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MATERIAL AND METHODS: This study was carried out on 20 male Wistar rats, of which 15 were injected with a bacterialfungal suspension into the cavity of the hock (tarsus) joint. Specifically, the rats were injected with KlebsiellaCandida suspension (1:1) with a microbial concentration of 106 CFU/ml prepared by using the museum strains Klebsiella pneumoniae ICIS-278 and Candida albicans ATCC 24433 under the control of an X-ray apparatus and digitizer FireCR+. Five intact animals served as the control. On day 15 of the experiment, a bacteriological and histological examination of the affected joints was carried out while maintaining topography after euthanasia. RESULTS: In rats infected with the KlebsiellaCandidiasis suspension, pain syndrome and functional changes typical of arthritis were noted on days 23. The bacteriological examination of the affected joints in 75% of rats isolated K. pneumonia cultures that were identical, according to polymerase chain reaction, to the original strain that was part of the bacterialfungal suspension. At the same time, fungi of the genus Candida, which was also used in the experiment to infect animals, were not detected in the affected joints of rats. The established model of KlebsiellaCandida arthritis was characterized by the pathomorphological signs of panarthritis accompanied by the purulent inflammation of all joint structures (synovial membrane, cartilage, and bone tissue) with the involvement of soft periarticular tissues. CONCLUSION: The model of Klebsiella and fungal arthritis presented by this work can be used to test new means of antimicrobial (antibacterial and antifungal) therapies, which are an integral component of the treatment of infection-associated arthritis.

Property relations between spouses are always in the focus of close attention of legal scholars. in recent years, the number of married couples using civil law forms of regulating the issues related to personal and conscientiously acquired property during marriage has been increasing. in family law, the legislator established a prenuptial agreement as this form. However, there is still no clear understanding of the legal nature of this agreement between spouses in the scientific literature; that is why the purpose of the work was to analyze the features of a prenuptial agreement as a type of civil law transactions. The main research methods were the private law method and comparative legal analysis. The authors conducted the study of existing theoretical and legal approaches regarding the specifics of a prenuptial agreement. in the scientific literature, there are three approaches to determining the legal nature of this legal fact: civilised, family-legal and mixed. Comparing the signs of transactions defined by the Civil Code of the Russian Federation and the prenuptial agreement as one of them, its essential signs are established. The paper shows that a prenuptial agreement is a bilateral (multilateral) consensual transaction that has a paid or gratuitous nature, carried out in a certain period or indefinitely, dependent or independent of any conditions. The conclusions note that a prenuptial agreement can be recognized as an independent civil transaction and a separate type of contract with a limited parties, the purpose of which is distribution of property interests between spouses. The conclusions note that the marriage contract can be recognized as an independent civil transaction and a separate type of contract with a limited subject composition, the purpose of which is the distribution of property interests between spouses.

Cytokines and chemokines, as well as gut microsymbionts, are sufficient participants in the intercellular communications, thus supporting homeostasis of gut mucosa. However, these components may be of key significance for intestinal inflammation and damage to epithelial barrier. This work expands the understanding of the relationships between intestinal microbial communities and the local cytokine network of the host. The paper presents the results of the correlation analysis between total microbial number of intestinal microsymbionts and the level of pro- (TNF, IFN, IL-8) and anti-inflammatory cytokines (IL-10, IL- 1ra) in coprofiltrates obtained from clinically healthy people examined for gut dysbiosis. Determination of cytokines in coprofiltrates was carried out by ELISA technique (JSC Vector-Best, Russia). The study of 65 microsymbiocenoses of the human gut was carried out by classical bacteriological methods. Identification of obligate-anaerobic, facultative-anaerobic bacteria and fungi was carried out by time-of-flight mass spectrometry using MALDI TOF-MS Microflex LT series (Bruker Daltonians, Germany). These studies have revealed the leading role of associations between enterobacteria, fungi and representatives of the Staphylococcus genus in gut dysbiosis. In general composition of the obligate-anaerobic association, we have observed a change of consortia from several types of Bifidobacteria and Lactobacilli in eubiotic state to a monoid variant in dysbiosis. At the same time, the number of associations that included Clostridia was increased. The analysis of correlations between cytokine indices and the number of gut microbiota showed persistance of significant associations established during eubiosis under dysbiosis conditions, with an increase in their correlation coefficient: Bifidobacterium spp., Enterobacteriaceae, Staphylococcus spp., Candida spp. and TNF. At the same time, in dysbiosis, the direction of the connections changed, and new correlations were determined: for Staphylococcus spp. and IFN; Staphylococcus spp. and IL-8; Enterobacteriaceae and IL-1ra, IFN. The established features of correlations between indices of microsymbiocenosis and quantitative changes in cytokines allow us to consider the number, composition of microsymbiocenosis and cytokine profile as factors that may affect the state of gut homeostasis in eu- and dysbiosis.

