Aqueous two-phase system based on benzethonium chloride and sodium dihexyl sulfosuccinate for extraction and ICP-OES determination of heavy metals.

An aqueous two-phase system (ATPS) based on benzethonium chloride (BztCl) and sodium dihexyl sulfosuccinate (NaDHSS) was proposed for the first time for liquid-liquid microextraction of Cd(II), Co(II), Cu(II), Mn(II), Ni(II), and Pb(II) followed by ICP-OES determination. The mixture of cationic and anionic surfactants, BztCl and NaDHSS, showed liquid-liquid phase separation at the molar ratio of 1:1, and the total surfactant concentration of 0.01-0.2 mol L-1 forming ATPS that was investigated in the extraction process. The extraction efficiency for Cd(II), Co(II), Mn(II), Ni(II), and Pb(II) was nearly 100 %, and for Cu(II) - not lower than 88 % in the presence of 8-hydroxyquinoline as a complexing agent. The surfactant-rich phase containing analytes was subjected to back-extraction with 0.2 M HNO3 before ICP-OES measurements. The preconcentration in the proposed BztCl-NaDHSS-H2O ATPS for 30 s and the high degree of back-extraction, which was achieved in 1 min, significantly reduced the sample preparation time, matrix effects and provided low LODs in the range of 0.04-1.0 µg L-1, the preconcentration factor was 120. The analysis of a certified reference material sample of surface water and the real samples of tap, sea, and waste water verified the method accuracy.

The Recycling of Substandard Rocket Fuel N,N-Dimethylhydrazine via the Involvement of Its Hydrazones Derived from Glyoxal, Acrolein, Metacrolein, Crotonaldehyde, and Formaldehyde in Organic Synthesis.

"Heptil" (unsymmetrical dimethylhydrazine-UDMH) is extensively employed worldwide as a propellant for rocket engines. However, UDMH constantly loses its properties as a result of its continuous and uncontrolled absorption of moisture, which cannot be rectified. This situation threatens its long-term usability. UDMH is an exceedingly toxic compound (Hazard Class 1), which complicates its transportation and disposal. Incineration is currently the only method used for its disposal, but this process generates oxidation by-products that are even more toxic than the original UDMH. A more benign approach involves its immediate reaction with a formalin solution to form 1,1-dimethyl-2-methylene hydrazone (MDH), which is significantly less toxic by an order of magnitude. MDH can then be polymerized under acidic conditions, and the resulting product can be burned, yielding substantial amounts of nitrogen oxides. This review seeks to shift the focus of MDH from incineration towards its application in the synthesis of relatively non-toxic and readily available analogs of various pharmaceutical substances. We aim to bring the attention of the international chemical community to the distinctive properties of MDH, as well as other hydrazones (such as glyoxal, acrolein, crotonal, and meta-crolyl), wherein each structural fragment can initiate unique transformations that have potential applications in molecular design, pharmaceutical research, and medicinal chemistry.

Novel Derivatives of Quinoxaline-2-carboxylic Acid 1,4-Dioxides as Antimycobacterial Agents: Mechanistic Studies and Therapeutic Potential.

The World Health Organization (WHO) reports that tuberculosis (TB) is one of the top 10 leading causes of global mortality. The increasing incidence of multidrug-resistant TB highlights the urgent need for an intensified quest to discover innovative anti-TB medications In this study, we investigated four new derivatives from the quinoxaline-2-carboxylic acid 1,4-dioxide class. New 3-methylquinoxaline 1,4-dioxides with a variation in substituents at positions 2 and 6(7) were synthesized via nucleophilic aromatic substitution with amines and assessed against a Mycobacteria spp. Compound 4 showed high antimycobacterial activity (1.25 µg/mL against M. tuberculosis) and low toxicity in vivo in mice. Selection and whole-genomic sequencing of spontaneous drug-resistant M. smegmatis mutants revealed a high number of single-nucleotide polymorphisms, confirming the predicted mode of action of the quinoxaline-2-carboxylic acid 1,4-dioxide 4 as a DNA-damaging agent. Subsequent reverse genetics methods confirmed that mutations in the genes MSMEG_4646, MSMEG_5122, and MSMEG_1380 mediate resistance to these compounds. Overall, the derivatives of quinoxaline-2-carboxylic acid 1,4-dioxide present a promising scaffold for the development of innovative antimycobacterial drugs.

