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Monoclonal antibodies (mAbs) of the IgG1 isotype obtained to the full-length recombinant pneumolysin (rPly) did not recognize or poorly recognized conformational epitopes of native pneumolysin in ELISA. At the same time, polyclonal antibodies (pAbs(rPly)) detected native pneumolysin in sandwich ELISA when they were used as capture and detecting antibodies simultaneously. All mAbs(rPly) inhibited erythrocyte hemolysis induced by native pneumolysin. The combined use of mAbs(rPly) did not reveal an increase in optical density in ELISA and an increase in inhibition hemolytic activity, which suggests that mAbs(rPly) are directed to the same or spatially closely located epitopes. pAbs(rPly) more effectively inhibited erythrocyte hemolysis than mAbs(rPly)., (© 2025. Springer Science+Business Media, LLC, part of Springer Nature.)

We report a spatial light interference microscopy (SLIM)-based methodology aimed at automatic monitoring and analysis of changes in cellular morphology within extended fields of view in cytological samples. The experimental validation was performed on HeLa cells in vitro subjected to localized photodynamic treatment. The performed long-term noninvasive monitoring using the SLIM technique allowed us to estimate quantitative parameters characterizing the dynamics of average phase shift in individual cells and to reveal changes in their morphology specific for different mechanisms of cell death. The results obtained evidenced that the proposed SLIM-based methodology provides an opportunity for identification of cell death type and quantification of cell death rate in an automatic mode. The major sources of potential errors that can affect the results obtained are discussed. The developed methodology is promising for automatic monitoring of large ensembles of individual cells and for quantitative characterization of their response to various treatment modalities.

We compared the immunogenicity of recombinant S. pneumoniae pneumolysin (rPly) when administered with and without Al(OH) 3 adjuvant, and evaluated the protective properties of recombinant protein in the active defense experiment. It was shown that double immunization with rPly+Al(OH) 3 increases the levels of IgG antibodies in comparison with the control (p<0.01), while triple immunization results in a more significant increase in IgG antibody levels (p<0.001). Double immunization with rPly without Al(OH) 3 does not induce a significant increase in antibody levels in comparison with the control, while triple immunization results in a slight but significant increase in antibody levels (p<0.05). The active defense test proved the protective activity of rPly against S. pneumoniae serotype 3 at intranasal infection., (© 2024. Springer Science+Business Media, LLC, part of Springer Nature.)

We report application of the fluorescence lifetime imaging microscopy (FLIM) for analysis of distributions of intracellular acidity using a chlorin-e6 based photosensitizer Radachlorin. An almost two-fold increase of the photosensitizer fluorescence lifetime in alkaline microenvironments as compared to acidic ones allowed for clear distinguishing between acidic and alkaline intracellular structures. Clusterization of a phasor plot calculated from fits of the FLIM raw data by two Gaussian distributions provided accurate automatic segmentation of lysosomes featuring acidic contents. The approach was validated in colocalization experiments with LysoTracker fluorescence in living cells of four established lines. The dependence of photosensitizer fluorescence lifetime on microenvironment acidity allowed for estimation of pH inside the cells, except for the nuclei, where photosensitizer does not penetrate. The developed method is promising for combined application of the photosensitizer for both photodynamic treatment and diagnostics., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Introduction: Although photophysical properties of Radachlorin photosensitizer (PS) were extensively studied in solutions and cells, no data is available on variations of its characteristics upon binding to serum albumins, which are major transporters in blood and nutrients in cell culture media., Objectives: The primary objective of this study was to analyze changes in photophysical properties of Radachlorin molecules upon their binding to human and bovine serum albumins at different microenvironment properties., Methods: Experiments were performed using time-resolved fluorescence spectroscopy and fluorescence recovery after photobleaching. Variations in fluorescence spectra and lifetime, fluorescence anisotropy, rotational and translational diffusion of PS molecules upon binding to albumins were studied in normal, basic and acidic conditions and at different concentrations of albumin and PS molecules., Results: Radachlorin molecules effectively bind to both types of serum albumins, which causes changes in photophysical properties of the PS. A minor red shift of the fluorescence spectrum, an increase in fluorescence lifetime and anisotropy and substantial decrease of translational and rotational mobility of PS molecules were observed upon their binding to albumins. The analysis of rotational diffusion time provided robust evaluation of the bound fraction of PS molecules. Both the highly acidic microenvironment and increase in alcohol concentration above 40% resulted in detachment of PS molecules from albumins. Photophysical properties of Radachlorin in complexes with BSA and HSA were found to be slightly different., Conclusions: Binding of Radachlorin photosensitizer to either BSA or HSA affects significantly its photophysical properties, which may also vary with microenvironment acidity and alcohol concentration., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

We present a thorough experimental investigation of fluorescence properties of Radachlorin photosensitizer in solutions of different acidity, viscosity and polarity. Experiments were performed using time-resolved fluorescence lifetime imaging and time-resolved analysis of polarized fluorescence. Variations of solution acidity resulted in considerable changes of Radachlorin fluorescence quantum yield and lifetime in the pH range from 4 to 7, but did not affect the rotational diffusion time, and almost did not change the quantum yield and characteristic times of singlet oxygen phosphorescence. Variations of solution polarity and viscosity were achieved by changing ethanol or methanol fraction in aqueous solution. The decrease of solution polarity resulted in nonlinear rise of Radachlorin fluorescence quantum yield and lifetime up to alcohol concentration of 50%-65%, as well as in considerable rise of singlet oxygen quantum yield and significant changes in characteristic times of its phosphorescence. Variations of solution viscosity resulted in changes of rotational diffusion time of Radachlorin molecules, which appeared to be in perfect correlation with methanol solution viscosity. Good correspondence with ethanol solution viscosity was observed only up to 50% alcohol fraction. Deviations of rotational diffusion time of Radachlorin molecules from direct proportionality with solution viscosity at higher ethanol concentrations were suggested to be due to different solvation conditions. The data obtained can give important reference points for analysis of microenvironment of Radachlorin molecules, their intracellular localization and performance in singlet oxygen generation., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Pneumolysin (Ply) is a target for the development of serotype-independent pneumococcal vaccines, an important condition for the efficacy of which is their ability to activate innate immunity with the subsequent formation of adaptive immunity. In this study, the ability of recombinant full-length Ply (rPly) of pneumococci to induce TLR expression and maturation of dendritic cells generated from mouse bone marrow was evaluated. It was shown that rPly in vitro increased the number of dendritic cells expressing Toll-like receptor 4 (TLR4) on the membrane. rPly caused maturation of dendritic cells generated from mouse bone marrow, which manifested in a decrease in the number of progenitor cells (CD34), an increase in the number of cells expressing the adhesion molecule CD38, costimulatory molecules CD80 and CD86, molecules of terminal differentiation of dendritic cells CD83, as well as molecules of antigenic presentation of the major histocompatibility complex class II., (© 2024. Springer Science+Business Media, LLC, part of Springer Nature.)

