Репозиторий Университета

© 2019 Elsevier Inc. Air pollution caused by vehicle emissions remains a serious environmental threat in urban areas. Sedimentation of atmospheric aerosols, surface wash, drainage water, and urbane wastewater can bring vehicle particle emissions into the aquatic environment. However, the level of toxicity and mode of toxic action for this kind of particles are not fully understood. Here we explored the aquatic toxic effects of particulate matter emitted from different types of vehicles on marine microalgae Porphyridium purpureum and Heterosigma akashiwo. We used flow cytometry to evaluate growth rate inhibition, changes in the level of esterase activity, changes in membrane potential and size changes of microalgae cells under the influence of particulate matter emitted by motorcycles, cars and specialized vehicles with different types of engines and powered by different types of fuel. Both microalgae species were highly influenced by the particles emitted by diesel-powered vehicles. These particle samples had the highest impact on survival, esterase activity, and membrane potential of microalgae and caused the most significant increase in microalgae cell size compared to the particles produced by gasoline-powered vehicles. The results of the algae-bioassay strongly correlate with the data of laser granulometry analyses, which indicate that the most toxic samples had a significantly higher percentage of particles in the size range less than 1 μm. Visual observation with an optical microscope showed intensive agglomeration of the particles emitted by diesel-powered vehicles with microalgae cells. Moreover, within the scope of this research, we did not observe the direct influence of metal content in the particles to the level of their aquatic toxicity, and we can conclude that physical damage is the most probable mechanism of toxicity for vehicle emitted particles.

© 2019 Elsevier Inc. Welding fumes are a major source of metal oxide particles, ozone, carbon monoxide, carbon dioxide, nitrogen oxides, and many other toxic substances. Hazardous properties and the level of toxicity of welding fumes depend mostly on the welding electrode type and the welding regime parameters. The specific objective of this study was to evaluate the aquatic toxicity of metal welding fume particles in vivo on microalga Heterosigma akashiwo. The quantity and size of particles were measured by flow cytometry using a scattering laser light with a wavelength of 405 nm. The number of microalgae cells after 72 h and 7 days exposition with welding fume particle suspensions was evaluated by flow cytometry. Morphological changes of the microalga were observed by optical microscopy. The toxic effect was demonstrated as a significant reduction of cell density after exposure of microalgae to welding fume particles. The greatest impact on the growth of microalga was caused by particles with high rutile content. It was shown that the adverse effect of metal oxide particles depends more on the chemical composition of particles in welding fume while the number and dispersity of particles had no noticeable toxic influence on microalgae. The findings of this research confirm the fact that the toxicity of welding fume particles can be significantly reduced by using rutile-cellulose coated electrodes.

© 2018, Allerton Press, Inc. The proportion of diploid cells in the G0/G1 cell cycle phases was estimated by flow cytometry in 60 samples of stage III serous ovarian cancer tissue. The tumor tissue shows considerable heterogeneity with regard to the content of this tissue fraction, which ranged from 27 to 95% with a median of 73%. Statistically significant differences in the size of this fraction were identified by comparing tumor subgroups sensitive and resistant to first-line platinum-taxane chemotherapy. Predictive significance of the G0/G1 fraction was concluded and quantitative evaluation of this fraction is recommended for clinical use.

© 2018 WILEY-VCH Verlag GmbH  &  Co. KGaA, Weinheim While noninvasive prenatal testing based on cell-free fetal DNA has recently revolutionized the field of aneuploidy screening in pregnancy, it remains limited to aneuploidy and microdeletion screening, and is unable to reliably detect single gene disorders. A number of recent studies have demonstrated the potential of circulating trophoblastic cells in providing cell-based noninvasive diagnosis with sequencing or array-based assays. However, considering the extreme rarity of these cells in blood, efficient, high-throughput, and clinically applicable enrichment technologies are yet to be developed. This study demonstrates for the first time the utility of inertial microfluidics for efficient isolation of trophoblastic cells from maternal peripheral blood. Under optimal operating conditions, high-recovery yields (79%) are obtained using a trophoblastic cell-line, which is subsequently confirmed with analysis of maternal blood. Feasibility of obtaining a diagnosis from cells isolated from a maternal sample is demonstrated in a case of confirmed fetal trisomy 21 in which six fetal cells are found in a 7 mL blood sample using fluorescence in situ hybridization. Finally, it is demonstrated that trophoblastic cells isolated using inertial microfluidics could be picked and subjected to a clinically validated sequencing assay, paving the way for further validation of this technology and larger clinical studies.

