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INVENTION
Russian Federation Patent RU2105310

METHOD OF TREATMENT acquired immune deficiency syndrome (AIDS)

METHOD OF TREATMENT acquired immune deficiency syndrome (AIDS)

Name of the inventor: Tkachenko Vitaliy V.
The name of the patentee: Vitaly V. Tkachenko
Address for correspondence:
Starting date of the patent: 1995.12.21

The method of treatment of AIDS, which consists of the patient's treatment by various viruses, including primarily on retrovirus - the AIDS pathogen (HIV), contaminated their blood cells and micro-organisms - Assiociants using plasma sorption when used individually manufactured for each patient immunosorbent, amplified covalently associated with him glyukoproteidov, which was isolated from human plantsety and that exhibits high affinity for the retrovirus AIDS. blood purification procedure involves the use of plasmapheresis directly after plasmasorption with compensation of blood plasma with an equal volume (1000-1200 ml) plasma expander as reopoliglyukina and re-use plasma sorption using fresh immunosorbent. blood purification procedures course interspersed exchange application-specific immune stimulation using autovaccine prepared in each case on the basis of UV-inactivated by blood culture of the patient. In the later stages of AIDS blood purification courses and specific immune stimulation using autovaccine complemented replenishing damaged in the AIDS macrophages and helper T-cells by intraosseous transplantation of umbilical cord blood from the cut-off at birth the umbilical cord, which contains all the necessary stem progenitor cells gemoleykopozza.

DESCRIPTION OF THE INVENTION

More than ten years are fruitless search for the creation of a specific therapeutic and prophylactic vaccine against a retrovirus HIV - the pathogen that is incurable AIDS, this "plague of the twentieth century" for all mankind.

Failures of such searches is not refuted by anyone explain the high mutable retrovirus HIV. However, on the other hand, the infectious process in AIDS in its genesis is closely associated with the manifestation of the activity of other viruses and microorganisms vary in the individual spectra of infection in patients with AIDS, referred to as "associants" and is generally regarded as the only causal factors 'opportunistic infections' or complications. Aware of endogenous latent micro and ultramikroflore, repressed in the human body it normally functioning immune system and reviving activate in terms of human infection with a retrovirus HIV, which is deposited and replicated first in macrophages, eventually causing them to mass death. Revives the latent ultramikromikroflora micro- and can not contribute to the pathogenesis and clinical manifestations of AIDS.

This convinces the hallmark of AIDS as an infectious process in comparison with the known viral and bacterial infections - double the incubation period for AIDS: first - before the predictive period, occurring in the form of a flu-like condition with a picture of the blood, similar to that in infectious mononucleosis, and due to the deposit of a retrovirus and HIV replication in macrophages in the mass destruction as a result of recent. The second incubation period for AIDS takes place after the prodromal (predictive) period due to breaking resistance immunobiological host retrovirus HIV and the connection to the previously suppressed infectious process and now activate latent micro and ultramikroflory. This does not exclude the possibility of subsequent accession of micro- and ultramikroflory exogenous origin of interkurretnyh infections and other complications, superimposed on the basic infection process, caused by the retrovirus HIV Association and the activated endogenous micro- and ultramikrofloroy.

Thus, the development of AIDS treatments have to bear in mind not only the pathogen retrovirus HIV infection source, but it Assiociants as various viruses, including some oncogenic DNA and RNA (retroviruses) viruses and various microorganisms (yeast fungi, bacteria, etc., including opportunistic) to which the retrovirus HIV interacts with the first moments of its invasion of the human body and further.

This interaction leads to massive damage clonal populations of antigen presenting first and specifically activated macrophages, and then the helper T-cells replicating retrovirus in their HIV and other lymphotropic previously latent now associants-active viruses. Thus there is a profound violation of prekrestov information pathways, vicious circles and so on. In the chain of immunological "processing" / antigen-presenting cells / macrophage and others. / helper cells / T 4 -limfotsit / B cell / plasma cell turning into producing specific antibodies / information on the antigenic characteristics of the antigens in the form of infectious agents (HIV retrovirus and other virusy- and micro-Assiociants) as the basis for the formation of an immune response to a specific antibody for the relevant genetic information program on principles of evolutionary bioinformatics and pathological processes [1-4].

