New Data on the Distribution of the Small Cave Bear (Ursus ex gr. savini-rossicus) in the Urals.

The lower molar (m1) of cave bears from Late Pleistocene localities of the Urals was studied employing the methods of traditional morphometry and geometric morphometrics. On the basis of the size and shape variation of m1, the small cave bear (Ursus ex gr. savini-rossicus) was found to have been a part of the faunas from the caves Skazka, Viasher, Dynamitnaya, Chudesnitsa, and Chernye Kosti. The small cave bear presence in faunas from the Medvezhya, Makhnevskaya Ledyanaya, Asha 1, Ignat'evskaya, and Barsuchii Dol caves was confirmed as well. The species range of the small cave bear encompassed the Northern, Middle, and Southern Urals in the Late Pleistocene. The ranges of the small cave bear and cave bear (Ursus kanivetz) overlapped from the beginning (marine isotope stage 5e) to the middle (middle marine isotope stage 3) of the Late Pleistocene.

[Preclinical and clinical trials of the new tuberculosis drug perchlozon].

The paper sets forth the stages of design and introduction of the new Russian tuberculosis (TB) drug perchlozon registered in the Russian Federation in 2012. Based on the results of Phases I-III clinical trials, the authors evaluate the efficacy and safety of the agent and consider the adverse effects of its treatment for respiratory TB. The use of perchlozon as a component of combination therapy versus standard chemotherapy regimens significantly reduces abacillation time in pulmonary TB caused by its drug-resistant pathogen. In terms of the higher prevalence of TB induced by its pathogen resistant to many drugs (with multiple and broad-spectrum drug resistance), perchlozon is an essential drug that has antituberculous activity mainly against multidrug-resistant Mycobacterium tuberculosis strains and gives patients with the severest and epidemiologically poor form of TB the chance to recover.

Effects of hydroxypyridine derivatives mexidol and emoxypin on the reparative processes in rabbit eye on the models of corneal epithelial defect and conjunctival ischemia.

Deepithelialization of the cornea (diameter 7 mm) was performed in rabbits and the rate of defect epithelialization was evaluated. Conjunctival ischemia was modeled by application of graduated alkaline burn. Antioxidant activity and content of nitrates and nitrites was measured in the tear fluid before and after burn by chemiluminescence and Griess methods, respectively. Emoxypin and mexidol promoted healing of corneal epithelial defect at the stage of epitheliocyte migration to the defect area and at the stage of their proliferation, respectively. After treatment with both agents, the area of conjunctival ischemia decreased more rapidly, but the efficiency of mexidol was higher. Antioxidant activity and content of products of NO metabolism in tear fluid decreased after burn. Mexidol, but not emoxypin, increased these parameters. Thus, mexidol and emoxypin have different effects on corneal epithelialization and conjunctival ischemia and effects of mexidol are more pronounced.

[Comparative efficacy of clinical use of reamberin, remaxol and ademethionine in patients with tuberculosis of the respiratory organs and liver drug-injury].

The efficacy ofreamberin, remaxol, S-adenosyl-L-methionine (ademethionine) and 5% glucose solution was estimated in the treatment of patients with tuberculosis of the respiratory organs and drug hepatotoxicity signs confirmed by higher activity of liver indicative enzymes and nitrogen oxide levels. Remaxol showed a pronounced positive effect on the cytolytic syndrome signs, evident from lower activity of alanine aminotransferase and aspartate aminotransferase. At the same time ademethionine was superior to remaxol in the effect on the cholestatic signs and inferior in the effect on the cytolytic signs. By the effect on the activity of alanine aminotransferase and aspartate aminotransferase, reamberin was inferior to remaxol and superior to ademe-thionine, its effect on the cholestasis markers level vs. the other drugs being superior only to that of 5% glucose solution. As compared to reamberin, ademethionine and 5% glucose solution, remaxol promoted higher integral indices of the host antioxidant protection (total antioxidant capacity and total antioxidant status), that partially explained the drug pronounced hepatoprotective effect.

[Hepatotropic therapy in treatment of liver injury].

