[Recovery of spinal cord functions after experimental complete crossection under the effect of chitosan polymeric compounds]. / Vozmozhnosti klinicheskogo vosstanovleniya funktsii spinnogo mozga pri ego polnom peresechenii v eksperimente pod vozdeistviem polimernykh soedinenii khitozana.

Currently, there is no effective method of treating complete spinal cord intersection. One of the promising experimental approaches is substances promoting repair and fusion of axonal membranes. OBJECTIVE: To study clinical and functional recovery in experimental animals with transected spinal cord after management with photo-cross-linked chitosan in a homogeneous mixture with polyethylene glycol. MATERIAL AND METHODS: We studied 20 rabbits with a model of complete spinal cord transection at the level of Th9. There were control and experimental groups (n=10; n=10). In the experimental group, we intraoperatively injected photo-cross-linked chitosan in a homogeneous mixture with polyethylene glycol. Neurological status was assessed using the modified Basso Beattie Bresnahan scale. Histological examination was performed after removing the animals from the experiment. RESULTS: In the experimental group, significant regression of neurological disorders was accompanied by partial recovery of movements, sensitivity and control of pelvic functions by the 30th day of the experiment. There was no mortality in the experimental group. Paraplegia and anesthesia persisted in the control group while mortality was 40% (n=4). Histological analysis in the main group revealed axonal «bridges¼ in the area of injury and spread of DiI dye through this area. CONCLUSION: These phenomena confirm the positive effect of chitosan and polyethylene glycol on functional recovery after experimental spinal cord injury. These data are consistent with histological findings.

EXPERIMENTAL STUDY OF NEW TECHNIQUES IN MODELLING OF ACUTE ADHESIVE SMALL BOWEL OBSTRUCTION.

The aim of our research was the elaboration of acute adhesive small bowel obstruction model and further study of pathomorphological changes of small bowel and peritoneum in experiment. The study was carried out on 40 rats which were done suturing of sero-serous small bowel stiches and modelling of angulations of small bowel with the use of mechanical and implantation factors of peritoneal adhesive disease development according to the worked out by us technique. The suggested experimental model allows modelling the acute adhesive small bowel obstruction just on the 3rd day of the experiment and specifying some aspects of adhesion formation in peritoneum under pathomorphological study.

[The influence of the drying method for food properties and hypolidemic potential of oyster mushrooms (Pleurotus ostreatus)].

The aim of this research was to study the influence of two drying methods: freeze-drying sublimation and dry-air drying on the selected nutritional properties and hypolipidemic potential of fruiting bodies of oyster mushroom (Pleurotus ostreatus). The criteria for evaluation of the food properties were the color, the morphological structure, regidratation capacity, the total level of soluble proteins, fats, polysaccharides, free amino acids and monosaccharides. Lipid-lowering potential of oyster mushroom was evaluated by the concentration of lovastatin and the level of antioxidant activity. It has been experimentally revealed that the value of optical density of hydro-alcohol extracts of dried oyster mushrooms at a wavelength of 295 nm most clearly characterized its color which intensity was almost twice less in sublimated mushrooms, than шт the sample dried by dry-air method. Histological data showed that dry-air drying lead to the destruction of the mushroom cells and to the formation of a dense layered structure. Sublimation drying preserved the ordered cell structure and provided less deformation and shrinkage of the tissues. Using X-ray microtomography it was reported that freeze-dried mushrooms had uniform pore volume distribution. Dry-air dehydration method lead to the formation of larger cavities. The average percentage of the open pores was: 29.41±0.52% (after dry-air method), 11.10±0.41% (after freeze-drying method). Respectively the number of closed pores, which reflected the true value of porosity, was 0.99±0.01 and 1.75±0.01%. Structural differences of the samples of the dry oyster mushroom combined with their unequal hydration ability. Indicator of rehydration for oyster mushroom dried by sublimation method was 5.4±0.1, and for samples obtained by dry-air method it was 3.2±0.1. Respectively the average time of maximum water absorption was 22.7±1.8 and 45.3±2.9 minutes. It was found that the freeze-drying sublimation conditions were more conducive for the preservation of the biologically active protein and polysaccharide components of oyster mushrooms and on the other hand dry-air drying method increased the nutritional value of oyster mushrooms due to the reactions of polysaccharides autohydrolysis. The number of proteins and polysaccharides of the Oyster mushrooms samples dried by dry-air method and freeze-drying method was 72.0% and 56.0% respectively. Concentrations of free amino acids and glucose in the samples dried by freeze-drying and dry-air methods were 11.60±0.31%; 175.20±6.10 mg% and 7.00±0.28%; 144.0±5.7 mg% respectively. It has been experimentally recorded that the conditions of freeze drying were optimal in terms of ensuring the preservation of the content of natural statin and the antioxidant capacity of oyster mushrooms that provided its hypolipidemic potential. The amount of lovastatin in an the freeze-dried samples was 342±9.0 mg/kg, and was significantly higher than in the samples received by dry-air method - 190±6.0 mg/kg. The level of antioxidant activity of the oyster mushrooms samples were respectively 3.83±0.02 against 2.0±0.03 mmol/100 g. The conducted researches proved that for the production of dry oyster mushroom as a potential biologically active feedstock for the functional food products with lipid-regulating directivity the choice of the drying method had a fundamental importance.