Solution of the problem of AIDS in terms of his approach to immunotherapy using the vaccine alone retrovirus HIV (even if any will be able to produce) virtually impossible to achieve. It is unlikely to be effective, and drugs that are targeted only at inhibiting the replication of the retrovirus HIV, which have, for example, antirevertaznoy activity. Finally, it is unlikely to justify himself attempts to date to use for the treatment of AIDS antibody administered to patients from the monkey-baboon and others. Animals with natural immunity to infection with the retrovirus HIV. As conclusion - the pathogenetic treatment of AIDS has not been developed to the present day, although the period of searching specific vaccine against AIDS (over 10 years) was infected with AIDS, and many of them have already died from AIDS. AIDS threatens to us in Russia. In the development of their way to treat AIDS guided me a long time ago known principles "to treat not a disease, and the patient" (Hippocrates) and "do no harm" (Paracelsus).

The technical result of the use of the invention: developed and tested on a limited kontigente patients, volunteers and AIDS virus carriers improved method for the treatment of acquired immune deficiency syndrome (AIDS). The method is safe, specific and effective according to the clinical condition of volunteers and according to serological tests. With this method, you can turn off the source of infection (AIDS virus carrier or sick) of the epidemiological chain of AIDS, and because the process may be seen as an important component of HIV prevention.

Method is as follows

I. In the course of carrying out blood purification of patients with AIDS or AIDS virus carriers of the virus, they contaminated blood cells and microorganisms Assiociants using plasma sorption, skompleksirovannoy with plasmapheresis in a single extracorporeal system. For plasma sorption column is used the adsorbent of FAS mark (furfural adsorbent spherical) with covalently attached to the sorbent (the existing method) immune antibodies are isolated in a purified gamma globulin fraction of the hyperimmune antiserum of rabbits under conditions of immunization blood cultures from this patient (patient or virus carrier) grown on embryonated hen eggs (according to standard procedures). Immunosvyazyvayuschie manufactured immunosorbent enhance properties covalently attached thereto (the existing method) as glucoproteins fraction by standard methods of salting out with ammonium sulfate (65% saturation) at the isoelectric point (pH 6.5) from human placenta (instead glyukoproteidnoy fraction isolated from mouthparts , and the salivary glands of female mosquitoes on the previously developed and used, but very time-consuming procedure). Isolation and use glyukoproteidnoy fraction (possibly cell receptor origin (based on the fact that the placenta performs, apparently as a kind of biological filter, retarding, in particular a retrovirus HIV because mother AIDS patients, give rise to quite healthy infants . this glucoproteins, showing an increased affinity for a retrovirus HIV has been found to improve the binding of the virus immunoabsorbent during plasma sorption described plasmasorption accompanied followed plasmapheresis:. patient blood (virus carrier) with plazmosorbtsionnoy column is fed to a plasmapheresis, in which may comprise a domestic . fractionator model "FS-0.5" is removed in the process of plasmapheresis liquid part of blood (plasma) containing "escaped" when plasma sorption viruses and micro-organisms, is replaced by an equal volume (1000-1200 ml) plasma expander as reopoliglyukina; plasmasorption then repeated using with her fresh immunosorbent. For half an hour prior to the blood-plasmasorption plasmapheresis plasmasorption-cleaning procedure to prevent blood clotting in the extracorporeal system are training the patient by intravenous administration of 300 international units (IU) of heparin per 1 kg of patient weight. At the same time treated with extracorporeal system solution of heparin (10,000 IU heparin in 400 ml physiological sodium chloride solution and connect it to the needle,. Introduced in the cubital vein of a patient all the preparatory operations and manipulations related to the plasma sorption and plasmapheresis procedures carried out in compliance with the absolute asepsis.

