Studies of Influenza A/H1N1 A/Tomsk/13/2010 Virus Topology during Development of Infectious Process in Mammals.

Influenza A/H1N1 A/Tomsk/13/2010 virus registered in Siberia in 2010 proved to be an extremely pathogenic strain. Dynamic study of the topology of this influenza virus strain in the lungs, liver, kidneys, lymph nodes, and great vessels of infected mice was carried out. Influenza A virus was detected by immunohistochemical methods in cells of different histogenesis in all the studied organs throughout the observation period (days 1-30 postinfection), which indicated effective replication and long persistence of influenza A/H1N1 A/Tomsk/13/2010 virus in mammalian cells.

[COMPARISON OF PHARMACOLOGICAL EFFECTS OF HEPTAPEPTIDE SELANK AFTER INTRANASAL AND INTRAPERITONEAL ADMINISTRATION TO BALB/c AND C57BL/6 MICE.]

Pharmacological effects of intraperitoneal (i.p.) and intranasal (i.n.) administration of heptapeptide selank (300 µg/kg/day for 5 days), known to possess anxiolytic and nootropic properties, were compared by studying the elevated-plus-maze behavior of inbred BALB/c and C57BL/6 mice and measuring the binding of markers to NMDA and GABA receptors of brain. The anxiolytic and nootropic efficiency of selank administered via both routes was observed only in BALB/c mice, which were characterized by initially reduced exploratory activity and higher levels of anxiety as compared to C57BL/6 mice. In BALB/c mice, i.p. selank increased the number of [G-(3)H]SR 95531 binding sites with GABA-receptors in the frontal cortex by 38%, without change in binding to NMDA receptors in the hippocampus. On the contrary, i.n. selank led to an increase in the density of [G-(3)H]MK-801 binding sites by 23% with no effect on GABA receptors. It is suggested that the differences in pharmacological spectra observed for the two routes of selank administration are determined by specific features of drug pharmacokinetics and biotransformation as well as by the dynamics of formation of the anxiolytic and nootropic effects of selank.

[Redox-dependent processes in blood plasma, neutrophils and erythrocytes of patients with ovary cancer after polychemotherapy by CAP scheme].

The dynamics of the redox-dependent processes in blood plasma, neutrophils and erythrocytes of the patients with ovary cancer of the IIIrd clinical stage by FIGO after polychemotherapy according to the CAP scheme is considered. In the blood plasma and erythrocytes there were estimated the values of protein oxidative modification: carbonyl derivatives at λ = 346 nm, 370 nm, 430 nm and 530 nm; the lipid peroxidation parameters: malonic dialdehyde, dienic conjugates, ketodiens, shiff's bases; the fermentative chain of the antioxidant system: activities of catalase, glutationtransferase and superoxide dismutase. In the peripheral blood neutrophils there were cytochemically determined the myeloperoxidase activity and the number of the active neutrophils in the spontaneous NBTR-test. After the polychemotherapy there were detected higher levels of the protein oxidative modification products and the products of the lipid peroxidation in the blood plasma and erythrocytes of the patients. Simultaneous increase of the activity of the antioxidant enzymes in the blood plasma could be evident of a high level of the lipid antioxidants peroxidation system functioning, whereas the simultaneous decrease of the activity of the antioxidant enzymes in the erythrocytes was indicative of passible development of oxidative stress in them. After the chemotherapy there was observed a significant and reliable decrease of the total number of the neutrophils. After the second course of the chemotherapy the activity of myeloperoxidase in them in the spontaneous NBTR-test as well decreased. Such a dynamics of the redox-depended processes in various components of the blood in the tumour carrier was characteristic of the tumor biological picture and required the use of differential multicomponent antioxidant therapy in patients with ovary cancer.

[Experimental study of dicholine succinate pharmacokinetics].

We have conducted for the first time an experimental study of pharmacokinetics of dicholine succinate (DCS) for different ways of its administration in rats The quantitative evaluation of DCS and its metabolites was performed by the radioactive isotope technique. Various parameters of DCS pharmacokinetics were estimated, including the dose dependence of drug content in the blood plasma, total bioavailability, distribution kinetics, and the main ways of DCS excretion.

