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Abstract Cellular senescence is considered an important tumour suppression mechanism in response to damage and oncogenic stress in early lesions. However, when senescent cells are not immune-cleared and persist in the tumour microenvironment, they can drive a variety of tumour-promoting activities, including cancer initiation, progression, and metastasis. Additionally, there is compelling evidence demonstrating a direct connection between chemo(radio)therapy-induced senescence and the development of drug resistance and cancer recurrence. Therefore, detection of senescent cells in tissues holds great promise for predicting cancer occurrence earlier, assessing tumour progression, aiding patient stratification and prognosis, and informing about the efficacy of potential senotherapies. However, effective detection of senescent cells is limited by lack of biomarkers and readout strategies suitable for in vivo clinical imaging. To this end, a nanoprobe composed of biocompatible polydopamine (PDA) nanoparticle doped with FDA-approved indocyanine green (ICG) dye, namely PDA-ICG, was designed as a contrast agent for senescence detection using photoacoustic imaging (PAI). In an in vitro model of chemotherapy-induced senescence, PDA-ICG nanoprobe showed an elevated uptake in senescent cells relative to cancer cells. In addition to its improved photostability, 2.5-fold enhancement in photoacoustic signal relative to ICG was observed. Collectively, the results indicate that the PDA-ICG nanoprobe has the potential to be used as a contrast agent for senescence detection of chemotherapy-induced senescence using PAI.

Abstract The Friendship Paradox is a simple and powerful statement about node degrees in a graph. However, it only applies to undirected graphs with no edge weights, and the only node characteristic it concerns is degree. Since many social networks are more complex than that, it is useful to generalize this phenomenon, if possible, and a number of papers have proposed different generalizations. Here, we unify these generalizations in a common framework, retaining the focus on undirected graphs and allowing for weighted edges and for numeric node attributes other than degree to be considered, since this extension allows for a clean characterization and links to the original concepts most naturally. While the original Friendship Paradox and the Weighted Friendship Paradox hold for all graphs, considering non-degree attributes actually makes the extensions fail around 50% of the time, given random attribute assignment. We provide simple correlation-based rules to see whether an attribute-based version of the paradox holds. In addition to theory, our simulation and data results show how all the concepts can be applied to synthetic and real networks. Where applicable, we draw connections to prior work to make this an accessible and comprehensive paper that lets one understand the math behind the Friendship Paradox and its basic extensions.

The failure of heart valve bioprostheses, requiring repeated intervention to replace them, is a serious drawback that narrows the scope of such devices. The present study demonstrates the experimental investigation of a medical device designed to partially solve the problem of repeated interventions on heart valves, primarily with an emphasis on reducing the duration and trauma of such procedures due to the sutureless balloon implantation method of the «valvein- valve» method.Material and methods. The paper presents a series of in silico, in vitro and in vivo experiments to evaluate various aspects of the device under development. Numerical modeling of the final shape setting to the supporting frame of the heart valve prosthesis to select the most promising concept for prototyping was carried out in the Abaqus/CAE (Dassault Systèmes, France) based on the finite element method. The selected optimal support frame model was prototyped in the form of a series of prosthesis samples of four standard sizes for hydrodynamic studies of quantitative characteristics in vitro. The study was carried out in the Vivitro Labs unit (Vivitro Labs, Canada), imitating the physiological mode of the heart, the prostheses were examined for the mitral position. The developed prototypes of the device were supplemented with related products, an implant holder and a balloon catheter, after which the proposed method of sutureless fixation was validated in an in vitro implantation procedure on a bovine heart model and a series of in vivo (n = 3) chronic experiments on animals.Results. In the course of numerical simulation, it was shown that in Model No. 3 of the supporting frame of the prosthesis, the smallest stresses occur – with an amplitude of up to 490 MPa. For other concepts (Models No. 1 and No. 2), this indicator was significantly higher, 543 and 514 MPa, respectively. Prototypes obtained on the basis of the selected shape of the support frame demonstrated satisfactory hydrodynamic characteristics: effective hole area 190-261 mm2, regurgitation volume 6–9 ml/cycle, average transprosthetic gradient 4.4–6.4 mmHg, depending on size. The study of the technology of sutureless balloon implantation in an in vitro model of the heart and the subsequent chronic experiment on sheep confirmed the main idea of the system – the possibility of a significant reduction in the duration of repeated prosthetics. It has been shown that the time of suture implantation of the “classic” frame prosthesis of the mitral valve is 23–29 minutes, with a total access time of 41–52 minutes. When implanting the experimental device, the sutureless prosthesis itself took 4–6 minutes, access time 24–29 minutes. At the same time, one-month results of an echocardiographic study of the operation of the prosthesis demonstrated satisfactory hemodynamics.Conclusions. This work demonstrates a consistent series of tests of the system being developed for repeated heart valve replacement, which substantiates some design solutions, confirms the effectiveness and viability of the chosen approach to sutureless minimally invasive implantation.