II. After each procedure, plasmapheresis plasma sorption-plasma sorption in order to soft-specific immune stimulation is administered parenterally autovaccine after each blood purification procedure using plasmapheresis and plasma sorption. Duration autovaktsinoterapii (number needed autovaccine injection), as well as blood purification rate (the amount of plasmapheresis and plasma sorption processes) is determined by the terms of AIDS and clinical manifestations of the disease. Autovaccine produce inactivated by ultraviolet radiation (UV) blood culture obtained from the patient (or the patient's AIDS virus carrier), grown in eggs. Each injection is determined by the need to introduce each time 5-6 ml autovaccine using 20-24 chick embryos in egg incubation conditions according to standard procedure. For sowing in alantois chick embryos 0.4-0.5 ml blood of a patient; incubation at 37 ° C of crops two or three days. After aspirating syringe of blood culture alantoisov combined blood culture volume adjusted to 5.6 mL with sterile isotonic sodium chloride solution, phosphate buffered (0.15 molar concentration, pH 7.1-7.2). In connection with the cultivation of blood culture and cooking from it autovaccine all manipulations produced under aseptic unconditional compliance with the rules.

III. In the later stages of the AIDS rate of blood purification (sm.I) and autovaktsinoterapii rate (see. II of) be sure to complete the replenishment of the pool of macrophages and helper T-cells, which permanently and irreversibly damaged in AIDS conditions. Produce is via transplantation of blood taken from the umbilical vein born infants (instead of the previously developed, but rather time-consuming methods of making and using hybridoma) and containing all the necessary stem progenitor cells gemoleykopoeza in the bone marrow of an AIDS patient. Thus, it provided the need to create at maternity hospitals blood banks from umbilical vein born babies (from cut-off at birth the umbilical cord from the placenta), with its screening for compatibility based on antigenic characteristics of blood (in the manner of those banks umbilical blood born babies that have long existed, for example, in USA). This takes into account the fact that such blood can be widely used for transplantation in the bone marrow at different diseases and injuries (radiation sickness, burns, etc.). In severe cases it is possible the direct use of the blood from the umbilical vein born infant intraosseous implantation of mothers with AIDS. Number and Periodicals intramedullary transplantation of umbilical blood from babies born with AIDS is determined by the severity of the process. All manipulations associated with taking the umbilical blood of infants and its use for transplantation is conducted in conditions of strict aseptic technique.

IV. Treatment of AIDS patients and virus carriers of AIDS using the method described (sm.I, II and III) is required to accompany the application of strengthening therapy (vitamin therapy, the introduction of trace minerals, leykogemopoeticheskih funds and so on.), Administered to a patient according to individual indications.

The process is strictly individualized combined treatment of AIDS in view of its improvement should be carried out under conditions of constant monitoring of the general state of the organism of patients, accompanying his mandatory clinical blood and urine tests, regular inspection of indications of specific serological tests on blood levels of the HIV retrovirus patients.

The proposed method of treating AIDS illustrated by the following examples.

Example 1 Preparation glyukoproteidnoy fraction ( "glyukoproteidov") of fresh (or stored in deep freeze at -40 o C for up to a week) human placenta.

Clipping placenta 50 g finely crushed with scissors and suspended in isotonic sodium chloride solution buffered sodium and potassium phosphate (0.15 molar concentration, pH 7.1-7.2) and is further divided into a tissue grinder using a teflon pestle. All these manipulations, but also the subsequent operations carried out in separation glyukoproteidov cold (+4 o C). Mode disintegration by impact crushed liver tissues in a conventional laboratory pulper model: 1600-1800 rpm for 3-5 minutes. The procedure of grinding and disintegration of the placenta is repeated to accumulate the raw material for the isolation glyukoproteidov reserve sufficient.

