Sequence analysis of the ribosomal DNA internal transcribed spacer 2 from populations of Anopheles nuneztovari (Diptera: Culicidae)

Sequence variation of the ribosomal DNA internal transcribed spacer 2 (ITS2) was examined for populations of the malaria vector Anopheles nuneztovari collected in Colombia, Venezuela, Bolivia, Suriname, and Brazil. Mosquitoes from Colombia and Venezuela had identical ITS2 sequences and were distinguished from sequences in other populations by three insertion/deletion events (indels) and by one transversion. The length of the ITS2 was 363-369 bp, and it had a G+C content of 55.3%- 55.7%. Variation in the length of the ITS2 between and within populations was due to indels in simple repeats. ITS2 consensus sequences were similar or identical for samples from the following three groups: (1) Colombia, Bolivia, and Venezuela; (2) Suriname and northern Brazil; and (3) eastern and central Brazil. The presence of two different consensus sequences from a single location near Manaus, Brazil, suggests that populations from eastern Brazil and those from Suriname converge in this region of the Amazon Basin. These data show that putative cryptic species of An. nuneztovari are distinguished by very minor differences in DNA sequence of the ITS2 region.      

Effects of cardiac myosin binding protein-C on the regulation of interaction of cardiac myosin with thin filament in an in vitro motility assay

Modulatory role of whole cardiac myosin binding protein-C (сMyBP-C) in regulation of cardiac muscle contractility was studied in the in vitro motility assay with rabbit cardiac myosin as a motor protein. The effects of cMyBP-C on the interaction of cardiac myosin with regulated thin filament were tested in both in vitro motility and ATPase assays. We demonstrate that the addition of cMyBP-C increases calcium regulated Mg-ATPase activity of cardiac myosin at submaximal calcium. The Hill coefficient for ‘pCa-velocity’ relation in the in vitro motility assay decreased and the calcium sensitivity increased when сMyBP-C was added. Results of our experiments testifies in favor of the hypothesis that сMyBP-C slows down cross-bridge kinetics when binding to actin.     

Physicochemical, toxicological and hygienic aspects of ethylene oxide application for the sterilization of medical products. III. Setting hygienic norms in sterilized medical products

Some of the drawbacks are discussed associated with hygienic norms for gaseous sterilizing agents in polymeric products for medical applications. A new approach is proposed for regulating ethylene oxide (EO) in such products. To this end, limiting types of biological effect and its pathways have been determined and substantiated as well as the reliability factor for establishing hygienic norms for EO in medical products. Daily threshold EO limit values were calculated for momentary and repeated exposure of humans. The duration of long-term and short-term exposure of the patient to sterilized products was evaluated on the basis of EO extraction kinetics from a variety of materials. Using daily threshold limit values, threshold residual values (TRV) of EO were calculated for different groups of polymeric products for medical applications.     

Protein interaction network for Alzheimer's disease using computational approach

Alzheimer''s disease (AD) is the most common form of dementia. It is the sixth leading cause of death in old age people. Despite recent advances in the field of drug design, the medical treatment for the disease is purely symptomatic and hardly effective. Thus there is a need to understand the molecular mechanism behind the disease in order to improve the drug aspects of the disease. We provided two contributions in the field of proteomics in drug design. First, we have constructed a protein-protein interaction network for Alzheimer''s disease reviewed proteins with 1412 interactions predicted among 969 proteins. Second, the disease proteins were given confidence scores to prioritize and then analyzed for their homology nature with respect to paralogs and homologs. The homology persisted with the mouse giving a basis for drug design phase. The method will create a new drug design technique in the field of bioinformatics by linking drug design process with protein-protein interactions via signal pathways. This method can be improvised for other diseases in future.     

The protective effect of prolil-glycil-proline peptide (PGP) under compound 48/80-induced anaphylactoid reaction in mice

The influence of PGP on compound 48/80-induced anaphylactoid reaction development in mice and on histamine secretion from rat peritoneal mast cells (RPMS) under their activation by compound 48/80 were investigated. Anaphylactoid reaction was caused by intraperitoneal injection of compound 48/80 into mice. The number of animals with manifestations of anaphylactoid reaction symptoms, the severity of these symptoms, the amount of died animals and the time of death were registering during an hour. Mast cells for in vitro investigations were obtained from rats’ peritoneal cavity. Secreted histamine was evaluated from formation of fluorescent product of it’s condensation with ortho-phthalaldehyde. The preventive injection of PGP in mice (15 min before compound 48/80) decreased the mortality rate of animals and intensity of anaphylactoid reaction symptoms. But PGP had no effect on histamine secretion from mast cells under their activation by compound 48/80 in vitro. Results show that there is a component in the mechanism of PGP protective effect under anaphylactoid reaction which is not connected with mast cells stabilization.     

