[Physiological Media in Studies of Cell Metabolism].

Changes in cell metabolism accompany the development of a wide spectrum of pathologies including cancer, autoimmune, and inflammatory diseases. Therefore, usage of inhibitors of metabolic enzymes are considered a promising strategy for the development of therapeutic agents. However, the investigation of cellular metabolism is hampered by the significant impact of culture media, which interfere with many cellular processes, thus making cellular models irrelevant. There are numerous reports that show that the results from in vitro systems are not reproduced in in vivo models and patients. Over the last decade a novel approach has emerged, which consists of adaptation of the culture medium composition to that closer to the composition of blood plasma. In 2017-2019, two plasma-like media were proposed, Plasmax and HPLM. In the review, we have summarized the drawbacks of common media and have analyzed changes in the metabolism of cells cultivated in common and plasma-like media in normal and pathological conditions.

[WAYS TO SOLVE THE PROBLEM OF HOME PARENTERAL NUTRITION IN PATIENTS WITH SHORT BOWEL SYNDROME].

The purpose of the work is to identify the most optimal ways to solve the problem of home parenteral nutrition for patients with short bowel syndrome.Basics. Discussion of the current situation will help optimize the work of the Orphan Diseases Center, lead to a greater introduction of online technology, and aim at the search and implementation of new methods of treating patients in this category.Conclusion. The most optimal strategy has been proposed to improve the interaction between various specialists and patients in need of long-term parenteral nutrition.

[Histological remission of ulcerative colitis with combined anti-cytokine and cell therapy].

AIM: To conduct comparative analysis of histological remission in patients with moderate and severe ulcerative colitis (UC), receiving biological therapy vedolizumab, mesenchymal stem cell (MSC) treatment and combined stem cells and vedolizumab therapy. MATERIALS AND METHODS: We studied biopsies of 75 patients with total or left-sided moderate and severe ulcerative colitis, divided into groups depending on treatment. The first group of UC patients (n=29) received stem cell therapy 2 mln per kg; the second group of UC patients (n=27) received vedolizumab and the third group (n=19) MSC and vedolizumab. The efficacy of treatment was assessed by C reactive protein (CRP), Mayo score (MS), fecal calprotectin (FC) and Geboes score (GS). RESULTS: We determined medium correlation between basic FC and MS before treatment (r=0.6605, p0.05). After 12 weeks of treatment in the first group of UC patients (n=29) CRP was 7.82.1 mg/l, FC 409.344.85 g/g, medium GS 1.20.1 points. After 12 weeks of treatment in the second group of UC patients (n=27) CRP was 8.41.4 mg/l, FC 435.547.3 g/g, medium GS 1.350.15 points. After 12 weeks of treatment in the third group of UC patients (n=19) CRP was 6.41.1 mg/l, FC 290.617.5 g/g, medium GS 0.90.1 points. We proved strong direct relationship between FC and GS after 12 weeks of treatment in UC patients, receiving MSC (r=0.8392, p0.05). The statistically significant majority of patients, achieved histological remission, have less than 5-year duration of disease. CONCLUSION: Our study showed that clinical and endoscopic remission in UC patients does not always correlate with histological remission. Combined anti-cytokine and stem cells therapy contributes to achieve deep remission and decrease mucosa inflammation rather than single MSC or vedolizumab treatment. Deep remission could be achieved by earlier start of biological therapy. FC could be a predictor and marker of mucosa healing and histological remission.

[Safety of enhanced recovery after surgery (ERAS) protocol in the treatment of patients undergoing pancreatoduodenectomy]. / Otsenka bezopasnosti protokola uskorennogo vosstanovleniya (ERAS) v lechenii bol'nykh, perenesshikh pankreatoduodenal'nuyu rezektsiyu.

OBJECTIVE: To study the effectiveness of enhanced recovery after surgery (ERAS) protocol versus traditional perioperative management in patients with hepatopancreatobiliary tumors undergoing pancreatoduodenectomy. MATERIAL AND METHODS: The study included 111 patients who have undergone pancreatoduodenectomy between January 2014 and December 2019. Patients were divided into 2 groups: perioperative ERAS protocol (85 patients) and traditional treatment (26 patients). Postoperative complications, length of hospital-stay and incidence of readmissions were analyzed. RESULTS: Mean length of hospital-stay for ERAS protocol was 13.4±7.6 days, conventional management - 16.5±7.5 days (p=0.004). Postoperative 30-day mortality was 8.24 and 7.7% in both groups, respectively (p=1.0). Intraoperative blood loss was significantly less in the ERAS group (248.24±214.0 vs. 321.15±155.0 ml, p=0.004). Overall incidence of postoperative complications was 56.5% and 65.4%, respectively (p=0.420). However, incidence of Clavien-Dindo grade IV complications was significantly higher in case of traditional treatment (19.2 vs. 4.7%, p=0.015). Readmission rate within 30 days was slightly less in the ERAS group (6.4 vs. 20.8%, p=0.052). CONCLUSION: Enhanced recovery after surgery protocol is safe, reduces the number of postoperative complications, length of hospital-stay and rate of readmissions.