Abstract The Friendship Paradox—the principle that “your friends have more friends than you do”—is a combinatorial fact about degrees in a graph; but given that many web-based social activities are correlated with a user’s degree, this fact has been taken more broadly to suggest the empirical principle that “your friends are also more active than you are.” This Generalized Friendship Paradox, the notion that any attribute positively correlated with degree obeys the Friendship Paradox, has been established mathematically in a network-level version that essentially aggregates uniformly over all the edges of a network. Here we show, however, that the natural node-based version of the Generalized Friendship Paradox—which aggregates over nodes, not edges—may fail, even for degree-attribute correlations approaching 1. Whether this version holds depends not only on degree-attribute correlations, but also on the underlying network structure and thus can’t be said to be a universal phenomenon. We establish both positive and negative results for this node-based version of the Generalized Friendship Paradox and consider its implications for social-network data.

Abstract Tumor-derived extracellular vesicles (EVs) are active contributors in metastasis and immunosuppression in tumor microenvironment. At least some of the EVs carry tumor surface molecules such as tumor-associated antigens (TAAs) and/or checkpoint inhibitors, and potentially could interact with T cells or CAR T cells. Upon contact with T cells, EVs could alter their phenotype and functions by triggering signaling through TCR or CAR reprogramming them to escape immune response. We hypothesize that EVs that possess TAA on the surface will probably interact with CAR T cells which can recognize and bind corresponding TAA. This interaction between EVs and CAR T cells may change the outcome of CAR T-based cancer immunotherapy since it should affect CAR T cells. Also, EVs could serve as adjuvants and antigenic components of antitumor vaccines. Herein, we isolated EVs from B cell precursor leukemia cell line (pre-B ALL) Nalm-6 and demonstrated that recognition and binding of CD19+EVs with CD19-CAR T cells strongly depends on the presence of CD19 antigen. CD19+EVs induce secretion of pro-inflammatory cytokines (IL-2 and IFN-y) and upregulated transcription of activation-related genes (IFNG, IFNGR1, FASLG, IL2) in CD19-CAR T cells. Tumor necrosis factor receptor superfamily (TNFRSF4 and TNFRSF9) and T-cell exhaustion markers (CTLA4, LAG3, TIM3 and PDCD1LG2) were also upregulated in CD19-CAR T cells after incubation with CD19+EVs. Long-term cultivation of CD19+ or PD-L1+EVs with CD19-CAR T cells led to increased terminal differentiation and functional exhaustion according to elevated expression of PD-1, TIGIT, CD57. In summary, our results suggest that chronic exposure of CD19-CAR T cells to CD19+EVs mediates activation and systemic exhaustion in antigen-specific manner, and this negative effect is accompanied by the impaired cytotoxic activity in vitro.

The results of morphology and topography of the greater palatine foramen and the greater palatine canal, and their practical application in clinical practice are presented. A systematic literature search was performed using PubMed, eLibrary, and HAC databases. Effective anesthesia in dentistry can be achieved using local anesthesia methods. The success of anesthesia in the posterolateral parts of the upper jaw depends on an accurate knowledge of the anatomy of this area. The review of the anatomy and morphology of the greater palatine foramen and the greater palatine canal as the target point for conducting analgesia, the anatomical structure of the neurovascular bundle passing in the greater palatine canal, the thickness of the mucous membrane of the palate in its various sections. Techniques of regional anesthesia through the greater palatine canal are considered. It is noted that when carrying out this type of anesthesia, it is necessary to take into account the length, shape of the greater palatine canal, the angle of inclination of the needle to the injection point, (taking into account individual characteristics and under the control of conebeam computed tomography). As a result of the analysis, it was found that the choice of a certain type of surgical intervention depends on the clinical situation. However, to date, neither in domestic nor in foreign literature there is no single point of view about the anatomy and morphology of the greater palatine foramen and the greater palatine canal, which emphasizes the extreme urgency of further research of these anatomical formations.

Chlamydia abortus is the etiological agent of abortion and fetal loss in sheep, goats and bovine cattle in many countries. Even though commercially available vaccines can reduce the incidence in sheep, the development of new, safe, and effective vaccines remains high on the agenda. In this study, an evaluation was made of the efficacy of a vaccine candidate, an inactivated antigen based on the extract of outer membrane proteins of a C. abortus strain known as Chlamydia VNITIBP-21, in combination with recombinant flagellin as an adjuvant. Pregnant sheep (n = 43) were divided into three groups: an experimental vaccinated group, a control infected group and a control non-infected group. The sheep were vaccinated twice, with an interval of 3 weeks, then infected with the homologous virulent strain of Chlamydia abortus on pregnancy day 75. The vaccine candidate reduced C. abortus shedding in vaginal swabs considerably, in comparison with the control group. In addition, ewes in the experimental group experienced no abortions, while those in the control group experienced instances of abortion, as well as births of weak and nonviable lambs. The findings show that the vaccine candidate proved itself to be promising in combatting the agent of ovine abortion and fetal loss.

