[The role of microbiota and flavonoids in maintaining the balance of helper and regulatory T-lymphocytes associated with the intestinal immune barrier].

The gastrointestinal tract is a barrier, represented by dynamic and mutually regulating components (microbial, chemical, physical and immune) for the selective penetration of luminal contents into the internal environment. From the point of view of immunologists, even in a physiological condition, the epithelium of the intestinal wall is in a state of mild inflammation, which is explained by the constant invasion of antigens (food, microbial) and, in turn, the constant readiness of the immune system to respond. The purpose of this review was to analyze information about the formation of microbial and immunological barriers, immunological tolerance to microbiota and the possible role of flavonoids in this. Material and methods. The literature search was carried out using PubMed, ResearchGate, Elibrary databases mainly for the last 10 years, using the following keywords: flavonoid, gut microbiome/microbiota, Th17, Treg, RORγt, immunity, segmented filamentous bacteria. Results. During the immune response, a significant role in maintaining the intestinal barrier function is assigned to helper T lymphocytes type 17 (Th17). The intestinal microbiome is a key element in the formation of the immune barrier. Th17 differentiation in the intestine is fully triggered by commensals (apparently, the main role belongs to segmented filamentous bacteria) after weaning and the start of complementary feeding. Pro-inflammatory Th17 effectors in the gut are controlled by anti-inflammatory regulatory T-cells (Treg). In recent years, it has been established that despite the opposing functions of regulatory cells and effector Th17 cells, their differentiation is similar and is characterized by the expression of the common transcription factor RORγt. The main part of the peripheral regulatory lymphocytes of the intestine is a population that stably expresses not only FOXP3, but also RORγt. Flavonoids, which are plant secondary metabolites of the polyphenolic structure, are able to inhibit intracellular kinases and, as a result, influence the activation and implementation of effector functions of immunocompetent cells. Some flavonoids promote RORγt expression and appear to be able to reprogram the effector phenotype of Th17 cells, reducing their pathogenicity. Conclusion. Understanding the interactions between the microbiota, immune cells, and factors involved in their regulation, which are critical for the maintenance of tolerance, may facilitate progress in the prevention and therapeutic approaches to treat immunoinflammatory and autoimmune diseases.

[Is it possible to detect surface antigen CD133 on patient-derived glioblastoma continuous cell cultures using fluorescent aptamers?] / Vozmozhna li detektsiya poverkhnostnogo antigena CD133 na perevivaemykh kul'turakh kletok glioblastomy patsientov s pomoshch'yu fluorestsentnykh aptamerov?

Theranostics combines diagnostics and therapeutic exposure. Regarding glioblastomas, theranostics solves the problem of detecting and destroying tumor stem cells resistant to irradiation and chemotherapy and causing tumor recurrence. Transmembrane surface antigen CD133 is considered as a potential marker of tumor stem cells. OBJECTIVE: To detect CD133 in patient-derived glioblastoma continuous cell cultures using fluorescence microscopy and modified aptamers (molecular recognition elements) anti-CD133. MATERIAL AND METHODS: To detect CD133, we used mousey fluorescence monoclonal antibodies anti-CD133 MA1-219, FAM-modified DNA aptamers anti-CD133 AP-1-M and Cs5. Non-aptamer DNA oligonucleotide NADO was used as a negative control. Detection was performed for three samples of patient-derived glioblastoma continuous cell cultures coded as 1548, 1721 and 1793. RESULTS: MA1-219 antibodies brightly stained cell culture 1548, to a lesser extent - 1721. There was diffuse staining of cell culture 1793. Cs5-FAM aptamer stained cells in a similar way, but much weaker. AP-1-M-FAM aptamer interacted with cells even weaker and diffusely stained only cell culture 1793. Non-aptamer NADO did not stain cell culture 1548 and very weakly diffusely stained cell culture 1793. CONCLUSION: For both molecular recognition elements (MA1-219 antibody and Cs5 aptamer), 3 cell culture samples can be arranged in the following order possibly reflecting CD133 status decrease: strong signal for cell culture 1548, much weaker for 1721, even weaker for 1793. Only cell culture 1548 can be considered CD133 positive with combination of Cs5+ and NADO signals. Cell culture 1793 is CD133 false positive with combination of Cs5+ and NADO+ signals.

[Bioregulation therapy in the treatment of age-related osteoarthropathy of the temporomandibular joint.]