The combined slurry was centrifuged placenta disintegrated in refrigerated centrifuge (3000hg, 30 min; +4 o C). The resulting pellet is frozen in liquid nitrogen and then freeze-dried in a conventional laboratory vacuum cabinet under model usual lyophilization. The freeze-dried preparation can be stored securely-closed containers in the freezer the average home refrigerator no more than a week.

To isolate placental glyukoproteidov lyophilized preparation was suspended in 10% ammonium sulfate solution buffered with phosphate (0.15 M; pH 7.1-7.2) at a ratio (g / ml) was 1:10. Fractionation placenta accompany drug periodic removal of sediment by centrifugation in the dropdown refrigeration supercentrifuge (30000h, 15 min; +4 o C) increase as the concentration of added batches of ammonium sulfate to a concentration of 65% saturation (while monitoring the pH value, which should be kept at 6 5 units). For fixing on the sorbent domestic brand FAS standard technique used precipitate isolated at 65% saturation of ammonium sulfate at pH 6.5 in a refrigerated centrifugation supercentrifuge (cm. Above), and subjected to lyophilization by the method described above. Simultaneously, the same sorbent of FAS fixed (by the same standard method) lyophilized preparation gamma globulin fraction isolated from rabbit hyperimmune antiserum by salting out by adding ammonium sulfate to the isoelectric point with fractional centrifugation and then lyophilized by standard methods. In the manufacturing method described immunosorbent reinforced covalent fixation thereon glucoproteins from human placenta, and to isolate glyukoproteidnoy gamma immunoglobulin fractions using all chemicals qualification "analytical grade" The resulting immunosorbent column used for the purification of blood plasma sorption AIDS or AIDS virus carriers of the retrovirus HIV, and other microorganisms, and virusov- Assiociants (see. Section I).

Example 2. Determination of HIV binding activity of human placenta glyukoproteidnoy fraction (in terms of its use to enhance the binding of HIV immunosorbent activity according to example 1). Weigh 100 mg of lyophilized placental glyukoproteidnoy fraction obtained according to Example 1 was dissolved in 100 ml of isotonic (0.85%) sodium chloride solution, phosphate buffer (0.15 m, pH 7.1-7.2) and to remove the ammonium sulphate was dialyzed against three changes of a physiological solution of sodium chloride in phosphate buffer at the indicated concentrations in the cold (+4 o C). To a solution prodializovannomu glucoproteins and one control sample containing only isotonic sodium chloride phosphate buffer (indicated by prescription) was added to 1 ml of the specific diagnostic kit used for serodiagnosis usually AIDS and 0.5 ml of chloroform to prevent germination tests. After thorough mixing of the sample containing 10 ml of solution glyukoproteidov diagnostic tools and chloroform, and a sample of the same volume of 1 control were incubated at 37 o C for one day under the cotton-gauze plugs; simultaneously incubated 10-mL sample containing only isotonic sodium chloride solution, phosphate buffer (recipe for the above) and chloroform, and subjected to all the manipulation which the sample is subjected glucoproteins and control sample 1 (control 2).

After incubation all samples heated in a water bath (40 o C) in a fume hood to remove chloroform to eliminate the smell of his trial. Then, after cooling in the cold (+4 o C) was added to all samples to a sample of ammonium sulfate saturation of 65% at pH 6.5. In samples with glyukoproteidov and control samples 1 flocculent precipitate falls, more pronounced in samples with glyukoproteidov. All samples were subjected to refrigerated centrifugation supercentrifuge (30000hg, 15 min; +4 o C). At the same time the sample is centrifuged out of control 2, wherein the precipitate formed in the presence of ammonium sulfate, ceteris paribus.

The resulting centrifugation precipitate from samples contained glucoproteins from placenta and specific AIDS diagnostic kit and to and from samples of control 1 which contained only a diagnostic kit, along with the supernatant, corresponding to all of these precipitates, and samples of control 2 was dialyzed as described above to remove the ammonium sulphate. Prodializovannye precipitates redissolved again in each case 10 ml of isotonic sodium chloride solution, phosphate buffer. All fractions obtained as a result of the dissolved precipitates and supernatants of the respective samples of sample glucoproteins and 1 control subject spectrophotometry versus control samples from two control for quantifying unbound specific diagnosticum for AIDS RNA using standard RNA detection methods polumikrospektrofotometricheskim method.