Diseases caused by Streptococcus pneumoniae, as well as antibiotic resistance of its serotypes, are the leading cause of death amongst children worldwide. To prevent pneumococcal infection, the population is immunised with conjugate vaccines containing different amounts of polysaccharides of certain serotypes. Development of a full-cycle Russian vaccine is vital because the active pharmaceutical ingredients for the vaccines registered in the Russian Federation are produced abroad, and only the final stages of production of vaccines of this group are performed in the territory of the Russian Federation. Considering the phenomenon of serotype replacement associated with the long-term widespread use of pneumococcal conjugate vaccines, it is necessary to carefully select the serotype composition for the new vaccine. The aim of this work was to analyse the serotype distribution of pneumococci in the Russian Federation and other countries in order to select optimal serotypes for the Russian vaccine for human use, taking into account vaccination schedules for each age group. This review presents an analysis of the pneumococcal serotype distribution in the Russian Federation in the pre-vaccination era, as well as after the introduction of routine vaccination. In addition, the review includes data on the serotype distribution in the Eurasian Economic Union countries. The authors described a model composition containing at least sixteen serotypes. It will increase effectiveness of immune protection of the population, providing a more complete coverage of serotypes, considering their prevalence in the Russian Federation. Based on the analysis, the serotype composition for the sixteen-valent pneumococcal conjugate vaccine is proposed for further production and preclinical and clinical trials. A new Russian pneumococcal conjugate vaccine will ensure vaccination of all population groups within the National Immunisation Schedule of the Russian Federation.

Vaccines are the cornerstone of infectious disease control and prevention. The outbreak of SARS-CoV-2 has confirmed the urgent need for a new approach to the design of novel vaccines. Plant viruses and their derivatives are being used increasingly for the development of new medical and biotechnological applications, and this is reflected in a number of preclinical and clinical studies. Plant viruses have a unique combination of features (biosafety, low reactogenicity, inexpensiveness and ease of production, etc.), which determine their potential. This review presents the latest data on the use of plant viruses with different types of symmetry as vaccine components and adjuvants in cancer immunotherapy. The discussion concludes that the most promising approaches might be those that use structurally modified plant viruses (spherical particles) obtained from the Tobacco mosaic virus. These particles combine high adsorption properties (as a carrier) with strong immunogenicity, as has been confirmed using various antigens in animal models. According to current research, it is evident that plant viruses have great potential for application in the development of vaccines and in cancer immunotherapy.

Background. The attention of many researchers is focused on studying the role of adipokines secreted by subcutaneous, visceral, epicardial, and perivascular adipose tissues in the pathogenesis of diseases of the cardiovascular system. At the same time, adipose tissue of retrosternal localization remains out of research focus. This pool of fat cells is formed at the site of the thymic involution and has a significant volume. However, their functional activity and participation in the development of cardiovascular pathology remain unexplored.Aim. To study the morphological characteristics of adipocytes of the retrosternal adipose tissue (RSAT) and their production of adipokines in comparison with epicardial (EAT) and subcutaneous adipose tissue (SCAT) and to investigate their relationships with arterial stiffness parameters in patients who underwent coronary artery bypass grafting.Materials and methods. The study included 17 patients (12 men/5 women aged 40–70 years) with the diagnosed coronary artery disease (CAD) who underwent coronary artery bypass grafting (CABG). Each patient underwent measurement of carotid-femoral pulse wave velocity (PWV) and aortic augmentation index (AIx) with the oscillometric device. Isolated adipocytes were obtained enzymatically from explants of SCAT, EAT and RSAT during coronary artery bypass grafting. The adipocytes were analyzed under the microscope at 200x magnification. The release of adiponectin, leptin and insulin was studied in the adipocyte supernatant after 1 hour incubation using ELISA.Results. It was found that adipocytes of the RSAT are smaller than adipocytes of SCAT: 83.96 ± 2.21 vs 98.62 ± 2.67 μm (p = 0.00002), respectively, and comparable in size to adipocytes of EAT: 86.65 ± 1.33 μm. The release of adiponectin by adipocytes of the RSAT turned out to be comparable to the production of this adipokine in SCAT and EAT, however, adipocytes of the RSAT produce less leptin than SCAT and EAT: 0.26 (0.19; 0.27) ng/l vs 0.37 (0.28; 0.55) (р = 0.01) and vs 0.32 (0.28; 0.44) (р = 0.006) ng/ml, respectively. Furthermore, RSAT produce less insulin than SCAT and EAT: 1.56 (1.03; 2.08) vs 1.70 (0.99; 2.18) ng/ml, (р = 0.0022) and 1.76 (1.16; 2.40) ng/ml (р = 0.006), respectively.A positive correlation was found between the secretion of leptin by adipocytes of the RSAT and the AIx (rs = 0.52, p = 0.046). An inverse relationship was found between insulin secretion by retrosternal adipocytes and PWV rs = –0.55, p = 0.035). There was no relationship between the size of the retrosternal adipocyte or hypertrophy of the thymic adipocytes (more than 100 μm) and the production of leptin and insulin and arterial stiffness parameters. Conclusions. The data of our pilot study show that adipocyte hypertrophy of the retrosternal AT is not a significant marker of adipokine production disturbance. The observed relationships suggest that an increase in leptin production and reduced insulin secretion by retrosternal AT may contribute to the formation of adipokine-related arterial stiffness. Based on the data obtained, it can be assumed that adipokines produced by the retrosternal AT can participate in the formation of arterial stiffness in patients with coronary artery disease.

