Cytokine Profile of CCR6+ T-Helpers Isolated from the Blood of Patients with Peptic Ulcer Associated with Helicobacter pylori Infection.

We previously found that the number of CCR6+ T-helpers with the phenotype of effector/effector memory T cells increases in the blood of patients with H. pylori-associated peptic ulcer. The mature phenotype and the expression of the chemokine receptor CCR6, which is involved in migration of lymphocytes to the inflamed mucous membrane of the gastrointestinal tract, suggests that these cells are involved in the immune response observed in this clinical condition. To better understand the pathogenetic role of these cells, it is necessary to study their functional activity, specifically, the production of pro-inflammatory cytokines involved in the pathogenesis of the disease. The aim of the study was to evaluate changes in the blood level of pro-inflammatory types of mature CCR6+ T-helpers in H. pylori-associated peptic ulcer disease. MATERIALS AND METHODS: CCR6+ T-helpers were isolated from the blood by using immuno-magnetic separation adapted to this study. The number of T-helpers of types 1 and 17 (Th1 and Th17) and cells with mixed properties of Th1 and Th17 (Th1/Th17) was determined by intracellular cytokine assay. RESULTS: Initially, we planned to activate unseparated peripheral blood mononuclear cells ex vivo and evaluate the number of cytokine producers among mature CCR6+ T-helper cells by gating them during the flow cytometry. However, dramatic changes in the phenotype of T-helpers upon activation did not allow us to reliably identify the cells of interest. Subsequently, we used a two-stage immunomagnetic separation procedure to obtain functionally active mature CCR6+ T-helpers with a purity of >90%. The quantitative yield of these cells from the blood of patients with gastric and duodenal peptic ulcer associated with H. pylori was 9 times higher than that from the blood of healthy donors. Activation of CCR6+ T-helpers purified from blood of ulcer patients revealed an increased content of Th1, Th17, and Th1/Th17. One ml of the patient's blood yielded 18.1 times more CCR6+ Th1, 19.4 times more CCR6+ Th17, and 21.1 times more CCR6+ Th1/Th17 compared with the blood of healthy subjects. CONCLUSION: The content of mature CCR6+ T-helper cells with pro-inflammatory activity significantly increases in the blood of patients with peptic ulcer associated with H. pylori infection.

[The changes of left ventricular longitudinal systolic function depending on hypertension and its control: analysis in a population].

Aim To evaluate changes in left ventricular (LV) systolic function by LV myocardial global longitudinal strain (GLS) and global strain rate (GSR) in patients with arterial hypertension (AH) and based on the effectiveness of blood pressure (BP) control in a Russian population sample of individuals older than 55 years.Materials and methods This cross-sectional study was a population-based cohort study (HAPIEE, Novosibirsk). LV myocardial GLS and GSR were studied by echocardiography in a random sample (n=1004, 55-84 years). Statistical analysis was performed with multivariate models of logistic regression.Results AH prevalence in the study sample was 78.4 %. Mean GLS was 19.1 % (SD, 4.07), which was less for men than for women (p=0.001). Mean GSR was 0.86 s-1 (SD, 0.19) and was not different between men and women. In individuals with AH, the GLS absolute value was lower than in normotensive people (18.8 %; SD, 4.04 vs. 20.2 %; SD, 4.03, p˂0.001); these differences remained irrespective of the age, gender, body weight index (BWI) (p=0.027), and LV mass index (p=0.05). When people with AH were divided into groups, the lowest GLS absolute values were observed among "ineffectively treated" or not receiving any therapy individuals (p<0.001 vs. normotensive group). AH 1.6 times increased the risk of LV GLS decrease. In individuals with AH, the GSR absolute value was lower than in normotensive people (- 0.85 s-1 (SD, 0.19) vs.- 0.92 s-1 (SD, 0.18), p<0.001); this difference remained in multivariate models. The lowest GSR absolute values were observed in the "ineffectively treated" group irrespective of the gender, age, and BWI (p=0.036 vs. normotensive group). AH doubled the risk of LV GSR decrease, which could be partially explained by the contribution of BWI and myocardial mass index.Conclusion In this population sample, LV GLS and GSR were independently associated with AH. The lowest GLS and GSR values were observed for ineffectively treated" individuals with AH, which may reflect an early decline of LV systolic function with inadequate control of AH.

Recombinant Human Erythropoietin Proteins Synthesized in Escherichia coli Cells: Effects of Additional Domains on the in vitro and in vivo Activities.