[Combined determination of spontaneous and antigen-induced production of cytokines for differential diagnostics of active tuberculosis of lungs and latent tuberculosis infection].

AIM: Choice of informative biomarkers for diagnostics of Mycobacterium tuberculosis infection and differential diagnostics of active tuberculosis (TB) of lungs and latent tuberculosis infection (LTBI). MATERIALS AND METHODS: 54 tuberculosis patients, 47 contact by TB, individuals and 43 healthy donors were examined. All the individuals included into the study had QuantiFERON-TB Gold In-Tube (Cellestis, Australia) carried out. Values of spontaneous (NIL) and antigen-induced (AG) production of 10 cytokines (EGF, MIP-1beta, VEGF, IL-2, IL-4, IL-6, IL-1alpha, IFN-alpha2, TGFalpha, TNFalpha) as well as sIL-2Ralpha and sCD40L using XMap technology were measured. IP-10 level was also determined in 48 individuals by using EIA. RESULTS: 6 out of 13 biomarkers distinguished active TB and LTBI. As a result of construction of a decision tree in JMP 9.0 program 3 most significant markers were selected. Use of combination of IFN(gammaAG-NIL), TGFalpha NIL and IL-6AG cytokines allowed to divide TB patients and contact individuals with sensitivity of 96.3% and specificity of 80.7% (AUC = 0.9). We also observed very high levels of IP-10 and IL-2 that correlated with IFN(gammaAG-NIL) (r = 0.71 and 0.79, respectively). CONCLUSION: IL-2 and IP-10 as well as IFNgamma may be used as helpful biomarkers as a first stage for diagnostics of (M.tb.) infection. At the second stage determination of IL-6, IFNgamma and TGFalpha for differential diagnostics of active TB and LTBI is proposed.

[Effects of nootropic drugs on behavior of BALB/c and C57BL/6 mice in the exploratory cross-maze test].

Exploratory behavior, locomotor activity, and anxiety in inbred mice of C57BL/6 and BALB/c strains subchronically treated with placebo or various types of nootropic (cognition enhancing) drugs (piracetam, phenotropil, noopept, semax, pantogam, nooglutil) have been evaluated using the exploratory cross-maze test. It was found that BALB/c mice in comparison to C57BL/6 mice are characterized by greater anxiety and lower efficiency of exploratory behavior in the previously unfamiliar environment. All tested drugs clearly improved the exploratory behavior in BALB/c mice only. In BALB/c mice, piracetam, phenotropil, noopept, and semax also reduced anxiety, while phenotropil additionally increased locomotor activity. Thus, the nootropic drugs displayed clear positive modulation of spontaneous orientation in the mice strain with initially low exploratory efficiency (BALB/c) in the cross-maze test. Some drugs (pantogam, nooglutil) exhibited only nootropic properties, while the other drugs exhibited both nootropic effects on the exploratory activity and produced modulation of the anxiety level (piracetam, fenotropil, noopept, semax) and locomotor activity (fenotropil).

[Studying specific effects of nootropic drugs on glutamate receptors in the rat brain].

The influence of nootropic drugs of different groups (piracetam, phenotropil, nooglutil, noopept, semax, meclofenoxate, pantocalcine, and dimebon) on the binding of the corresponding ligands to AMPA, NMDA, and mGlu receptors of rat brain has been studied by the method of radio-ligand binding in vitro. It is established that nooglutil exhibits pharmacologically significant competition with a selective agonist of AMPA receptors ([G-3H]Ro 48-8587) for the receptor binding sites (with IC50 = 6.4 +/- 0.2 microM), while the competition of noopept for these receptor binding sites was lower by an order of magnitude (IC50 = 80 +/- 5.6 microM). The heptapeptide drug semax was moderately competitive with [G-3H]LY 354740 for mGlu receptor sites (IC50 = 33 +/- 2.4 microM). Dimebon moderately influenced the specific binding of the ligand of NMDA receptor channel ([G-3H]MK-801) at IC50 = 59 +/- 3.6 microM. Nootropic drugs of the pyrrolidone group (piracetam, phenotropil) as well as meclofenoxate, pantocalcine (pantogam) in a broad rage of concentrations (10(-4)-10(-10) M) did not affect the binding of the corresponding ligands to glutamate receptors (IC50 100 pM). Thus, the direct neurochemical investigation was used for the first time to qualitatively characterize the specific binding sites for nooglutil and (to a lower extent) noopept on AMPA receptors, for semax on metabotropic glutamate receptors, and for dimebon on the channel region of NMDA receptors. The results are indicative of a selective action of some nootropes on the glutamate family.