Background: Older primary central nervous system lymphoma (PCNSL) patients have an inferior prognosis compared to younger patients because available evidence on best treatment is scarce and treatment delivery is challenging due to comorbidities and reduced performance status. High-dose chemotherapy and autologous stem cell transplantation (HCT-ASCT) after high-dose methotrexate (MTX)-based immuno-chemotherapy has become an increasingly used treatment approach in eligible elderly PCNSL patients with promising feasibility and efficacy, but has not been compared with conventional chemotherapy approaches. In addition, eligibility for HCT-ASCT in elderly PCNSL is not well defined. Geriatric assessment (GA) may be helpful in selecting patients for the best individual treatment choice, but no standardized GA exists to date. A randomized controlled trial, incorporating a GA and comparing age-adapted HCT-ASCT treatment with conventional chemotherapy is needed., Methods: This open-label, multicenter, randomized phase III trial with two parallel arms will recruit 310 patients with newly diagnosed PCNSL > 65 years of age in 40 centers in Germany and Austria. The primary objective is to demonstrate that intensified chemotherapy followed by consolidating HCT-ASCT is superior to conventional chemotherapy with rituximab, MTX, procarbazine (R-MP) followed by maintenance with procarbazine in terms of progression free survival (PFS). Secondary endpoints include overall survival (OS), event free survival (EFS), (neuro-)toxicity and quality of life (QoL). GA will be conducted at specific time points during the course of the study. All patients will be treated with a pre-phase rituximab-MTX (R-MTX) cycle followed by re-assessment of transplant eligibility. Patients judged transplant eligible will be randomized (1:1). Patients in arm A will be treated with 3 cycles of R-MP followed by maintenance therapy with procarbazine for 6 months. Patients in arm B will be treated with 2 cycles of MARTA (R-MTX/AraC) followed by busulfan- and thiotepa-based HCT-ASCT., Discussion: The best treatment strategy for elderly PCNSL patients remains unknown. Treatments range from palliative to curative but more toxic therapies, and there is no standardized measure to select patients for the right treatment. This randomized controlled trial will create evidence for the best treatment strategy with the focus on developing a standardized GA to help define eligibility for an intensive treatment approach., Trial Registration: German clinical trials registry DRKS00024085 registered March 29, 2023., (© 2023. BioMed Central Ltd., part of Springer Nature.)

Intracellular localization of photosensitizer molecules is influential on cell death pathway at photodynamic treatment and is thus an important aspect in achieving enhanced efficacy of photodynamic therapy. In this paper we performed thorough studies of the distribution of Radachlorin photosensitizer in three established cell lines: HeLa, A549, and 3T3 with fluorescence lifetime imaging microscopy through the analysis of lifetime distributions. Experiments carried out in Radachlorin solutions in phosphate buffered saline revealed the pronounced dependence of the fluorescence quantum yield and lifetime on solution pH. This finding was used for analysis of lifetime images of living cells and their phasor plot representations and allowed us to suggest that Radachlorin localized predominantly in lysosomes, known to have acidic pH values. Experiments on co-localization of Radachlorin fluorescence lifetimes and LysoTracker fluorescence intensity supported this suggestion. The results obtained show that the inhomogeneity of fluorescence quantum yield within a cell can be significant due to lower pH values in lysosomes than in other intracellular compartments. This finding suggests that the actual amount of accumulated Radachlorin can be underestimated if being evaluated solely by comparison of fluorescence intensities., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

In this paper we compare the response of cells of established lines of different origin: HeLa, A549 and 3T3 to photodynamic treatment with Radachlorin photosensitizer. The analysis was performed on different aspects of the treatment procedure including photosensitizer accumulation, localization and photobleaching in cells and post-treatment dynamics of changes in cellular morphology at different treatment doses. It was shown that in the three cell lines Radachlorin accumulated in lysosomes to much greater extent than in mitochondria. The cells' response to treatment was analyzed by identification of their death pathways and evaluation of average phase shift dynamics using digital holographic microscopy. The analysis performed on the three cell lines allowed us to evaluate treatment doses specific for each pathway in each line. Among the three lines HeLa cells were found to be the most susceptible to treatment while 3T3 cells the most resistant. The comparison of these results with the data on Radachlorin accumulation, localization and photobleaching rates showed that the observed higher sensitivity of HeLa cells to photodynamic treatment correlated with higher photosensitizer uptake and more intensive photobleaching while lower sensitivity of 3T3 cells correlated with lower uptake and less intensive photobleaching., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Introduction: Limited availability of individual standards is a bottleneck for quality control of functional foods and natural medicines. The use of standard mixtures or secondary standards is a possible alternative in this case. Earlier, an approach known as standardised reference extract (RE) strategy was introduced for HPLC-UV analysis of different plant materials; however, its application in HPLC-MS analysis has not been investigated., Objective: To establish an HPLC-MS-based RE method for determination of ginsenoside content in ginseng infusions using commercially available extract reference material of Panax quinquefolius L., Results: The developed HPLC-MS method was validated as precise (1.1%-9.4% intra-day variation; 1.6%-12.8% inter-day variation) and highly sensitive [limit of detection (LOD): 1-40 ng/mL; limit of quantification (LOQ): 4-120 ng/mL]. The stability of samples was satisfactory (5.7%-16.3%). The RE quantification method was compared with the external standard method, and the obtained difference was not significant, mostly in the range of 5%-10%. Matrix effects for the diluted samples of RE and ginseng infusions, determined via the standard addition method, were in the range of 85%-115% and 80%-126%, respectively, and were also positively correlated with the ginsenoside concentration. Eleven batches of ginseng infusions from different manufacturers were analysed using the established method., Conclusion: The method for HPLC-MS-based ginsenoside quantification using RE as a secondary standard was established for the first time. The results of this study demonstrate that the application of the standardised RE strategy in HPLC-MS can minimise the matrix effect-related error in addition to the cost-effective quality control of herbal products, foods, and traditional medicines., (© 2022 John Wiley & Sons Ltd.)