Abstract Homophily—the tendency of nodes to connect to others of the same type—is a central issue in the study of networks. Here we take a local view of homophily, defining notions of first-order homophily of a node (its individual tendency to link to similar others) and second-order homophily of a node (the aggregate first-order homophily of its neighbors). Through this view, we find a surprising result for homophily values that applies with only minimal assumptions on the graph topology. It can be phrased most simply as “in a graph of red and blue nodes, red friends of red nodes are on average more homophilous than red friends of blue nodes”. This gap in averages defies simple intuitive explanations, applies to globally heterophilous and homophilous networks and is reminiscent of but structually distinct from the Friendship Paradox. The existence of this gap suggests intrinsic biases in homophily measurements between groups, and hence is relevant to empirical studies of homophily in networks.

The vast majority of SARS-CoV-2 vaccines which are licensed or under development focus on the spike (S) protein and its receptor binding domain (RBD). However, the S protein shows considerable sequence variations among variants of concern. The aim of this study was to develop and characterize a SARS-CoV-2 vaccine targeting the highly conserved nucleocapsid (N) protein. Recombinant N protein was expressed in Escherichia coli, purified to homogeneity by chromatography and characterized by SDS-PAGE, immunoblotting, mass spectrometry, dynamic light scattering and differential scanning calorimetry. The vaccine, formulated as a squalane-based emulsion, was used to immunize Balb/c mice and NOD SCID gamma (NSG) mice engrafted with human PBMCs, rabbits and marmoset monkeys. Safety and immunogenicity of the vaccine was assessed via ELISA, cytokine titer assays and CFSE dilution assays. The protective effect of the vaccine was studied in SARS-CoV-2-infected Syrian hamsters. Immunization induced sustainable N-specific IgG responses and an N-specific mixed Th1/Th2 cytokine response. In marmoset monkeys, an N-specific CD4+/CD8+ T cell response was observed. Vaccinated Syrian hamsters showed reduced lung histopathology, lower virus proliferation, lower lung weight relative to the body, and faster body weight recovery. Convacell® thus is shown to be effective and may augment the existing armamentarium of vaccines against COVID-19.

Reproductive disorders, presumably caused by Chlamydia abortus, are common among the ovine population of the Mari El Republic, Russia. C. abortus infection was determined by serologic testing or isolation and detection of the organism by PCR and direct immunofluorescence in tissue samples. Rams, ewes, and lambs (10 individuals each) were randomly chosen for serological testing by the complement fixation test and 7 of 30 (23%) animals tested were positive. Tissue samples were collected from ewes and aborted fetuses for isolation by inoculating chicken embryo yolk sacs (n = 41). The same samples were analyzed by PCR using commercial and in-house PCR kits and by direct immunofluorescence. C. abortus was detected in 58.5% of samples using PCR and in 60.9% of the samples by direct immunofluorescence. Five Chlamydia isolates were cultured in egg yolk sacs and adapted for growth in cell cultures. Phylogenetic analysis showed no substantial difference between Russian isolates and those from other parts of the world. The results of the study further demonstrate the usefulness of PCR for detection of C. abortus as a faster, simpler, and more reliable approach in comparison to culturing the organism and underscoring the necessity of screening for chlamydiosis as a cause of ovine abortion.

The urgency of the development and improvement of methods for studying the functional state of users of computer technology is growing in parallel with the growth of their negative impact on the main targets - visual and neuropsychic functions. Therefore, the aim of the study was to study the processes of perception of rhythmic light incentives and induced electrical activity of the visual cortex during a computer load. The study was conducted with the participation of clinically healthy students ( n = 182), who determined the critical fusion frequency of flashes and recorded rhythmic cortical evoked visual potentials during a computer-assisted visual load. To assess the nature and duration of use of computer students, as well as determining the presence and severity of computer dependency, a survey was conducted. Various types of perception of light flashes and changes in the rhythm assimilation reaction under the influence of a computer load are revealed. The data are discussed taking into account the rhythmic processes of the brain. A conclusion is drawn about the weakened, stable and strengthened types of response of oscillatory neural mechanisms to computer visual loads.