Application of laser interference microscopy (LIM) for investigation of the protective effect of prolyl-glycyl-proline (Pro-Gly-Pro) on mast cells

The effect of peptide prolyl-glycyl-proline (Pro-Gly-Pro) on morphometric parameters of mast cells upon their activation by compound 48/80 or synacten was investigated. Cell image, obtained by the method of laser interference microscopy (LIM), is a distribution of the optical path difference of light (OPD). It evaluates the changes of the individual components of cytoplasm (maxOPD) and the total distribution of OPD (“dry mass”). The changes of “dry mass” in cytoplasm correlate with the changes of the secreted histamine amount (?0.86). Preliminary incubation of mast cells with Pro-Gly-Pro (6 × 10^?5 M) did not change the area, the state of the individual components of the cytoplasm (nucleus) and “dry mass” (histamine vesicles) in cells. The “dry mass” (histamine vesicles) and maxOPD decreased while the release of histamine increased upon the activation of mast cells by compound 48/80 (0.02 mg/mL). Preincubation of cells with Pro-Gly-Pro had no effect. Activation of cells by synacten (2 and 20 μM) led to the increase of the cell area and the reduction of maxOPD and “dry mass” (histamine vesicles). Preincubation of the cells with Pro-Gly-Pro prevented these changes. So, the protective effect of Pro-Gly-Pro was observed in the case of the activation of mast cells by synacten but not by compound 48/80.     

Study of regulatory effect of tropomyosin on actin-myosin interaction in skeletal muscle by in vitro motility assay

The interaction between myosin and actin in striated muscle tissue is regulated by Ca²⁺ via thin filament regulatory proteins. Skeletal muscle possesses a whole pattern of myosin and tropomyosin isoforms. The regulatory effect of tropomyosin on actin-myosin interaction was investigated by measuring the sliding velocity of both actin and actin-tropomyosin filaments over fast and slow skeletal myosins using the in vitro motility assay. The actin-tropomyosin filaments were reconstructed with tropomyosin isoforms from striated muscle tissue. It was found that tropomyosins with different content of α-, β-, and γ-chains added to actin filaments affect the sliding velocity of filaments in different ways. On the other hand, the sliding velocity of filaments with the same content of α-, β-, and Γ-chains depends on myosin isoforms of striated muscle. The reciprocal effects of myosin and tropomyosin on actin-myosin interaction in striated muscle may play a significant role in maintenance of effective work of striated muscle both during ontogenesis and under pathological conditions.     

Prevention of the formation of drug resistance in bacterial populations by using biologically active substances

The patients with infected wounds of the extremities were treated with kanamycin electrophoresis in combination with chlorhexidine bigluconate, an antiseptic. As compared to the patients treated with kanamycin alone, the rate of the wound size decrease in such patients was 2 times higher. The levels of microbial contamination in these patients were much lower. The contamination level with the aerobic flora was 4.8 times lower, including staphylococci, the level of contamination with which was 5.9 times lower. The contamination level with the kanamycin-resistant bacteria was 22 times lower. The treatment with kanamycin alone resulted in a 2.6-fold increase in the number of the antibiotic-resistant variants in the microbial populations of the wounds. In 48.2 per cent of the patients, this was accompanied by development of resistance to kanamycin in the whole microbial population of the wound. The development of the kanamycin resistance in the staphylococcal populations of 18.1 per cent of the patients was associated with changed sensitivity of the initial strains and in 81.9 per cent of the patients, with superinfection by the resistant strains. No changes in the kanamycin sensitivity of the initial gram-negative organisms during the treatment were observed. The use of chlorhexidine bigluconate, as a biologically active substance in combination with kanamycin potentiated the action of the antibiotic, prevented development and accumulation of the antibiotic-resistant variants in the microbial populations of the wounds and development of the drug resistance in these populations.     

A comparative analysis of the distribution of glyprolines after their administration by different ways

The distribution of the glyprolines, Pro-Gly-Pro and Thr-Lys-Pro-Arg-Pro-Gly-Pro (Selanc), was analyzed and compared in tissues of rat organs after different ways of their administration using the peptides uniformly labeled with tritium. Comparative data on changes of concentrations of the peptides in the rat organs after their intraperitoneal, intranasal, intragastric, and intravenous administration are given. The intranasal administration of both peptides was shown to be optimal for delivery of glyprolines molecules in the CNS. A high affinity of the studied glyprolines for gastric tissues was found for all the ways of their administration. We suggest that high efficacy of action of glyprolines on homeostasis of the gastric mucosa was partially provided by accumulation of these peptides (to high concentrations) in gastric tissues.