The rapid expansion of food and nutrition information requires new ways of data sharing and dissemination. Interactive platforms integrating data portals and visualization dashboards have been effectively utilized to describe, monitor, and track information related to food and nutrition; however, a comprehensive evaluation of emerging interactive systems is lacking. We conducted a systematic review on publicly available dashboards using a set of 48 evaluation metrics for data integrity, completeness, granularity, visualization quality, and interactivity based on 4 major principles: evidence, efficiency, emphasis, and ethics. We evaluated 13 dashboards, summarized their characteristics, strengths, and limitations, and provided guidelines for developing nutrition dashboards. We applied mixed effects models to summarize evaluation results adjusted for interrater variability. The proposed metrics and evaluation principles help to improve data standardization and harmonization, dashboard performance and usability, broaden information and knowledge sharing among researchers, practitioners, and decision makers in the field of food and nutrition, and accelerate data literacy and communication., (© The Author(s) 2022. Published by Oxford University Press on behalf of the American Society for Nutrition.)

The rapid development of nutrition science is embracing digital transformation to generate large amounts of data. Precision nutrition and "Big Data" place increasing demand for data repositories and visualization, which enhances the digital transformation. We defined the need for an integrated nutrition data platform as a web-based platform that can collect, store, track, analyze, monitor, and visually display key metrics in nutrition and health while allowing users to interact with visuals and download data provided in the platform. Interactive dashboards create new opportunities for scholars and practitioners to generate and test hypotheses. We present the development and implementation of the Global Nutrition and Health Atlas (GNHA; https://sites.tufts.edu/gnha/), an open-access online platform covering nutrition and health data with 26 themes and 500+ indicators from 190+ countries up to 30 y. We view GNHA as an interactive tool aiming to share information and perspectives and foster collaborations and innovations., (© The Author(s) 2022. Published by Oxford University Press on behalf of the American Society for Nutrition.)

The paper presents steady-state and time-resolved experiments on photophysical processes associated with photodynamic inactivation of infections provided by nebulization of Radachlorin photosensitizer solution. As models of surfaces subjected to photodynamic inactivation we used glass, plant leaf, mushroom cap peel and superficial fascia of chicken and salmon skin flaps. The oxygen content in the photosensitizer solution was varied by blowing with atmospheric air and with pure oxygen. It was shown that singlet oxygen was generated efficiently in the aerosol jet and that its amount increased noticeably at higher oxygen concentrations. The kinetics of photosensitizer photobleaching on different surfaces were found to be significantly different with characteristic decay times varying from seconds for leaf and glass to minutes for fascial flaps. This observation was attributed to much faster oxygen depletion on rough crumbly surfaces of biological samples due to effective oxidation reactions occurred. The singlet oxygen generation and degradation times, and the relative quantum yield were determined on different surfaces by recording time-resolved phosphorescence at about 1270 nm under normoxic and hyperoxic conditions and analyzed on the basis of the set of master equations. The results obtained provide reference marks for choosing optimal irradiation durations for photodynamic inactivation of pathogenic infectious agents (bacteria, mycobacteria, fungi, viruses) on mucous membranes, including the tracheobronchial tree., (Copyright © 2022 Elsevier B.V. All rights reserved.)

The investigation of in-vitro response of cell cultures derived from tumor material of individual patients with similar tumor localizations to photodynamic treatment is presented. Tumor types included in the research were renal cell carcinoma, melanoma and alveolar, synovial, lypo- and osteo- sarcomas. Long-term observations of treatment-induced morphological changes in cells were performed by means of digital holographic microscopy. A substantial variance in response of cells of individual patients with similar tumor types and localizations to photodynamic treatment with the same dose has been observed. These peculiarities are indicative of the demand to personalized protocols of photodynamic treatment. The elevated resistance of cells of some patients to treatment at high doses highlights potential limitations of photodynamic therapy for some patients. Digital holographic microscopy is shown to be an informative label-free noninvasive tool allowing for long-term monitoring of cell samples in vitro and providing quantitative information on necrosis rate and loss of cellular dry mass. The developed methodology can be generalized for analysis of cellular response to various therapies., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Microtubules (MTs) often form a polarized array with minus ends anchored at the centrosome and plus ends extended toward the cell margins. Plus ends display behavior known as dynamic instability-transitions between rapid shortening and slow growth. It is known that dynamic instability is regulated locally to ensure entry of MTs into nascent areas of the cytoplasm, but details of this regulation remain largely unknown. Here, we test an alternative hypothesis for the local regulation of MT behavior. We used microsurgery to isolate a portion of peripheral cytoplasm from MTs growing from the centrosome, creating cytoplasmic areas locally depleted of MTs. We found that in sparsely populated areas MT plus ends persistently grew or paused but never shortened. In contrast, plus ends that entered regions of cytoplasm densely populated with MTs frequently transitioned to shortening. Persistent growth of MTs in sparsely populated areas could not be explained by a local increase in concentration of free tubulin subunits or elevation of Rac1 activity proposed to enhance MT growth at the cell leading edge during locomotion. These observations suggest the existence of a MT density-dependent mechanism regulating MT dynamics that determines dynamic instability of MTs in densely populated areas of the cytoplasm and persistent growth in sparsely populated areas.

Immunogenic and protective activity of recombinant pneumolysin was studied in experiments on male BALB/c mice. The mice were immunized intraperitoneally with recombinant pneumolysin sorbed on Al(OH) 3 (200 μg per mouse). In 2 weeks after immunization, the isotypes of antibodies to recombinant pneumolysin in the serum of immunized mice were determined by ELISA. The animals were infected with Streptococcus pneumoniae serotype 3. Immunization with recombinant pneumolysin induced the production of anti-pneumolysin antibodies, mainly of IgG1 subisotype. On day 21 after intraperitoneal infection with S. pneumoniae serotype 3 in a dose of 10 6 microbial cells, the survival rate of animals immunized with recombinant pneumolysin in a dose of 25 μg/mouse was 67% vs. 0% in the control (p<0.001). Recombinant pneumolysin could be considered as a promising protective antigen for inclusion in the serotype-independent vaccine against S. pneumoniae.

Digital holographic microscopy supplemented with the developed cell segmentation and machine learning and classification algorithms is implemented for quantitative description of the dynamics of cellular necrosis induced by photodynamic treatment in vitro. It is demonstrated that the developed algorithms operating with a set of optical, morphological, and physiological parameters of cells, obtained from their phase images, can be used for automatic distinction between live and necrotic cells. The developed classifier provides high accuracy of about 95.5% and allows for calculation of survival rates in the course of cell death.

Temporal dependence of changes in the morphological characteristics of cells of two cultured lines of cancer origin, HeLa and A549, induced by photodynamic treatment with Radachlorin photosensitizer, have been monitored using digital holographic microscopy during first two hours after short-term irradiation. The observed post-treatment early dynamics of the phase shift in the transmitted wavefront indicated several distinct scenarios of cell behavior depending upon the irradiation dose. In particular the phase shift increased at low doses, which can be associated with apoptosis, while at high doses it decreased, which can be associated with necrosis. As shown, the two cell types responded differently to similar irradiation doses. Although the sequence of death scenarios with the increase of the irradiation dose was the same, each scenario was realized at substantially different doses. These findings suggest that the average phase shift of the transmitted wavefront can be used for quantitative non-invasive cell death characterization. The conclusions made were cofirmed by commonly used test assays using confocal fluorescent microscopy., Competing Interests: The authors declare that there are no conflicts of interest related to this article., (© 2019 Optical Society of America under the terms of the OSA Open Access Publishing Agreement.)