[Production of Methane in Anaerobic Digestion of Organic Waste of Vegetable Processing Facilities].

The article concerns converting waste from vegetable processing facilities into methane in anaerobic reactors with a small amount of inoculum (8.4%). Anaerobic digestion of vegetable waste with a high content of organic acids and carbohydrates makes it possible to achieve a methanogenesis productivity of 273­436 L CH4/kg of volatile solidis, which is comparable to or higher than the productivity of such reactors in the world (according to the literature). The contents of ammonia nitrogen and soluble phosphorus in the form of on undiluted substrate basis in the digested vegetable wastes ranged from 3.39 to 5.06 and from 0.78 to 1.03 g/L respectively. Thus, mineralized vegetable waste can be used as an organic fertilizer with a high nutrient content. The results show the feasibility of the technology of conversion of organic waste from vegetable processing facilities into methane and organic fertilizer in anaerobic fermenters (digesters).

[RETROSPECTIVE ANALYSIS OF BIOLOGICAL AND MOLECULAR-GENETIC PROPERTIES OF STRAINS - CAUSATIVE AGENTS OF CHOLERA - ISOLATED IN UKRAINE IN 1994 - 2011].

AIM: Retrospective analysis of biological and molecular-genetic properties of strains - cau- sative agents of cholera - isolated in the period of epidemics in Ukraine in 1994 - 2011. MATERIALS AND METHODS: Phenotypic and molecular-genetic properties of 5 strains of cholera vibrios, biovar El Tor isolated from cholera patients and 4 strains from the environmental samples were studied using traditional bacteriological and genetic methods. Detection of DNA for toxigenicity genes and genes characteristic for El Tor and classic biovar were carried out by PCR method using rea- gent kits <> and <> an experimental test system). Sequencing of genomes of 4 strains of causative agents of cholera was carried out in genetic analyzer Ion Torrent Personal Genome Machine. RESULTS: Strains of cholera vibrios identified in Ukraine in 1994 and 2011 such as a typical toxigenic biovar El Tor (V cholerae Ο1, El Tor, Ogawa, Hly-, ctxA+, tcpA+) contain genes ofthe classic cholera vibrio in their genome and are genetically altered (hybrid) variants of cholera vibrio biovar El Tor producing enterotoxin CTI and having increased virulence, that was clinically manifested in predominance of severe forms of cholera in Mariupol of Donetsk region in 2011. Genome sequences ofthe 4 studied strains were deposited into the international database DDBJ/EMBL/GenBank. CONCLUSION: The studied isolates were established to belong to a clade of strains associated with cholera outbreaks in Haiti and Asian continent, from where genetically altered strains of cholera vibrios biovar El Tor were introduced to Haiti in 2010, based on results of comparison of genomic sequences of the studied strains with genomes of V cholera strains from the international database GenBank.

[FEATURES OF MASS-SPECTROMETRIC PROTEIN PROFILES OF STRAINS OF BRUCELLOSIS CAUSATIVE AGENT DURING PREPARATION OF CULTURE ON VARIOUS NUTRIENT MEDIA].