Bacterial Lux Biosensors in Genotoxicological Studies.

The aim of this study was to assess the applicability of the bacterial lux biosensors for genotoxicological studies. Biosensors are the strains of E. coli MG1655 carrying a recombinant plasmid with the lux operon of the luminescent bacterium P. luminescens fused with the promoters of inducible genes: recA, colD, alkA, soxS, and katG. The genotoxicity of forty-seven chemical compounds was tested on a set of three biosensors pSoxS-lux, pKatG-lux and pColD-lux, which allowed us to estimate the oxidative and DNA-damaging activity of the analyzed drugs. The comparison of the results with the data on the mutagenic activity of these drugs from the Ames test showed a complete coincidence of the results for the 42 substances. First, using lux biosensors, we have described the enhancing effect of the heavy non-radioactive isotope of hydrogen deuterium (D2O) on the genotoxicity of chemical compounds as possible mechanisms of this effect. The study of the modifying effect of 29 antioxidants and radioprotectors on the genotoxic effects of chemical agents showed the applicability of a pair of biosensors pSoxS-lux and pKatG-lux for the primary assessment of the potential antioxidant and radioprotective activity of chemical compounds. Thus, the results obtained showed that lux biosensors can be successfully used to identify potential genotoxicants, radioprotectors, antioxidants, and comutagens among chemical compounds, as well as to study the probable mechanism of genotoxic action of test substance.

Aqueous Two-Phase Systems Based on Cationic and Anionic Surfactants Mixture for Rapid Extraction and Colorimetric Determination of Synthetic Food Dyes.

In this study, aqueous two-phase systems (ATPSs) containing a cationic and anionic surfactants mixture were used for the preconcentration of the synthetic food dyes Allura Red AC, Azorubine, Sunset Yellow, Tartrazine, and Fast Green FCF. A rapid, simple, low cost, affordable, and environmentally friendly methodology based on microextraction in ATPSs, followed by spectrophotometric/colorimetric determination of the dyes, is proposed. The ATPSs are formed in mixtures of benzethonium chloride (BztCl) and sodium N-lauroylsarcosinate (NaLS) or sodium dihexylsulfosuccinate (NaDHSS) under the molar ratio close to equimolar at the total surfactant concentration of 0.01-0.20 M. The density, viscosity, polarity, and water content in the surfactant-rich phases at an equimolar ratio BztCl:NaA were determined. The effects of pH, total surfactant concentration, dye concentration, and time of extraction/centrifugation were investigated, and the optimum conditions for the quantitative extraction of dyes were established. The smartphone-based colorimetric determination was employed directly in the extract without separating the aqueous phase. The analytical performance (calibration linearity, precision, limits of detection and quantification, reproducibility, and preconcentration factor) and comparison of the spectrophotometric and smartphone-based colorimetric determination of dyes were evaluated. The method was applied to the determination of dyes in food samples and food-processing industrial wastewater.

Mapeamento Potencial de Superfície Corporal durante a Despolarização Ventricular em Ratos Após Exercício Exaustivo Agudo / Body Surface Potential Mapping during Ventricular Depolarization in Rats after Acute Exhaustive Exercise

Resumo Fundamento O exercício físico exaustivo pode causar alterações significantes nas propriedades elétricas do miocárdio. Objetivo Avaliar, através do mapeamento potencial de superfície corporal, a atividade elétrica do coração de ratos durante a despolarização ventricular após exercício exaustivo agudo. Métodos Ratos machos com doze semanas de idade foram submetidos a exercício agudo em esteira a 36 m/min até a exaustão. Eletrocardiogramas unipolares (ECGs) da superfície do tronco foram registrados em ratos anestesiados com zoletil três a cinco dias antes (Pré-Ex), 5 e 10 minutos após exercício exaustivo (Pós-Ex 5 e Pós-Ex 10, respectivamente) simultaneamente com ECGs nas derivações dos membros. Os mapas potenciais de superfície corporal instantâneos (BSPMs, body surface potential maps ) foram analisados durante a despolarização ventricular. Os valores de p <0,05 foram considerados estatisticamente significantes. Resultados Comparado com o Pré-Ex, uma conclusão precoce da segunda inversão de distribuições de potencial, uma conclusão precoce da despolarização ventricular, bem como uma diminuição na duração da fase média e a duração total da despolarização ventricular nos BSPMs foram reveladas no Pós-Ex5. Além disso, em comparação com o Pré-Ex, um aumento na amplitude do extremo negativo do BSPM no pico da onda R no ECG na derivação II (pico RII) e uma diminuição na amplitude do extremo negativo do BSPM a 3 e 4 ms após o pico RII foram demonstrados no Pós-Ex 5. No Pós-Ex 10, os parâmetros dos BSPMs não diferiram daqueles do Pré-Ex. Conclusão Em ratos, o exercício exaustivo agudo causa alterações reversíveis nas características temporais e de amplitude dos BSPMs durante a despolarização ventricular, provavelmente relacionadas a alterações na excitação da massa principal do miocárdio ventricular.