The article is devoted to the study of the implementation of the power holding strategies, as well as the stylistic devices of the American political discourse based on of D. Trump’s public statements. It gives the definition of strategy, tactics of political discourse. The author points out the most frequently used tactics of the strategies of persuasion during the pre-election race by D. Trump, as well as the lexical stylistic devices embodying these tactics. The author points to the variety of tactics used by the subject of research, psychologically directed towards the addressee's emotions, describes the key features of each of the tactics. As an example of the implementation of power holding strategies, the author considers the election campaign of the politician in Cleveland, Ohio. The author identifies many cases of application of the informational and interpretational strategy and the strategy of forming the emotional mood of the addressee, which are part of the strategies for holding power.

The present work addresses the thermal remodelling of flexible plant viruses with a helical structure and virus-like particles (VLPs). Here, for the first time, the possibility of filamentous Alternanthera mosaic virus (AltMV) virions' thermal transition into structurally modified spherical particles (SP) has been demonstrated. The work has established differences in formation conditions of SP from virions (SPV) and VLPs (SPVLP) that are in accordance with structural data (on AltMV virions and VLPs). SP originate from AltMV virions through an intermediate stage. However, the same intermediate stage was not detected during AltMV VLPs' structural remodelling. According to the biochemical analysis, AltMV SPV consist of protein and do not include RNA. The structural characterisation of AltMV SPV/VLP by circular dichroism, intrinsic fluorescence spectroscopy and thioflavin T fluorescence assay has been performed. AltMV SPV/VLP adsorption properties and the availability of chemically reactive surface amino acids have been analysed. The revealed characteristics of AltMV SPV/VLP indicate that they could be applied as protein platforms for target molecules presentation and for the design of functionally active complexes.

Previously conducted studies showed endocrine-disruptor properties of pyrethroids when exposed to mammals, causing disturbances in the reproductive system. The purpose of these studies was to identify the hazard of lambda cyhalothrin (LCT) reproductive toxicity, as well as to study the reversibility and/or irreversibility of the resulting damage during the recovery period. Research methods. Lambda-cyhalothrin 98.06% of purity was administered ex tempore daily, except Saturday and Sunday, by oral gavage to three groups of animals in doses 0,3; 3,0 and 10 mg/kg of body weight for 11 weeks. After the expiry of the exposure period, part of the males was selected to study the parameters of sperm and blood serum testosterone levels, while the remaining males were used for a recovery period without exposure for one full cycle of spermatogenesis (70 days). Morpho-functional indicators of the gonad state and the level of total testosterone in the blood serum were studied in all males after exposure and recovery period. The total amount and the absolute and relative number of motile sperm were determined, the percent of sperm abnormal forms was calculated. Morphometric parameters of testes and epididymis were recorded. Results. Investigated LCT causes antiandrogenic effect which characterized by impaired of spermatogenesis and oligospermia, as well as a change in the testosterone content in the blood serum of experimental animals compared to the control. Dose dependence of the severity of oligospermia and spermatozoa adynamia is linear in nature both before and after the recovery period, increasing markedly at the end of the recovery period. While the response level of testosterone to increase of the dose is non-monotonic.The most pronounced statistically significant (P

The purpose of the study is an analysis of the consumption of antibacterial drugs in the therapy of salpingoophoritis in hospital settings and evaluation of changes in the real clinical practice of antibiotic therapy after the introduction of the “Order of medical care for the profile “obstetrics and gynecology...”.Materials and methods. In a retrospective study, an assessment was made of the prescriptions of antibacterial drugs in the therapy of salpingoophritis in specialized treatment and prophylactic institutions of the city of Tomsk in the period 2010–2014. The analysis included 160 case histories. The structure of prescriptions was evaluated on the basis of the application of anatomical-therapeutic-chemical classification and international non-proprietary names (INN), the structural improvement factor K. Gatev was calculated. To quantify the consumption of medications, the calculated daily dose (PDD) was used.Results. Comparative analysis of antibacterial therapy of salpingoophoritis in hospital before and after the introduction of the “Order of medical care for the profile of «obstetrics and gynecology...”. Leading groups of antibacterial drugs were identified. А comparative analysis of the frequency of assignment of various INN was made. The К. Gate’s coefficient was 0.281. The calculation of the average prescribed daily doses of antibacterial drugs was performed.Conclusion. The review of the consumption of antibacterial drugs for salpingoophoritis therapy in hospital settings before and after the introduction of the “Order of medical care for the profile “obstetrics and gynecology...”, showed that the range of antibacterial drugs used increased from 18 to 24 INN, mainly due to the inclusion of modern medicines. In the structure of appointments in 2013–2014, the proportion of drugs not recommended for use by the «Order of medical care for the profile of «obstetrics and gynecology» was 37.5%. It was found that PDD clarithromycin, clindamycin and cefotaxime did not meet the recommended dosing regimens specified in the clinical recommendations for antibiotic therapy. The gap between actual clinical practice and existing clinical recommendations determines the need for in-depth pharmacoepidemiological evaluation to identify the causes and develop measures of an organizational and educational nature to optimize the use of medications.