The results obtained by evaluating polumikrospektrofotometricheskoy RNA retrovirus HIV-specific binding of the diagnostic kit for AIDS:

a) reconstituted precipitate fraction of placenta samples glyukoproteidov comprises 75-80% of the initial concentration added to these RNA samples diagnosticum, whereas the corresponding fraction of the supernatant (supernatant) from the same samples contain only 20-25% of the initial concentration of the same RNA diagnosticum;

b) changes in the content of added diagnosticum precipitate redissolved in case one of the control (sample not containing placental glucoproteins) and the supernatant corresponding method are within the errors and are, respectively, 5.2 and 95-98% of the initial concentration of added RNA diagnosticum.

These data indicate that the isolated fraction glyukoproteidov from human placenta contains (connects) the bulk of the substance responsible for binding to specific AIDS diagnostic kit. Allocating from placenta of human fraction binding specific to AIDS diagnosticum, in the standard process conditions based on the salting-out with ammonium sulfate at the isoelectric point and commonly used in analytical and industrial and pharmaceutical practice fractionation to isolate the substance (s) of protein nature, indicating protein the nature of the substance (s) responsible for the binding of the diagnostic kit. The use of existing quality of analytical samples for various proteins has allowed to conclude that the precipitate separated fraction is represented in the main substance (s) glycoprotein nature - glyukoproteidov (glyukoproteidov), in all probability, a cell receptor of origin. Warming glyukoproteidnoy drug faction, connected the particular diagnostic kit, at 100 o C for 5 min did not significantly affect the results of the analysis, which indicates a sufficiently strong binding of antigenic structure of HIV diagnostic kit used in the composition.

Example 3. Application improved method of treatment of AIDS in its early stages. For blood purification procedure by plasmapheresis, plasma sorption-plasma sorption column with plasma sorption (in extracorporeal plasmapheresis and plasma sorption system) used immunoabsorbent produced in accordance with Example 1 and verified in accordance with Example 2. The purification of blood plasma sorption procedure, plasmapheresis, plasma sorption is carried out in a the course of two or three procedures for a duration of 1.5-2 hours once a week for a total volume of 1000-1200 ml perfusion liquid portion of blood (plasma) followed by substitution with an equal volume plasma expander (reopoliglyukina). Immediately after each blood purification treatments by plasma sorption-plasmapheresis, plasma sorption produce parenteral (intramuscular) introduction of autovaccine.

autovaktsinoterapii course consists of four to five injections: two or three injections autovaccine made during the course of blood purification and the final one or two injections after the blood purification rate at weekly intervals (intervals). Each injection autovaccine (in terms of use 20-24 chick embryos for sowing in their alantois at 0.4-0.5 ml of blood of the patient (see. II) is determined by intramuscular injection of 5-6 ml autovaccine.

Example 4. Use of a method of virus with HIV. The course of treatment is made in accordance with Example 3. In the case of persistent virus infection general treatment should be repeated.

Example 5. Use of a method when worn and asymptomatic forms of AIDS, identified on the basis of epidemiological data. To prevent the development of acute course of infectious process or the development of virus infection treatment should be carried out in accordance with Example 1.

Example 6. Use of a method in the later stages of AIDS (with the obvious manifestations of AIDS clinic:. Lymphadenopathy, etc.) and in complicated AIDS: the presence of opportunistic infections and other complications).. Treatment is carried out in accordance with Example 3, but complement weekly administration of the bone marrow (e.g., intrasternally) blood from umbilical vein baby given antigenic compatibility (sm.III) for each week during the main treatment (cleaning-blood plasma sorption plasmapheresis -plazmosorbtsii and autovaktsinoterapiya according to example 3). Following the general course of treatment, patients should be monitored with periodic audits of serological tests for HIV. This was repeated a total treatment cycles (according to the above): the first year, 3-4 months, the second year - after 5-6 months, third year - once during the year, in order to avoid recurrence of AIDS. Only after three years after the establishment of infection or AIDS virus infection provided a full course of treatment in accordance with Examples 3, 4, 5 or 6, patients can be withdrawn from the account and observation.

