THE RESULTS OF A 30-YEAR RESEARCH IN MOLECULAR BIOLOGY

Is the battle against cancer and AIDS already lost? Resignation and hopelessness: is that all the patients have left? Considering the therapeutic results obtained so far, this is what we might rightfully think As far as AIDS is concerned, the millions of dollars poured out into research projects did not bear fruit The drugs offered either are inefficient, or exhibit severe side effects. As to cancer, the optimistic speeches delivered by some researchers, and Intended for the general public, are offset by darker, more truthful statements published in medical or scientific journals. Efficiency burdened with too much toxicity seems to be the rule in the field (see annex 1).

It is in this context that a man dares to say pessimism is out of place and that it is now possible to treat successfully cancer and AIDS patients. If the subject allowed it, one would smile at such a naive attitude. By the way, who dares to say that?

An official scientist of the highest level: Mirko Beljanski, former Research Director at CNRS (Centre National de Ia Recherche Scientifique)! Most of his career was dedicated to research in molecular biology at the Institute Pasteur (from 1948 to 1978), then at the Faculty of Pharmacy at Chatenay Malabry for ten years, until his retirement in August 1988. His scientific papers (more than a hundred) were published in important French and international scientific journals; he took part in conferences all over the world. This should lend credit to what he says, even though it is quite amazing.

A nonconformist researcher

Before revealing the results of his research in the biomedical field, lets go back a few decades. There is a leading strand in this researcher's scientific work and It is important to understand Its continuity.

In the 50s, molecular biology was preparing for battle and researchers took an interest essentially in DNA, considered as the only pillar of living matter. Mirko Beljanski preferred to investigate RNA, which interacts with DNA. His efforts were soon crowned with success since, between 1958 and 1961, he discovered a peptide synthesis process involving a particular type of RNA: his work received the Charles Leopold Mayer Prize, awarded by the Academy of Sciences in 1961.

But his subsequent observations were much more fundamental, and questioned the "central dogma" of DNA supremacy, an unforgivable sacrilege which raised hostile reactions... and caused serious complications in his professional activity. By studying the carcinogenic activity of the bacterium Agrobacledum tumefacjens in plants, he discovered transforming RNA, which may totally or partly suppress the carcinogenic properties of this bacterium. This was a major observation, since for the first time in the history of biology, proof was given that RNA could induce a stable hereditary genetic transformation in bacteria. Conscious that this was an extraordinarily rich field to exploit. Beljanski continued his work and made one of his most fundamental discoveries: the existence of a reverse transcriptase in bacteria.

In 1970, Howard Temin, an American scientist, discovered in an RNA virus an enzyme capable of copying RNA into DNA. This process was contrary to the scientific knowledge of the time. It was thought that information could only go from DNA to RNA, the latter being meaningfully called messenger RNA. The enzyme discovered by Temin was called reverse transcriptase and Tem in was awarded a Nobel Prize, shared with Baltimore, in 1975.

In 1972, Beljanski published for the first time a paper on the existence of a reverse transcriptase in bacteria, which gave a coherent explanation to his previous work Since then, reverse transcriptase has often been observed in bacteria and plants.

Having observed the importance of RNA activity, Beljanski wondered whether this molecule might not be involved in plant cancer formation caused by Agrobactedum tumefaciens. This proved to be the case. Yet the special tumor-inducing RNA involved here cannot induce these plant tumors unless a certain amount of auxin (a plant hormone) is also present. We must insist on the fact that there are many kinds of RNAs, both large and small, some of which are beneficial and other dangerous.

In 1974: new important premiere concerning RNA. Beljanski and his co-workers achieved the enzymic synthesis of an RNA, acting as primer in DNA replication (primer RNA). Professor Pierre Lepine, Director of the Virology Department at the Institut Pasteur, announced this result, far from keeping with the traditional doctrine, to the Academy of Sciences.

From fundamental research to anticancer therapy.

Today, one may say that this period of research was a turning point in Professor Beljanski's scientific work It had two characteristics which are only apparently contradictory: great unity and radical change.