The study consists of clinical and radiological parallels between the severity of functional pathology of the temporomandibular joint, bone density of the joint elements and the effect of bioregulatory therapy on it at different ages. Male patients (35-74 years old) with functional pathology of the temporomandibular joint were under observation. The study showed a significant decrease in bone density and correlation of this index with the severity of the disease course in all patients with temporomandibular joint dysfunction compared with the control group of healthy subjects. After a course of bioregulatory therapy the bone density and functional pathology index of the temporomandibular joint indicate a change from the severe course to a form of moderate severity in the group of elderly patients. The correlation correlation between the severity of the course of functional pathology and the decrease in the density of bone structures of the temporomandibular joint has been revealed, increasing with age. Use of bioregulatory therapy in complex treatment of functional pathology has favorable influence on a course of functional pathology of a temporomandibular joint at elderly patients.

[Geriatric aspects of teaching maxillofacial surgery and surgical dentistry.]

Competence-based approach plays an important role in the training of a modern doctor. Considering that the strategy of dental care for older age groups is determined not only by the demographic situation and its development, but also by the justified need for effective dental care, it is important to implement the teaching of gerontostomatology in a medical university based on the results of evidence-based medicine and a personalized approach. The aim of the work was to develop a methodology for teaching geriatric aspects according to the curriculum of a specialist in the discipline of maxillofacial surgery and surgical dentistry. Based on the study of scientific works on various aspects of gerontostomatology, the author's position is indicated in the work and methodological foundations for modern teaching of the gerontostomatology section are presented according to the curriculum of a specialist in the main areas of training in the field of maxillofacial surgery and surgical dentistry. Particular attention is paid to the methodology of teaching pyoinflammatory diseases and injuries of the maxillofacial region, salivology, pathology of the temporomandibular joint, masticatory muscles and oncostomatology.

Generation of an induced pluripotent stem cell line, ICGi029-A, by reprogramming peripheral blood mononuclear cells of a patient suffering from hypertrophic cardiomyopathy and carrying a heterozygous p.N515del mutation in MYBPC3.

Hypertrophic cardiomyopathy (HCM) is a common cardiovascular disease. However, effective methods of its therapy have not been developed so far. To date patient-specific induced pluripotent stem cell-derived cardiomyocytes are supposed to be a useful tool for studying HCM molecular mechanisms and to help find new approaches to HCM therapy. Using non-integrating episomal vectors, we generated an iPSC line from peripheral blood mononuclear cells of an HCM patient carrying a heterozygous p.N515del mutation in MYBPC3. The iPSC line expressed pluripotency markers, gave rise to derivatives of three germ layers during spontaneous differentiation, had normal karyotype, and retained the patient-specific mutation.

[Stage wise treatment of uterine arteriovenous malformation complicated by recurrent haemorrhage]. / Étapnoe lechenie arteriovenoznoi mal'formatsii matki, oslozhnennoi retsidiviruiushchimi krovotecheniiami.

Uterine arteriovenous malformation is a rarely encountered disease threatening with massive haemorrhage. The article describes a clinical case report regarding a 37-year-old woman presenting with this pathology and previously hospitalized twice with severe posthaemorrhagic complications at a 5-month interval due to refusal from timely hysterectomy. A vascular formation in the womb was detected at ultrasonography, however its pattern was identified only by computed tomography of small pelvis organs with intravenous contrasting. However, the complete picture of the architectonics of uterine arteriovenous malformation and extension of the pathology was obtained by selective subtraction angiography, making it possible not only to perform diagnosis but also, if necessary, to immediately perform selective embolization of the supplying vessels. Due to massive uterine bleeding on the background of womb malformation, the woman was twice subjected to roentgenoendovascular embolization of afferent vessels, with the achievement of persistent haemostasis. Hysterectomy was performed after stabilization of the state. Thus, an extensive angiomatous uterine lesion accompanied by recurrent bleedings, along with roentgenoendovascular methods of treatment there is a need of additional surgical resection with the removal of the angiodysplasia focus.

[Geriatric aspects of modern plastic surgery.]