Finally I have to note that the list of drugs offered by the world for the treatment of AIDS is constantly updated with new safeguards cure for AIDS. Unfortunately, the testing of such drugs in the clinic guarantees are only safeguards. The last thing offered in this area - it's AIDS treatment using extracts from the bark and leaves of the mango tree at the request of the Indian researchers. These extracts were tested in the University of California (USA) with a positive opinion on the elimination of HIV retrovirus infecting doses in model experiments on animals under the influence of such extracts. The positive effect of these extracts explain immunostimulating action on an organism infected animals. However, unfortunately, the model experiments with infected animals retrovirus HIV is not reflected something with which we have to deal with human disease doctor AIDS, and therefore expect the cure of AIDS only with the help of these drugs, it is obvious, is not necessary.

The proposed improved method for the treatment of AIDS undeniably cumbersome and expensive, but such is currently a cumbersome problem of AIDS since it is too costly for human decision, while taking lives, which is nothing more in the world. It is possible that further development of the treatment of AIDS with the planned position will facilitate the proof. The next step - the resolution of cancer and flu problems.

LITERATURE

1. Tkachenko VV The operation of the reverse data communication "protein gene "as a precondition for the survival of organisms in their habitat. / New scientific concept /. In Proc. Cheget Forum '89. Intellectual resources of scientific and technical progress. M., Proc. Scientific Research. Institute of Patent Information, 1989 part II, pp. 418-422.

2. Tkachenko VV Membrane hypothesis of opposite (protein-gene "information link as a new conception in Science In Constituen Congress, International Society for Pathophysiologi, Moscow, May 28 -. June 1, Abstracts, 9.2.20, 1991, pp 223. -223. Ibid., Problems of immunology as applied aspects of a conception of opposite (protein-gene) information link. Abstracts 7.1.36, 1991, p. 181

3. Tkachenko, VV A new code: A new approach to treating protein biosynthesis as a self-regulating system molecular-biological mechanisms. In Sov. Mtd. Rev. , Section 6, Hematology Revs. Problems in Molecular Biology and Hemostasiology, Harwood Academic Publishers GmbH. 1991, vol. VIII, pt.5, pp. 33-48

4. Tkachenko, VV Immunogenesis as a substantion and an applicative aspect of information feedback during the biosynthesis in Lymphocytes. In Russian Med. Rev. , Section 6, Hematology Revs. Thrombophilia, Cytokines, Immunogenesis, 1995, vol. VII, pt. 3, pp. 115-131r

CLAIM

1. A method of treating acquired immune deficiency syndrome (AIDS) comprising the immunogenic transplant patient cell suspension, wherein the treatment is carried out on the basis of the principles of biological science, while the patient previously extracorporeal blood purification is carried out using plasma sorption on the sorbent mark the FAS, which is covalently linked antibodies obtained using blood culture from a given patient, and is covalently bonded to glyukoproteidov isolated from human plantsety then carried immunostimulation course using autovaccine made based on inactivated blood culture from a given patient, then carry out the transplantation to the bone marrow of cord blood taken cut-off from the umbilical cord at birth with plantsetoy.

2. The method of claim. 1, characterized in that the plasmasorption, from plasmapheresis skompleksirovannuyu.

3. The method of claim. 1 and 2, characterized in that use plasma sorption glyukoproteidov isolated from human plantsety by the standard method of salting out with ammonium sulfate at 65% saturation per cent isoelectric point pI of 6.5.

4. The method of claim. March 1, characterized in that said autovaccine made based on blood culture inactivated by ultraviolet irradiation of the patient.

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Publication date 06.01.2007gg