Abstract Background Exhaustive physical exercise can cause substantial changes in the electrical properties of the myocardium. Objective To evaluate, using body surface potential mapping, the electrical activity of the heart in rats during ventricular depolarization after acute exhaustive exercise. Methods Twelve-week-old male rats were submitted to acute treadmill exercise at 36 m/min until exhaustion. Unipolar electrocardiograms (ECGs) from the torso surface were recorded in zoletil-anesthetized rats three to five days before (Pre-Ex), 5 and 10 minutes after exhaustive exercise (Post-Ex 5 and Post-Ex 10, respectively) simultaneously with ECGs in limb leads. The instantaneous body surface potential maps (BSPMs) were analyzed during ventricular depolarization. P values <0.05 were considered statistically significant. Results Compared with Pre-Ex, an early completion of the second inversion of potential distributions, an early completion of ventricular depolarization, as well as a decrease in the duration of the middle phase and the total duration of ventricular depolarization on BSPMs were revealed at Post-Ex 5. Also, compared with Pre-Ex, an increase in the amplitude of negative BSPM extremum at the R-wave peak on the ECG in lead II (RII-peak) and a decrease in the amplitude of negative BSPM extremum at 3 and 4 ms after RII-peak were showed at Post-Ex 5. At Post-Ex 10, parameters of BSPMs did not differ from those at Pre-Ex. Conclusion In rats, acute exhaustive exercise causes reversible changes in the temporal and amplitude characteristics of BSPMs during ventricular depolarization, most likely related to alterations in the excitation of the main mass of the ventricular myocardium.

Body Surface Potential Mapping during Ventricular Depolarization in Rats after Acute Exhaustive Exercise. / Mapeamento Potencial de Superfície Corporal durante a Despolarização Ventricular em Ratos Após Exercício Exaustivo Agudo.

BACKGROUND: Exhaustive physical exercise can cause substantial changes in the electrical properties of the myocardium. OBJECTIVE: To evaluate, using body surface potential mapping, the electrical activity of the heart in rats during ventricular depolarization after acute exhaustive exercise. METHODS: Twelve-week-old male rats were submitted to acute treadmill exercise at 36 m/min until exhaustion. Unipolar electrocardiograms (ECGs) from the torso surface were recorded in zoletil-anesthetized rats three to five days before (Pre-Ex), 5 and 10 minutes after exhaustive exercise (Post-Ex 5 and Post-Ex 10, respectively) simultaneously with ECGs in limb leads. The instantaneous body surface potential maps (BSPMs) were analyzed during ventricular depolarization. P values <0.05 were considered statistically significant. RESULTS: Compared with Pre-Ex, an early completion of the second inversion of potential distributions, an early completion of ventricular depolarization, as well as a decrease in the duration of the middle phase and the total duration of ventricular depolarization on BSPMs were revealed at Post-Ex 5. Also, compared with Pre-Ex, an increase in the amplitude of negative BSPM extremum at the R-wave peak on the ECG in lead II (RII-peak) and a decrease in the amplitude of negative BSPM extremum at 3 and 4 ms after RII-peak were showed at Post-Ex 5. At Post-Ex 10, parameters of BSPMs did not differ from those at Pre-Ex. CONCLUSION: In rats, acute exhaustive exercise causes reversible changes in the temporal and amplitude characteristics of BSPMs during ventricular depolarization, most likely related to alterations in the excitation of the main mass of the ventricular myocardium.