Aging of the human body is a natural physiological process. In the last century, the main reasons for people turning to plastic surgeons were congenital and acquired deformities, as well as age-related changes in the skin of the face. Nowadays, a person's appearance has become the main factor in his success in society. The work determines the motivation of elderly and senile people to perform plastic surgeries, and also analyzes the work of clinics in performing plastic surgeries for people of older age groups. It was found that 96,4% of men and 86,9% of women in older age groups have a fairly complete understanding of plastic surgery. Among the elderly and old people, 27,7% of men and 17,2% of women have a desire to perform plastic surgery for themselves, however, women are 10,9 times more likely to go to plastic surgeons. Analysis of the plastic surgery clinics showed a steady trend of increasing the number of elderly and senile people, regardless of gender, to plastic surgeons to perform operations aimed mainly at improving the aesthetics of the face, neck and body shaping by eliminating gravitational ptosis and removing excess subcutaneous fat. deposits on the trunk. The main reason for refusal to perform the operation desired by the patient and recommended by the plastic surgeon is the lack of funds.

Generation of induced pluripotent stem cell lines ICGi021-A and ICGi022-A from peripheral blood mononuclear cells of two healthy individuals from Siberian population.

ICGi021-A and ICGi022-A iPSC lines were obtained by reprogramming PBMCs of two healthy women of the Siberian population using episomal non-integrating vectors expressing Yamanaka factors. iPSC lines expressed pluripotency markers, had a normal karyotype and demonstrated the ability to differentiate into derivatives of the three germ layers. Clinical exome sequencing data of the original biosamples of the donors are available in the NCBI SRA database. The generated cell lines are useful as "healthy" control in biomedical studies.

Generation of induced pluripotent stem cell line ICGi018-A from peripheral blood mononuclear cells of a patient with Huntington's disease.

Huntington's disease (HD) is an autosomal dominant neurodegenerative disease caused by CAG repeat expansion in the HTT gene. HD patient-specific induced pluripotent stem cells (iPSCs) represent an excellent model for the disease study. We generated iPSC line from blood mononuclear cells of HD patient with 38 CAG repeats in the HTT exon 1 using integration free episomal plasmids expressing Yamanaka factors. The iPSC line retained the disease causing mutation and expressed pluripotency markers. It also displayed a normal karyotype and the ability to differentiate into derivatives of three germ layers.

[Allergic rhinitis: the modern aspects of therapy]. / Allergicheskii rinit: sovremennye aspekty terapii.

The main objectives of the present article were to systematize the modern views of the causes and risk factors of allergic rhinitis, to clarify the manner of its development, to define the leading etiological mechanism underling the pathogenesis of this condition, to consider the methods used for the diagnostics and the treatment of this disease, and to sum up the clinical experience with the use of Levocetirizine (Allerwey) for the management of allergic rhinitis. Special attention is given to the achievement and the maintenance of control of the persistent or intermittent forms of allergic rhinitis and to approaches to its treatment based on the medications registered in the Russian Federation.

Low-burden TP53 mutations in chronic phase of myeloproliferative neoplasms: association with age, hydroxyurea administration, disease type and JAK2 mutational status.

The multistep process of TP53 mutation expansion during myeloproliferative neoplasm (MPN) transformation into acute myeloid leukemia (AML) has been documented retrospectively. It is currently unknown how common TP53 mutations with low variant allele frequency (VAF) are, whether they are linked to hydroxyurea (HU) cytoreduction, and what disease progression risk they carry. Using ultra-deep next-generation sequencing, we examined 254 MPN patients treated with HU, interferon alpha-2a or anagrelide and 85 untreated patients. We found TP53 mutations in 50 cases (0.2-16.3% VAF), regardless of disease subtype, driver gene status and cytoreduction. Both therapy and TP53 mutations were strongly associated with older age. Over-time analysis showed that the mutations may be undetectable at diagnosis and slowly increase during disease course. Although three patients with TP53 mutations progressed to TP53-mutated or TP53-wild-type AML, we did not observe a significant age-independent impact on overall survival during the follow-up. Further, we showed that complete p53 inactivation alone led to neither blast transformation nor HU resistance. Altogether, we revealed patient's age as the strongest factor affecting low-burden TP53 mutation incidence in MPN and found no significant age-independent association between TP53 mutations and hydroxyurea. Mutations may persist at low levels for years without an immediate risk of progression.

Femtosecond laser written waveguides deep inside silicon.