[METABOLIC COMPLICATIONS IN PARENTERAL-ENTERAL NUTRITION].

Analysis and prevention of complications of parenteral and enteral nutrition recently receiving more attention. This is because a full infusion-nutritive correction ensures shorter postoperative period and can be used as the protector of metabolic damage, aggravating, and sometimes determining operating.

[GENETIC ASPECTS OF INFLAMMATORY BOWEL DISEASE AND TREATMENT USING SURGICAL AND NUTRITIONAL CORRECTION].

Currently nutrigenetics is an independent high technology. Patients with genetically caused and complicated ulcerative colitis needs earlier surgery treatment than patient without genetically caused ulcerative colitis. Patients with no genetic predisposition can be treated conservatively over a long time.

[THE CLINICAL ORGANIZATIONAL SUBSTANTIATION OF NEW TECHNOLOGY OF HOSPITAL PSYCHIATRIC CARE].

The new technology of hospital psychiatric care, developed and implemented in the Mordovia republican clinical hospital, permits resolving problems of hospitalism, lethality, pharmaceutical resistance and others. The essence of this technology is in staging of hospital care under condition of intensification and standardization of curative diagnostic process, implementation of complex approach to treatment of psychiatric disorders. The patient sequentially passes through three stages: intensive diagnostics and intensive treatment (intensive care department, intensive therapy department), supportive therapy (general psychiatric department); rehabilitation measures (curative rehabilitative department). The concentration of resources at the first stage, application of intensive therapy techniques permit in the shortest period to arrest acute psychotic symptomatic. The described new technology of hospital psychiatric care permits enhancing effectiveness of treatment, significantly shorten period of hospitalization (37.5 days), to obtain lasting and qualitative remission, to rehabilitate most fully social working status of patient and to significantly decrease lethality.

[CHANGE OF ANTICOAGULANT EFFECT OF RIVAROXABAN DURING A DAY].

The article presents an assessment of anticoagulant effect of rivaroxaban in 35 patients with thromboembolic diseases. The results of expressed anticoagulant effect of rivaroxaban were obtained during a day (or more). The data of increase in sensitivity to thrombomodulin indicated about work enchancement of protein C system against the background of rivaroxaban therapy.

[Comparative analysis of the immune response to DNA constructions encoding hepatitis C virus nonstructural proteins].

A promising approach to construction of antiviral vaccines consists in activation of cellular immunity with the DNA vaccines. The goal of this work was to evaluate the efficacy of genetic immunization of mice with DNA pcNS3-NS5B encoding five hepatitis C virus (HCV) nonstructural proteins: NS3, NS4A, NS4B, NS5A, and NS5B in comparison with plasmids containing genes of same individual nonstructural proteins. The DNA constructions were injected intramuscularly in DBA mice three times. The humoral immune response was assessed with ELISA; cellular immune response--in blast transformation reaction, by quantitation of CD4+ and CD8+ T cell proliferation using flow cytofluorometry, by intracellular synthesis and secretion of IFN-gamma and IL-2 in ELISpot and ELISA. It was found that the functionally active T cell response was achieved to antigens presenting NS3, NS4, NS5A, and NS5B epitopes of different HCV genotypes in response to pcNS3-NS5B plasmid and was stronger than that to plasmids carrying individual genes. A high proliferation rate of CD4+ T cells, secretion of IL-2 and IFN-gamma, induction of anti-NS3 and anti-NS5B IgG2a were demonstrated. These findings indicate that DNA construction pcNS3-NS5B is one of promising candidates for anti-HCV vaccine developing.

[Prospects for nutritional support of patients with ulcerative colitis].

The article presents the data on the assessment of the degree of severity of ulcerative colitis and the selection of nutritional support with the account of the violations of the nutrition status with ulcerative colitis.

[Solved and unsolved problems of nutritional support in surgical gastroenterology].

The article has revealed the relevance of nutriciology as a science, classification and types of nutritional support. Nutritional support--diagnostics and the metabolic correction of arising violations of various kinds of homeostasis, aimed at ensuring the functioning of the systems of protein-energy synthesis and intermediate exchange of nutrients, vitamins, macro--and microelements, or a temporary replacement therapy of their failure. The notions of "lack of food", "artificial" food, protein-energy insufficiency with the exception of the syndromes of malabsorption, alimentary anemia, the effects of protein-energy malnutrition, consumptive disease, starvation have been interpreted. It was noted that nutriciology has taken place as a science. The problems of prognostification of nutritional deficiency, assessment of nutritional risk, the organization of nutritional support in the hospital and at home have been presented. Some fundamental bases of enteral probe feeding have been described.