At present, the conception of the use, efficacy and safety of hepatotropic agents in treatment of drug-induced liver injury, in particular due to antituberculosis drugs is not yet final, which is conditioned by extremely rare clinical trials on the subject adequate to the up-to-date principles of the conclusive medicine. The review presents data on the hepatotoxic effect of antituberculosis drugs, analysis and systematization of the data on the use of hepatotropic agents in liver injury induced by antituberculosis drugs, the principles and characteristics of their clinical use. The mechanism of action of remaxol, a new original hepatotropic agent and the indications of its use are discussed. The experimental findings on the remaxol ability to decrease the antituberculosis drug-induced liver injury through lowering the carbohydrate, albuminous and fatty degeneration and activating the organ reduction are presented. The clinical trials are evident of the most efficient action of remaxol on the signs of toxemia, as well as cytolysis and cholestasis, which along with its antiasthenic and antidepressant action allows to use remaxol as an universal hepatotropic agent in the treatment of diverse drug-induced liver injuries in both the therapeutic and prophylactic schemes.

[The clinical laboratory comparison in the assessment of treatment prognosis in patients with infiltrative tuberculosis of lungs].

The comprehensive prospective examination of 66 patients with first established non-treated infiltrative tuberculosis of lungs was used to analyze the possibility of optimization of assessment of course of specific inflammatory process on the basis of levels of proteins-reactants of acute phase. The characteristics of dynamics of clinical roentgenologic data and terms of coming of abacillarization as a result of three months anti-tuberculosis therapy has been used as grouping factors. It is established that the constellation of 3 out of 12 basic (before treatment) analyzed level indicators are the most prognostic informative--haptoglobin, ceruloplasmin and blood albumin. Their combined application according the proposed decisive rule provides 90.6% of effectiveness of prognosis of the results of treatment in the discussed category of patients.

[Peculiarities of the circulating phagocytes functional activity in patients with different forms of drug-resistant pulmonary tuberculosis].

Functional activity of circulating phagocytes (macrophages--Ms and neutrophils--Ns) was studied in 30 patients with infiltrative (I) and 30 patients with fibro-cavernous (FC) pulmonary tuberculosis (PT). Difference of the functional activity of both types of cells depending on the PT form was revealed: more significant increase in the oxygen-depending activity in FCPT while bactericide potential estimated with a zymosane induced NST-test was more pronounced in IPT patients. These data correlate with the blood levels of neopterin and elastase, the markers of the M and N activity, respectively. Participation of intracellular ADA in realization of oxygen-depending processes was demonstrated. Results of the multivariant analysis of the whole complex of the studied phagocyte characteristics, reflect their different roles in ther pathological process a prevailing role of Ms in the firstly diagnosed acute tuberculosis process (IPT) and Ns in the chronic progressive process (FCT).

[Localization of human cytomegalovirus immediate-early protein IE-1 pp72 in the juxtanuclear inclusion at the late stage of infection].

Immediate-early protein IE-1 pp72 is one of the most abundant proteins at the early stage of human cytomegalovirus infection and has a number of intranuclear activities. This paper gives immunocytochemical and ultrastructural data on IE-1 pp72 accumulation in the juxtanuclear inclusion at the late stage of low-multiplicity infection. Detection of a new localization site infers that this protein may participate in the final steps of virus morphogenesis and play a functional role in the pathogenesis of cytomegalovirus infection.

In silico analysis of the structure of variable domains of mouse single-chain antibodies specific to the human recombinant interferon beta1b.

In silico analysis of the DNA encoding single-chain Fv antibodies (ScFv) specific to the human recombinant interferon beta1b and alpha2b (rhIFN-beta1b, rhIFN-alpha2b) has been carried out. The V-, D- and J-gene segments, the complementarity-determining (CDR) and framework (FR) regions, n-nucleotides as well as mutation rates which take place during the affinity maturation of the examined sequences have been determined. For the panel of ScFv against rhIFN-alpha1b isolated from an immune combinatorial cDNA library uniqueness of the CDRH3 loop by the length and amino acid composition has been shown. Multiple alignments with the nearest homologies from the NCBI databases have revealed that the sequences of ScFv obtained are new.

[Clinical course of arterial hypertension in young subjects with vibration disease].