Photonic devices that can guide, transfer, or modulate light are highly desired in electronics and integrated silicon (Si) photonics. Here, we demonstrate for the first time, to the best of our knowledge, the creation of optical waveguides deep inside Si using femtosecond pulses at a central wavelength of 1.5 µm. To this end, we use 350 fs long, 2 µJ pulses with a repetition rate of 250 kHz from an Er-doped fiber laser, which we focused inside Si to create permanent modifications of the crystal. The position of the beam is accurately controlled with pump-probe imaging during fabrication. Waveguides that were 5.5 mm in length and 20 µm in diameter were created by scanning the focal position along the beam propagation axis. The fabricated waveguides were characterized with a continuous-wave laser operating at 1.5 µm. The refractive index change inside the waveguide was measured with optical shadowgraphy, yielding a value of 6×10-4, and by direct light coupling and far-field imaging, yielding a value of 3.5×10-4. The formation mechanism of the modification is discussed.

[The application of the acupuncture techniques for the combined treatment of neurotic tics in the children]. / Ispol'zovanie metodov refleksoterapii v kompleksnom lechenii nevroticheskikh tikov u detei.

At present, the frequency of neurosis in the children is growing under the influence of the increasingly intensive stressful factors. The neurotic ticks are one of the manifestations of neurosis in the children characterized by involuntary movements; they are often accompanied by hyperactivity, deficit of attention, fatigue, and headaches. AIM: The objective of the present study was to evaluate the effectiveness of acupuncture used for the treatment of neurotic tics in the children. MATERIAL AND METHODS: The study was carried out at the basis of the department of children's rehabilitation of the Moscow city polyclinic #39. All the children were counselled by a neurologist, psychologist, and acupuncturist. An electroencephalogram was obtained from each patient and a questionnaire study involving the patients' parents was conducted. The patients were treated with the application of reflexotherapy and were given recommendations of a psychologist. A total of 30 children at the age from 6 to 12 years were available for the examination. The participants of the study were allocated to two groups depending on the strategy of the outpatient care. Group 1 was comprised of the patients (n=17) who were given 3 courses of reflexotherapy in addition to pharmacotherapy and psychologist's recommendations. The patients included in group 2 served as controls (n=13) and received only medications and psychotherapy without acupuncture. The effectiveness of the treatment was evaluated based on the dynamics of the patients' complaints. RESULTS AND DISCUSSION: The resullts of the study give evidence of the higher effectiveness of the treatment of the children suffering from neurotic ticks with the application of acupuncture therapy compared with the conventional approach. A few courses of reflexotherapy provided in the combination with pharmacotherapy and the compliance with the recommendations of a psychologist resulted in the more efficient elimination of the neurotic tics together with the associated hyperactivity, deficit of attention, fatigue, and headaches than the use of medication therapy and psychotherapy without acupuncture.

Antiproliferative Effects of Modified Bioflavonoid in Ex Vivo Model.

We studied the effect of modified bioflavonoid and reference drug quercetin dihydrate on proliferation of mononuclears triggered by T- and B-cell mitogens. Lymphocytes were in vivo pretreated with the examined agents followed by their explantation and in vitro activation with T- and B-cell mitogens in cell culture. Intraperitoneal injection of modified bioflavonoid and quercetin dihydrate produced a dose-dependent inhibition of proliferation of the in vitroactivated splenocytes; modified bioflavonoid demonstrated higher antiproliferative activity.

[Long-term survival in working-age patients after cerebrovascular accident]. / Otdalennaya vyzhivaemost' patsientov trudosposobnogo vozrasta posle tserebral'nogo insul'ta.

AIM: To assess the long-term survival in working-age patients after cerebrovascular accident (CVA) and to define basic medical and social factors determining the survival rates. MATERIAL AND METHODS: A continuous retrospective study included 756 working-age patients (<60 years) after CVA. The life-table method and Kaplan-Meier analysis were used to calculate a survival rate. RESULTS: The highest risk of death was observed in the first year of CVA. One-year survival after CVA was 81.7%, falling to 74.7% at 3 years and to 57.9% at 7 years. The five-year survival was lower among men (60%) compared to women (74.5%). The figures were higher in working population compared to non-working population (78% vs. 56%, respectively). Long-term survival rate depends on the type of CVA: the five-year survival rate after unspecified stroke was 83.0%, after ischemic stroke - 69.0%, after hemorrhagic stroke - 43.0%. The direct dependence between long-term survival and patient age has been identified: older patients have worse long-term prognosis. CONCLUSION: The seven-year survival rate in working-age patients after cerebrovascular accident was 58%. The health and social factors affecting the long-term survival rates in working-age patients are as follows: hemorrhagic stroke, unemployment, male gender and advanced age.