AIM: Carry out comparative analysis using time-of-flight mass-spectrometry with matrix laser desorption/ionization (MALDI-TOF MS) of protein profiles of brucellosis causative agents (Brucella melitensis Rev-1 and Brucella abortus 19BA), cultivated in various nutrient media: Albimi agar, brucellagar and erythrit-agar. MATERIALS AND METHODS: Vaccine,strains: Brucella melitensis Rev-1 and Brucella abortus 19BA. Protein profiling in linear mode on Microflex "Bruker Daltonics" MALDI-TOF mass-spectrometer. RESULTS: A number of characteristic features of brucella mass-spectra was detected: in particular, preservation of the total qualitative composition of protein profiles of cultures and significant differences in the intensity of separate peaks depending on the nutrient medium used. CONCLUSION: Based on the analysis of the data obtained, use of Albimi agar as the nutrient medium for preparation of brucella culture samples for mass-spectrometric analysis was shown to be optimal.

[New single nucleotide polymorphisms of androgen receptor gene (AR) in the Russian breed of Dzhalginsky Merino sheep].

This paper investigates the structure of androgen receptor gene (AR) in the Russian breed of Dzhalginsky Merino sheep. Polymorphisms of the gene were detected using NimbleGen sequencing technology (Roche, United States). Eight single nucleotide polymorphisms (SNPs) and two insertions were detected. Five of these SNPs (c.335T>G, c.339G>A, c.342T>C, c.2491-327T>A, and c.2491-325A>T) and both insertions were identified for the first time. Three SNPs and the insertions are located in the coding part of exon. Insertion c.336_337 is found in most of the animals of this breed and can be used as a genomic marker of the breed. Animals with mutant variant of SNP c.1496+15T>C have significantly lower live weight and body size compared with the wild type genotype. This SNP can be used as a genetic marker of meat production in marker-assisted selection.

[On the classification of the failure to render medical assistance to the patient].

The authors propose the classification of the failure to render medical assistance to the patient. At present, much attention is given to the analysis of drawbacks in the provision of medical care to the victims of various human activities. A variety of classifications of these disadvantageous practices have been proposed. However, they either only casually deal the problem of failure to render medical assistance to the patient or totally disregard it. The necessity to develop the new classification of the failure to render medical assistance to the patient arises from the formulation of article 124 of the Criminal Code of the Russian Federation that requires to evaluate the severity of harm to the health of the patient resulting from the inaction of the person(s) obliged to provide the necessary medical aid.

[The immunohistochemical study of the structures of the cardiac conduction system in the case of death from alcoholic cardiomyopathy].

The objective of the present study was to study the morphological criteria for toxic cardiopathy with the use of histological and immunohistochemical methods. The results of immunohistochemical studies of the sinoatrial node---???---(SAN) and the working myocardium in the patients presenting with alcoholic cardiomyopathy (ACMP) are presented. It was shown that vimentin expression in the SAN structures and the contractile myocardium is slightly increased whereas the expression of sarcomeric actin is decreased and that of fibrinogen is increased too. The authors put forward an assumption about the role of lesions in the membrane apparatus of the pacemaker cells in the development of arrhythmia characteristic of tanatogenesis associated with alcoholic cardiomyopathy.

[The characteristics of the notions used in forensic medicine and their logical basis].

The authors briefly characterize the formerly poorly defined notions used by forensic medical experts for the completion of the conclusions based on the results of primary forensic medical examinations of the corpses. The main errors made in the use of these notions encountered in the materials of primary forensic medical expertises of the corpses, the recommendations to avoid these mistakes are proposed. The present paper continues the series of specialized articles dealing with the main types of logical fallacies encountered in the expert conclusions with a view to further publication of the results of the in-depth analysis of logical mistakes in these documents.

[The definition of a failure to render assistance to the patient in the forensic medical practice].

The authors consider the necessity of determining the severity of harm inflicted to human health by the inaction of the officials and other persons responsible for rendering assistance to a patient in accordance with article 124 of the Criminal Code of the Russian Federation. The absence of the normative formulation of the forensic medical criteria for the assessment of the degree of the harm to human health as a result of inaction of the subjects responsible for rendering medical aid creates difficulties for the enforcement of article 124 of the Criminal Code of the Russian Federation in its present wording. This problem awaits further studies for the possible adoption of the uniform normative and regulatory documents for the consistent application of the definition of inaction with reference to the assessment of not rendering medical assistance.