[Immunological criteria for evaluation of nutritional support in ulcerative colitis].

The article presents the data on the assessment of nutritional status, taking into account humoral immunologic criteria of patients with ulcerative colitis.

[Use of thrombin generation assay for the evaluation of coagulation and anticoagulant activity of hemostasis system in patients with abdominal sepsis].

Generally recognized factor, which complicates the course of sepsis, is the development of hypercoagulation syndrome. The increase of thrombin coagulation indicates on the elevation of risk of thrombus formation in microcirculation vessels, which could cause the formation of multiple organ failure. The thrombin generation assay is a new method of the evaluation of homeostasis system status. The test reflects the fermentation activity of thrombin and shows the functional condition, which arises in the interaction of procoagulant and anticoagulant. The diagnosis of generalized peritonitis had 30 patients (18 men and 12 women, aged 61+/-18,3 years) and they were included in the research. It was shown, that the use of thrombin generation assay in patients with the abdominal sepsis could give the well-timed analysis of hypercoagulation changes and the assessment of protein C system investment in the thrombin generation.

[Assessment of endogenous thrombin potential and influence on it of different regimens of heparin therapy in patients with abdominal sepsis].

Abdominal sepsis (AS) is a systemic inflammatory reaction of organism in response to the development of infectious process in organs of the abdominal cavity. The course of AS is complicated by hypercoagulation syndrome which facilitates progression of endogenous intoxication. The investigation included 26 patients (14 men and 12 women, mean age 65.5 +/- 16.4 years) with AS. The test of thrombin generation (TTG) used in patients with AS allows assessment of changes in the hemostasis system and control of the heparin dose. It was established that TTG revealed elevated endogenous thrombin potential of blood practically in all patients with AS. In the patients given 25 000 IU of heparin the TTG indices showed a reliably decreased endogenous thrombin potential and peak thrombin concentration as compared with a group of patients given 10 000 IU of heparin.

Physiological Media in Studies of Cell Metabolism.

Changes in cell metabolism accompany the development of a wide spectrum of pathologies including cancer, autoimmune, and inflammatory diseases. Therefore, usage of inhibitors of metabolic enzymes are considered a promising strategy for the development of therapeutic agents. However, the investigation of cellular metabolism is hampered by the significant impact of culture media, which interfere with many cellular processes, thus making cellular models irrelevant. There are numerous reports that show that the results from in vitro systems are not reproduced in in vivo models and patients. Over the last decade a novel approach has emerged, which consists of adaptation of the culture medium composition to that closer to the composition of blood plasma. In 2017‒2019, two plasma-like media were proposed, Plasmax and HPLM. In the review, we have summarized the drawbacks of common media and have analyzed changes in the metabolism of cells cultivated in common and plasma-like media in normal and pathological conditions.

[Comparison of measles incidence in different age groups with the intensity of specific humoral immunity in healthcare workers in Moscow].

INTRODUCTION: The study of the intensity of humoral immunity to the measles virus (Paramyxoviridae: Morbillivirus) (MV) remains relevant due to the ongoing registration of the cluster cases of measles (including nosocomial cases).The aim of the study was to analyze correlation between the intensity of the humoral immunity to measles virus and the measles incidence in different age groups of the Moscow residents. MATERIAL AND METHODS: A simple analytic study was carried out under the framework of measles prevention improvement program conducted in the subjects of the Russian Federation. The study included the analysis of the reasons for the increase in the proportion of seronegative individuals and the comparison of the results of seromonitoring with the data of age-related measles incidence. Throughout 2018, total 1,855 blood serum samples obtained from employees of a large hospital center in Moscow who were previously vaccinated against measles were tested for IgG antibodies (ABs) to MV. The subjects were divided into 11 groups according to their age. Statistical analysis of the obtained data was performed using nonparametric criteria. RESULTS AND DISCUSSION: The proportion of seronegative individuals among the staff of the medical facility decreased with age, from 38.5% in persons aged 19-23 to 0% among employees aged 64 and over. Persons aged 19-43 years were the most susceptible to measles infection, which was confirmed also by the highest incidence rate at these age groups (from 6.93 to 14.03 per 100,000 population). The age group most protected against measles was those over 64 years of age, who were all seropositive and had the lowest rates of incidence. This is probably due to the fact that by this age most of them have had a measles infection and have formed a sufficient immune response. At the same time, younger individuals living in the post-elimination era have only vaccine-induced ABs to MV, the level of which declines rapidly to non-protective values in the absence of boosterization due to the exposure to the wild-type strain of the pathogen. CONCLUSION: The monitoring of the MV-specific IgG- AB levels is required for employees in the medical institutions under the age of 35. Administration of the single dose of the appropriate vaccine is recommended for seronegative individuals.