The growth of urbanization is one of the confirmations of the global evolutionary process acceleration. Sustainable cities and towns is one of the UN Sustainable Development Goals for the period up to 2030 (No. 11). One of the key tasks of modern education and enlightenment is the formation of a sustainable lifestyle among the urban population and the reduction of anthropogenic pressure on the environment. However, the corresponding psychological and pedagogical direction, which can be conditionally called educational eco-urban studies, is only being formed today. The purpose of the research is to identify the prerequisites and basic approaches for the formation of a new direction in pedagogical theory and practice – education for urban residents. The research methods are: analysis of international and Russian documents relating to various aspects of urban development; study of existing experience – educational initiatives in the field of formal, non-formal and informal education in the city and for the city; pedagogical observations and conversations were held with schoolchildren and students of Moscow, as well as a number of other regions. These methods have made it possible to conduct a comparative analysis and identify the specifics of ecological culture (as a way of interacting with the immediate environment – the city) of the inhabitants of the metropolis and small towns. The article discusses possible methodological approaches, content and forms of implementation (training courses, summer schools, competence centers, etc.) of education for sustainable urban development. The main conclusions and results of the work include the need to use an interdisciplinary approach in the development of the main provisions of educational eco-urban studies as an independent direction of pedagogical theory and practice since the complex synthetic nature of the object under study – education for the city and the formation of the ecological culture of urban residents cannot be fully described and studied from the standpoint of any particular science, theory or concept.

BACKGROUND: Dental implant stability depends on the mechanical bone properties in the area of the planned surgical intervention and the correctness of the implant embedded in the bone tissue. A significantly decreased bone tissue mineral density of the jaws was found in patients with osteoporosis, which complicates or even prevents the restoration of the integrity of the dentition by the dental implantation method. AIM: This study aimed to conduct a comparative assessment of the effectiveness of the treatment of dentition defects by dental implantation in patients with a normal state of bone tissue, osteopenia, and osteoporosis, to assess the stability of the installed implants using the resonance frequency analysis. MATERIALS AND METHODS: All patients underwent a densitometric study using double X-ray photon absorptiometry, in addition to the basic examination, in dental implantation operation preparation, which made it possible to identify systemic and regional changes in bone mineral density. Dental implant stability and osseointegration were controlled by the resonant frequency analysis method. RESULTS: The parameters of resonance frequency analysis in patients with normal bone mass, osteopenia, and osteoporosis vary depending on the area where the implants are installed when studying the implant stability in patients with different bone mineral densities. The overall index of resonance frequency analysis values differed from those of the group with osteopenia and osteoporosis, regardless of the area of implants installed in patients with normal bone tissue, and was respectively higher by 7.1% (p 0.001) and 10.1% (p 0.001), respectively. The overall index in the group with osteopenia was higher and differed from that in the group with osteoporosis by 5.7% (p 0.001). CONCLUSION: The resonance frequency analysis results provide significant information about the state of the implant-bone interface at different treatment stages and during follow-up examinations of patients. The measurement technique makes it possible to observe the dynamics of the osseointegration process, and, if necessary, make timely adjustments to the functional load. Resonance frequency analysis is an important method for documenting the clinical outcome of implantation. This is of particular importance from a medical-legal point of view.