The aim of the work was to study prevalence of arterial hypertension (AH) in young subjects and its clinical features in patients with vibration disease. The analysis included 652 medical histories of patients at the age of 22-72 years treated for AH and vibration disease in 1997-2007. AH and CHD patients under 40 year of age comprised 18.3% (somewhat more than in the general population) and 9.1% respectively. Grade I and II AH was diagnosed in 48.7 and 40% of the patients. Capillaroscopy revealed predominance of angiospastic syndrome (54.2%) and rheovasography decreased peripheral vascular tone and radial artery pulse. Plasma cholesterol level in young patients was 5.68 +/- 0.99 mmol/l, hypercholesterimenmia occurred in 69.4%. Vibration was shown to be an additional risk factor of AH development and progression. It is concluded that early diagnosis and treatment of AH is paramount for the prevention of cardiovascular complications in young patients.

[Effects of novel antiviral agents on HIV-1 replication].

Two groups of the antiviral agents: 1) adamantane- and norbornen-containing compounds with in-built cholesterol to potentiate the membranotropic properties and 2) synthetic matrix protein peptides (peptides A and B) were found to have effects on HIV replication. The agents of the former group produced antiviral activity only when added in combination with the virus. Peptide A (matrix protein 43-60 amino acids) inhibited viral replication when added in both the early and late periods. Fluorescein-labeled peptide A was detectable in the cytoplasm and nucleus (although adsorption of a portion of the peptides cannot be excluded onto the cell surface). Peptide A was shown to inhibit Gag precursor p55 transport from the nuclei to the plasma membrane, the site of virus assembly. Peptide B had no antiviral activity.

[Monitoring of chemotherapy in patients with pulmonary tuberculosis by the blood bacteriostatic activity-determining test].

The paper gives the results of studies to determine blood bacteriostatic activity (BBA) in the use of a patient's autostrain and semiliquid medium versus the clinical and laboratory parameters of the course of a process in 101 patients with pulmonary tuberculosis. There is evidence for the relationship of the BBA to the sensitivity to isoniazid and the structure of drug resistance. The zero values of BBA correspond to the severest course of the disease. The efficiency of treatment is much higher in patients with high and moderate BBA. The latter's determination using the semiliquid medium permits an objective evaluation of the efficiency of chemotherapy, identification of patients with a poor prognosis, and then choice of an individual treatment regimen on day 7 after the test just before obtaining the data on drug sensitivity.

[Construction and characterization of immune combinatorial cDNA library of mouse variable immunoglobuline genes].

A cDNA combinatorial antibody library of mouse variable immunoglobulin fragments has been constructed from mice immunized with rhIFN-beta1b. For this purpose, cDNAS of immunoglobulin variable heavy (V(H)) and variable light (V(L)) chains genes amplified from splenocytes were joined with linker DNA to form ScFv's (single-chain Fv-antibodies). The obtained ScFv-DNA pool was cloned into a phagemid vector and used for Esherichia coli transformation. Using the phage display technique, bacterial clones producing single-chain antibodies specific to rhIFN-beta1b were selected. The following characteristics of the combinatorial library were determined in this work: abundance, functional size, and the initial ScFv-DNA diversity in the library constructed. High specificity of interaction between phage displayed ScFv's and rhIFN-beta1b has been demonstrated.

[Influence of membrane-active polyanions on various stages of the human cytomegalovirus life cycle in vitro].

Human cytomegalovirus (CMV), an agent of infection (CMVI), lethally dangerous for immune deficient neonates and adults was investigated in vitro as a target for a therapeutic effect of new membrane-active polyanionic compounds (MPC). Previous studies on the alicycle- and sulfate-modified carboxy-MPCs revealed a well-defined tendency of the anti-CMV activity amplification in parallel with increasing of the content of sulfate groups, enhancing the negative charge of the macromolecule. The dominating role of the electrostatic factor was confirmed by the highest activity of AS-688, compound with maximum sulfation among the tested MPCs. Its selectivity index (SI) of the CMVI inhibition in human diploid fibroblast cells reached 5450, 7500, 250 and 4286 in the microbicidal, viricidal, prophylactic and therapeutic schemes of the experiment respectively. The antiviral activity at the first, second and third schemes was explained by the polyanion-typical potential of electrostatic neutralization of the countercharged virions and prevention of the virus adsorption on the cell membranes (in competition with heparin sulfate, a cellular receptor of CMV), whereas the therapeutic effect required the ability of MPC to influence the intracellular stages of the CMV life cycle. The PCR and immunochemical assays revealed an inhibitory action of AS-688 on replication of the viral DNA and the following synthesis of the late viral protein gB with efficiency similar to that of gancyclovir (GCV). However, in contrast to GCV, acting as inhibitor of enzyme (viral RNA-polymerase) factor of the biosynthesis, the therapeutic activity of MPC could be interpreted by competition with viral RNA/DNA due to the specific character of the MPC molecular basis, initially constructed on the principle of nucleic acids backbone and charge adjustable imitation. This mechanism assuming reduction of the cytotoxicity risks, explained the experimentally observed fact of low cytotoxicity of MPCs and possible achievement of high SI. The MPC ability to penetrate into the cells without disruption of cellular membrane permeability was confirmed in experiments with the fluorescent-labeled derivate AS-679, structurally and functionally related to AS-688. In the light of the previously described HIV inhibiting properties of AS-688, AS-679 and MPC analogous, the results could be considered prospective in development of new highly effective agents for combined antiviral protection.