A POSSIBLE CYTOGENETIC ANALOGY TO GENOMIC «SPECIATION ISLANDS¼ AS REVEALED BY CHROMOSOME STUDY OF A NATURAL HYBRID VOLE.

Chromosome analysis in mammals over the last half century has largely focused on species identification. A growing number of hybrid zones of karyotypically differentiated cryptic taxa have been described. A good example is provided by two karyoforms of the 46-chromosome common vole (long known as «arvalis¼ and «obscurus¼) that make contact longitudinally from the north to the south of European Russia. The hybrid F1 karyotype displays genomic markers which distinguish arvalis and obscurus and which are cytogenetically detectable as minor variants. The apparent insignificance of the genomic region of heterozygosity associated with these markers perhaps does not prevent hybrid chromosome pair formation at meiosis but may reflect a site resistant to gene flow. A cytogenetic analogy with so-called «speciation islands¼, based, for example, on the study of the corvine hybrid zone and represented by a small number of limited genomic sites (occupying less than 1 % of the genome) (Poelstra et al., 2014), may be relevant.

Therapy of Chronic Cardiosclerosis in WAG Rats Using Cultures of Cardiovascular Cells Enriched with Cardiac Stem Cell.

We developed a protocol for preparing cardiac cell culture from rat heart enriched with regional stem cells based on clonogenic properties and proliferation in culture in a medium with low serum content. Experiments on WAG rats with experimental ischemic myocardial damage showed that implantation of autologous regional stem cells into the left ventricle reduced the volume of cicatricial tissue, promoted angiogenesis in the damaged zone, and prevented the risk of heart failure development.

Evaluation of the Effect of Modified Bioflavonoid and Quercetin Dihydrate on Cytokine Secretion by Mitogen-Activated Mononuclear Cells.

An experimental study revealed the effect on modified bioflavonoid on the inhibition of secretion of IFN-γ and IL-2 by ConA-stimulated mononuclear cells. These changes were accompanied by an increase in the secretion of IL-17 and IL-6. Our results suggest the differentiation of CD4(+) T helper cells into Th1 and Th17 subpopulations. The reference drug quercetin dihydrate induced an insignificant change in the level of IL-2 and IL-6 and small increase in IFN-γ content. The content of IL-17 was shown to decrease above the detection limit.

Detailed analysis of therapy-driven clonal evolution of TP53 mutations in chronic lymphocytic leukemia.

In chronic lymphocytic leukemia (CLL), the worst prognosis is associated with TP53 defects with the affected patients being potentially directed to alternative treatment. Therapy administration was shown to drive the selection of new TP53 mutations in CLL. Using ultra-deep next-generation sequencing (NGS), we performed a detailed analysis of TP53 mutations' clonal evolution. We retrospectively analyzed samples that were assessed as TP53-wild-type (wt) by FASAY from 20 patients with a new TP53 mutation detected in relapse and 40 patients remaining TP53-wt in relapse. Minor TP53-mutated subclones were disclosed in 18/20 patients experiencing later mutation selection, while only one minor-clone mutation was observed in those patients remaining TP53-wt (n=40). We documented that (i) minor TP53 mutations may be present before therapy and may occur in any relapse; (ii) the majority of TP53-mutated minor clones expand to dominant clone under the selective pressure of chemotherapy, while persistence of minor-clone mutations is rare; (iii) multiple minor-clone TP53 mutations are common and may simultaneously expand. In conclusion, patients with minor-clone TP53 mutations carry a high risk of mutation selection by therapy. Deep sequencing can shift TP53 mutation identification to a period before therapy administration, which might be of particular importance for clinical trials.

Characteristics of cardiac cell cultures derived from human myocardial explants.

Primary cell cultures derived from human myocardial explants were obtained and characterized. The explant cultures contained cardiac stem cells (c-kit(+); ≈ 4%), microvascular cells (endothelial cells and pericytes), fibroblasts, and myofibroblasts. It was demonstrated that culturing of cardiac cells in cardiospheres did not promote enrichment of the cell culture with stem cells. MACS-sorted c-kit(+) cells from the explant culture were characterized by limited proliferative capacity and were capable of cardiomyogenic differentiation. The presence of microvascular cells determined general angiogenic potential of the culture.