The aim of this study was to investigate the metal content in the hair of children living near abandoned nonferrous metal mines located in the rural settlements Tubinsk, Ishmurzino and Semenovsk the South Urals (Russia), and in the soils obtained from the same area. The hair and soils samples were examined for the presence of the following metals: copper (Cu), zinc (Zn), iron (Fe), cobalt (Co), nickel (Ni), manganese (Mn), lead (Pb) and cadmium (Cd). The results of this study showed that soil from this territory contained an increased level of Cu, Fe, Zn, Mn, Pb, Cd, exceeding the maximum permissible concentration (MPC) 38.3, 16.8, 4.5, 1.5, 1.4, 1.5 - fold, respectively. These values are indicative of severe soil pollution in the studied locality, thus calling for further detailed investigations. Mineral analysis of the hair samples was performed using a combination of atomic emission spectrometry and inductively coupled plasma mass spectrometry. In the hair samples, positive correlations between the following pairs of the studied metals were observed (p < 0.05): Pb-Cd (r= 0.73), Mn-Cd (r= 0.79), Fe-Cd (r= 0.56), Fe-Co (r= 0.85), Mn-Co (r= 0.65), Ni-Co (r= 0.67), Mn-Fe (r= 0.78), Ni-Fe- (r= 0.45), Pb-Fe (r= 0.46); Pb-Mn (r= 0.58). The comparison between Zn and Pb showed a negative linear correlation for the hair samples (r= -0.50; p<0.05) and for the soil samples (r= -0.68; p<0.05) suggesting that the presence of Zn and Pb in the hair could correlate with the content of these two metals in the soil. Comparison of the data on hair from this study with the average reference data for Russia showed the presence of an excess of Fe, Mn and Zn in the hair samples of the majority of the studied population: 100, 72.5 and 54.5 percent, respectively. We also observed an excess of Ni, Cd and Pb in a small number of children (in 18.2, 13.6 and 9.1 percent of the studied population, respectively). A deficiency of Cu and Co, the essential microelements for human health, was found in 27.3 percent of the studied population. The results of this study could be used to analyze the internal toxic metal burden in contaminated areas of Bashkortostan and could also serve as reference for further studies., (Copyright © 2018 Elsevier GmbH. All rights reserved.)

The development of new express methods for the analysis of the efficacy of anti-cancer therapy on the cellular level is highly desirable for the analysis of chemotherapeutic agent performance. In this paper we suggest the use of parameters of cell morphology determined by holographic microscopy and tomography for the effective label free quantitative analysis of cell viability under antitumor chemotherapy and thus of cytostatic agent efficacy. As shown, measured phase shifts and cell morphology change dramatically as a result of chemotherapy and depend strongly on the cell type and agent applied. Experimentally, a comparative analysis of the antitumor efficacy of the two cytostatics, cisplatin and dioxadet, that are commonly used for chemotherapy of disseminated ovarian carcinoma has been performed. The experiments were carried out on the Wistar rat model. An essential difference in the morphology of cells, both normal (erythrocytes) and cancerous, present in ascitic fluid taken from the non-treated group of rats and the groups treated with either dioxadet or cisplatin, has been observed. The results obtained can be interpreted as an indication of the antitumor performance of both cytostatics at the cellular level and as a demonstration of the higher efficacy of therapy with dioxadet as compared to that with cisplatin. Differences in cell morphology are suggested to be applied as quantitative markers of cell viability and cytostatic agent efficacy. The conclusions made are supported by a comparison with the results of recent experiments based on survival rates of laboratory animals treated with these agents.., Competing Interests: The authors declare that there are no conflicts of interest related to this article.

Microtubule (MT)-based transport can be regulated through changes in organization of MT transport tracks, but the mechanisms that regulate these changes are poorly understood. In Xenopus melanophores, aggregation of pigment granules in the cell center involves their capture by the tips of MTs growing toward the cell periphery, and granule aggregation signals facilitate capture by increasing the number of growing MT tips. This increase could be explained by stimulation of MT nucleation either on the centrosome or on the aggregate of pigment granules that gradually forms in the cell center. We blocked movement of pigment granules to the cell center and compared the MT-nucleation activity of the centrosome in the same cells in two signaling states. We found that granule aggregation signals did not stimulate MT nucleation on the centrosome but did increase MT nucleation activity of pigment granules. Elevation of MT-nucleation activity correlated with the recruitment to pigment granules of a major component of MT-nucleation templates, γ-tubulin, and was suppressed by γ-tubulin inhibitors. We conclude that generation of new MT transport tracks by concentration of the leading pigment granules provides a positive feedback loop that enhances delivery of trailing granules to the cell center., (© 2017 Semenova et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).)

We report results of spectral- and time-resolved study of Radachlorin photosensitizer luminescence in water in the spectral range of 950-1350nm and for determination of the photosensitizer triplet state and the singlet oxygen lifetimes responsible for singlet oxygen generation and degradation. At any wavelength within the explored spectral range the luminescence decay contained two major contributions: a fast decay at the ns time scale and a slow evolution at the μs time scale. The fast decay was attributed to electric dipole fluorescence transitions in photosensitizer molecules and the slow evolution to intercombination phosphorescence transitions in singlet oxygen and photosensitizer molecules. Relatively high-amplitude ns peak observed at all wavelengths suggests that singlet oxygen monitoring with spectral isolation methods alone, without additional temporal resolution can be controversial. In the applied experimental conditions the total phosphorescence signal at any wavelength contained a contribution from the photosensitizer triplet state decay, while at 1274nm the singlet oxygen phosphorescence dominated. The results obtained can be used for optimization of the methods of singlet oxygen monitoring and imaging., (Copyright © 2017 Elsevier B.V. All rights reserved.)