[Antiviral activity of polycarboxylic substances modified by cage hydrocarbon and sulfoacid pharmacophors against cytomegalovirus infection in vitro].

Human cytomegalovirus (CMV) infection (CMVI) results in lethal risks at the immunodeficiency status, including the HIV co-infection. Carboxy-mimickers of the polymeric backbone of nucleic acids, potential agonists and antagonists of the virus genome were developed as promising candidates for the antiviral protective agents. In parallel with stimulation of antiviral immunity the mimickers derived membrane potent compounds (MPC), were shown to be able to prevent directly and efficiently the cell infection by various strains of the human immunodeficiency virus (HIV) [Antibiotics and Chemother 2003; 48: 2:29-41; 5:7-15]. The paper presents new data and discussion of the results on investigation of the MPC, modified by the previously designed adamantane or norbornene and by the recently applied sulfoacidic pharmacophores in the experimental model of CMVI in vitro (human diploid fibroblast cells). Eight substances with various ratios of theabove mentioned cage-hydrocarbon and/or anion pharmacophores in the macromolecule were tested and active MPC modifications were detected which efficiently inhibited the CMVI with high indexes of selectivity up to 250, 4286 and 7500 in prophylactic, therapeutic and viricidal experimental schemes respectively. Modulating influence of the lipotropic (cage-hydrocarbon) pharmacophores on the anti-CMV activity was observed only in the viricidal and prophylactic experimental schemes, in which the lipid membranes of the cells and/or virus envelopes were involved. Still, the dominant role in the antiviral activity of MPC in all the experimental schemes was played by the sulfoacid-anionic chemical structure modulation. By increasing the density of the negative charge of the macromolecules to the levels comparable with the charge of the genome molecules, theanionic modification evidently amplified the potential of the antagonistic competition of the synthetic MPC with the virus genome, thus impairing the virus-specific interactions. The most promising compounds AS-688 and AS-678/-679 were selected for further investigation of the mechanisms of the anti-CMV and anti-HIV activity.

[Characterization of a panel of mouse single-chain antibodies against recombinant human interferon beta1b].

A panel of single-chain antibodies (ScFv's--single-chain Fv-antibodies) against recombinant human interferon beta 1b (rhIFN-beta1b) has been obtained from immune and naive combinatorial cDNA libraries of the mouse variable immunoglobulin genes. ScFv's were expressed into Escherichia coli cells. For producers isolated from the immune library a difference in production yield of ScFv's in periplasm and incubation medium as well as their expression stability in passages and storage stability have been demonstrated. After sequencing of target DNA the multiple alignment and structural analysis of ScFv's sequences with different primary structures were carried out and significant difference in both complementarity-determining (CDR) and framework (FR) regions of their variable domains has been shown. For the ScFv's isolated from the immune library, specificity of their binding with native and denatured rhIFN-beta1b in ELISA and Western-blotting as well as their high storage stability have been shown. The affinity constants for each representatives of the ScFv's panel were in the range from 1.96 x 10(-8) to 1.69 x 10(-9) M.

[Drug sensitivity in Mycobacterium tuberculosis versus its viability, cytotoxicity, genotype, and the course of the process in patients with pulmonary tuberculosis].