FUNDAMENTO: O exercício físico exaustivo pode causar alterações significantes nas propriedades elétricas do miocárdio. OBJETIVO: Avaliar, através do mapeamento potencial de superfície corporal, a atividade elétrica do coração de ratos durante a despolarização ventricular após exercício exaustivo agudo. MÉTODOS: Ratos machos com doze semanas de idade foram submetidos a exercício agudo em esteira a 36 m/min até a exaustão. Eletrocardiogramas unipolares (ECGs) da superfície do tronco foram registrados em ratos anestesiados com zoletil três a cinco dias antes (Pré-Ex), 5 e 10 minutos após exercício exaustivo (Pós-Ex 5 e Pós-Ex 10, respectivamente) simultaneamente com ECGs nas derivações dos membros. Os mapas potenciais de superfície corporal instantâneos (BSPMs, body surface potential maps ) foram analisados durante a despolarização ventricular. Os valores de p <0,05 foram considerados estatisticamente significantes. RESULTADOS: Comparado com o Pré-Ex, uma conclusão precoce da segunda inversão de distribuições de potencial, uma conclusão precoce da despolarização ventricular, bem como uma diminuição na duração da fase média e a duração total da despolarização ventricular nos BSPMs foram reveladas no Pós-Ex5. Além disso, em comparação com o Pré-Ex, um aumento na amplitude do extremo negativo do BSPM no pico da onda R no ECG na derivação II (pico RII) e uma diminuição na amplitude do extremo negativo do BSPM a 3 e 4 ms após o pico RII foram demonstrados no Pós-Ex 5. No Pós-Ex 10, os parâmetros dos BSPMs não diferiram daqueles do Pré-Ex. CONCLUSÃO: Em ratos, o exercício exaustivo agudo causa alterações reversíveis nas características temporais e de amplitude dos BSPMs durante a despolarização ventricular, provavelmente relacionadas a alterações na excitação da massa principal do miocárdio ventricular.

DNA Environment of Centromeres and Non-Homologous Chromosomes Interactions in Mouse.

Although the pericentromeric regions of chromosomes that are enriched in tandemly repeated satellite DNA represent a significant part of eukaryotic genomes, they remain understudied, which is mainly due to interdisciplinary knowledge gaps. Recent studies suggest their important role in genome regulation, karyotype stability, and evolution. Thus, the idea of satellite DNA as a junk part of the genome has been refuted. The integration of data regarding molecular composition, chromosome behaviour, and the details of the in situ organization of pericentromeric regions is of great interest. The objective of this work was a cytogenetic analysis of the interactions between pericentromeric regions from non-homologous chromosomes in mouse spermatocytes using immuno-FISH. We analysed two events: the associations between centromeric regions of the X chromosome and autosomes and the associations between the centromeric regions of the autosomal bivalents that form chromocenters. We concluded that the X chromosome forms temporary synaptic associations with different autosomes in early meiotic prophase I, which can normally be found until the pachytene-diplotene, without signs of pachytene arrest. These associations are formed between the satellite-DNA-rich centromeric regions of the X chromosome and different autosomes but do not involve the satellite-DNA-poor centromeric region of the Y chromosome. We suggest the hypothetical model of X chromosome competitive replacement from such associations during synaptic correction. We showed that the centromeric region of the X chromosome in association remains free of γH2Ax-dependent chromatin inactivation, while the Y chromosome is completely inactivated. This finding highlights the predominant role of associations between satellite DNA-rich regions of different chromosomes, including the X chromosome. We suppose that X-autosomal transient associations are a manifestation of an additional synaptic disorder checkpoint. These associations are normally corrected before the late diplotene stage. We revealed that the intense spreading conditions that were applied to the spermatocyte I nuclei did not lead to the destruction of stretched chromatin fibers of elongated chromocenters enriched in satellite DNA. The tight associations that we revealed between the pericentromeric regions of different autosomal bivalents and the X chromosome may represent the basis for a mechanism for maintaining the repeats stability in the autosomes and in the X chromosome. The consequences of our findings are discussed.

Age-Specific Etiology of Severe Acute Respiratory Infections and Influenza Vaccine Effectivity in Prevention of Hospitalization in Russia, 2018-2019 Season.