• Great unity because, as a persistent, methodical researcher, Beljanski advanced step by step in his research work, each discovery raising new questions and hypotheses, that entailed original experiments and observations.

• Radical change because this was the time when Mirko Beljanski shifted from fundamental to applied research. He had enough material in his possession to consider a concrete application of his works. Totally convinced that the fundamental mechanisms of living matter are similar in animals and in plants, he would show that the real difference between healthy cell and cancer cell.

DNAs lies in their secondary structure. This was a major element in what he thought to be a feasible fight against several serious diseases.

The reward brought to Beljanski and his team was to be in keeping with the obstinate work performed. The same year, 1974, he demonstrated in vitro the efficient role of RNA as primer for DNA replication; this was also the time when he discovered several RNAs capable of stopping DNA virus multiplication in animals. Once more, Professor Lepine, who took a close interest In Beijanski's work, announced this result to the Academy of Sciences on 16 December1974. As Ms C.G. Nordau, a biologist and scientific writer, put it' "Beilanskj had just discovered the first efficient and non-toxic antiviral drug".

In spite of the fundamental significance of this discovery, Jacques Monod, Director of the Institut Pasteur, who worked obstinately on DNA, told Beijanski that the project he presented "found no place in the applied research programme"! Non-scientific considerations had undoubtedly motivated these words.

Contrary to the generally admitted opinion, scientific activity is not always devoid of ideological a prioris or personal ambitions. However, when people's healths and even lives are at stake, such an attitude is unacceptable.

But for all that, Beljanski was not discouraged. Under difficult working conditions, he went on with his research. In collaboration with physicians, he observed the efficiency of one of his substances in several cases of intractable herpes.

A priority: healthy cell protection

Besides, in the second half of the 70's, Beljanski was deeply concerned with the high toxicity of anticancer therapies (chemo or radiotherapy). They have two serious consequences: chromosome breakage and the destruction of bone marrow cells, which results in the decrease of white and red cell and of platelet counts, weakening the body's defences against infections and inducing risk of hemorrhage. These therapies may also induce secondary cancers. For Beljanskl, it was then urgent to develop substances that would protect healthy cells while toxicity would affect cancer cells only.

In fact, the word "develop" so often used in scientific research, is not appropriate here. The term "discover" is more accurate. Beljanski had always been convinced that the best way to help the body to resist a disease is to use natural substances rather than synthetic chemical molecules. The discovery of new RNA fragments would confirm this intuition. Obtained from an essential host of the human intestinal tract, Escherlchia coil, they stimulate normal bone marrow and spleen DNA replication, without affecting that of cancer DNA.

The experiments revealed the astonishing efficiency of these RNA fragments. After very high doses of antimitotic have been injected to rabbits, leucocyte and platelet counts collapse dramatically, jeopardizing the animals' lives (Fig. 2).. But the injection of RNA fragments induces a spectacular rise in these counts within a few days. Tested on patients undergoing radiotherapy or intensive chemotherapy, these molecules allow them to live normally, without having to interrupt their work Further studies even showed that these RNA fragments favour the appearance of killer lymphocytes that attack cancer cells. Emphasizing the fact that he used natural substances, Beljanski asked: "Why are molecules which are made artificially outside the body preferred, at the risk of disorganizing cell life, to natural molecules which stimulate hemopoletic cell formation".

Beijanski called these substances "BLRs", i.e. "Beljanski~Leucocyte-Restorers". He had brought to the fore a revolutionary principle, namely the capacity of a molecule to distinguish between normal cells and cancer cells, and to act on one type rather than on the other. Until then, the scientific community was convinced---and unfortunately, this opinion is still widely admitted---that it was impossible to act efficiently on tumor cells without affecting the surrounding healthy cells. In spite of the importance of this principle, he ended very simply his publication on that subject in the Bulletin de l'Academli nationale de medecine (1978): "When blood liucocyte and platelet counts have been depleted by the side effects of chemotherapy, these counts may be safely restored to their normal values by RNA fragments, the BLRs."

