theperthgroup.com/CONTINUUM/PapadopolousReallyAchieved1996.pdf
theperthgroup.com/CONTINUUM/pgvsduesbergreward.html
6.1 MINIMUM EVIDENCE REQUIRED TO PROVE THE EXISTENCE OF HIV DNA
If "HIV DNA" is the genome of a unique retroviral particle then the most basic requirement is proof for the existence of a unique molecular entity "HIV DNA", that is, unique fragments of DNA identical in both composition and length in all infected individuals. The claim that a stretch of RNA (cDNA) is a unique molecular entity which constitutes the genome of a unique retrovirus can be accepted if and only if it is shown that the RNA belongs to a particle with the morphological, physical and replicative characteristics of a retroviral particle. Proof of these properties can only by obtained by isolating the putative viral particles, that is, by obtaining them separate from everything else, extracting the nucleic acids and demonstrating that such particles are identical (their constituents including their nucleic acids are identical) and infectious. The correct procedures, now having been used for over half a century to achieve this proof, require demonstration that:
1. In "infected" cell cultures (cocultures) there are particles with a diameter of 100-120nM containing "condensed inner bodies (cores)" and surfaces "studded with projections (spikes, knobs)";(82)
2. In sucrose density gradients the particles band at a density of 1.16 gm/ml;
3. At the density of 1.16 gm/ml these is nothing else but particles with the morphological characteristics of retroviral particles;
4. The particles contain only RNA and not DNA and that the RNA consistently has the same length (number of bases) and composition no matter how many times the experiment is repeated;
5. When the particles are introduced into secondary cultures, but mindful of the critical caveat discussed below:
(a) the particles are taken up by the cells;
(b) the entire RNA is reverse transcribed into cDNA;
(c) the entire cDNA is inserted into the cellular DNA;
(d) the DNA is transcribed into RNA which is translated into proteins.
6. As a result of 5 the cells in the secondary cultures release particles into the culture medium;
7. The particles released in the secondary cultures have exactly the same characteristics as the original particles, that is, they must have identical morphology, band at 1.16 gm/ml and contain the same RNA and proteins.
The caveat is that while the introduction of the majority of infectious particles into cell cultures and subsequent release of similar particles is proof that such particles are indeed infectious, this is not the sufficient case for retroviruses. The basis of this exception is the fact that "one of the most striking features that distinguishes retroviruses from all other animal viruses is the presence in the chromosomes of normal uninfected cells, of genomes with those of infectious viruses".(83) In fact, a cell may contain the genome of many retroviruses. As far back as 1976 retrovirologists recognised that "the failure to isolate endogenous viruses from certain species may reflect the limitations of in vitro cocultivation techniques".(84) In other words, the finding of a retrovirus in both the primary and secondary "infected" cultures/cocultures is not proof that the cells have been infected with an exogenous retrovirus.
One way which will suggest but will not prove that the cells acquired virus from the outside (exogenously acquired retrovirus, infectious retrovirus) and have not assembled a retrovirus from information already existing in normal cells (endogenous retrovirus) is to conduct experiments that use controls, that is, to run in parallel with test cultures/cocultures control cultures/cocultures. The only difference between the test and control cultures should be the introduction of particles into the test cultures. In other words, apart from the introduction of particles, in every other respect control cultures must be dealt with identically. For example:
(a) because detection of RT and retroviral genetic sequences and release of retroviral particles depends on the metabolic state of the cells, the physiological state of the cells used in the control cultures should be as close as possible to those of AIDS patients;
(b) because the mere act of co-cultivation alone may lead to release of endogenous retroviral particles, if test cells are cocultured, so should the cells used in control experiments; (85)
(c) extracts, even from normal unstimulated cells, when added to the cultures may increase endogenous retroviral expression. (86) Because of this, when cells are cultured with "HIV" (supernatant or material which bands at 1.16 gm/ml), the controls must be cultured with similar material from cell cultures originating from sick individuals with illnesses similar to AIDS, that is, matched individuals who are immunosuppressed;
(d) the appearance of endogenous retrovirus can be accelerated and the yield increased a million fold by stimulating the cultures with mitogens,(87) mutagens, chemical carcinogens and radiation.(88,89) If test cultures are exposed to or employ such agents so should the controls;
(e) since AIDS patients and those at risk of developing the syndrome are exposed to strong oxidising agents,(79,90) the control cells should also originate from such patients;
(g) to avoid observer bias and in the best interests of science, blind examination of test and control cultures/cocultures should be performed.
79. Papadopulos-Eleopulos E. Reappraisal of AIDS: Is the oxidation caused by the risk factors the primary cause? Med Hypotheses 1988;25:151-162.
82. Gelderblom HR, ™zel M, Hausmann EHS, et al. Fine Structure of Human Immunodeficiency Virus (HIV), Immunolocalization of Structural Proteins and Virus-Cell Relation. Micron Microscopica 1988;19:41-60.