Activation of the stimulator of interferon genes (STING) pathway promotes antitumor immunity but STING agonists have yet to achieve clinical success. Increased understanding of the mechanism of action of STING agonists in human tumors is key to developing therapeutic combinations that activate effective innate antitumor immunity. Here, we report that malignant pleural mesothelioma cells robustly express STING and are responsive to STING agonist treatment ex vivo. Using dynamic single-cell RNA sequencing of explants treated with a STING agonist, we observed CXCR3 chemokine activation primarily in tumor cells and cancer-associated fibroblasts, as well as T-cell cytotoxicity. In contrast, primary natural killer (NK) cells resisted STING agonist–induced cytotoxicity. STING agonists enhanced migration and killing of NK cells and mesothelin-targeted chimeric antigen receptor (CAR)-NK cells, improving therapeutic activity in patient-derived organotypic tumor spheroids. These studies reveal the fundamental importance of using human tumor samples to assess innate and cellular immune therapies. By functionally profiling mesothelioma tumor explants with elevated STING expression in tumor cells, we uncovered distinct consequences of STING agonist treatment in humans that support testing combining STING agonists with NK and CAR-NK cell therapies.

Supplementary Data from Activation of Tumor-Cell STING Primes NK-Cell Therapy

PDF file - 1.3MB, Supplementary data on MAPK1 copy number and expression, combination treatment with WZ4002 and PI3K inhibitors, impact of MEK1 K57N on WZ4002 sensitivity, Mapk1 copy number analyses and NF1 and dusp expression in drug sensitive and resistant mouse tumors

KRAS mutation is present in approximately 30% of human lung adenocarcinomas. Although recent advances in targeted therapy have shown great promise, effective targeting of KRAS remains elusive, and concurrent alterations in tumor suppressors render KRAS-mutant tumors even more resistant to existing therapies. Contributing to the refractoriness of KRAS-mutant tumors are immunosuppressive mechanisms, such as increased presence of suppressive regulatory T cells (Treg) in tumors and elevated expression of the inhibitory receptor PD-1 on tumor-infiltrating T cells. Treatment with BET bromodomain inhibitors is beneficial for hematologic malignancies, and they have Treg-disruptive effects in a non–small cell lung cancer (NSCLC) model. Targeting PD-1–inhibitory signals through PD-1 antibody blockade also has substantial therapeutic impact in lung cancer, although these outcomes are limited to a minority of patients. We hypothesized that the BET bromodomain inhibitor JQ1 would synergize with PD-1 blockade to promote a robust antitumor response in lung cancer. In the present study, using Kras+/LSL-G12D; Trp53L/L (KP) mouse models of NSCLC, we identified cooperative effects between JQ1 and PD-1 antibody. The numbers of tumor-infiltrating Tregs were reduced and activation of tumor-infiltrating T cells, which had a T-helper type 1 (Th1) cytokine profile, was enhanced, underlying their improved effector function. Furthermore, lung tumor–bearing mice treated with this combination showed robust and long-lasting antitumor responses compared with either agent alone, culminating in substantial improvement in the overall survival of treated mice. Thus, combining BET bromodomain inhibition with immune checkpoint blockade offers a promising therapeutic approach for solid malignancies such as lung adenocarcinoma. Cancer Immunol Res; 6(10); 1234–45. ©2018 AACR.

PD-L1 immunohistochemical staining does not always predict whether a cancer will respond to treatment with PD-1 inhibitors. We sought to characterize immune cell infiltrates and the expression of T-cell inhibitor markers in PD-L1–positive and PD-L1–negative malignant pleural mesothelioma samples. We developed a method for immune cell phenotyping using flow cytometry on solid tumors that have been dissociated into single-cell suspensions and applied this technique to analyze 43 resected malignant pleural mesothelioma specimens. Compared with PD-L1–negative tumors, PD-L1–positive tumors had significantly more infiltrating CD45+ immune cells, a significantly higher proportion of infiltrating CD3+ T cells, and a significantly higher percentage of CD3+ cells displaying the activated HLA-DR+/CD38+ phenotype. PD-L1–positive tumors also had a significantly higher proportion of proliferating CD8+ T cells, a higher fraction of FOXP3+/CD4+ Tregs, and increased expression of PD-1 and TIM-3 on CD4+ and CD8+ T cells. Double-positive PD-1+/TIM-3+ CD8+ T cells were more commonly found on PD-L1–positive tumors. Compared with epithelioid tumors, sarcomatoid and biphasic mesothelioma samples were significantly more likely to be PD-L1 positive and showed more infiltration with CD3+ T cells and PD-1+/TIM-3+ CD8+ T cells. Immunologic phenotypes in mesothelioma differ based on PD-L1 status and histologic subtype. Successful incorporation of comprehensive immune profiling by flow cytometry into prospective clinical trials could refine our ability to predict which patients will respond to specific immune checkpoint blockade strategies. Cancer Immunol Res; 4(12); 1038–48. ©2016 AACR.