Such results obviously had to be followed by medical applications. Cancer patients were given BLRs simultaneously with chemo- or radiotherapy. The following case, among others, shows the beneficial effect of BLRs

(Fig. 3)..

From then on, three complementary rules will dominate Beljanski's research strategy:

· selectivity of action,

· absence of toxicity for healthy cells,

· defence and promotion of healthy cells.

Beijanski thinks that "non selective therapy is nonsense. Destroying as many healthy cells as unhealthy ones is a hazardous policy, of a poor biomedical level".

Recent confirmations abroad

Recent publications confirmed the remarkable protective effect of some of the products developed by Mirko Beijanski.

Professor Maurizio Grandi, member of the Academy of Sciences of Rome, and Director of the Centre for Study, Research and Therapy of Neoplasms, in Turin, published in 1990 the results obtained with 30 cancer patients1 who were administered several of Mirko Beijanski's products.

1) BLRs

To begin with, Prof. Grandi notices that "no efficient substance has yet been discovered to prevent blood cell depletion or to restore leucocyte and/or platelet counts during or after chemotherapy" But he underlines that interesting results have been observed for several years in chemotherapy-treated animals treated simultaneously with BLRs..

Then, everyone knows that each chemo- or radiotherapy must be followed by a period of rest, to limit damages and allow the body to recover. in the present case, all patients could undergo several consecutive chemo-therapeutic treatments without interruption and BLRs were very well tolerated.

For Prof. Grandi, this study "proves that the perlingual administration of BLRs to chemo or radiotherapy-treated cancer patients, efficiently prevents leucocyte and platelet depletion, and quickly restores normal cell counts after chemotherapy".

2) Anticancer drugs

In another study2 by the same author, 8 patients with malignant glioma underwent radiotherapy and were given a specific anticancer drug prepared by Beljanski (see below) simultaneously. This drug potentialized destruction of tumor cells during radiation, without side effects on normal cells. The alkaloid selectively bound to cancer cell DNA only, causing destruction of malignant cells. Prof. Grandi confirmed Mirko Beijanski's earlier works by saying that "the synergy of action with radiotherapy Is due to the fact that radiation opens DNA chains and the alkaloid can thus bind more easily to cancer cell DNA".

In a third research, also by Prof. Grandi 3 and coll., 11 patients with prostatic cancer were studied. This disease generally causes psychological problems since the only available therapies lead to sterilization (following chemical or surgical treatment). Some men prefer not to be treated rather than be mutilated. In this study, all patients had refused treatment. They accepted to undergo radiotherapy while taking alstonine-derived substances developed by Mirko Beljanski. After 18-80 months, 2 complete remissions, 4 partial remissions and 4 stabilizations were observed. The conclusion of the report mentioned that "It was obvious that the alstonine therapy could theoretically represent a valuable alternative to the hormone treatment" Prof. Grandi and his coworkers say they prefer the aistonine treatment: "In fact, theoretically, whereas the effect o fhormone treatments is cytostatic, i.e. the neoplastic cells go Into mitotic rest, the effect of the alkaloid (alstonine) on the neo-plastic cells is cytotoxic, which makes it potentially able to kill (diseased) staminal cells; moreover, It Is potentially active in all patients with metastases".

Recently, a medical team from a C.H.U. (University Hospital) in Montpellier (France) has demonstrated the efficiency of BLRs in cancer patients undergoing chemotherapy 4.

The Oncotest, a revolutionary technique!

Beljanski went deeper into his research and developed the Oncotest, a technique that allows precise measurement of a substance's carcinogenic potential (Fig. 4).. Another test, developed by Ames and frequently used, is based on the belief that cancer formation is induced by mutation which transforms normal DNA into tumor DNA. But there are too many exceptions to this mutagenetic theory: Beljanski offered a completely new interpretation of cancer formation (see annex 2).

