Two-component covalent inhibitor

Inhibitors that covalently damage proteins or nucleic acids offer great potency, but are difficult to rationally design and suffer from poor specificity. Here we outline a general concept for constructing covalent inhibitors, called the two-component covalent inhibitor (TCCI). The approach takes advantage of two ligand analogs equipped with pre-reactive groups. Binding of the analogs to the adjacent sites of a target biopolymer brings the pre-reactive groups in close proximity and causes their interaction followed by covalent damage of the target. In the present study we used light-activated pre-reactive groups to inactivate a DNA polymerase. It was found that the efficiency of a traditional single-component inhibitor was greatly reduced in the presence of a non-target protein, while the TCCI was not significantly affected. Our findings suggest that TCCI approach has advantages in inactivation of biopolymers in complex multi-component systems.     

Effects of support stimulation on human soleus fiber characteristics during exposure to "dry" immersion

In this study the model of 7-day dry immersion (DI) was used. 17 male volunteers (23-29 years old) were divided in 2 groups: (i) 7-day DI without support (DI, n=9), (ii) 7-day DI using support stimulation (DIS, n=8). Support stimulator device exerted pressure of 0.2 +/- 0.15 kg/cm2 upon the plantar support zones simulating the walking pattern 6 times a day for 20 minutes of every hour: 10 minutes at a speed of 75 steps/min and 10 minutes at a speed of 120 steps/min. M. soleus biopsy was performed before and immediately after DI. The m. soleus fiber myosin heavy chain (MHC) profile, myofiber cross-sectional area (CSA) and total protein concentration were analyzed in frozen serial sections. In addition, NO-synthase 1 (NOS1) levels indicative of normal muscle cell signaling were analyzed by western blotting in 4 persons in each group. After dry immersion, percentage of muscle fibers containing type I MHC decreased by 6% (p<0.05) in group DI, but was not changed significantly in group DIS. Percentage of the type IIa fibers was significantly altered in none of the groups. Type I fiber CSA decreased by 24.4% (p<0.05) in group DI. No significant changes of type I fiber CSA were found in group DIS. CSA of the type IIa fibers significantly altered in none of the groups. The total protein concentration was found increased by 17.6% in group DI and by 21% in group DIS. The increased total protein content in group DI suggests a diminution of fiber CSA attributed to the loss of non-protein component of fibers. NOS1 decreased by 35.6% in group DI and increased by 58.1% in group DIS. We conclude that 7 days in dry immersion lead to reduction in the type I muscle fiber percentage, loss of the non-protein component and decline in NOS1. These changes were clearly prevented by the support stimulation protocol applied during the DI period.     

Survival of civilian and prisoner drug-sensitive, multi- and extensive drug- resistant tuberculosis cohorts prospectively followed in Russia

Objective and Methods

A long-term observational study was conducted in Samara, Russia to assess the survival and risk factors for death of a cohort of non-multidrug resistant tuberculosis (non-MDRTB) and multidrug resistant tuberculosis (MDRTB) civilian and prison patients and a civilian extensive drug-resistant tuberculosis (XDRTB) cohort.

Results

MDRTB and XDRTB rates of 54.8% and 11.1% were identified in the region. Half (50%) of MDRTB patients and the majority of non-MDRTB patients (71%) were still alive at 5 years. Over half (58%) of the patients died within two years of establishing a diagnosis of XDRTB. In the multivariate analysis, retreatment (HR = 1.61, 95%CI 1.04, 2.49) and MDRTB (HR = 1.67, 95%CI 1.17, 2.39) were significantly associated with death within the non-MDR/MDRTB cohort. The effect of age on survival was relatively small (HR = 1.01, 95%CI 1.00, 1.02). No specific factor affected survival of XDRTB patients although median survival time for HIV-infected versus HIV-negative patients from this group was shorter (185 versus 496 days). The majority of MDRTB and XDRTB strains (84% and 92% respectively) strains belonged to the Beijing family. Mutations in the rpoB (codon 531 in 81/92; 88.8%), katG (mutation S315T in 91/92, 98.9%) and inhA genes accounted for most rifampin and isoniazid resistance respectively, mutations in the QRDR region of gyrA for most fluroquinolone resistance (68/92; 73.5%).