[Video-assisted surgery of Hirschsprung's disease in children].

Results of the laparoscopic treatment of 36 patients with Hirschsprung's disease were analyzed. The majority of patients were the newborns or before 12 months old. The use of laparoscopy permitted a full-fledged topical diagnostics, aganglionosis zone identification, gut mobilization and voiding. Laparoscopic method also allowed to decrease the ICU stay in 3,1 times and start breastfeeding earlier, lessen the pelvic floor intraoperative traumatization and decrease the level of commissural and inflammatory complications. Good and excellent functional results were achieved in 93,7% of children, stenosis of the anastomotic area was registered postoperatively in 2 patients.

Effect of long-term constant illumination of female rats on the parameters of brain development in their 40-day-old progeny.

We studied the progeny from female rats exposed to constant illumination for 1 month and mated with intact males 14 days after this exposure. At the age of 40 days, the progeny from experimental rats differed from the control by lower body weight, thickness of the adrenal cortex, and diameter of convoluted seminal tubules. The weight of the brain was similar in both groups. The thickness of the cortex in the parietal lobe, and especially, in the frontoparietal lobe was decreased; the neuronal density in these lobes (layers II and V) was reduced. In layer II and V neurons of the parietal lobe, the size of neuronal nuclei reduced, the concentration of RNA and activities of NADH and NADPH dehydrogenases in the cytoplasm were considerably increased. Elevated concentration of lipids was found in layer I of cerebral and cerebellar cortex and in the white matter, which attests to higher myelinization degree compared to the control.

[Perturbations of the vesicle cycle in reticulospinal synapses of axons in lamprey after presynaptic microinjections of GTPgammaS].

The synaptic vesicle cycle sustains neurotransmission and keeps pace between exo- and endocytosis in synapses. GTP-binding proteins function as key regulators of this cycle. The large GTPase dynamin is implicated in fission of clathrin-coated vesicles from the presynaptic membrane during endocytosis. The present study addresses the effect of the non-hydrolysable GTP analog, GTPgammaS, on the assembly of the dynamin fission complex in situ. Intraaxonal microinjections of GTPgammaS induced distinct ultrastructural changes in synapses: the number of synaptic vesicles at active zones was reduces, and the number of docked vesicles was increased; at the same time the number of clathrin-coated intermediates at the synaptic endocytic zone was increased, indicating that synaptic vesicle recycling was inhibited. Clathrin-coated intermediates with unusual shape were found. At low concentrations of GTPgammaS they were represented by long tubules decorated by spirals containing dynamin and clathrin-coated vesicles on the top. At high concentrations of GTPgammaS the tubulular structures were shorted and branched. The pitch of the spiral and tubule's diameter were significantly reduced (23.1 +/- 0.4 and 19.0 +/- 0.5 nm, respectively, as compared to those at low concentration of GTPgammaS, 26.6 +/- 0.4 and 23.3 +/- 0.4 nm; P < 0.001). We suggest that these structural changes correspond to distinct steps in the fission reaction. A model is proposed. It implies that the fast GTP hydrolysis leads to an increase in length of the spiral due to the straightening of the dynamin dimmers, composing the spiral. This leads to a fast increase both in the pitch and the diameter of the helix. The shift in diameter breaks the local hydrophobic interactions between the inner and the outer leaflets of the lipid membrane at the sites of dynamin binding. Stretching of the spiral leads to an expansion of the neck in the longitudinal direction and promotes severing of the membrane that subsequently results in the release of the clathrin-coated vesicle.

[Mitotic activity in serous adenocarcinoma of the uterine body].