83. Weiss R, Teich N, Varmus H, et al. RNA Tumor Viruses. Cold Spring Harbor, New York: Cold Spring Harbor Laboratory, 1982.
84. Todaro GJ, Benveniste RE, Sherr CJ. Interspecies Transfer of RNA Tumour Virus Genes: Implications for the search for "Human" Type C Viruses. In: Baltimore D, Huang AS, Fox CS, eds. Animal Virology. New York: Academic Press Inc., 1976: 369-384.
85. Hirsch MS, Phillips SM, Solnik C. Activation of Leukemia Viruses by Graft-Versus-Host and Mixed Lymphocyte Reactions In Vitro. Proc Natl Acad Sci U S A 1972;69:1069-1072.
86. Toyoshima K, Vogt PK. Enhancement and Inhibition of Avian Sarcoma Viruses by Polycations and Polyanions. Virol 1969;38:414-426.
87. Aaronson SA, Todaro GJ, Scholnick EM. Induction of murine C-type viruses from clonal lines of virus-free BALB/3T3 cells. Science 1971;174:157-159.
88. Minassian A, Merges M, Garrity R, et al. Induction of a SMRV-like retrovirus from a human T-cell line after treatment with the mutagen ethyl-methyl-sulfonate. J Acquir Immun Defic Syndr 1993;6(No 6):738.
89. Papadopulos-Eleopulos E, Turner VF, Papdimitriou JM. Is a Positive Western Blot Proof of HIV Infection? Bio/Technology 1993;11(June):696-707.
90. Turner VF. Reducing agents and AIDS--Why are we waiting? Med J Aust 1990;153:502.
virusmyth.com/aids/perthgroup/geneva/printable.htm
To claim isolation of HIV by this method one must show:
1. Firstly, that the culture cell-free fluids contain particles having morphological characteristics of retrovirus the principle of which are:
- a diameter of 100-120nm
- condensed inner cores
- surfaces studded with spikes
2. When these fluids are banded in sucrose gradients the particles accumulate, that is, band, at the density of 1.16g/ml.
3. At that density there is nothing but retroviral-like particles.
Once this is proven, one proceeds to characterise the particle proteins and nucleic acids. If the particles are retrovirus, they should contain only RNA and not DNA, as well as an enzyme, reverse transcriptase, that copies RNA into DNA.
Because not all particles which look like retrovirus are infectious, that is are viruses, one must then prove that the particles which band at 1.16g/ml are indeed infectious.
5. When the particles are introduced into secondary cultures:
- the particles are taken up by the cells
- the entire RNA is reverse transcribed into cDNA
- the entire cDNA is inserted into cellular DNA
- the DNA is transcribed into RNA which is translated into proteins
6. The cells in the secondary cultures release particles into the culture medium.
7. The particles in the secondary cultures have exactly the same characteristics as the original particles, that is, they must have identical morphology, band at 1.16 gm/ml and contain the same RNA and proteins.
Indeed this is the method which Luc Montagnier and his colleagues claimed to have used to isolate HIV and thus to prove its existence.
theperthgroup.com/the12points.doc
1. Isolation of particles from cultures containing tissue from AIDS patients. The particles should be separated from everything else. That is, they must be pure and have all the morphological characteristics of retroviruses.
2. All the particles are similar; that is, they have the same morphology and constituents.
3. The particles contain RNA and not DNA. Because small genetic differences lead to significant phenotypic differences (the difference between the human and chimpanzees is less than 2%), the RNA in particles isolated from different AIDS patients should vary by no more than the RNAs from other RNA viruses.
4. The particles are infectious.
5. Similar particles cannot be isolated from cultures under the same conditions containing tissue from non-AIDS patients but nonetheless sick people. For example, individuals with autoimmune diseases or those treated with chemotherapeutic agents.
virusmyth.com/aids/award.htm
The rules for isolation of a retrovirus ... are the logical minimum requirements for establishing the independent existence of HIV. They are:
1. Culture of putatively infected tissue.
2. Purification of specimens by density gradient ultracentrifugation.
3. Electron micrographs of particles exhibiting the morphological characteristics and dimensions (100-120 nm) of retroviral particles at the sucrose (or percoll) density of 1.16 gm/ml and containing nothing else, not even particles of other morphologies or dimensions.
4. Proof that the particles contain reverse transcriptase.
5. Analysis of the particles' proteins and RNA and proof that these are unique.
6. Proof that 1-5 are a property only of putatively infected tissues and can not be induced in control cultures. These are identical cultures, that is, tissues obtained from matched, unhealthy subjects and cultured under identical conditions differing only in that they are not putatively infected with a retrovirus.
7. Proof that the particles are infectious, that is when PURE particles are introduced into an uninfected culture or animal, the identical particle is obtained as shown by repeating steps 1-5.