Conclusions

Alarmingly high rates of XDRTB exist. Previous TB treatment cycles and MDR were significant risk factors for mortality. XDRTB patients'' survival is short especially for HIV-infected patients. Beijing family strains comprise the majority of drug-resistant strains.     

Localization of catecholamines, serotonin and histamine in the cells and mucus of bronchial washings from apparently healthy subjects

By means of luminescent-histochemical methods localization of catecholamines, serotonin, histamine has been studied in cells and mucus of bronchial washings of 30 practically healthy persons. Cytofluorometry is performed in mucus, pulmonary epithelium, alveolar macrophages, lymphocytes and neutrophils of the bronchial washings. In the alveolar macrophages a high level of the compounds studied is detected.     

Critical Role of S1PR1 and Integrin β4 in HGF/c-Met-mediated Increases in Vascular Integrity

Vascular endothelial cell (EC) barrier integrity is critical to vessel homeostasis whereas barrier dysfunction is a key feature of inflammatory disorders and tumor angiogenesis. We previously reported that hepatocyte growth factor (HGF)-mediated increases in EC barrier integrity are signaled through a dynamic complex present in lipid rafts involving its receptor, c-Met (1). We extended these observations to confirm that S1PR1 (sphingosine 1-phosphate receptor 1) and integrin β4 (ITGB4) are essential participants in HGF-induced EC barrier enhancement. Immunoprecipitation experiments demonstrated HGF-mediated recruitment of c-Met, ITGB4 and S1PR1 to caveolin-enriched lipid rafts in human lung EC with direct interactions of c-Met with both S1PR1 and ITGB4 accompanied by c-Met-dependent S1PR1 and ITGB4 transactivation. Reduced S1PR1 expression (siRNA) attenuated both ITGB4 and Rac1 activation as well as c-Met/ITGB4 interaction and resulted in decreased transendothelial electrical resistance. Furthermore, reduced ITGB4 expression attenuated HGF-induced c-Met activation, c-Met/S1PR1 interaction, and effected decreases in S1P- and HGF-induced EC barrier enhancement. Finally, the c-Met inhibitor, XL880, suppressed HGF-induced c-Met activation as well as S1PR1 and ITGB4 transactivation. These results support a critical role for S1PR1 and ITGB4 transactivation as rate-limiting events in the transduction of HGF signals via a dynamic c-Met complex resulting in enhanced EC barrier integrity.     

Selection of a community of acidochemolithotrophic microorganisms with a high oxidation rate of pyrrhotite-containing sulphide ore flotation concentrate

A community of acidochemolithotrophic microorganisms with a high oxidation rate of pyrrhotite-containing sulphide ore flotation concentrate was selected. The Acidithiobacillus caldus OP-1 and Ferroplasma acidiphilum OP-2 cultures were identified to be dominating members. The presence of the Acidithiobacillus ferrooxidans OP-3, Leptospirillum ferriphilum OP-4, and Sulfobacillus thermosulfidooxidans OP-5 cultures in the community’s composition was also mentioned. The analysis results of solid residues of the process showed a greater elemental sulfur oxidation level and gold recovery when the initial pH value in tank I was maintained at a level of 1.8–2.0 (90.5%) rather than 1.6–1.8 (86.3%).     

Fauna of bumblebees (Hymenoptera: Apidae: Bombus Latreille) in the mainland part of Arkhangelsk Region,NW Russia

The focus of this study is to present bumblebee fauna of Arkhangelsk Region (north-western Russia). This research is based on the museum collections and materials collected by the authors. Collecting localities and data of faunistic records are given in the species list. We include 34 species in the fauna of bumblebees of Arkhangelsk Region. The regional fauna is dominated by Transpalaearctic species. Bumblebee fauna in the study region has low specificity. The recent distribution of bumblebee species in the study region is a result of post-glacial immigration.     

Synthesis and Antioxidant Activity of Carane and Pinane Based Sulfenimines and Sulfinimines

The synthesis of sulfenimines and sulfinimines has been carried out with 10‐hydroxyisocamphylthiol. The configuration of the compounds has been deduced by methods of NMR, DFT calculations and X‐ray diffraction analysis. The cytotoxic, antioxidant and membrane‐protective activity of the synthesized compounds as well as of the previously obtained sulfenimines and sulfinimines based on 4‐caranethiol have been determined.