© 2018 The Union. BACKGROUND: Tumour necrosis factor-alpha (TNF-α) and its inhibitors are involved in both defence against tuberculosis (TB) and damage to the host by TB. Notably, the change in receptor expression on cell density is a key mechanism in regulation of the biological properties of cytokines. OBJECTIVE : To study the differences in TNF-α receptor (TNFR) expression in patients with active pulmonary tuberculosis (aPTB) in correlation with the parameters of disease severity. METHODS : TNFR1/2 levels on peripheral blood mononuclear cells (PBMCs) from 45 patients with aPTB and 150 healthy controls were analysed by flow cytometry using monoclonal antibodies and QuantiBRITE beads. Soluble TNFR1/2 and TNF-α in serum were measured using an enzyme-linked immunosorbent assay. RESULT S : TNFR1 expression in aPTB patients was increased in the main populations of immune cells. Patients who were Mycobacterium tuberculosis culturepositive on bronchoscopy had higher levels of the soluble forms of TNFR1 (sTNFR1) than M. tuberculosisnegative patients. CONCLUS ION: Active TB was shown to cause activation of different immune cell types by increasing TNFR1 expression on cells and reducing sTNFR1 expression compared with healthy controls. M. tuberculosis-positive patients with disseminated infection had the highest sTNFR1 serum level compared with other patients, but did not differ in receptor expression on PBMCs.

© 2018 Ima-Press Publishing House. All rights reserved.  The clinical efficacy and safety of interleukin-6 (IL-6) receptor blockade have been well studied, but the data on the impact of therapeutic inhibition of IL-6 on B cells are scarce and contradictory. Preliminary reports have shown that B cell function and a humoral immune response may be modulated by an IL-6 receptor inhibitor. Objective: to assess the effect of 12-month tocilizumab (TCZ) therapy on B-cell phenotype and gene expression in RA and to analyze the association between B-cell subsets and RA activity. Subjects and methods. Examinations were made in 24 active RA patients (20 women and 4 men) (median age, 55 [49; 64] years; disease duration, 72 [24; 108] months; DAS28 5.8 [5.3; 6.3]; the patients were seropositive for rheumatoid factor (RF) (100%) and for anti-cyclic citrullinated peptide antibodies (87.3%). The patients received TCZ 8 mg/kg every 4 weeks. After 12 months of therapy, 54% of patients were categorized as good responders, 46% as moderate responders according to the EULAR response criteria. A control group consisted of 29 volunteers (21 women and 8 men; median age, 58.5 [53.0; 62.0] years). Peripheral blood lymphocytes were immunophenotyped at the time of enrollment and after 12 months. The absolute and relative counts of CD19+B lymphocytes, memory B cells (CD19+CD27+), non-switched memory B cells (CD19+IgD+CD27+), switched memory B cells (CD19+IgDCD27+), naive (CD19+IgD+CD27-), double-negative (CD19+IgD-CD27-), transitional (CD19+IgD+CD10+CD38++CD27) B cells, plasma cells (CD19+CD38+), and plasmablasts (CD19+CD38+++IgD-CD27+CD20-) were estimated using multicolor flow cytometry. Results and discussion. The relative and absolute counts of memory B cells (CD19+CD27+) (1.3 [0.9; 1.7]%, 0015 [0.001; 0.003]•10 9 /l), switched memory B cells (CD19+IgD-CD27+) (6.8 [3.6; 11.6]%, 0.01 [0.005; 0.02]•10 9 /l), and the absolute number of transitional B cells (CD19+CD38++CD10+IgD+CD27-) (0.00009 [0; 0.00028]•10 9 /l) were found to be lower in RA patients than in donors: 2.2 [1.1; 3.0]%, 0.003 [0.001; 0.007]•10 9 /l; 12.8 [9.3; 17.0]%, 0.02 [0.01; 0.04]•10 9 /l; 0.0001 [0; 0.0003]•10 9 /l, respectively (p<0.05 for all cases). After 12 months of TCZ therapy initiation, there were decreases in the relative and absolute counts of plasmablasts (CD19+CD38+++CD27+IgD-CD20-) from 0.15 [0.1; 0.3] to 0.1 [0.01; 0.1]% and from 0.0003 [0.00007; 0.004]•10 9 /l to 0.0001 [0; 0.0003]•10 9 /l, respectively (p<0.05). At the same time, the relative and absolute counts of memory B cells (CD19+CD27+) and switched memory B cells (CD19+CD27+IgD-) remained lower in RA patients than in donors: 1.0 [0.7; 1.2] and 2.2 [1.1; 3.0]%; 0.001 [0.006; 0.003]•10 9 /l and 0.003 [0.001; 0.007]•10 9 /l; 3.1 [1.1; 4.2] and 12.8 [9.3; 17.0]%; 0.003 [0.002; 0.006]•10 9 /l and 0.02 [0.01; 0.04]•10 9 /l, respectively (p<0.05 for all cases). Following 12 months of TCZ therapy, the numbers of other B-cell subpopulations were not considerably altered. When included in the study, the patients with RA showed correlations between the absolute count of memory B cells (CD19+CD27+) and the level of C-reactive protein (r=0.50; p<0.05); between the absolute count of plasmablasts (CD19+CD38+++CD27+IgD-CD20-) and the level of RF (r=0.41 and r=0.52; p<0.05). There were no correlations of B cell subsets with clinical and laboratory findings after 12 months of TCZ initiation. Conclusion. Immunophenotyping of peripheral blood B lymphocyte subsets showed the lower relative and absolute counts of memory B cells (CD19+CD27+) and switched memory B cells (CD19+CD27+IgD-) in RA patients than in healthy donors. The found correlations between the counts of memory B cells and plasmablasts and the values of laboratory parameters in patients with high RA activity may suggest that B lymphocytes are involved in the pathogenesis of RA. There was a decline in plasmablast levels after 12 months of TCZ therapy.