The Oncotest is based on the observation that well known carcinogens strongly stimulate cancer DNA synthesis and very slightly that of normal DNA (Fig. 5). Thus if a drug is carcinogenic, cancer DNA synthesis is significantly more active than that of normal DNA. The reliability of the Oncotest is remarkable; it allows the detection of very low doses of carcinogens, and the response times are very short (1-3 hours, while the Ames test requires a week).

Besides, the Oncotest was used to detect natural plant substances. capable of selectively recognizing cancer DNAs. of binding to them and of stopping their replication. By trying out various plants, Beljanski discovered the remarkable properties of some alkaloids: their restabilising "bolt" molecules specifically act on cancer DNA and not on normal DNA. To this day, half a dozen specific anticancer compounds have been found, that are highly active in vivo. Three alkaloids In particular (alstonine. serpentine and sempervirine) very efficiently inhibit cancer cell proliferation (Fig~6 and 7).. These anti-cancer substances cause either death of the malignant cell, which cannot multiply or express its genes, or recovery of cells, in which the malignant process has not gone too far. In contrast, they have no effect on normal DNA replication 5.These results have been confirmed by several research teams (In Belgium, Switzerland and France).

Specific anticancer drugs, enzymatic regulator, selective antiviral drugs...

a) BG-8 was developed from one of these alkaloids. Given to tumor bearing mice, it leads to 60-80% survival without toxic side effects, and even 90% survival under certain conditions. Used in combination with conventional therapies, this compound is even more efficient The explanation given by Beljanaki is straightforward. Radiation and chemical molecules break and destabilize DNA, that is why they are dangerous. But this favours binding of anticancer drugs, such as BG-8, specifically to the damaged areas of DNA.

Statistical results obtained in animals unquestionably proved the validity of this combination, the effects of which are not additive, but synergistic. Similarly, in man, BG-8 proved its efficiency. Sometimes used together with BLRs, it allows patients to lead a normal life, while undergoing chemo- or radiotherapy, and without experiencing side effects such as hair loss, nausea, or blood cell depletion. Patients, considered incurable, were in remission. It must be noted that BG-8 is also efficient on plant crown-gall tumors; another proof of the biological similarity between plants and animals.

b) Furthermore, for several years, Beljanski has been particularly interested in cell regulation; he developed several substances intended to restore the normal functions of unhealthy cells. He thus developed BIOPARYL, a plant preparation stimulating cell regeneration by correcting various enzymic abnormalities, corresponding to the abnormal expression of a gene. A famous French cancerologist, Prof. Lucien Israel, has used BIOPARYL for severe fibroses induced by anticancer radiotherapies, with very satisfactory results; recoveries were obtained in two out of three cases.

The validity of this approach, i.e. trying to reverse cancer cells to normal rather than killing them, has recently been confirmed by Prof. Degos and Wang Zeng Yi's results. These two researchers have succeeded in transforming cancer cells into normal cells by treating acute promyelocytic leukemia patients with retinoic acid (derived from vitamin A). Prof. Degos' comment is significant: "A new road opens. We have now a new weapon at our disposal. We have the tangible proof that reversibility is possible, that a malignant cell can be transformed in vivo into a normal cell. The end of a dogma..." 6

c) Besides, and this is a major point, Beljanski developed plant-derived therapeutic drugs (PB1OO, PB 400...) that act on several enzymes (among them reverse transcriptase), preventing RNA and DNA virus multiplication. Moreover, these molecules destroy virus particles; this was demonstrated by Beljanski, Le Goff and Aaron with plant RNA viruses and spectacularly confirmed by researchers from Berne (Switzerland) to whom Beljanski had given his products. Then, with a whole range of selective, non toxic substances, each of them acting at a specific stage in cell development, Beljanski felt he was ready to take up the challenge of AIDS and many other severe diseases (annex 3).

Can AIDS patients be cured?

AIDS: a disease that frightens so much nowadays, seems to yield to no treatment. Every new drug, given wide publicity in the media, is forgotten after a few months, or accused to have side effects more harmful than the disease itself. Here again, Beljanski asserts he has discovered new substances, capable of fighting AIDS.