The adipose tissue is considered today as an endocrine organ in which tissue-specific regulation of gene expression plays a key role in the processes of development of obesity and comorbidities, such as diabetes and cardiovascular disease. The present study is focused on ITLN1, PPARã and TNFá gene expression in intra-abdominal adipose tissue and its effect on the serum levels of omentin 1 and TNFa in individuals with different body mass. It has been shown that serum TNFa level is significantly higher in the subgroup of patients with overweight and obesity (BMI ≥ 25 kg/m2) as compared to individuals with normal body weight (BMI < 25 kg/m2)( p < 0.03). We have demonstrated that the expression level of the PPARã gene is positively correlated with the ITLN1 gene expression level in the intra-abdominal adipose tissue (r = 0.516, p = 0.020). Serum level of omentin 1 positively correlates with PPARã mRNA and protein levels in intra-abdominal adipose tissue (r = 0.550, p < 0.05 and r = 0.581, p < 0.03, respectively). For the subgroup of patients with overweight and obesity, we have shown negative correlation of the level of TNFá mRNA with PPARã and ITLN1 mRNA levels was shown (r = –0.549, p < 0.05 and r = –0.475, p < 0.05, respectively). This study is the first to show a correlation relationship between PPARã gene expression level in the intra-abdominal adipose tissue and the expression and secretion levels of omentin 1.

Introduction: The most common reason of chronic pancreatitis is liver and bile ducts disease: functional disorders, chronic cholecystitis, cholelithiasis and cholecystectomy in medical history. All these changes are associated with the colloidal structure of bile, increased lithogenicity, gallstones formation, Oddi's sphincter dysfunction, dysmotility and inflammation in the bile ducts., The Aim: to study the effectiveness of using medicine Liveria IC (metadoxine) in standard therapy as well as effect on spectrum of blood serum lipids and structural condition of liver (stiffness) and pancreas in patients with chronic biliary pancreatitis combined with obesity., Materials and Method: 115 patients suffering from chronic biliary pancreatitis and obesity were the subjects of the study. They were compared to etiological factor socioeconomic conditions and nutrition (regular food 5 times a day without aggressive food (fatty, spicy, sour, fried products)). Also the effect of the alcohol factor was excluded., Results: The obtained decrease in stiffness of the liver and pancreas indicates an improvement of their structural state., Conclusions: Using medication LiveriaIC (metadoxine) as the part of the complex therapy for the patients who are suffering from CBP combined with obesity gives some improvement of the lipid profile indices and the structural condition of liver and pancreas (according to the data of SWE) (р<0.05).

Aim: To determine whether there is a correlation of the composition of circulating lymphocyte subpopulations, the serum concentrations of interferon (IFN)-α, IFN-γ, and IFN-λ3, and the lymphocyte expression of types I and II IFN receptors with the species of a disease pathogen and the degree of liver fibrosis (LF) in patients with chronic hepatitis B (CHB) and in those with chronic hepatitis C (CHC)., Subjects and Methods: The investigation enrolled 44 patients with CHC, 9 patients with CHB, and 13 clinically healthy donors. The degree of LF in the patients was determined using transient elastography. The composition of peripheral blood lymphocyte subpopulations was examined; the concentrations of IFN-α, IFN-γ, and IL-28B were estimated., Results: Lymphocyte counts were higher in patients with CHC and in those with CHB than those in healthy donors; and the number of neutrophils was lower. There were no differences between the groups in the composition of lymphocyte subpopulations with the exception of the number of CD3 + CD4 + cells, which in patients with CHC was larger than in those with CHB. In CHC and CHB patients, the counts of CD118 + lymphocytes were higher than those in healthy donors. Patients with CHB and those with CHC did not differ between themselves and from healthy donors in the expression of CD119 on the lymphocytes. In CHC patients, the relative CD119 + cell counts were higher between CD4 + lymphocytes than those in healthy donors. The serum levels of IFN-α and IFN-γ in CHC and CHB patients were similar, but higher in healthy donors. The concentration of IL-28B genotype in patients with CHC was twice as high as in those with CHB, but the differences were statistically insignificant. The number of lymphocytes increased with the progression of fibrosis; that of neutrophils decreased. There was an inverse relationship between platelet counts and LF severity. Multiple comparisons of the clusters of patients with different degrees of LF revealed no differences in the number of major lymphocyte subpopulations. However, the number of CD3 + CD16 + CD56 + natural killer-like T (NKT) cells correlated with fibrosis severity. Patients with different degrees of LF showed no differences in the proportion of CD118 - and CD119 cells between lymphocytes and in the serum levels of IFN-α, IFN-γ, and IL-28B levels. Patients with grade IV LF displayed a higher proportion of CD4 + CD119 + lymphocytes between CD45 + cells than did those with grade III LF., Conclusion: Several new clinical and laboratory trends were identified and the nature and extent of previously described hematological and immunological changes were clarified in CHC or CHB patients with various degrees of LF. Some indicators may be used as additional criteria for the prognosis of the above forms of hepatitis, and a number of newly described facts suggest that it is necessary to revise the protective/phlogogenic value of types I, II, and III IFNs in chronic viral hepatitis C and B.

A methodology providing noninvasive monitoring and evaluation of the effect of photodynamic treatment on live cells in vitro is presented. Variations in morphological characteristics of cells in the course and after treatment are recorded by means of digital holographic microscopy. High-precision measurements of phase shift gained by probe radiation in HeLa and human endometrial mesenchymal stem cell cultures demonstrate for the first time changes of their volume occurred in response to treatment.

Bidirectional transport of membrane organelles along microtubules (MTs) is driven by plus-end directed kinesins and minus-end directed dynein bound to the same cargo. Activities of opposing MT motors produce bidirectional movement of membrane organelles and cytoplasmic particles along MT transport tracks. Directionality of MT-based transport might be controlled by a protein complex that determines which motor type is active at any given moment of time, or determined by the outcome of a tug-of-war between MT motors dragging cargo organelles in opposite directions. However, evidence in support of each mechanisms of regulation is based mostly on the results of theoretical analyses or indirect experimental data. Here, we test whether the direction of movement of membrane organelles in vivo can be controlled by the tug-of-war between opposing MT motors alone, by attaching a large number of kinesin-1 motors to organelles transported by dynein to minus-ends of MTs. We find that recruitment of kinesin significantly reduces the length and velocity of minus-end-directed dynein-dependent MT runs, leading to a reversal of the overall direction of dynein-driven organelles in vivo. Therefore, in the absence of external regulators tug-of-war between opposing MT motors alone is sufficient to determine the directionality of MT transport in vivo., (© 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Purine receptors are located on immune and somatic cells of animal and human organisms. Summation of signals from purine and TOLL-like receptors takes place on the level of inflammasome formation and results in summation of the first and second signals of innate immunity. The first signal--from PAMPs (pathogen associated molecular patterns), the second--from DAMPs (danger associated molecular patterns). Adenosine triphosphate (ATP) is the most studied DAMP. ATP connects with purine receptors which include P2 (P2X7 receptors are the best described), that results in opening of channels of these receptors and transit of ATP into the cell. In parallel exit of K⁺ from cells and entrance of Ca²⁺ and Na⁺ into the cells is observed, that is associated with activation of the immune competent cell. Damaged cells dying via necrosis or apoptosis are the source of extracellular ATP, as well as activated immunocytes. Signals from P2 and TOLL-like receptors are summarized in effectors of immune response, and activation of P2 receptors in lymphocytes makes a contribution into activation of cells, mediated by T-cell receptor. Negative side of purine receptor activation is a stimulating effect on proliferation and metastasis of malignant cells. The practical output of knowledge on functioning of purine receptors for clinical immunology is the application of agonists and antagonists of purine receptors, as well as explanation of effect of immune modulators from the position of launch of K⁺/Na⁺-pump; resulting in prolonged activation of immune competent cells.