The expansion and standardization of clinical trials, as well as the use of sensitive and specific molecular diagnostics methods, provide new information on the age-specific roles of influenza and other respiratory viruses in development of severe acute respiratory infections (SARI). Here, we present the results of the multicenter hospital-based study aimed to detect age-specific impact of influenza and other respiratory viruses (ORV). The 2018-2019 influenza season in Russia was characterized by co-circulation of influenza A(H1N1)pdm09 and A(H3N2) virus subtypes which were detected among hospitalized patients with SARI in 19.3% and 16.4%, respectively. RSV dominated among ORV (15.1% of total cases and 26.8% in infants aged ≤ 2 years). The most significant SARI agents in intensive care units were RSV and influenza A(H1N1)pdm09 virus, (37.3% and 25.4%, respectively, of PCR-positive cases). Hyperthermia was the most frequently registered symptom for influenza cases. In contrast, hypoxia, decreased blood O2 concentration, and dyspnea were registered more often in RSV, rhinovirus, and metapneumovirus infection in young children. Influenza vaccine effectiveness (IVE) against hospitalization of patients with PCR-confirmed influenza was evaluated using test-negative case-control design. IVE for children and adults was estimated to be 57.0% and 62.0%, respectively. Subtype specific IVE was higher against influenza A(H1N1)pdm09, compared to influenza A(H3N2) (60.3% and 45.8%, respectively). This correlates with delayed antigenic drift of the influenza A(H1N1)pdm09 virus and genetic heterogeneity of the influenza A(H3N2) population. These studies demonstrate the need to improve seasonal influenza prevention and control in all countries as states by the WHO Global Influenza Strategy for 2019-2030 initiative.

Repertoire of Rearranged Immunoglobulin Heavy Chain Genes in Russian Patients With B-Cell Lymphoproliferative Diseases.

INTRODUCTION: Immunoglobulin heavy chain variable region (IGHV) repertoire narrowing could be an evidence for the involvement of a limited set of antigens in the development of lymphomas. For chronic lymphocytic leukemia (CLL) the existence of more than 200 subgroups of tumor IGHV antigen-binding sites, so called "stereotypical" antigen receptors (SAR) has been shown. For others lymphomas the possibility of SARs is also suggested. The aim of this study is to compare the tumor IGHVs and possible SARs in various B-cell malignancies in Russia and other countries. MATERIALS AND METHODS: The study included samples of 1800 CLL patients, 52 patients with mantle cell lymphoma, 48 patients with hairy cell lymphoma and 37 patients with splenic marginal cell lymphoma. The nucleotide sequences of the IGHV genes were determined according to ERIC protocol. RESULTS: In CLL most common IGHV genes were IGHV1-69, IGHV1-2, IGHV3-30 and IGHV4-34. The most common SARs were CLL#1, CLL#6, CLL#2, CLL#3. In MCL the most common genes were IGHV4-34, IGHV3-21, IGHV3-23. In 5 MCL patients CDR3 sequences were identified matching definitions of a stereotyped. In the half of SMZL patients was identified gene IGHV1-2. Other IGHV genes were much less common. Two pairs of SMZL patients have motives similar to each other. In HCL IGHV repertoire was the most variable, no trends for antigen receptor stereotypy were observed. It was found that SARs are highly disease-specific both at the level of nucleotide and amino acid sequences. CONCLUSION: Our results suggest that antigens crucial for the pathogenesis of B-cell malignancies could be disease-specific. Further studies on extended samples of non-CLL patients concerning the role of SARs in pathogenesis of these diseases may also contribute to the development of new diagnostic and prognostic markers.

Inhibitor of BRAFV600E Mutation as a Treatment Option for Hairy Cell Leukemia With Deep Neutropenia and Infectious Complications.