A study made by Professor Jachertz, former Director of the Medical institute for Microbiology and Hygiene, University of Berne 7, demonstrated the remarkable In vitro efficiency of a molecule developed by M. Beljanski on the AIDS virus. The Swiss researcher prepared in parallel healthy culture cells, infected culture cells without therapeutic substance, and various infected culture cells with different doses of the therapeutic substance. He observed that the substance did not affect non-infected cell multiplication. Besides, he said: "Whereas virus particles increase in untreated infected cell In function of time, we were not able to detect the presence of Infectious units beyond 3,000 units in Infected cells treated with the "H" compound (30 or 60 microgrammes). This proves that at least over 99% Inhibition of Infection was obtained".

Two remarks about these results.

· Virus destruction was perhaps complete, as, with the method used, precision is limited to 99%; this explains the last sentence quoted.

• But above all, one must not forget the significance of the double result obtained, clearly summed up by the study title: "In vitro Destruction of HIV (AIDS virus) in Cell Cultures In vitro without Effect on Normal Cells".

It is to be noted that several compounds can destroy HIV1 in vitro, chlorine for instance. But they also destroy healthy cells; this makes them useless for therapy. Thus the outstanding property of the substance studied by Prof. Jachertz: is Its capacity not only to destroy HIV1 but above all to destroy it without damaging healthy cells. Again, this is the principle of selectivity of action, a priority for Mirko Beljanski.

He thinks that anti-AIDS strategy has to be multifocal. In addition to virus destruction, "the solution to the problem lies either In the control of the replication of DNA into DNA (for DNA multiplication), or, forRNA viruses, in the control of the transcription of viral RNA into DNA (by reverse transcriptase)". HIV1 is precisely a retrovirus, i.e. a virus that requires reverse transcriptase for its multiplication.

Furthermore, this virus preferentially attacks T4 lymphocytes, the "bandleaders" of the immune system; this system then collapses, leaving the way open to various so-called opportunistic diseases (herpes, pneumocystosis, cytomegalovirus, mycosis, tuberculosis, Kaposi sarcoma, etc.). The ideal solution would be to tackle the problem of AIDS from different angles, I.e.: REST MISSING

ANNEX 1

WHAT FAMOUS SPECIALISTS HAVE SAID

"The new discoveries made in cancerology, with their novel approaches. have made obsolete all we have done for the last thirty years, i.e. using lots of surgery, radiotherapy, chemotherapy in a totally empirical manner.. It Is obvious that we have used too many toxic and Intensive chemotherapies; to continue in this way is deplorable". Prof. Lucien Israel, Le Monde de Ji Mddecine, 16 December1987.

-The global death rate due to cancer continually increases".-----Prof. Georges Mathe, Le Monde de la Medecine, 4 May. 1988.

"Hematologlsts are still making mistakes. Present therapies will perhaps appear wrong to our successors. Thus the destruction of leukemic cells, the only metn#od used today, and the probably fallacious dogma of the complete eradication of leukemic cells". Prof. Jean Bernard, Actualites: hematologiques:, 20.1987, pp.7-8.

"ln 95% of cases. chemotherapies do not improve the prognosis of malignant tumors".----Prof. Bernard Plerqu in, La Pratique medicale quotidienne, 17 July, 1986.

"Between 1975 and 1983, the increased use of chemotherapy did not induce a decrease of the death rate due to breast cancer". General Accounting Office Report, quoted In La Recherche, June 1989.

"The Incapacity of surgery to cure cancer is confirmed by striking figures: 20% of the patients die of local recurrence, 80% die of distant metastases".--- Prof. Lucien Israel, Cancer d'aujourd'hui.

"The knowledge acquired from the observation of leukemic cells, the maturation and death of cultured leukemic cells, allows us to hope for the development, in the near future, of treatments that will no more destroy, but correct, cure, leukemic cells. Destructive therapies will then appear dangerous, pathetic and erroneous".----Prof. Jean Bernard, Actualites: hematologlques:, 20,1987, PP.7-8.