Tissue specific expression of genes encoding cholesterol transporters ABCA1 and ABCG1 as well as genes encoding the most important transcriptional regulators of adipogenesis - LXRa, LXRb, PPARg and RORa has been investigated in intraabdominal adipose tissue (IAT) samples.A direct correlation between the content of ABCA1 and ABCG1 proteins with RORa protein level (r=0.480, p<0.05; r=0.435, p<0.05, respectively) suggests the role of the transcription factor RORa in the regulation of IAT ABCA1 and ABCG1 protein levels. ABCA1 and ABCG1 gene expression positively correlated with obesity indicators such as body mass index (BMI) (r=0.522, p=0.004; r=0.594, p=0.001, respectively) and waist circumference (r=0.403, p=0.033; r=0.474, p=0.013, respectively). The development of obesity is associated with decreased IAT levels of RORa and LXRb mRNA (p=0.016 and p=0.002, respectively). These data suggest that the nuclear factor RORa can play a significant role in the regulation of cholesterol metabolism and control IAT expression of ABCA1 and ABCG1, while the level of IAT LXRb gene expression may be an important factor associated with the development of obesity.

This paper explores the concept of image-plane holographic tomography applied to the measurements of laser-induced thermal gradients in an aqueous solution of a photosensitizer with respect to the reconstruction accuracy of three-dimensional variations of the refractive index. It uses the least-squares estimation algorithm to reconstruct refractive index variations in each holographic projection. Along with the bitelecentric optical system, transferring focused projection to the sensor plane, it facilitates the elimination of diffraction artifacts and noise suppression. This work estimates the influence of typical random and systematic errors in experiments and concludes that random errors such as accidental measurement errors or noise presence can be significantly suppressed by increasing the number of recorded digital holograms. On the contrary, even comparatively small systematic errors such as a displacement of the rotation axis projection in the course of a reconstruction procedure can significantly distort the results.

Dumps of quarries for the extraction of copper pyrite in Urals region of the Bashkortostan Republic are the problem object, which represents a main technogenic source of environmental pollution. Soils of Sibaisky quarry dumps are characterized by the elevated content of heavy metals relatively to maximum permissible concentrations and regional geochemical background. In sanitary hygienic evaluation of the pollution by studied technosols according to the total pollution index (Zc) it was revealed that according to the content of total forms studied technosols are referred to moderately hazard, while in regards of the same index concerning the content of the mobile forms technosols are referred to the category of hazardous. In this regard, in plant products grown in the area adjacent to the dumps, there was revealed an excess on cadmium. The biological activity of soils is very low, due to the low microbial community development. Soil formation in the dumps is almost not expressed, appearing in the formation of low-thick underdeveloped humus horizon with a little diversity of species and projective cover. Sanitary-hygienic situation in conditions in the quarry dump area is unfavorable, that does not allow newly formed soils to carry out environmental functions effectively

Aim: To determine the stages of development of cardiovascular diseases and the evolution of their pattern in the veterans of the Great Patriotic War during 70 postwar years., Subjects and Methods: The investigation used the 1946-2014 annual reports on the hospital's work; analysis of the pattern of therapeutic and cardiovascular diseases over time; the results of laboratory and functional tests in 148 patients who developed in-hospital myocardial infarction (cardiospecific enzymes, electrocardiography, echocardiography (EchoCG), and, if clinically indicated, coronary angiography) and in 112 cardiopulmonary patients who were detected to have a chronic multimorbid heart phenomenon (EchoCG, radiological examination, blood gas tests, and statistical processing of the findings)., Results: The postwar evolution of therapeutic and cardiovascular diseases was ascertained to consist of 5 stages, each of which averaged about 15 years. At the same time, the nosological entities typical of the subsequent stage developed in the pattern of those at the preceding stage and gradually became predominant. The form of in-hospital myocardial infarction, as well as the chronic multimorbid heart phenomenon in the presence of comorbidity (chronic obstructive pulmonary disease and hypertensive disease) at Stage IV was described., Conclusion: With allowance made for the stages of development of therapeutic diseases and evolution of its pattern in the veterans of the Great Patriotic War, the clinical departments of the hospital, its laboratory and functional diagnosis departments have been reorganized for 70 postwar years in order to render adequate specialized medical care to the patients.

Aim: Study protective activity of protein-containing antigens of pneumococcus, obtained from serotypes 6B, 10A, 14, 19F, 23F and 36R, against infection with heterologous strains of S. pneumoniae., Materials and Methods: S. pneumoniae strains of serotypes 3, 6B, 10A, 14, 19F, 23F and 36R, obtained from the collection of pneumococcus strains of Mechnikov RIVS, were used in the study. Protein-containing antigens of S. pneumoniae were isolated by acetone precipitations of supernatant fraction of culture medium. Protective activity of preparations of protein-containing antigens of pneumococcus as studied in experiments of active protection of BALb/c line mice., Results: The data obtained give evidence, that protein-containing antigens of pneumococcus, isolated from serotypes 6B, 10A, 14, 19F and 23F, effectively protect animals from subsequent infection with a heterologous S. pneumoniae strain of serotype 3 No. 11/56. Protection was noted at a level from 80 to 100% (p ≤ 0.05). Similar protective effect was detected in another experiment in a group of mice, immunized with preparations of protein-containing antigens of pneumococcus, obtained from serotypes 6B and 36R, against infection with a heterologous S. pneumoniae strain of serotype 3 No. 11/56. Protection was noted at a level of 90% (p ≤ 0.05)., Conclusion: The results of the experiments carried out allow to assume, that the main role in formation of cross-protection in experiments in animals is played by pneumococcus, proteins, that are a part of the studied preparations, and not polysaccharide antigens.