[Cellular defense systems against oxidative and ER stresses: mechanisms of regulation and influence of hepatitis C virus].

Hepatitis C virus is one of the most spread and dangerous human pathogens. In most cases hepatitis C develops into chronic diseases which in many cases escape antiviral therapy and is associated with contributed to progression of various virus-associated organ damage and disorders including liver fibrosis, steatosis and hepatocellular carcinoma. Many of these diseases are currently linked to oxidative and ER stresses induced by the viral proteins. At the same time, hepatitis C virus disturbs systems protecting cells from these stresses, thus avoiding their effect on processes of the virus life cycle. Here, we have analyzed recent data on mechanisms of the cellular defense system functioning in infected and uninfected cells. In addition, major data on the hepatitis C virus genome structure and main functions of the virus proteins have been summarized briefly.

[DNA immunization with a plasmid containing gene of hepatitis C virus protein 5A (NS5A) induces the effective cellular immune response].

In spite of extensive research, no effective vaccine against hepatitis C virus (HCV) has been developed so far. DNA immunization is a potent technique of vaccine design strongly promoting the cellular arm of immune response. The genes encoding nonstructural HCV proteins (NS2-NS5B) are promising candidates for vaccine development. NS5A is a protein involved in viral pathogenesis, in the induction of immune response, and probably in viral resistance to interferon treatment. The objective of this study was to construct a DNA vaccine encoding NS5A protein and evaluate its immunogenicity. A plasmid encoding a full-size NS5A protein was produced using the pcDNA3.1 (+) vector for eukaryotic expression system. The expression of the NS5A gene was confirmed by immunoperoxidase staining of the transfected eukaryotic cells with anti-NS5A monoclonal antibodies. Triple immunization of mice with the plasmid vaccine induced a pronounced cellular immune response against abroad spectrum of NSSA epitopes as assessed by T-cell proliferation andsecretion of antiviral cytokines IFN-gamma and IL-2. In in vitro T-cell stimulation experiments, NS5A-derived antigens were modeled by synthetic peptides, recombinant proteins of various genotypes, and phages carrying exposed NS5A peptides. A novel immunomodulator Immunomax showed high adjuvant activity in DNA immunization. The data obtained indicate that the suggested DNA construct has a strong potential in the development of the gene vaccines against hepatitis C.

Response of Human Small Intestinal Epithelium to Fractionated Irradiation: Dynamical Modeling Approach.

A biologically motivated mathematical model of the dynamics of the small intestinal epithelium in humans treated with fractionated radiotherapy has been developed and is further investigated here. This model, originating from our previous work, is implemented as a system of nonlinear ordinary differential equations, in which the variables and parameters have a clear biological meaning. The model also includes, as input, the key parameters of fractionated irradiation. The modeling results on the dynamical response of the human normal small intestinal epithelium to fractionated radiation therapy regimens were in agreement with the corresponding empirical data, which, in turn, demonstrates the capability of the developed model for predicting the dynamics of this vital body system in humans receiving fractionated radiotherapy. It is also revealed that the cumulative damage effects of hypofractionated radiation therapy regimens on the human normal small intestinal epithelium are somewhat less pronounced than those of conventional fractionated radiation therapy regimens with the same total doses.

[New furano- and pyrrolo[2,3-d]pyrimidine nucleosides and their 5'-triphosphates: synthesis and biological properties].

Bicyclic furano[2,3-d]pyrimidine ribonucleosides were synthesized by Pd(0)- and CuI-catalyzed coupling of 5-iodouridine with terminal alkynes. The treatment of the resulting nucleosides with ammonia or methylamine solution in aqueous alcohol resulted in pyrrolo- and N(7)-methylpyrrolo[2,3-d]pyrimidine nucleosides. 5'-O-Triphosphates of bicyclic nucleosides were obtained by the treatment of the nucleosides with POCl3 in the presence of a "proton sponge." The 5'-O-triphosphates are not substrates for HCV RNA-dependent RNA polymerase, but are effective substrates for HCV RNA helicase/NTPase and did not inhibit ATP hydrolysis. Only 3-(beta-D-ribofuranosyl)-6-decyl-2,3-dihydrofuro-[2,3-d]pyrimidin-2-one showed a moderate anti-HCV activity in the HCV replicon system and efficiently inhibited replication of bovine viral diarrhea virus (BVDV) in KCT-cells, other compounds being inactive. None of the compounds were cytotoxic within the tested range of concentrations.