Objective. To analyze the relationship between allelic variants of angiogenesis factor genes and development of genital endometriosis (GE), its clinical evidence and treatment efficacy.Material and methods. 417 women with laparoscopically verified GE (main group) and 112 women without GE (control group) were examined. Among 358 women in both groups (251 with GE and 107 without GEregions of polymorphic angiogenesis factor genes G-1154A VEGF, 405C VEGF, T-604C KDR and G-735A Ang2 were identified by RFLP-analysis.Results. It has been established that carriage of separate polymorphic angiogenesis factor genes VEGF G-1154A (allele А and genotypes GA, АА) and G-405C (allele С and genotype GC), KDR T-604C (allele С and genotypes ТС and СС), Ang2 G-735A (allele А and genotypes СА and GА) and their combinations predispose to GE development in women of reproductive age. Carriage of genotype GA of polymorphism G-1154A of VEGF gene predisposes to the development of dysmenorrhea during GE. Pelvic pains and infertility during GE as well as efficacy of hormone treatment for these problems are not associated with carriage of separate polymorphisms of angiogenesis factor genes G-405C and G-1154A of VEGF gene, T-604C of KDR gene, G-735A of Ang2 gene. The most potent combination of polymorphisms of angiogenesis factor genes which predisposes to the development of GE is VEGF1154GА/KDRTС/Ang2АA, while VEGF1154АА/ KDRСС/Ang2GG combination predisposes to the development to endometriosis-associated infertility, and VEGF1154GG/KDRТC/Ang2AA combination predisposes to efficient treatment of infertility during GE.Conclusion. Polymorphic variants of angiogenesis factor genes G-1154A and 405C VEGF, T-604C KDR, G-735A Ang2 (separately or in their combinations) predispose to GE development. They are associated with its clinical evidence and treatment efficacy. For the formation of risk groups of GE development for primary prevention of GE and effective treatment of endometriosis-associated infertility it is advisable to identify combinations VEGF1154GА/KDRTС/Ang2АA, VEGF1154АА/KDRСС/Ang2GG, and VEGF1154GG/KDRТC/ Ang2AA of angiogenesis factors.

Exosomes are a type of extracellular vesicles (EVs) secreted by multiple mammalian cell types and involved in intercellular communication. Numerous studies have explored the diagnostic and therapeutic potential of exosomes. The key challenge is the lack of efficient and standard techniques for isolation and downstream analysis of nanovesicles. Conventional isolation methods, such as ultracentrifugation, precipitation, filtration, chromatography, and immune-affinity-based approaches, rely on specific physical properties or on surface biomarkers. However, any of the existing methods has its limitations. Various parameters, such as efficacy, specificity, labor input, cost and scalability, and standardization options, must be considered for the correct choice of appropriate approach. The isolation of exosomes from biological fluids is especially challenged by the complex nature and variability of these liquids. Here, we present a comparison of five protocols for exosome isolation from human plasma: two chemical affinity precipitation methods (lectin-based purification and SubX™ technology), immunoaffinity precipitation, and reference ultracentrifugation-based exosome isolation method in two modifications. An approach for the isolation of exosomes based on the phenomenon of binding and aggregation of these particles via clusters of outer membrane phosphate groups in the presence of SubX™ molecules has been put forward in the present study. The isolated EVs were characterized based upon size, quantity, and protein content.

Short term storage of extracellular vesicle (EV) solutions at +4°C is a common practice, but the stability of EVs during this procedure has not been fully understood yet. Using nanoparticle tracking analysis, we have shown that EVs isolated from the conditioned medium of HT-29 cells exhibit a pronounced concentration decrease when stored in PBS in ordinary polypropylene tubes within the range of (0.5-2.1) × 1010 particles/ml. EV losses reach 51±3% for 0.5 ml of EVs in Eppendorf 2 ml tube at 48 hours of storage at +4°C. Around 2/3 of the observed losses have been attributed to the adsorption of vesicles onto tube walls. This result shows that the lower part (up to at least 2 × 1010 particles/ml) of the practically relevant concentration range for purified EVs is prone to adsorption losses at +4°C. Total particle losses could be reduced to 18-21% at 48 hours by using either Eppendorf Protein LoBind tubes or ordinary tubes with the surface blocked with bovine serum albumin or EVs. Reduction of losses to 15% has been shown for isolated EVs dissolved in the supernatant after 100 000 g centrifugation as a model of conditioned medium. Also, a previously unknown feature of diffusion-controlled adsorption was revealed for EVs. In addition to the decrease in particle count, this process causes the predominant losses of smaller particles.