Standard therapy in hairy cell leukemia (HCL) is often impossible at the time of deep neutropenia/agranulocytosis with or without infectious complications; it is thus a complex therapeutic problem. Vemurafenib has been used to treat resistant HCL since 2012. Because vemurafenib does not have a myelotoxic effect, we thought that it could be used to treat HCL associated with deep neutropenia/agranulocytosis with or without the development of infectious complications as a preliminary stage before treatment with cladribine. We conducted a retrospective analysis of treatment with vemurafenib followed by a standard course of cladribine provided to 22 patients with deep neutropenia/agranulocytosis with or without infectious complications at diagnosis. Vemurafenib was provided to 22 patients with HCL. The response to therapy was evaluated by complete blood cell count (absolute neutrophil count [ANC], hemoglobin concentration, platelet count, absence of hairy cells), spleen size (assessed by ultrasound), and reduce infectious complications. After that, a standard course of cladribine was provided. Among the 22 patients, the male/female sex ratio was 2:1, and median (range) age was 52 (24-78) years. There were 7 patients with severe infectious manifestations admitted to the intensive care unit, including 1 patient during extracorporeal membrane oxygenation. The median (range) ANC at diagnosis was 0.3 (0.04-0.7) × 109/L. Vemurafenib was provided at a dosage of 240 mg 1 or 2 times a day. In 20 patients, vemurafenib was provided for 3 months or more. In 1 case, the effect was not obtained during 1 month of treatment, and the patient died from severe infectious complications during prolonged agranulocytosis. In 21 patients treated with vemurafenib, an increase of ANC was observed and the infectious complications resolved, thus allowing the application of cladribine therapy. After a standard course (0.1 mg/kg per day for 7 days) of cladribine chemotherapy, 18 patients (90%) experienced complete clinical remission and 2 patients (10%) experienced partial remission with residual splenomegaly. In 1 patient, vemurafenib therapy was still ongoing 2 months after initiating therapy. In cases of proven BRAFV600E mutation, vemurafenib can be successfully used as an effective preliminary therapy in patients with deep neutropenia/agranulocytosis with or without infectious complications before standard therapy with purine analogs.

Extraction and ICP-OES determination of heavy metals using tetrabutylammonium bromide aqueous biphasic system and oleophilic collector.

Preconcentration of Cd(II), Co(II), Cu(II), Ni(II), Pb(II), and Zn(II) in aqueous biphasic system tetrabutylammonium bromide (TBAB) - H2O - (NH4)2SO4 followed by ICP-OES determination is reported for the first time. All metal ions extracted quantitatively with 4-(2-pyridylazo)-resorcinol. Nonwoven polypropylene collector was used to capture the extract and to assist phase separation. The concentrate was eluted with 0.3 mol L-1 nitric acid and introduced directly into ICP-OES sampling line. Detection limits for cadmium, cobalt, copper, nickel, lead, and zinc were 0.012, 0.2, 0.17, 0.11, 0.2, and 0.4 µg L-1, respectively; preconcentration factor was 150. The method accuracy was verified by the analysis of certified reference materials samples of waste waters and surface water, as well as the real samples of spring and well water. The use of oleophilic polypropylene collector allowed to handle small volume extract, attain high preconcentration factor and significantly decrease detection limits for ICP-OES determination of heavy metals.

Effect of the C-reactive protein gene on risk and clinical characteristics of schizophrenia in winter-born individuals.

C-reactive protein (CRP) levels are elevated in a subset of schizophrenia patients and correlated with more severe symptoms, which makes CRP a potential theranostic biomarker for the disease. However, genotypes associated with higher CRP concentrations have the protective effect against schizophrenia. To resolve this discrepancy, more research on the role of CRP in schizophrenia is needed. The present study aimed to investigate the effects on schizophrenia of the CRP gene in combination with season of birth (SOB), the known risk factor for the disease. We first examined the impact of seasonality on schizophrenia risk in the Russian population, using samples of 2452 patients and 1203 controls, and then assessed the CRP rs2794521 polymorphism × SOB interaction effect on the disease risk, age-of-onset and symptoms severity in 826 patients and 476 controls. An excess of winter births in patients was not significant. At the same time, we found that winter-born patients carrying the CRP GG genotype, which is associated with low transcriptional activity, had an earlier age at onset than the other patients. The findings are in line with the protective role of high active CRP genetic variants in the development of schizophrenia and provide support for the hypothesis that this effect of CRP takes place early in life.

High Incidence of Clonal CD8+ T-cell Proliferation in Non-malignant Conditions May Reduce the Significance of T-cell Clonality Assay for Differential Diagnosis in Oncohematology.