PDF file - 83K

PDF file - 74K, Table describing clinical and molecular characteristics of NSCLC patients

figure legends for supplementary Table S1-S3 and supplementary Figure S1-S13

Supplemental Figure 1. Difference in CD3+ immune cell infiltrates by histologic subtype of mesothelioma. Supplemental Figure 2. Comparison of immune cell infiltrates among patients who received or did not receive neoadjuvant chemotherapy prior to surgical resection of their tumor. Supplemental Figure 3. Quantification of monocyte subtypes in tumor samples. Supplemental Figure 4. Comparison of immune cell infiltrates by mutational subtype in mesothelioma samples. Supplemental Figure 5. Statistical analysis of t-SNE maps from Figure 6. Supplemental Figure 6. Kaplan-Meier overall survival analysis for patients according to histologic subtype, PD-L1 immunohistochemical status, and CD3 infiltration as assessed by either flow cytometry or immunohistochemistry.

DFCI358 signaling with respect to mTOR; crizotinib+trametinib combination;RTK activation

Supplemental Figure 1

Detailed description of methodology and reagents

Summary of EGFR inhibitor resistant patient-derived xenografts.

Antibodies used in the study

Epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKI) are the standard-of-care treatment for EGFR-mutant non–small cell lung cancers (NSCLC). However, most patients develop acquired drug resistance to EGFR TKIs. HER3 is a unique pseudokinase member of the ERBB family that functions by dimerizing with other ERBB family members (EGFR and HER2) and is frequently overexpressed in EGFR-mutant NSCLC. Although EGFR TKI resistance mechanisms do not lead to alterations in HER3, we hypothesized that targeting HER3 might improve efficacy of EGFR TKI. HER3–DXd is an antibody–drug conjugate (ADC) comprised of HER3-targeting antibody linked to a topoisomerase I inhibitor currently in clinical development. In this study, we evaluated the efficacy of HER3–DXd across a series of EGFR inhibitor–resistant, patient-derived xenografts and observed it to be broadly effective in HER3-expressing cancers. We further developed a preclinical strategy to enhance the efficacy of HER3–DXd through osimertinib pretreatment, which increased membrane expression of HER3 and led to enhanced internalization and efficacy of HER3–DXd. The combination of osimertinib and HER3–DXd may be an effective treatment approach and should be evaluated in future clinical trials in EGFR-mutant NSCLC patients.Significance:EGFR inhibition leads to increased HER3 membrane expression and promotes HER3–DXd ADC internalization and efficacy, supporting the clinical development of the EGFR inhibitor/HER3–DXd combination in EGFR-mutant lung cancer.See related commentary by Lim et al., p. 18

Antibodies, compounds, oligonucleotides, and other reagents used in the study

Allelic Frequencies in PC-9 vs PC-9 T790M/C797S cells.

Purpose:The RET proto-oncogene encodes a receptor tyrosine kinase that is activated by gene fusion in 1%–2% of non–small cell lung cancers (NSCLC) and rarely in other cancer types. Selpercatinib is a highly selective RET kinase inhibitor that has recently been approved by the FDA in lung and thyroid cancers with activating RET gene fusions and mutations. Molecular mechanisms of acquired resistance to selpercatinib are poorly understood.Patients and Methods:We studied patients treated on the first-in-human clinical trial of selpercatinib (NCT03157129) who were found to have MET amplification associated with resistance to selpercatinib. We validated MET activation as a targetable mediator of resistance to RET-directed therapy, and combined selpercatinib with the MET/ALK/ROS1 inhibitor crizotinib in a series of single patient protocols (SPP).Results:MET amplification was identified in posttreatment biopsies in 4 patients with RET fusion–positive NSCLC treated with selpercatinib. In at least one case, MET amplification was clearly evident prior to therapy with selpercatinib. We demonstrate that increased MET expression in RET fusion–positive tumor cells causes resistance to selpercatinib, and this can be overcome by combining selpercatinib with crizotinib. Using SPPs, selpercatinib with crizotinib were given together generating anecdotal evidence of clinical activity and tolerability, with one response lasting 10 months.Conclusions:Through the use of SPPs, we were able to offer combination therapy targeting MET-amplified resistance identified on the first-in-human study of selpercatinib. These data suggest that MET dependence is a recurring and potentially targetable mechanism of resistance to selective RET inhibition in advanced NSCLC.

