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The aim of the research was to study the mechanism of intermolecular interaction of allicin and lecithin with pancreatic lipase, and developing the composition, contributing to the inactivation process of lipolysis, and reduce the absorption of fats in the organism. Methods of computer chemistry have been used for modeling spatial structures of allicin and lecithin. Geometric optimization was carried out, quantum chemical characteristics and the distribution of charge density of the molecules of the studied biologically active substances and pancreatic lipase were studied. In the study of the molecular properties of the lipase of pancreatic juice before and after molecular docking, it was found that one molecule of lecithin didn't fully block the active site of the enzyme. For complete inactivation of lipase, two molecules of lecithin or one molecule of allicin were required (Epot. = -412.36 and -159.4 kcal, respectively). An optimal composition of supplement to blocking pancreatic lipase has been set: allicin containing additive - 95% (75% sunflower oil, 25% chopped garlic), lecithin - 5%. The efficacy of lipase inactivation by supplement stored at 2-4 °C in fat-water mixture has been studied. It was found that after storage for three days the developed composition retained its properties. Acid number of fat, subjected to enzymatic treatment varies slightly (0.1 to 0.25 mg KOH/g). Peroxide value and microbiological characteristics of the investigated fraction also did not exceed the permissible norms. When evaluating the biological value of the developed composition in the experiment on white BALB/c mice (with initial body weight of 20-30 g), it was found that its administration on the background of high fat (19%) diet at a dose of 6% of the diet fat from the 15th to the 40th day was accompanied by 1.5 fold reduce in body weight increasing in comparison with animals which received no additives. Their blood levels of total lipids, cholesterol, triglycerides and glucose reduced while the level of total protein and urea increased to the level of the control group (without obesity).

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The authors validate the efficiency of enzyme-linked immunoassay (ELISA) for rapid diagnosis of rabies and titration of specific antibodies and of the method of isolation of field rabies strains in the rat Gasser's node neurinoma cells. Highly active specific diagnostic agents for ELISA and immunofluorescent test based on sheep immunoglobulins have been created for detection of the rabies agent in pathological material from different animals and humans.

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The present work summarizes the results of 11 groups of experiments carried out with the aim to complexly quantify the residual virulence of a cold mutant of the Vnukovo-32/107 rabies virus vaccination strain intended for the preparation of an oral rabies vaccine (Kamark) for the immunization of free-living carnivores. According to WHO prescriptions, residual virulence was quantified in experiments on carnivores, mainly red foxes (Vulpes vulpes)--the presumed target species, and farm-bred polar foxes (Alopex lagopus)--a related species. Further experiments were carried out in cats, dogs, non-target autochthonous micromammals, predatory birds (Microtus arvalis, Apodemus flavicollis, Falco tinnunculus) and in a large number of laboratory animals--white mice. At oral administration (including extremely high doses) the strain Vnukovo-32/107 proved to be apathogenic to the target carnivores--Vulpes vulpe and Alopex lagopus as well as cats, dogs and the autochthonous micromammals. For Falco tinnunculus the strain proved to be apathogenic even at intramuscular and intracerebral administration. The residual virulence of the Vnukovo-32/107 vaccination strain, also quantified by comprehensive model experiments on white mice of different weight categories that had been infected orally, subcutaneously, intramuscularly, intracerebrally, by contact, with ingestion of rabic material or by modelled immune suppression, proved to be extremely low-levelled. The strain under investigation revealed a high level of attenuation and a low level of residual virulence and proved to be suitable for the preparation of non-reactogenic oral vaccine intended for foxes, an extremely susceptible target species.

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The glycoprotein gene of the rabies virus vaccine strain Vnukovo-32 was sequenced and the deduced protein sequence was analyzed and compared with that of various laboratory and street strains. The amino acid sequence homologies of strain Vnukovo-32 were compared with fixed strains ERA, SAD B19, PV, HEP-Flury, CVS and two street strains, canine and CXX89-1, were 98.9% (6 replacements), 98.3% (9), 96.2% (20), 91.4% (45), 87.0% (68), 93.5% (34) and 91.4% (45), respectively. Sequence alignments of the proteins revealed that the most conserved region is the ectodomain, whereas the transmembrane and cytoplasmic domains showed significant divergence.

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271 strains of rabies virus, isolated in different regions of the former USSR, were studied with the use of monoclonal antibodies P-41 (Germany). All strains isolated in the arctic regions, as well as some of the strains isolated in nonarctic regions situated 300-1700 km south of the polar circle (the Baltic countries, Central Yakutia, the eastern part of the Transbaikal region), gave positive reaction with these antibodies. Cases of hydrophobia caused by virus with the 41 (+) marker were described. Information on the spread such strains in other parts of the world is presented.