Background: Extracellular vesicles are small membrane particles which are produced by the cells and released into the surrounding space. They can transfer biomolecules into recipient cells. The typical vesicle size is 100-1000 nm, so transmission electron microscopy (TEM) is the most suitable tool for their visualization. This is especially true for the smallest vesicles – the exosomes with the size below 200 nm. Methods: We isolated exosomes produced by MCF-7 cancer cells and the two drug-resistant MCF-7 sublines (Semina et.al., Molecules, 2018). The exosomes were isolated from the conditioned medium by ultracentrifugation, and their size was measured by nanotracking analysis (NTA) and TEM. Immunogold labelling was carried out on exosomes deposited onto carbon TEM grids. It involved the following principal steps: the vesicle deposition, sample treatment by the anti-CD9 antibodies, sample treatment by the 10 nm colloidal gold nanoparticles conjugated with protein A, staining with uranyl acetate. Washing and blocking steps were done between the principal ones. The samples were imaged using a JEM-1011 microscope at 80 kV. Results: NTA and TEM yielded similar results – the exosomes produced by the three studied cell lines had the almost equal size in the range from 50 to 200 nm. When imaged by TEM, most exosomes demonstrated the typical cup-shaped morphology. We used immunogold labelling to verify that the exosomes carried the CD9 marker on them (Fig. 1). We achieved the labelling specificity in the range of 8-10; this value was calculated as the ratio of the surface densities of the exosome-bound labels (colloidal gold nanoparticles) and the non-specific background ones. Conclusion: We hope that the sample preparation and imaging procedures that we used could be useful for the investigation of other extracellular vesicles.

Previously, we have reported that spherical particles (SPs) are formed by the thermal remodeling of rigid helical virions of native tobacco mosaic virus (TMV) at 94°C. SPs have remarkable features: stability, unique adsorption properties and immunostimulation potential. Here we performed a comparative study of the amino acid composition of the SPs and virions surface to characterize their properties and take an important step to understanding the structure of SPs. The results of tritium planigraphy showed that thermal transformation of TMV leads to a significant increase in tritium label incorporation into the following sites of SPs protein: 41-71 а.a. and 93-122 a.a. At the same time, there was a decrease in tritium label incorporation into the N- and C- terminal region (1-15 a.a., 142-158 a.a). The use of complementary physico-chemical methods allowed us to carry out a detailed structural analysis of the surface and to determine the most likely surface areas of SPs. The obtained data make it possible to consider viral protein thermal rearrangements, and to open new opportunities for biologically active complex design using information about SPs surface amino acid composition and methods of non-specific adsorption and bioconjugation.

92 patients with genital endometriosis and diagnosed infertility were clinically examined and went through molecular-genetical testing and therapy. Complex therapy included laparoscopic surgery and the course of hormonal therapy (gonadotrophin releasing hormone agonists, combined oral contraceptives). The effect of the therapy increases as increases the possibility of pregnancy in the presence of IL2 gene T-330G TT polymorphism variant and IL4 gene C-590T CC polymorphism variant in the genotype of a woman. The decreased effect of infertility treatment with the diagnosed endometriosis is associated with the presence of the hazardous genotype combination: IL2 gene T-330G GG polymorphism and IL4 gene C-590T TT polymorphism.

In this article clinical indexes of the dynamics of mineral bone density according to both densitometry and marker of resorption as well as bone formation and steroid blood hormons in patients in postovarioectomy are analysed. These patients state was complicated by cancer of the corpus and colli uterus after etiopathogenetic treatment including total husterectomy, radiation therapy and polychemotherapy. According the results of the comparative analysis in patients with postvarioectomic syndrome developed after surgical treatment for benign internal genitalia diseases we may make the conclusion on pathogenetic role of postovarioectomic syndrome in the development of bone disturbances in female patients which underwent total hysterectomy and special therapy.

In article the disturbance problem of blood flow the vaginas of the woman in a state of a medicamental and surgical menopause after spent этиопатогенетического the treatment including total husterectomy, radiation therapy and polychemotherapy by results of an expression Cd34 and bloods steroid sexual hormonesis, in particular aestradiolum. Comparison with bunch of women in a state of the postovariectomy which have educed after surgical treatment concerning good-quality diseases of internial genital. The pathogenetic role of surgical and radial influences on a vagina of the woman is proved.

In an effort to diagnose inflammatory diseases of small pelvis organs among women there was used the scintigraphy with 199Tl-chloride that is nonspecific marker of inflammation. There were studied 97 patients and 8 volunteers of female sex. There was found statistically important interconnections of inflammation clinic-laboratory criteria with scintigraphy results. Scintigraphy with 199Tl-chloride showed to be the effective method of diagnostic and activity of inflammatory diseases of small pelvis organs among women. Sensibility, specificity, exactness of diagnostics of presence of inflammation comprised 92,0; 88,9; 91,7, and its activity – 87,8; 82,3; 85,0%. The most diagnostic effectiveness of method was determined when studying patients with acute inflammatory processes, the least – among patients with exacerbation of chronic processes or different kinds of obesity.Scintigraphy with 199Tl-chloride is the perspective radionuclide method of diagnostics of nonspecific diseases of inflammatory diseases of small pelvis organs among women .