A comparative study of uterine serous (36) and endometrial (32) adenocarcinoma was undertaken. The former pathology revealed a higher mitotic index (18.2%) than the latter (11.4%) (p < 0.01). Predominance of metaphases among the variety of mitotic phases as well as high metaphase-prophase index (73.6% and 69.2% in serous and endometrial adenocarcinoma, respectively) were typical of malignant tumors in general. The share of patholological mitoses in serous adenocarcinomas was 1.6 times (61.8%) that in endometrial adenocarcinomas. Moreover, serous adenocarcinomas showed a wide range of pathological mitoses which were often responsible for aneuploidy development. The above differences in uterine serous and endometrial adenocarcinoma were in full conformity with the well known aggressive nature of the former pathology and may be used as an additional criterion of differential diagnosis.

[Diagnostic role of organ-specific isoforms of lactate dehydrogenase in blood plasma].

Changes in plasm isoenzymic spectrum of lactate dehydrogenase (LDG), a glycolytic enzyme, were studied in death of cardiovascular diseases, ethanol poisoning and a mechanical injury. The tests used electrophoresis in 1% solution of agarose. Isoenzymic composition differed between the groups with acute coronary failure, chronic coronary heart disease, alcohol cardiomyopathy and a mechanical injury. Ethanol poisoning caused two types of characteristic shifts in LDG isoenzymic spectrum. Nomograms were designed for practical application in forensic medical expertise of sudden death.

[Serous adenocarcinoma of the uterus: criteria of morphological diagnosis and immunohistochemistry].

The analysis of the histopathologic features in 138 patients with uterine serous adenocarcinoma in comparison with 146 patients with uterine endometrioid papillary adenocarcinoma revealed morphological specificities of these carcinomas. Immunohistochemical study found that 66.7% uterine serous adenocarcinomas were negative both to estrogen and progesterone receptors and 86.7% uterine serous adenocarcinomas showed p53 oncoprotein overexpression. The data support the hypothesis that uterine serous adenocarcinoma is a hormone-negative tumor and that mutation of p53 tumor suppressor gene may play a leading role in its carcinogenesis.

[The synthesis and antiviral activity of glycyrrhizic acid conjugates with alpha-D-glucosamine and some glycosylamines]. / Sintez kon''iugatov glitsirrizinovoi kisloty s al'fa-D-gliukozaminom i nekotorymi glikozilaminami i ikh protivovirusnaia aktivnost'.

Glycyrrhizic acid and its 30-methyl ester were conjugated with 2-amino-1,3,4,6-tetra-O-acetyl-2-deoxy-alpha-D-glucopyranose, 2,3,4,6-tetra-O-acetyl-beta-D-glucopyranosyl amine, 2,3,4-tri-O-acetyl-apha-L-arabinopyranosyl amine, 2-acetamido-2-deoxy-beta-D-glucopyranosyl amine, and beta-D-galactopyranosyl amine using N,N'-dicyclohexylcarbodiimide and its mixtures with N-hydroxybenzotriazole. Structures of the conjugates were confirmed by IR, UV, 1H, and 13C NMR spectroscopy. The glycoconjugate with the residues of 2-acetamido-2-deoxy-beta-D-glucopyranosyl amine in the carbohydrate part of its molecule exhibited antiviral activity (ID50 4 microg/ml) toward the herpes simplex type 1 virus (HSV-1) in the VERO cell culture. Two compounds demonstrated anti-HIV-1 activity (50-70% inhibition of p24) in a culture of MT-4 cells at concentrations of 0.5-20 microg/ml. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2004, vol. 30, no. 3; see also http://www.maik.ru.

Effect of testosterone derivatives administered to pregnant rats on the brain of their offspring (delayed consequences).

We examined the offspring of rats receiving sustanon with testosterone derivatives on day 19 of pregnancy. At the age of 1 month, these rats differed from control animals in the indexes of higher nervous activity. The relative weight of the brain increased in 54-day-old rats receiving the hormone. Female rats were characterized by increased sudanophilia of the white matter in cerebellar hemispheres and high RNA content in layer V neurons of the parietal lobe, increased volume of nuclei in hippocampal neurons, weight of the ovaries (tendency), size of the largest follicles, and high activity of 3beta-ol-steroid dehydrogenase. The weight of the adrenal glands and 3beta-ol-steroid dehydrogenase activity in adrenocortical cells of the fascicular zone increased in male rats.