The authors studied drug sensitivity, mutations in the katG, in-hA, alpC, rpoB genes, virulence via the cytotoxicity test on THP-1 cells, and the viability and genetic affiliation of 53 clinical M. tuberculosis isolates versus data on the form and dynamics of a process. Sensitive and resistant strains did not significantly differ in viability and cytotoxicity. The highest death of infected macrophages was observed was seen with infection of M. tuberculosis of the Beijing B0 genotype, the least one seen with that of LAM with the similar rate of multiple drug resistance. There was a correlation of the changes in the count of lymphocytes in patients with the genetic affiliation of a causative agent. The severest course of the tuberculous process was observed in baseline lymphopenia (before treatment) in combination with multidrug resistance of mycobacteria, high and moderate cytotoxicity and high viability. Ser-Leu 531 mutation resulted in cross resistance to rifampicin and mycobutin in most cases.

[Immunogenetic aspects of the course and treatment of pulmonary tuberculosis].

The paper presents the clinical, X-ray, and laboratory characteristics of patients with pulmonary tuberculosis. Both general regularities and differences in the frequency of alleles of the HLA-DQB1* locus have been revealed in the groups of patients with pulmonary tuberculosis as compared with healthy individuals. There are specificities associated with the risk of pulmonary tuberculosis and the variants of the course of infection; thus, allele 05 of the HLA-DQB1* locus is positively associated with the incidence of tuberculosis. Specificity 03 of the HLA-DQB1* locus has been ascertained to be associated with the poor course of the disease. The most pronounced immunological changes have been observed in patients with the poor course of the disease, who are the carriers of specificity 05 of the HLA-DQB1* locus. The totality of immunological parameters and the data of genetic studies provide a basis for using the selective immunomodulator rIL-2 (roncoleukin) in the most seriously ill patients who are carriers of specificity 05.

[A role of the HLA-DRB1* genotype in the complex therapy of pulmonary tuberculosis].

It is as for now evident that in-depth researches of the genotype of a patient are needed to enhance the efficiency of therapeutic measures. The purpose of the present study was to enhance the efficiency of complex antituberculous therapy, by genetically identifying the allelic polymorphism of the HLA-DRB1* gene in a person who had ill with tuberculosis. The material of the study was the results of a followup and treatment of 100 patients with pulmonary tuberculosis. The subject of a special study was the molecular typing of the HLA genes of the DRB1* locus by polymerase chain reaction (PCR-SSP). The findings suggest that an individual approach to choosing treatment regimens (chemotherapy and pathogenetic therapy) for patients with pulmonary tuberculosis, by taking into account the HLA-DRB1* genotype enables one to enhance the efficiency of treatment in the major clinical and X-ray parameters.

[Carboxy mimetic derivatives of nucleic acid polymeric backbones inhibiting human cytomegalovirus. 1. In vitro microbicidal effect].

The artificial polycarboxyacidic compounds (PC), imitating the principle of furan-derived and negatively charged structures alternating in the polymeric backbone of nucleic acids, previously explored as interferon inductors and stimulators of antiviral immunity in vivo, were modified by the side groups to amplify the direct antiviral potency in vitro and investigated in the cell culture model of human diploid fibroblasts infected with human cytomegalovirus (CMV) in a microbicidal scheme. Reconstruction from the PC to membrane potent compounds (MPC) was carried out by covalent modification with lipotropic pharmacophores (of cage-hydrocarbon structures similar to rimantadine or camphor-like terpenoids), as well as by conversion of the carboxy groups to sulfate-anionic derivates, related to the CMV sensitive heparansulfate receptor (HSR) of the cells. Both the factors of the MPC structure-functional modulation (lipotropic and anionic) were found to be effective tools for amplification of the microbicidal activity. The maximum inhibitory effect against CMV and minimum cytotoxicity (with the best selectivity, the chemotherapeutic index of > or = 3000-5000) were achieved mainly through increasing the anionic groups content, elevating the MPC negative charge to the level comparable with one of the like charged viral genome and HSR. In relation with the previously found anti-HIV efficacy of the same MPCs in analogous experimental models and in view of the fact that CMV is one of the most dangerous opportunistic co-factors of HIV/AIDS pathogenesis, the obtained data can be used as a basis for further development of new generation microbicides, promising for combined prevention of sexually transmitted infections.