Supplementary Methods

Supplementary Figures and legends

Introduction. The introduction of amendments and additions to the anti-smoking Federal Law No. 15-FZ of 23.02.2013 in 2020 contributed to the cardinal revision of the name of this law and the introduction of a ban on consumption of any nicotine-containing products in public places. There are no separate, specially allocated isolated premises provided for the consumption of tobacco products and other types of nicotine-containing products by law, even though specially conducted studies have shown significantly different levels of additional inhalation risk arising from air pollution when consuming tobacco products and electronic nicotine delivery systems or heating tobacco. The aim of the study: based on experimental data, to justify the multiplicity of air exchange in unique isolated rooms designed to protect against the effects of ambient tobacco smoke, the consequences of tobacco consumption or nicotine-containing products. Materials and methods. The study was subject to 3 types of nicotine-containing products: tobacco cigarettes (cigarettes), an electronic nicotine delivery system (ENDS) and an electronic tobacco heating system (ETHS). In the course of the study were carried out: a series of experiments on the consumption of nicotine-containing products in various ways in a specially equipped indoor room, air sampling and laboratory studies were performed, the gross and specific emissions of chemicals entering the air environment from one unit of production per hour were calculated, the selection of priority chemicals for calculating the air removed from the room when using nicotine-containing products of each type was carried out, the necessary level of air exchange of premises was calculated, intended for the consumption of tobacco or nicotine. Results. The indoor air quality assessment showed that the main components of the gross intake into the air environment are suspended particles and carbon monoxide when smoking cigarettes. However, taking into account the values of the maximum permissible concentrations, the most significant contribution to the integral indicator is made by acetaldehyde and suspended particles PM10 and PM2.5. Using ENDS, the most significant pollutants were acetaldehyde and carbon monoxide, using ETHS - acetaldehyde and suspended particles PM10 and PM2.5. For all the investigation types of products, the priority substance for calculating the removed air is acetaldehyde, its share in the structure of all detected pollutants was: for cigarettes - 56.27%, for ENDS - 62.7%, for ETHS - 82.51%, in connection with which its specific values can be used to calculate the consumption of the removed air when modelling a specially allocated room intended for the consumption of tobacco products and other types of nicotine-containing products. Conclusion. The obtained findings demonstrated an obvious, more than 10-fold difference in the requirements for air exchange of premises intended for cigarette smoking and premises intended for the consumption of ENDS or ETHS, at the same intensity of consumption. Therefore, when organizing specially designated places for smoking or the consumption of nicotine-containing products, it is advisable to separate them depending on the type of products consumed: tobacco smoking separately, ENDS and ETHS consumption separately.

PURPOSE: PARP inhibitor resistance may be overcome by combinatorial strategies with agents that disrupt homologous recombination repair (HRR). Multiple HRR pathway components are HSP90 clients, so that HSP90 inhibition leads to abrogation of HRR and sensitisation to PARP inhibition. We performed in vivo preclinical studies of the HSP90 inhibitor onalespib with olaparib and conducted a Phase 1 combination study. PATIENTS AND METHODS: Tolerability and efficacy studies were performed in patient-derived xenograft(PDX) models of ovarian cancer. Clinical safety, tolerability, steady-state pharmacokinetics and preliminary efficacy of olaparib and onalespib were evaluated using a standard 3 + 3 dose-escalation design. RESULTS: Olaparib/onalespib exhibited anti-tumour activity against BRCA1-mutated PDX models with acquired PARPi resistance and PDX models with RB-pathway alterations(CDKN2A loss and CCNE1 overexpression). Phase 1 evaluation revealed that dose levels up to olaparib 300 mg/onalespib 40 mg and olaparib 200 mg/onalespib 80 mg were safe without dose-limiting toxicities. Coadministration of olaparib and onalespib did not appear to affect the steady-state pharmacokinetics of either agent. There were no objective responses, but disease stabilisation ≥24 weeks was observed in 7/22 (32%) evaluable patients including patients with BRCA-mutated ovarian cancers and acquired PARPi resistance and patients with tumours harbouring RB-pathway alterations. CONCLUSIONS: Combining onalespib and olaparib was feasible and demonstrated preliminary evidence of anti-tumour activity.