Polymerase chain reaction (PCR) analysis of rearranged T-cell receptor (TCR) genes is a valuable diagnostic tool for differential diagnosis of T-cell large granular lymphocytic (T-LGL) leukemia and reactive lymphocytosis. Age-related narrowing of T-cells repertoire and expansion of immune or autoimmune clones may lead to false-positive results. The objective of this study was to evaluate the specificity and positive predictive value of PCR-based clonality assessment for a differential diagnostics of T-LGL leukemia. Rearrangements of TCRG and TCRB genes using the BIOMED-2 protocol were assessed in healthy individuals including the elderly (n = 62) and patients with rheumatic diseases (n = 14), transitory reactive CD8+ lymphocytosis (n = 17), and T-LGL leukemia (n = 42). Monoclonal TCRG/TCRB rearrangements in blood were identified in 11.3%/4.8% (7/3 of 62) of healthy individuals; 21.4%/14.3% (3/2 of 14) of patients with rheumatic diseases, and 17.6%/11.8% (3/2 of 17) of patients with reactive lymphocytosis. Immunomagnetic selection of lymphocytes in healthy individuals (31 of 33) revealed that clonal T-cells belong to CD8+ and CD57+ population. No clonal Vß-Jß TCRB rearrangements were found in the control group, only Dß-Jß TCRB and TCRG. Given the high detectability (96.7%) of Vß-Jß TCRB monoclonal rearrangements in patients with αß-T-LGL leukemia, this marker had the greatest specificity and positive predictive value (100%; 99.2%). The presence of clonal CD8+CD57+ cells in blood is common for healthy individuals and patients with reactive conditions and may not associate with any malignancy. Different specificity of TCRG/ Dß-Jß TRB/ Vß-Jß TCRB PCR reactions should be taken into account for T-cell clonality data interpretation.

Multielement Determination of Trace Heavy Metals in Water by Microwave-Induced Plasma Atomic Emission Spectrometry after Extraction in Unconventional Single-Salt Aqueous Biphasic System.

For the first time liquid-liquid extraction was used for the preconcentration of heavy metals prior to their determination in water by microwave-induced plasma atomic emission spectrometry (MP-AES). Extraction of Pb(II), Cd(II), Co(II), Ni(II), Zn(II), and Cu(II) was performed in unconventional aqueous biphasic system (ABS) formed by addition of hydrophobic solid salt, namely, tetrahexylammonium bromide, to aqueous sample, with neither organic solvents nor salting-out agents being used. The metal ions were quantitatively recovered with 4-(2-pyridylazo)-resorcinol (PAR). The extract was diluted with ethanol/HCl and introduced directly into an MP-AES instrument. The factors influencing extraction (pH, reagent concentration, phase contact time, etc.) and MP-AES detection parameters were studied and optimized. For the developed method, limits of detection of 1.3, 4.9, 0.06, 1.2, 4.2, and 3.2 µg L-1 were obtained for cadmium, cobalt, copper, nickel, lead, and zinc, respectively, providing from 60- to 500-fold improvement as compared with the analysis without preconcentration. The method was applied for the analysis of two certified reference materials (CRM) of wastewater and surface water as well as the samples of well and seawater. Coupling MP-AES with ABS extraction significantly extends the capabilities of the method, especially for the analysis of high salinity waters.

Phylogeography of Crimean Congo Hemorrhagic Fever Virus.

Crimean Congo hemorrhagic fever virus (CCHFV) is one of the most severe viral zoonozes. It is prevalent throughout Africa, Asia and southern Europe. Limited availability of sequence data has hindered phylogeographic studies. The complete genomic sequence of all three segments of 14 Crimean Congo hemorrhagic fever virus strains isolated from 1958-2000 in Russia, Central Asia and Africa was identified. Each genomic segment was independently subjected to continuous Bayesian phylogeographic analysis. The origin of each genomic segment was traced to Africa about 1,000-5,000 years ago. The virus was first introduced to South and Central Asia in the Middle Ages, and then spread to China, India and Russia. Reverse transfers of genomic segments from Asia to Africa were also observed. The European CCHFV genotype V was introduced to Europe via the Astrakhan region in South Russia 280-400 years ago and subsequently gradually spread westward in Russia, to Turkey and the Balkans less than 150 years ago. Only a few recombination events could be suggested in S and L genomic segments, while segment reassortment was very common. The median height of a non-reassortant phylogenetic tree node was 68-156 years. There were reassortment events within the European CCHFV lineage, but not with viruses from other locations. Therefore, CCHFV in Europe is a recently emerged zoonosis that represents a spillover from the global gene pool.

Genetic dissection of host immune response in pneumonia development and progression.