"Researchers should try and develop biological regulating therapies, rather than chemotherapies (that prevent only the proliferation of cancer cells. and degrade, instead of improving their relation with the organism). After the chemotherapeutic war, the time has come for biological diplomacy."----Prof. Georges Mathe, Le Panorama du Medecln, 7 November 1984.

"Biological therapies are rapidly gaining ground. The hope we have in some of them is based on concrete results. Their role is likely to increase in the years to come".----- Prof. claude Jasmin, impact Medecin, 25 March 1989.

ANNEX 2

One or several gene mutations are generally thought to be at the origin of the transformation of a normal cell Into a cancer cell. But this theory has a major flaw In most cases, no significant difference can be found between the nucleotide sequences (ie. the primary structure) of cancer DNA and normal DNA. These nucleotide sequences make up the strands of the double-stranded DNA molecule, which is coiled into a helix.

In Beljanski's view, the difference between normal and cancer DNA does not lie in the nucleotlide sequences themselves:, but in the way the two strands: of the molecule are held together by hydrogen bonds (this: is: called DNA secondary structure). These hydrogen bonds normally open, but only locally and briefly, for DNA replication or for gene expression. Bond breakage gives replication or transcription (gene expression) enzymes access: to so-called "intiation sites" along the temporarily isolated strands:.

Bejlanski demonstrated that in a cancer DNA molecule many hydrogen bonds are permanently broken. Over large areas the DNA strands are further apart than normal; the DNA double chain is looser partly uncoiled, while this Is compensated by coil tightening on either side of these areas. In the loose areas many initiation sites are now exposed. How does this happen ? What carcinogens do is precisely to bind to a DNA molecule and induce hydrogen bond breakage, which progressively spreads along the molecule as more carcinogen finds its way to the damaged regions. Hydrogen bond breakage destabilizes the DNA molecule which then becomes an easy prey not only for many carcinogens, but for all sorts: of damaging substances: from the cell Itself or the environment. In addition, replication and transcription enzymes: can now bind to normally unused initiation sites. In the loose or "open" areas. while losing access to normally used initlation sites in the tightened areas. This explains: why: 1) cancer cells: multiply fast; 2) gene expression is: dysregulated; 3) persistently increasing strand separation may eventually lead to chromatid or chromosome breakage and mutations may arise from strand damage.

Convincing proof of this theory has been put forward and published by BeIjanski. It has moreover resulted In his:discovery of therapeutic molecules. This is the way he reasoned: if carcinogens: preferentIally "open'" DNA double chains, there should also exist molecules: which can act in the opposite way, that is, bind to DNA and "close" the abnormally open chains by bringing the strands: closer together so that hydrogen bonds can form once more. This proved indeed to be the case.

PROFESSOR ISRAEL TESTIFIES

"REAL SUCCESSES"

'We have used a product recommended by M. Bejanski in the treatment of post-radiation fibroses, in my department, openly, and with real success in 2 cases out of 3. We never had the opportunity nor the possibility of testing anticancer products that should first have undergone toxicological trials, and should receive experts consents before we can simply have the right to offer them .(Lucien Israel, in a French radio programme. (France Inter) -23 October 1989)

ANNEX 3

Beljanski and the members at COBRA are often asked the following question: "How can you assert that it is possible to fight simultaneously against so many different diseases: cancer, aids, fibrosis, degenerative diseases"?

He explains that "an agent involved in a pathology, according to the organ in which it is confined, may provoke various symptoms:. The same causal agent may then induce various disorders that require different complementary treatments:.

Of an antibiotic drug that is used in raging toothache, cystisis:, furoncle or septicemy, no one would say it cures all diseases:. It acts on the causal agent, right from the start of bacterial multiplication, and this: Is how It can be efficient In the treatment of different diseases:. As antibiotic drugs, molecular biology, biomedicine, as we see it, goes to the root of the disease.