Aim: Study epitopic specificity of synthetic disaccharide, recurring link of serotype 3 S. pneumoniae, conjugated with bovine serum albumin (BSA)., Materials and Methods: Conjugate of the synthetic disaccharide with BSA was obtained by squarate method. Antigenic activity of the conjugate was studied in competitive EIA. Titers of IgG against capsule polysaccharide of serotype 3 S. pneumoniae were determined in EIA by using sera of mice immunized twice with disaccharide conjugate sorbed onto aluminum hydroxide., Results: Disaccharide conjugate used as a well-covering antigen (4 µg/well) in EIA was characterized by a high degree of specificity and interacted only with IgG against serotype 3 S. pneumoniae in antimicrobial sera of animals without reacting with antibodies (ABs) against other pneumococcus serotypes (6B, 10A, 19A, 19F, 23F). Disaccharide conjugated with BSA was determined in competitive EIA to inhibit bonding of ABs to disaccharide by 78.8%, bacterial capsule polysaccharide by 56.9%, BSA did not inhibit the sera activity. The study of sera of mice immunized by serotype 3 S. pneumoniae disaccharide conjugate in EIA, where capsule polysaccharide was used as a plate-sorbed antigen, has established the presence of IgG against capsule polysaccharide at a titer of 1:1600., Conclusion: The disaccharide that is a single recurring link of serotype 3 S. pneumoniae contains a key epitope of capsule polysaccharide. The synthetic disaccharide could be used as a component of multivalent conjugated pneumococcal vaccines and for development of diagnostic test-systems.

Twenty-four hour blood pressure monitoring in groups of patients of elderly age with bronchial asthma and hypertension was conducted. The study showed an increased variability of blood pressure and insufficient reduction in night time that leads to development of cardiovascular complications.

The paper presents a novel algorithm based on digital holographic interferometry and being promising for evaluation of phase variations from highly noisy or modulated by speckle-structures digital holograms. The suggested algorithm simulates an interferogram in finite width fringes, by analogy with classical double exposure holographic interferometry. Thus obtained interferogram is then processed as a digital hologram. The advantages of the suggested approach are demonstrated in numerical experiments on calculations of differences in phase distributions of wave fronts modulated by speckle structure, as well as in a physical experiment on the analysis of laser-induced heating dynamics of an aqueous solution of a photosensitizer. It is shown that owing to the inherent capability of the approach to perform adjustable smoothing of compared wave fronts, the resulting difference undergoes noise filtering. This capability of adjustable smoothing may be used to minimize losses in spatial resolution. Since the method allows to vary an observation angle of compared wave fields, an opportunity to compensate misalignment of optical axes of these wave fronts arises. This feature can be required, for example, when using two different setups in comparative digital holography or for compensation of recording system displacements during a set of exposures in studies of dynamic processes.

The expediency of Vitrum Calcium 600 + D400 and Risendros using in the complex treatment of the patients with a chronic pancreatitis and osteodeficiency syndrome has been proved. As a result the status of the bone mineralization is improved, the mineral density of the bone tissue is increased.

Microtubule (MT)-based transport of organelles driven by the opposing MT motors kinesins and dynein is tightly regulated in cells, but the underlying molecular mechanisms remain largely unknown. Here we tested the regulation of MT transport by the ubiquitous protein MAP4 using Xenopus melanophores as an experimental system. In these cells, pigment granules (melanosomes) move along MTs to the cell center (aggregation) or to the periphery (dispersion) by means of cytoplasmic dynein and kinesin-2, respectively. We found that aggregation signals induced phosphorylation of threonine residues in the MT-binding domain of the Xenopus MAP4 (XMAP4), thus decreasing binding of this protein to MTs. Overexpression of XMAP4 inhibited pigment aggregation by shortening dynein-dependent MT runs of melanosomes, whereas removal of XMAP4 from MTs reduced the length of kinesin-2-dependent runs and suppressed pigment dispersion. We hypothesize that binding of XMAP4 to MTs negatively regulates dynein-dependent movement of melanosomes and positively regulates kinesin-2-based movement. Phosphorylation during pigment aggregation reduces binding of XMAP4 to MTs, thus increasing dynein-dependent and decreasing kinesin-2-dependent motility of melanosomes, which stimulates their accumulation in the cell center, whereas dephosphorylation of XMAP4 during dispersion has an opposite effect., (© 2014 Semenova et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).)

The content of S100b protein, HLDF24 peptide, and autoantibodies to these factors in blood serum was measured in healthy individuals (35-64-year-old men and women) with various levels of "normal" BP. Significant differences in the amount of these molecular factors were found in individuals with various categories of BP. We revealed age-related and gender differences in the content of molecular factors in the serum. Our results indicate that variations in the concentration of S100b, HLDF24, and autoantibodies to these factors in blood serum from adult people can serve as a reliable criterion for the risk of arterial hypertension.

SUMMARY The expediency of Risendros using in the complex treatment of the patients with a chronic pancreatitis and osteodeficiency syndrome has been proved. As a result the status of the bone mineralization is improved, the mineral density of the bone tissue is increased.

Aim: Study antigenic and immunogenic activity of a conjugate of synthetic hexasaccharide related to a S. pneumoniae serotype 14 capsule polysaccharide chain fragment with bovine serum albumin (BSA)., Materials and Methods: Synthetic glucoconjugate based on BSA protein carrier and hexasaccharide ligand reflecting capsule polysaccharide chain fragment was obtained by using squarate method. Natural polysaccharide-protein complex from S. pneumoniae serotype 14 strain was obtained from cultural fluid supernatant by acetone precipitation. IgG titer against hexasaccharide/capsule polysaccharide was determined in antimicrobial sera and sera of mice immunized with glucoconjugate by EIA method., Results: Immunogenic activity ofglucoconjugate based on BSA protein carriers and synthetic hexasaccharide reflecting S. pneumoniae serotype 14 capsule protein chain fragment was established. After 2 immunizations antibodies against hexasaccharide ligand and BSA were determined in blood sera of mice. Antibody titers against hexasaccharide exceeded the level in intact mice by 4.2 times. BSA in the conjugate did not have effect on production of antibodies against hexasaccharide., Conclusion: The developed experimental test-system based on synthetic glucoconjugate is useful for evaluation of level of antibodies against S. pneumoniae serotype 14 in infected and, probably, carriers of bacteria.

A conjugate of a synthetic hexasaccharide fragment of the Streptococcus pneumoniae type 14 capsular polysaccharide with bovine serum albumin (BSA) has been prepared. The antigenic activity and specificity of this conjugate are comparable with those of natural antigens of S. pneumoniae type 14. The data suggest that the resulting synthetic conjugate can be used as a coating antigen in an experimental test system (based on enzyme immunoassay) for evaluating the antigenic activity and specificity of synthetic oligosaccharide ligands and for testing specimens of natural capsular polysaccharides and immune sera.