The role of host genetic variation in pneumonia development and outcome is poorly understood. We studied common polymorphisms in the genes of proinflammatory cytokines (IL6 rs1800795, IL8 rs4073, IL1B rs16944), anti-inflammatory cytokines (IL10 rs1800896, IL4 rs2243250, IL13 rs20541) and toll-like receptors (TLR2 rs5743708 and rs4696480, TLR4 rs4986791, TLR9 rs352139, rs5743836 and rs187084) in patients with community-acquired pneumonia (CAP) (390 cases, 203 controls) and nosocomial pneumonia (355 cases, 216 controls). Experimental data were included in a series of 11 meta-analyses and eight subset analyses related to pneumonia susceptibility and outcome. TLR2 rs5743708 minor genotype appeared to be associated with CAP/Legionnaires' disease/pneumococcal disease. In CAP patients, the IL6 rs1800795-C allele was associated with severe sepsis/septic shock/severe systemic inflammatory response, while the IL10 rs1800896-A allele protected against the development of these critical conditions. To contribute to deciphering of the above results, we performed an in silico analysis and a qualitative synthesis of literature data addressing basal and stimulated genotype-specific expression level. This data together with database information on transcription factors' affinity changes caused by SNPs in putative promoter regions, the results of linkage disequilibrium analysis along with SNPs functional annotations supported assumptions about the complexity underlying the revealed associations.

Wildlife forensic examination in Russia: the practice and perspectives.

Two case studies are described which show application of forensic expertise to establish important circumstances related to the investigation of the crimes against wildlife flora and fauna. Forensic study of the animal hair is a method for investigation of the crimes against wildlife objects which is used more frequently during the recent years. The perspectives of development of the new branch of forensic research in Russia are formulated, and the proposals and recommendations for developing the Eurasian wildlife flora and fauna network are formulated in this article.

Hsp70-Bag3 interactions regulate cancer-related signaling networks.

Bag3, a nucleotide exchange factor of the heat shock protein Hsp70, has been implicated in cell signaling. Here, we report that Bag3 interacts with the SH3 domain of Src, thereby mediating the effects of Hsp70 on Src signaling. Using several complementary approaches, we established that the Hsp70-Bag3 module is a broad-acting regulator of cancer cell signaling by modulating the activity of the transcription factors NF-κB, FoxM1, Hif1α, the translation regulator HuR, and the cell-cycle regulators p21 and survivin. We also identified a small-molecule inhibitor, YM-1, that disrupts the Hsp70-Bag3 interaction. YM-1 mirrored the effects of Hsp70 depletion on these signaling pathways, and in vivo administration of this drug was sufficient to suppress tumor growth in mice. Overall, our results defined Bag3 as a critical factor in Hsp70-modulated signaling and offered a preclinical proof-of-concept that the Hsp70-Bag3 complex may offer an appealing anticancer target.

Achnanthidium minutissimum (Bacillariophyta) valve deformities as indicators of metal enrichment in diverse widely-distributed freshwater habitats.

In the presence of different environmental stressors, diatoms can produce frustules presenting different types of deformities. Metals and trace elements are among the most common causes of these teratological forms. Metal enrichment in water bodies can be attributed to the geological setting of the area or to pollution. The widespread benthic diatom Achnanthidium minutissimum (ADMI) is one of the most metal-tolerant species. In the present study, ADMI teratologies were defined from samples taken from eight very diverse, widely-distributed inland-water habitats: streams affected by active and abandoned mining areas, a metal-contaminated stream, a spring in an old chalcopyrite mine, a mineral-water fountain, and a sediment core taken from a lake affected by metal contamination in the past. Deformed frustules of ADMI were characterised mainly by one (sometimes two) more or less bent off ending, conferring to the specimens a cymbelloid outline (cymbelliclinum-like teratology, CLT). Marked teratologies were distinguished from slight deformities. Hydrochemical analyses, including metals and trace elements, were carried out and enrichment factors (EF) relative to average crustal composition were calculated. To improve our knowledge on the potential of different metals and trace elements to trigger the occurrence of ADMI CLT, we carefully selected 15 springs out of 110 (CRENODAT dataset) where both ADMI and above-average metal or metalloid concentrations occurred, and re-analysed these samples. The results from the eight widely-distributed core sites as well as from the 15 selected CRENODAT springs led to the hypothesis that two metals (copper and zinc) and a metalloid (antimony) were the most likely triggers of ADMI CLT formation. From a quantitative point of view, it is worth noting that the lowest concentrations triggering ADMI CLT can be fairly low, particularly in the case of copper contamination. The antimony-rich site was characterised by a marked-teratology variant where both ends of ADMI were bent off.