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A combined vaccination schedule using commercial antirabies immunoglobulin G and experimental vaccine from strains Vnukovo-32 or Yuli beginning 2 hr before intracerebral (i.c.) challenge with a high dose of Yuli virus conferred no protection to Cercopithecus aethiops monkeys. In monkeys inoculated into lip with a middle dose of Yuli virus, administration of large amounts of antirabic IgG (up to 5000 national units, NU/kg) had a clearcut effect. The disease in Yuli virus-infected monkeys showed typical signs of acute encephalitis with lethal outcome, although one animal which developed typical encephalitis recovered as evidenced by increased virus-neutralizing antibodies in its serum. Inflammatory and degenerative lesions developed in the CNS of animals with signs of acute encephalomyelitis; their intensity was less prominent in those monkeys which underwent the combined treatment. In the cytoplasm of brain neurons of monkeys infected with Yuli virus relatively small Babes-Negri bodies with more or less apparent internal structure were detected.

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Two strains (UB-1 and UB-2) of rabies-related viruses were isolated from the brain of Nyctalus noctula and Vespertilio murinus captured from the hollows of tall trees on the left bank of Pripyat river in the Volynsky region of Ukrainian S.S.R. The viruses were isolated by means of intracerebral inoculation to white mice. The isolates were identified as rabies-related viruses of Duvenhage type in an indirect test of fluorescent antibodies with the panels of nucleocapsid monoclonal antibodies (NC Mab) provided by Wistar Institute (Philadelphia) and by Central Veterinary Laboratory (CVL, Weybridge). During the typing with the Wistar panel of NC Mab complete antigenic similarity was established between the newly isolated strain and Yuli virus. The reaction with CVL NC Mab revealed group-specific antigenic similarity between Yuli virus on one hand, Duvenhage-6 and Duvenhage-66 on the other hand, as well as between UB-1 and UB-2 and Duvenhage-26. The reaction with antibodies to clones DB-3,4,6,9, and 10 detected antigenic similarity between the viruses of chiropteric origin isolated in the U.S.S.R., North-West Europe as well in Africa, although some differences were discovered. Yuli, UB-1, and UB-2 viruses isolated in the U.S.S.R. were proved to belong to Duvenhage group of viruses (serotype 4).

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98 rabies virus strains isolated from different species of wild and domestic animals have been studied by means of 36 monoclonal antibodies obtained from the Wistar Institute (USA) and 3--of the Federal Research Center on Viral Diseases of Animals (FRG). The antigenic variants determined in this study have been analyzed in comparison with the data obtained in other regions of the world, thus establishing the spread of these variants and their relationship with different species of animals.

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Yuli virus was isolated by intracerebral (i.c.) inoculation of suckling mice with a 10% brain suspension from 11-year-old patient who died under signs of atypical hydrophobia after a bat bite into lower lip. Identification with a panel of monoclonal antibodies (MoAb) to nucleocapsid protein (NP) confirmed that Yuli virus belongs to Lyssavirus genus, as an antigenic variant of the European Duvenhage virus.

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Tissue culture rabies vaccine has been used for subcutaneous immunization of 158 subjects according to official instructions and also for intramuscular immunization of 128 subjects according to a short schedule with booster inoculations. All 286 subjects were either bitten or contaminated with saliva of rabid animals or animals suspected of having rabies. The 1168 serum samples were tested by neutralization test (NT) in mice, by radial haemolysis (RH) and by indirect haemagglutination (IHA). The highest, earliest and longest active post-vaccination immunity was registered after the most intensive subcutaneous vaccination course at a dose of 5 ml for 25 days with 3 booster inoculations. Subcutaneous inoculation of 3 ml vaccine for 12 days (36 ml) failed to produce a satisfactory elevation of antibody titre. After 2 to 4 booster inoculations, however, a satisfactory level of antibody was observed. The tissue culture vaccine was shown to have good prospects for clinical vaccination by intramuscular route. On intramuscular vaccination at 1.5 ml for 9 days with 6 booster inoculations on days 16, 23, 30, 37, 67 and 97 (initial vaccine volume 45 ml) the mean geometric antibody titres (MGT) reached 93, 160, 322 and 165 on days 30, 60, 90 and 112, respectively. The economically efficient and rapid IHA and RH tests were confirmed to be specific and suitable for titration of antirabies antibody.

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In a coded experiment on rabies virus antibody detection in the blood sera of humans immunized with rabies vaccines, coincidence of the results of indirect ELISA, neutralization test and radial hemolysis test (RHT) was observed in 83% and 90% of the cases. The correlation coefficient of antibody titres in ELISA and NT was r = +0.75, ELISA and RHT r = +0.81. Instances of discrepancy of the results were observed with sera of low titres, no more than 1:50 by neutralization test.

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Continuous cells of green monkey kidney 4647 were found to be highly sensitive to three variants of the Vnukovo-32 vaccine strain of rabies virus preliminarily adapted to the primary line of Syrian hamster kidney cells. The virus litre increased with passages in the 4647 cell culture. The resulting virus variants had sufficient immunogenic potency. The time course of virus replication at a temperature of 32 degrees C was studied. The heteroploid cell line 4647 was found to be promising for manufacture of medical and veterinary cultural rabies vaccines.

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