Aim: Determine protein specter that Staphylococcus aureus synthesizes and secretes at early growth phase--the exponential phase., Materials and Methods: Proteins secreted by S. aureus strain 6 into cultivation medium at the end of exponential growth phase (4.5 hours) were studied. 11 proteins were identified by liquid chromatography--mass-spectrometry method., Results: Only in 3 of these proteins the presence of signal peptides was predicted, which indicates their extracellular localization; the rest of the proteins were localized predominantly in bacterial cytoplasm. 5 of 11 proteins function as enzymes of carbohydrate metabolism. Other extracellular proteins that could indicate its contamination with proteins from disrupted bacterial cells were not detected in S. aureus cultural liquid filtrate. It has been suggested that enzymes of carbohydrate metabolism can provide bacterial cells with energy necessary for passage from lag-phase into exponential growth phase. Superoxide dismutase enzyme probably provides the course of oxidation-reduction processes. Synthesis of other proteolytic enzymes and toxins is carried out at later stages of development of bacterial population. Immunization of mice with a mixture of 11 identified proteins showed their protective properties after infection by S. aureus 6 strain., Conclusion: Based on the above-mentioned, the complex of isolated proteins may be perspective in development of a new strategy of prophylaxis and therapy of staphylococcus infections.

Aim: Study protective activity of S. pneumoniae protein-containing antigen complex obtained from T3No.3 strain against infection by homologous pneumococcus strain., Materials and Methods: S. pneumoniae T3No.3 (serotype 3) strain obtained from collection of pneumococcus strains of Mechnikov Research Institute of Vaccines and Sera was used in the study. S. pneumoniae protein-containing antigen complex was isolated by precipitation by 2 volumes of acetone of supernatant fraction of cultural medium used for pneumococcus cultivation. Molecular mass of proteins contained in S. pneumoniae antigen complex was determined by SDS electrophoresis in polyacrylamide gel. Protective activity of S. pneumoniae protein-containing antigen complex was studied in BALB/c line mice active protection experiments. Activity of mice immune sera obtained against whole-cell pneumococcus culture (T3No.3 strain) was determined in vitro by solid phase indirect EIA., Results: The data obtained give evidence that the isolated protein-containing antigen complex from S. pneumoniae T3No.3 strain effectively protects mice from consequent infection by a homologous S. pneumoniae strain. S. pneumoniae protein-containing antigen complex sorbed on solid phase at 5 microg dose was established by using EIA to interact with homologous mice immune sera., Conclusion: The results of the carried out studies allow to move to studies of cross-activity of S. pneumoniae protein-containing antigen complex isolated from T3No.3 strain.

Clinical efficacy of sodium nucleospermate (SNS) has shown in the treatment of 26 female patients with chronic endometritis, unsuccessful attempts at in vitro fertilization, and persistent viral infection. Cells from endometrium were processed by liquid cytology and studied by immunocytochemistry techniques. It has been clearly established that, after SNS treatment, the frequency of CD20- and CD56-positive cells was significantly decreased (p < 0.05) and the frequency of TLR4 and TLR9 expressing cells in endometrium has been significantly improved (p < 0.05). In addition, the human papilloma virus (HPV) load was reduced as manifested by the real-time PCR test. The obtained results show good prospects for successful use SNS as a means of pathogenetic therapy in patients with this diagnosis.

We studied the interactions between sulfated polysaccharides, fucoidans from sea brown algae Laminaria japonica, Laminaria cichorioides, and Fucus evanescens, with human Toll-like receptors (TLR) expressed on membranes of cultured human embryonic kidney cells (HEK293-null, HEK293-TLR2/CD14, HEK293-hTLR4/CD14-MD2, and HEK293-hTLR5). Fucoidans interacted with TLR-2 and TLR-4, but not with TLR-5, and were nontoxic for the cell cultures. L. japonica fucoidan (1 mg/ml), L. cichorioides fucoidan (100 μg/ml and 1 mg/ml), and F. evanescens fucoidan (10 μg/ml-1 mg/ml) activated transcription nuclear factor NF-ϰB by binding specifically to TLR-2. L. japonica fucoidan (100 μg/ml and 1 mg/ml), L. cichorioides fucoidan (10 μg/ml-1 mg/ml), and F. evanescens fucoidan (1 μg/ml-1 mg/ml) activated NF-ϰB via binding to TLR-4. These results indicated that fucoidans could induce in vivo defense from pathogenic microorganisms of various classes.

Aim: Study effect of fucoidans from brown algae on proliferative and cytotoxic activity of mice splenocytes., Materials and Methods: Proliferative and cytotoxic activity of mice splenocytes in vitro and ex vivo were studied in lymphocyte blast transformation reaction and in cytotoxic MTT-test on K562 human erythroblast leucosis cell line. Microphotography and microscopy were performed by using Axiocam HS photosystem and computer program AxioVision 4 (Germany)., Results: Fucoidans from brown algae Fucus evanescens, Laminaria cichorioides and Laminaria japonica in vitro and ex vivo systems were established to increase proliferative activity of mice splenocytes which is evidenced by an increase of stimulation index. Results of in vitro and ex vivo cytotoxic activity studies demonstrate that fucoidans which are various by chemical structure stimulate activity of NK-cells and facilitate an increase of splenocyte cytotoxic potential level against NK sensitive K562 cell line., Conclusion: The data obtained from the study demonstrate an ability of fucoidans to stimulate splenocyte proliferation and NK-cell killer activity, and studies of relation between structure and functions of sulfated polysaccharides facilitate a more detailed understanding of aspects of their mechanism of action on innate immunity system, thus providing the basis for development of new immunobiologic preparations - modifiers/agonists of innate immunity.

The interaction of sulfated polysaccharides--fucoidans from brown seaweeds Laminaria japonica, Laminaria cichorioides and Fucus evanescens with Toll-like receptors (TLRs) expressed on membranes of embryonic human kidney epithelial cells (HEK293-null, HEK293-TLR2/CD14, HEK293-hTLR4/CD14-MD2 and HEK293-hTLR2/6) was investigated. In vitro fucoidans specifically interacted with TLR-2, TLR-4, and the heterodimer TLR-2/6 resultated in activation of transcription nuclear factor NF-kappaB. Analysis of composition the hydrolyzed fucoidan from F. evanescens was carried out by gas-liquid chromatography and chromatography-mass spectrometry. Results indicated the absence of 3-3-hydroxytetradecanoic acid (3-OHC14), the basic component of lipopolysaccharides in the preparation. Thus, the obtained results suggested that fucoidans from brown seaweeds possessing immunotropic activity are independent ligands for TLRs, and are able to induce genetically determined biochemical processes of protection organisms against pathogenic microorganisms.

The present clinical immunological study included 90 women of reproductive age presenting with mycoplasma infection of the lower urogenital tract. Low-frequency ultrasonic radiation was shown to successfully correct the disbalance in the system of congenital immunity that manifested itself as marked dysfunction of neutrophil granulocytes. The local application of low-frequency ultrasonic radiation normalized concentration of neutrophil defensins in cervical secretion as well as the number and function of neutrophils themselves.