[Building questions in forensic medicine and their logical basis].

The authors characterize in brief the requirements to the correct formulation of the questions posed to forensic medical experts with special reference to the mistakes made in building the questions and the ways to avoid them. This article actually continues the series of publications of the authors concerned with the major logical errors encountered in expert conclusions. Further publications will be dedicated to the results of the in-depth analysis of the logical errors contained in the questions posed to forensic medical experts and encountered in the expert conclusions.

[Improvement of laboratory diagnostics of cholera due to genetically altered (hybrid) variants of cholera Vibrio biovar El Tor].

AIM: Improvement of laboratory diagnostics of cholera taking into the account appearance of hybrid variants of cholera vibrio El Tor biovar in the 1990s. MATERIALS AND METHODS: Phenotypic and molecular-genetic properties of typical toxigenic (151 strains) and hybrid (102 strains) variants of El Tor biovar cholera vibrios, isolated in the Caucuses in 1970-1990 and 1993-1998, respectively, were studied. Toxigenicity gene DNA fragments, inherent to El Tor biovars or classic, were detected by using a reagent kit "Genes of Vibrio cholerae variant ctxB-rstR-rstC, REF" developed by us. RESULTS: Reagent kit "Genes of V. cholerae variant ctxB-rstR-rstC, REF" is proposed to be used for laboratory diagnostics of cholera during study of material from humans or environmental objects and for identification of V. cholerae 01 on genome level in PCR-analysis as a necessary addition to the classic scheme of bacteriological analysis. CONCLUSION: Laboratory diagnostics of cholera due to genetically altered (hybrid) variants of cholera vibrio El Tor biovar is based on a complex study of material from humans and environmental objects by routine bacteriologic and PCR-analysis methods with the aim of detection of gene DNA fragments in the studied material, that determine biovar (classic or El Tor), identification of V. cholerae O1 strains with differentiation of El Tor vibrios into typical and altered, as well as determination of enterotoxin, produced by the specific cholera vibrio strain (by the presence ctxB(El) or ctxB(Cl) gene DNA fragment, coding biosynthesis of CT-2 or CT-1, respectively).

[Comparative study of pharmacokinetics of ofloxacin in a free and niosomal forms in experiments on white mice when administered per os].

AIM: To study features of pharmacokinetics of ofloxacin as a part of anion PEGylated niosomes on a basis of sorbitan monostearate (Span 60) to experimental white mice per os. MATERIALS AND METHODS: Ofloxacin was entrapped in niosomes consisting of Span 60, cholesterol, PEG 4000 and dicetylphosphate. Sizes of niosomes estimated by means of probe microscopy. Efficiency of inclusion of an antibiotic in niosomes defined after removal of free drug by a centrifugation. The analysis of the quantitative contents of ofloxacin in samples carried out a method of a high performance liquid chromatography. RESULTS: We studied the main pharmacokinetic parameters of ofloxacin when used free and niosomal forms of antibiotic to experimental white mice per os. It is shown that use of oral niosomal forms leads to decrease of maximal concentration in serum and increase of ofloxacin half-life by 7,4 times in average compared to the free form. It is determined that bioavailability of ofloxacin in the niosomal form is 154% relative to the free form of the antibiotic. CONCLUSIONS: Niosomal microcontainers are perspective technology of encapsulation and the directe transport of antibacterial preparations through biological barriers. Using of niosomal formulation of ofloxacin is able to afford to increase considerably efficiency of treatment in comparison with a free form and significantly decrease negative effects of antibiotic therapy.

[Effect of new derivatives of natural nitrogen-containing compounds on the level and metabolism of biogenic amines in brain structures and the life span of rats].

Effects of the analogs of transmitter amino acids on the level of biogenic amines and their metabolism in various structures of brain and on the life span of rats have been studied.

[Use of multiple locus variable number tandem repeats analysis for the Brucella systematization].