© 2018 Park-media, Ltd.  Common marmosets are small New World primates that have been increasingly used in biomedical research. This report presents efficient protocols for assessment of the parameters of adaptive cell-mediated immunity in common marmosets, including the major subpopulations of lymphocytes and main markers of T- and B-cell maturation and activation using flow cytometry with a multicolor panel of fluorescently labelled antibodies. Blood samples from eight common marmosets were stained with fluorescently labeled monoclonal antibodies against their population markers (CD45, CD3, CD20, CD4, CD8) and lymphocyte maturation and activation markers (CD69, CD62L, CD45RO, CD107a and CD27) and analyzed by flow cytometry. Within the CD45 + population, 22.7±5.5% cells were CD3 - CD20 + and 67.6±6.3% were CD3 + CD20 - . The CD3 + subpopulation included 55.7±5.5% CD3 + CD4 + CD8 - and 34.3±3.7% CD3 + CD4 - CD8 + cells. Activation and maturation markers were expressed in the following lymphocyte proportions: CD62L on 54.0±10.7% of CD3 + CD4 + cells and 74.4±12.1% of CD3 + CD8 + cells; CD69 on 2.7±1.2% of CD3 + CD4 + cells and 1.2±0.5% of CD3 + CD8 + cells; CD45RO on 1.6±0.6% of CD3 + CD4 + cells and 1.8±0.7% of CD3 + CD8 + cells; CD107a on 0.7±0.5% of CD3 + CD4 + cells and 0.5±0.3% of CD3 + CD8 + cells; CD27 on 94.6±2.1% of CD3 + cells and 8.9±3.9% CD20 + cells. Female and male subjects differed in the percentage of CD3 + CD4 + CD45RO + cells (1.9±0.5 in females vs 1.1±0.2 in males; p < 0.05). The percentage of CD20 + CD27 + cells was found to highly correlate with animals' age (r = 0.923, p < 0.005). The basal parameters of adaptive cell-mediated immunity in naïve healthy marmosets without markers of systemic immune activation were obtained. These parameters and the described procedures are crucial in documenting the changes induced in common marmosets by prophylactic and therapeutic immune interventions.