[Determination of the severity of endogenous intoxication at the level of average molecular peptides]. / Opredelenie tiazhesti éndogennoi intoksikatsii po urovniu srednemolekuliarnykh peptidov.

A possibility was demonstrated of using the method related with determination of the level of average-weight molecules or average molecules (AM) for the diagnosis of sudden death in forensic medical expertise of cadaver. Blood plasma was investigated according to the method of M.I. Gabriekyan modified by Yu. V. Pervushin, which involves deproteinization of samples with subsequent spectrophotometry at 254 and 280 nm. The AM levels were studied in different pathologies including cardiovascular diseases, alcoholic and narcotic intoxications and pneumonia, caused by a variety of agents, as well as the syndrome of sudden children's death. An analysis of the obtained results denotes a possibility of using the postmortem determination of the AM level concurrently with determination of an endogenous intoxication degree in the forensic medical practice for establishing the cause and mechanisms of nonviolent death.

[Expression of Hsp70 and Hdj1 chaperone proteins in human tumor cells]. / Analiz ékspressii belkov-shaperonov, Hsp70 i Hdj1, v opukholevykh kletkakh cheloveka.

Heat shock proteins (Hsp), or "stress proteins", play an important role in maintenance of cellular homeostasis both under normal conditions and during cellular stress. We tested malignant female urogenital and breast tumors for Hsp70 and Hdj1 (Hsp40) chaperones. Immunoenzyme procedure (based on Hsp70 and ATP interaction) was used to assay Hsp70 levels. Hsp70 and Hdj1 expression was higher in malignant cells than in benign ones. We were the first to demonstrate the feasibility of using Hdj1 expression as a novel prognostic factor for neoplastic disease.

[Synthesis and immunomodulating activity of new diglycopeptides of of glycyrrhizinic acid and it's 30-methyl ester]. / Sintez i immunomoduliruiushchaia aktivnost' novykh diglikopeptidov glitsirrizinovoi kisloty i ee 30-metilovogo éfira.

New amino acid derivatives of glycyrrhizic acid and its methyl ester were selectively synthesized using active N-succinimide esters. The compounds with residues of glycine ethyl ester and alanine methyl and butyl esters increased the level of agglutinins and hemolysins in blood serum of mice two- to threefold in comparison with the control upon parenteral administration at a dose of 2 mg/kg for 14 days. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2004, vol. 30, no. 2; see also http://www.maik.ru.

[Acetaldehyde and some biochemical parameters in alcoholic intoxications]. / Atsetal'degid i nekotorye biokhimicheskie parametry pri alkogol'nykh intoksikatsiiakh.

The need in comprehensive gas chromatography and biochemistry examinations is grounded for cadaver expertise in order to cope with issues related with alcoholic intoxication. Descriptions of 3 examination methods of biological fluids are elucidated, i.e. gas chromatography, electrophoresis and fixing of a degree of endogenous intoxication. The concentration of acetaldehyde in 3 body media (blood, urine and liquor) are analyzed in detail; the isoenzyme spectra of lactate-, alcohol- and aldehyde dehydrogenase as well as the contents of medium molecules in death of alcohol poisonings and due to mechanical trauma are also in the focus of attention.

[Synthesis and immunostimulating activity of cysteine-containing glycopeptide derivatives of glycyrrhizic acid]. / Sintez i immunostimuliruiushchaia aktivnost' tsisteinsoderzhashchikh glikopeptidnykh proizvodnykh glitsirrizinovoi kisloty.

New cysteine-containing derivatives of glycyrrhizic acid were synthesized by its coupling with Cys(Bzl) esters or the Cys(Bzl)-Val-OBu(t) dipeptide by the active ester method (DCC/HOSu) or by Woodward's reagent K. The derivatives with Cys(Bzl) and Cys(Bzl)-Val residues attached to the carbohydrate part of the molecule stimulated the primary immune response and the reaction of delayed-type hypersensitivity in mice at a dose of 2 mg/kg. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2004, vol. 30, no. 1; see also http://www.maik.ru.