For more effectiveness of pelvic inflamatory desease treatment there were observed 61 patients with different forms of this patology and saggested a new complex method of treatment, including the usage of magnetolazer therapy after endoscopical organokeeping operations in early after operation period. The dynamics of proinflamation citokins change content (interleikin-1 beta and alfa tumor necrosis factor) in blood serum of patients with pelvic inflamatory desease confirmed the effectiveness of magnetolazer therapy. Resalts of the invesnigation proved that saggested complex method of treatment for patients with pelvic inflamatory desease allows to make organokeeping operations for reproductive women and to improve results of treatment.

Now, as before, transmission electron microscopy (TEM) is a widely used technique for the determination of virions size. In some studies, dynamic light scattering (DLS) has also been applied for this purpose. Data obtained by different authors and using different methods could vary significantly. The process of TEM sample preparation involves drying on the substrate, which can cause virions to undergo morphology changes. Therefore, other techniques should be used for measurements of virions size in liquid, (i.e. under conditions closer to native). DLS and nanoparticle tracking analysis (NTA) provide supplementary data about the virions hydrodynamic diameter and aggregation state in liquid. In contrast to DLS, NTA data have a higher resolution and also are less sensitive to minor admixtures. In the present work, the size of non-enveloped icosahedral viruses of different nature was analyzed by TEM, DLS and NTA: the viruses used were the encephalomyocarditis virus (animal virus), and cauliflower mosaic virus, brome mosaic virus and bean mild mosaic virus (plant viruses). The same, freshly purified, samples of each virus were used for analysis using the different techniques. The results were compared with earlier published data and description databases. DLS data about the hydrodynamic diameter of bean mild mosaic virus, and NTA data for all examined viruses, were obtained for the first time. For all virus samples, the values of size obtained by TEM were less than virions sizes determined by DLS and NTA. The contribution of the electrical double layer (EDL) in virions hydrodynamic diameter was evaluated. DLS and NTA data adjusted for EDL thickness were in better agreement with TEM results.

The aim of the research is the study of efficiency of early postoperative rehabilitation of patients with purulent inflammatory formations of uterine appendages with microwave therapy. Postoperative treatment of 36 women-patients with the use of combined skin and intravaginal microwave exposure was done. The clinical characteristics of postoperative period and the results of the therapy in 18 month were studied. Dynamical changes of quantitative characteristics of cells at leukemia, of C-reactive protein, content in blood serum, in cervical canal and endometrium of tumor necrosis factor-alpha, interleukin-1 beta and interleukin-6 were estimated. If comparing with the patients who didn’t get physiotherapy exposure the use of microwave therapy at the patients with purulent tubo-ovarian formations was followed by earlier disappearance of inflammation, restoration of quantitative characteristics of leucocytes in blood, normalization of C-reactive protein concentration in blood serum and normal characteristics of erythrocyte sedimentation rate. Diminution of tumor necrosis factor-alpha in blood serum, interleukin-1 beta and interleukin-6, reduction of concentration in cervical canal and endometrium interleukin-6 were observed. In the postoperative period under the microwave therapy womenpatients had pain in pelvis, disturbance of menstrual cycle, 26,5% of women had the reproductive function to be restored.

At pregnant women with gestosis the increasing of concentration in blood serum of soluble molecules of adherence sVCAM-1 and sICAM-1 is discovered. The same changes are revealed at pregnant with bronchial asthma, the period of gestosis being 18 weeks. With it there is a positive correlation between the substance of soluble molecules of adherence and the concentration of cytokines of atopic inflammation in blood serum.

In a prospective study the group of 60 women with AIT and thyroidological uneventful control women, matched for age and parity (n = 30) were tested at 12 and 32 weeks gestation for percentages of peripheral blood lymphocyte subsets (CD3, CD4, CD8, CD16, HLA-DR major histocompatibility complex (MHC)-class II), IL-2, IFN-gamma, TPO-Abs, TSH and thyroxin. Twenty five pregnant women with AIT (41,7%) developed preeclampsia (against 16,7% in control). Pregnancyrelated CMI alterations consisted of low percentages of CD16+ NK cells and all subpopulations of T-cells at 12 weeks gestation. The development of preeclampsia in women with AIT was accompanied with increasing percentage of HLADR+ T- lymphocytes. The high levels of TPO-Abs and activated T-lymphocytes in pregnant women with autoimmune thyroiditis has been suggested as a predictor for later preeclampsia development.

In the work, the impact of general air cryotherapy on a water homeostasis in healthy volunteers is estimated. It is revealed that along with the duration of exposure the redistribution of water forms with an increase of associated fraction took place. This character of metabolic processes testifies to adaptive impact of cold stress on rheologic properties of blood.