societal aspects of the " AIDS " myth. http://www.aliveandwell.com A more rational, informed perspective on " AIDS " . http://www.virusmyth.com Exposing the erroneous science behind current " HIV/AIDS " thinking. http://www.aidsmythexposed.com Forum with a " WAR On AIDS Science " conscious focus. Sites of " ultimate concern " ; http://www.lucistrust.org/links/index.shtml http://www.lucistrust.org/goodwill/links.shtml Quote Link to comment Share on other sites More sharing options...
Guest guest Posted September 25, 2003 Report Share Posted September 25, 2003 Hi there! AIDS is an Industry, and many patients are religiously compliant in taking their meds. in often complicated regimes. The money involved in the current AIDS strategy is shocking. Interesting how no mention of side effects emerges, and more interesting how accepting many patients are of the disastrous effects. Hydroxurea is known to cause death by heart attacks. One of the protease inhibitors causes the death of the blood supply to the large joints. I know one patient, who has had one hip replacement, is due to have the other hip replaced, as well as needing both knees and a shoulder replaced, because the bone died in those areas. Very strangely, some drugs are causing rectal tears that result in infections, and are usually dealt with surgically -- but can recur. Cosmetic effects are facial wasting, a skeletal effect, and large deposits of fat around internal organs, resulting in " the paunch " . Explosive diarhea is common. The expenses of the most recent conference on AIDS would have likely provided some basic necessities like food and simple health care in the Third World, for persons with AIDS. The whole system is unbeleivably corrupt, self-sustaining, with as much intention of curing AIDS , as the Cancer Industry has of curing cancer and ending the careers and empires built on the suffering of others. Left to the 'Experts', all will continue as it has, with newer drugs and more research money to keep the whole system viable. Steve Frank wrote: >http://www.nytimes.com/2003/09/23/health/23IMMU.html?th > >September 23, 2003Trying to Kill AIDS Virus by Luring It Out of HidingBy DONALD G. McNEIL Jr. > > >ho knows what evil lurks in the lymph nodes of men? > >The immunologist knows. > >But the body may not even suspect it. > >That evil is the AIDS virus, which has the power to hibernate, virtually forever, even in patients taking their triple-therapy cocktails with religious devotion. > >Many AIDS specialists are working on ways to tease the virus out of hiding so it can be killed, and real progress has been made. A laboratory at the University of California at Los Angeles recently reported 80 percent success in mice. > >Even that, however, cannot stop the virus from roaring back. " Eighty percent is close, " said Dr. Roger J. Pomerantz, an AIDS researcher at Thomas Jefferson University in Philadelphia. " But close only counts in horseshoes and hand grenades. " > >Even if the virus, battered by the combination punching of antiretroviral therapy, is found in just one dormant T cell in a million, repeated tests in humans have shown that it will re-emerge if the therapy stops. > >Simply hunting it is a ticklish business, because the virus hides in the same memory T cells that act as triggers for the immune system. Failing to " light up " the virus in them will keep it hidden. But activating too many T cells means a cascade of immune reactions like those in toxic shock syndrome, possibly killing the patient. > >Since beginning their quest 19 years ago with the realization that infected T cells somehow slept in " reservoirs " in the lymph glands, blood and perhaps in parts yet unknown, researchers have experimented with immune-boosting drugs borrowed from cancer therapy and with tiny " smart bombs " that use antibodies to hunt for one cell among billions and kill it with a minuscule dose of poison. > >Despite regular advances, some prominent AIDS researchers worry that the task may be hopeless. > > " It's a very, very, very difficult situation, " Dr. Anthony S. Fauci, director of the National Institute of Allergy and Infectious Diseases, said. " You can get a reservoir down to an undetectable level, and then if someone gets even a little blip of viremia, it can reseed the area, and you're back to Square 1. " > >That " blip of viremia, " a jump in virus circulating in the bloodstream, can be set off even by a bout of flu or a rusty nail, because