UNLABELLED: The methods of molecular-genetic differentiation to strain level acquire increasing significance in the current system of struggle with brucellosis. MLVA (multiple locus variable number tandem repeats analysis) was selected for molecular-genetic differentiation to strain level and simultaneous establishment of the genetic relationship of investigated Brucella strains. The goal of this work was MLVA typing of three pathogenic Brucella species strains with the analysis of stability of chosen loci, discrimination power and concordance to conventional phenotypic methods of the Brucella differentiation for use in systematization of brucellosis causing agents. MATERIALS AND METHODS: Twenty six Brucella strains representing reference (n = 15), vaccine (n = 2) and field strains of three pathogenic Brucella species were tested: B. melitensis (n = 3), B. abortus (n = 2), B. suis (n = 2), and isolates (n = 2) with unidentified taxonomic position using MLVA with 9 pairs primers on known variable loci of Brucella genome. The analysis of the stability of chosen loci, discrimination power on Hunter-Gaston discrimination index (HGDI) and consistency to phenotypic methods of identification was performed. RESULTS: MLVA was confirmed for the results of phenotypic methods of identification, stability of the chosen loci in majority reference, and vaccine strains with a high index of variability HGDI 0.9969 for all loci. A dendrogram was plotted on the basis of MLVA data on distributed Brucella strains in related clusters according to its taxonomic species and biovar positions and construction of 25 genotypes. B. melitensis strains formed cluster related to the reference strain of B. melitensis 63/9 biovar 2. Australian isolates of Brucella 83-4 and Brucella 83-6 isolated from rodents formed a cluster distant from other strains of Brucella. CONCLUSION: MLVA is a promising method for differentiation of Brucella strains with known and unresolved taxonomic status for their systematization and creation of MLVA genotype catalogue that will promote qualitative improvement of brucellosis surveillance system in Russia.

[Effect of new adenine derivative on emotional status of rats under conditions of "voluntary alcoholization" model].

In continuation of the study of pharmacological properties of the new drug VMA-99-82 (9-[2-(4-isopropylphenoxy)ethyl]adenine), the influence of this drug on the psycho-emotional status of rats was evaluated using a voluntary alcoholization model. It is established that VMA-99-82 administered at doses of 10 and 50 mg/kg produces a positive effect on impaired cognitive functions in alcoholized rats and eliminated the depression caused by the consumption of excess ethanol.

[Antidepressant effect and mechanism of action of a novel aminouracil derivative].

The psychotropic activity of a series of new aminouracil derivatives was studied on the widely applied models of pharmacological screening in rats. The most expressed antidepressant effect at the stage of primary pharmacological screening was been revealed for compound PIR-03-52 in a dose of 10 mg/kg. A profound study of the antidepressant activity on models of pathological states, PIR-03-52 exhibited a pronounced antidepressant effect that was comparable with that of a reference drug (imipramine). The given effect is related to the positive influence of PIR-03-52 on the serotoninergic and dopaminergic systems.

[Circumstances of the longevity of bats (based on the studies of bats over the territory around Saint-Petersburg)].

According to Medawar's concept on the accumulation of harmful mutations, bat longevity should be explained by the fact, that they have no enemies: since the organisms were not being exterminated at the young age, the selection towards longevity took place at their ancestors; the specific mutations causing senescence have not been accumulating. However bats have natural enemies, and in natural environment bats usually die long before maximum lifespan. This is confirmed by direct observations and, especially, by the fact, that the bat populations increase slowly. In 1950-60s the bat populations declined over the territory around Saint-Petersburg because of inappropriate scientific research (mass sampling, unsuccessful banding and experiments), and they still cannot reach the past condition. The bat longevity is caused by long periods of the inactive condition, slow reproduction rate, little light exposure. Concept of mutations accumulations have to be rejected. The following concept is proposed instead: senescence results from any change in organism, i. e. the changes of an organism are inevitable, are not always healthy, and are possible in definite directions, that is why they become unhealthy in a moment.

[Neurochemical mechanisms of antidepressant action of new adenine derivative].

We have studied the influence of the new adenine derivative VMA-99-82 on the exchange of monoamines and their metabolites in the brain of Wistar rats. In addition, the effect of VMA-99-82 on binding of 5-HT1A and 5-HT2A serotonin receptors and the system of 3H-serotonin reuptake in the brain synaptosomes has been studied in vitro. It is established that VMA-99-82 at a dose of 10 mg/kg has no affinity to 5-HT1A and 5-HT2A serotonin receptors and produces no effect on the reuptake of [3H]-5-HT. It is suggested that VMA-99-82 has a modulating effect on ion channel NMDA receptor complex of the glutamatergic system, which leads to the manifestation of antidepressant activity.