Activation of the stimulator of interferon genes (STING) pathway promotes antitumor immunity but STING agonists have yet to achieve clinical success. Increased understanding of the mechanism of action of STING agonists in human tumors is key to developing therapeutic combinations that activate effective innate antitumor immunity. Here, we report that malignant pleural mesothelioma cells robustly express STING and are responsive to STING agonist treatment ex vivo. Using dynamic single-cell RNA sequencing of explants treated with a STING agonist, we observed CXCR3 chemokine activation primarily in tumor cells and cancer-associated fibroblasts, as well as T-cell cytotoxicity. In contrast, primary natural killer (NK) cells resisted STING agonist–induced cytotoxicity. STING agonists enhanced migration and killing of NK cells and mesothelin-targeted chimeric antigen receptor (CAR)-NK cells, improving therapeutic activity in patient-derived organotypic tumor spheroids. These studies reveal the fundamental importance of using human tumor samples to assess innate and cellular immune therapies. By functionally profiling mesothelioma tumor explants with elevated STING expression in tumor cells, we uncovered distinct consequences of STING agonist treatment in humans that support testing combining STING agonists with NK and CAR-NK cell therapies.

Автором проанализированы нормы действующего законодательства об основаниях осуществления досмотра в ходе производства по делам об административных правонарушениях. Определены условия обеспечения законности и соблюдения прав и свобод граждан в ходе указанной процедуры.

Retinal vein occlusion (RVO) is the most common retinal vascular disease, it ranks second in terms of severity of retinal damage and prognosis after diabetic retinopathy. The main cause of visual impairment in patients with RVO is the manifestation of macular edema (ME). Persistent ME may block the recovering of visual acuity. The development of new methods of diagnosis and treatment of this pathology has changed approaches to the management of patients with RVO. This review presents the advantages and promising treatment modalities of retinal venous occlusions, it also demonstrates their advantages and disadvantages. Administration of drug treatment in the therapy of RVO is advisable only in the acute period of the disease. Laser coagulation is still the standard treatment for neovascular complications associated with RVO. The necessity of surgical repair remains controversial, since its use can result in severe complications, such as damage to the central retinal artery, perforation of the eye, retinal detachment and traumatic neuropathy. The use of steroids for the treatment of ME is associated with their ability to reduce capillary permeability, however, there is high risk of cataract. Antiangiogenic therapy is effective against pathogenesis of ME in RVO and is a first-line treatment for this disorder. The use of subthreshold microimpulse laser treatment is promising. Possibilities of further improvement and use of combined techniques in the treatment of this pathology are considered.

BACKGROUND AND OBJECTIVE: A computational model has predicted parameters for using a navigated microsecond pulsing laser system to treat central serous chorioretinopathy (CSC). PATIENTS AND METHODS: A prospective, single-center, interventional case series was conducted for patients with acute CSC who were enrolled following screening and informed consent. Treatment involved laser pulse duration of 50 μs, 2.4% Duty Cycle, 100 μm spot size, and 10 ms pulse duration. RESULTS: Average best-corrected visual acuity (decimal) of 12 patients improved from 0.86 ± 0.03 at baseline to 0.97 ± 0.01 at 3 months. Baseline central retinal thickness decreased from 452.58 μm ± 24.53 μm at baseline to 249.25 μm ± 2.92 μm at 3 months. Retinal sensitivity improved from 24.1 dB ± 1.09 dB at baseline to 28.98 dB ± 0.23 dB at 3 months. In all cases, subretinal fluid was resorbed. CONCLUSION: The parameter sets derived from the computer model can be applied safely and effectively for CSC treatment using the navigated microsecond pulsing laser system. [ Ophthalmic Surg Lasers Imaging Retina . 2020;51:512–520.]