the AIDS virus can hide in a T cell that normally attacks the flu virus or tetanus. > >Dr. Robert C. Gallo, director of the Institute of Human Virology at the University of Maryland, said many labs had attacked latent virus, with no breakthrough success yet. > >The newest work, at U.C.L.A.'s AIDS Institute, was in mice bred without immune systems in which human thymus cells were implanted and infected. Although he praised the research, Dr. Gallo said the mouse-thymus model was " a very artificial system. " > > " If you had this in a monkey model, with demonstrable safety data, " he said, " that would merit highlighting it. " > >The U.C.L.A. team, led by Dr. Jerome A. Zack and Dr. David G. Brooks, used interleukin-7 and prostratin to light up infected cells. If humans were given big enough doses to reach the most hidden cells, " I don't know that it wouldn't be terribly dangerous, " Dr. Gallo said. > >The AIDS virus is frequently described as wily for its ability to mutate drug-resistant forms. But it is equally wily in its hibernation style, sitting as silent as a nuclear submarine on the ocean floor. Incorporated into the DNA of a shrunken, inactive T cell, the virus is a mere speck of genetic code, 10,000 base pairs out of the 6 billion in each human nucleus. > > " It exists as pure information, " Dr. Robert F. Siliciano, an AIDS researcher at the Johns Hopkins University, said. " The immune system can't see it, because the system sees proteins, and it's not making proteins. The drugs don't touch it, because they stop replication, and it's not replicating. " > >Dr. Siliciano recently demonstrated that the numbers of infected dormant cells remained steady over 7 years and might take 70 years to die off under normal antiretroviral assault. Trying to defeat that, researchers try ever more complicated regimens. > >Dr. Fauci described treating 10 patients with multiple cycles of interleukin-2 to " flush the virus out. " > > " We thought the cells would spit out their virus and die, which would take care of those suckers, and then antiretrovirals would stop proliferation, " he said. > >After a moment of jubilation when he found no virus even in biopsies from his patients' lymph nodes, he went for " the proof of the pudding " and took the patients off their AIDS drugs. > > " Everyone rebounded, " he said. " We failed. " > >Mike Flanagan, 41, an architect, was in an even harsher pharmacological study led by Dr. Pomerantz. He was chosen, he said, because he knew the exact date when he was infected and was very good at taking his pills on time. > >He was hospitalized to begin receiving a mix of eight AIDS and cancer drugs, including antiretrovirals, interleukin-2, hydroxyurea and OKT-3. > > " It was pretty rough, " Mr. Flanagan said. " I got really paranoid, and I had trouble sleeping. " > >After another year as an outpatient, he was told to stop all drugs and wait. > >In three months, " it came back, " he said, conceding disappointment. " A lot of people put a lot of energy into that test, and I felt like I let them down. I thought there was a chance it would work. " > >In the U.C.L.A. study, released last week by the journal Immunity, Dr. Zack described choosing his drugs — prostratin and interleukin-7, to " tickle the cell, just turn the virus on without turning on the cell. " > >A " turned-on " virus, Dr. Zack explained, displays some of its proteins on the surface of the T cell, but does not prod the cell to divide or to wake up other T cells to begin an immune system assault on a foreign invader. > > " Then we come in with the immunotoxin, " he said. It is a molecule-size smidgen of bacterial poison attached to an antibody that gloms onto the T cell and injects the toxin. > >Immunotoxins are too poisonous to use alone against AIDS, but they may work in combination with antiretrovirals, he said. If all goes well, he will try the tactic in monkeys and then humans. > >But, Dr. Zack conceded, killing even 80 percent of the latent virus is probably not enough to finish the job. He may try more " tickling " drugs in combination or he may give the drugs in pulses. " Maybe, " he said, " we can do better. " > >Copyright 2003 The New York Times Company > > >NEW WEB MESSAGE BOARDS - JOIN HERE. >Alternative Medicine Message Boards.Info >http://alternative-medicine-message-boards.info > > > > > > Quote Link to comment Share on other sites More sharing options...
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