The aim of this work was to compare biological activities of three variants of bacterially expressed human recombinant erythropoietin (EPO) with additional protein domains: 6His-s-tag-EPO protein carrying the s-tag (15-a.a. oligopeptide from bovine pancreatic ribonuclease A) at the N-terminus and HBD-EPO and EPO-HBD proteins containing heparin-binding protein domains (HBD) of the bone morphogenetic protein 2 from Danio rerio at the N- and C-termini, respectively. The commercial preparation Epostim (LLC Pharmapark, Russia) produced by synthesis in Chinese hamster ovary cells was used for comparison. The EPO variant with the C-terminal HBD domain connected by a rigid linker (EPO-HBD) possesses the best properties as compared to HBD-EPO with the reverse domain arrangement. It was ~13 times more active in vitro (i.e., promoted proliferation of human erythroleukemia TF-1 cells) and demonstrated a higher rate of association with the erythropoietin receptor. EPO-HBD also exhibited the greatest binding to the demineralized bone matrix (DBM) and more prolonged release from the DBM among the four proteins studied. Subcutaneous administration of EPO-HBD immobilized on DBM resulted in significantly more pronounced vascularization of surrounding tissues in comparison with the other proteins and DBM alone. Therefore, EPO-HBD displayed better performance with regard to all the investigated parameters than other examined EPO variants, and it seems promising to study the possibility of its medical use.

[On the Issue of Register of Patients of Gynecological Profile with Anemia Syndrome].

The abundant menorrhagia is the most prevalent cause of iron-deficient conditions in women. Though diagnostic of anemia is a simple task deficiency of knowledge in the area of hematology favors spreading and aggravation of anemia processes. The implementation of the register of patients with anemia syndrome into practical activities of physicians of all specialties is a modern direction of studying the given pathology. The proposed system of management of treatment diagnostic process permitted to optimize data collection related to disease, to ameliorate diagnostic search and to rationalize treatment of patients.

[ANEMIA OF CHRONIC DISEASES AS A SYSTEMIC MANIFESTATION OF CHRONIC PULMONARY OBSTRUCTIVE DISEASE].

Anemia of chronic disease (ACD) is one of the most frequent forms of anemia is often observed in patients with infections, cancer and chronic inflammatory or autoimmune diseases. The underlying mechanisms are complex and include dysregulation of iron homeostasis and erythropoietin production, impaired proliferation of erythroid progenitor cells and reduced life span of red blood cells. Moreover, ACD is often superimposed by malnutrition, bleeding and renal failure. ACD is mediated through inflammatory cytokines and characterized by low serum iron (hypoferremia) and often increased reticuloendothelial stores of iron. ACD is usually normocytic, normochromic anemia, but it can become microcytic and hypochromic as the disease progresses. Hepcidin, the main regulator of iron homeostasis and its synthesis, is inhibited by iron deficiency and stimulated by inflammation. In many patients the disease is associated with several extrapulmonary manifestations regarded as the expression of the systemic inflammatory state of chronic obstructive pulmonary disease (COPD). Recent studies showed that anemia in patients with COPD is more frequent than expected, with its prevalence ranging from 8 to 33%. Systemic inflammation may be an important pathogenic factor, but anemia in COPD can also be the result of a number of factors, such as the treatment with certain drugs (angiotensin-converting enzyme inhibitors or theophylline), endocrine disorders, acute exacerbations and oxygen therapy. Anemia in COPD patients is strongly associated with increased functional dyspnea, decreased exercise capacity and is an independent predictor of mortality. Treatment options to correct anemia used in other chronic diseases, such as congestive heart failure, cancer or chronic kidney disease have not been explored in COPD (i.e. erythropoietic agents, iron supplements or combined therapy). It is not known whether treating the underlying inflammation could improve hematological characteristics. It is important to develop basic diagnostic modalities for this group of patients and formulate methods of anemia correction.

Design of a stable cell line producing a recombinant monoclonal anti-TNFα antibody based on a CHO cell line.

Recombinant monoclonal antibodies (mAbs) against tumor necrosis factor alpha are widely used in the biopharmaceutical therapy of autoimmune diseases. Currently, a large number of drugs based on these antibodies are available. Accordingly, the development of these products for the Russian market is an important goal. The aim of the current study is to describe the development of one such technology. CHO-DG44-derived cell lines producing mAb were developed using two strategies, one based on individual clones and the other based on cell pools. To obtain recombinant cell lines with highly amplified genes of interest, the clones underwent dihydrofolate reductase-mediated gene amplification. Using the best strategy for the selection and amplification of mAb-producing clones, we achieved the production of more than 1 g/L in small scale, non-optimized conditions.

[ANEMIC SYNDROME IN PATIENTS WITH COMMUNITY-ACQUIRED PNEUMONIA].

Community-acquired pneumonia remains a most widespread acute infectious disease of socio-economic significance all over the world. Up to 30% of the patients present with anemia responsible for the unfavourable prognosis and elevated mortality. Not infrequently, anemia is not diagnosed during the hospital stay und therefore remains uncorrected. Severe anemia results in enhanced hypercapnia and slowed maturation of red blood cells in the bone marrow which facilitates the development of ischemic syndrome. Hepcidin, a mediator of inflammation and iron-regulatory hormone, plays an important role in the clinical course of community-acquired pneumonia. Hepsidin production increases during inflammation; it suppresses erythtropoiesis and depletes the iron depot leading to so-called anemia of inflammation. Hypoxia and anemia activate erythtropoiesis, and the released erythropoietin inhibits hepsidin production. During pneumonia resolution, hepsidin promotes recovery from anemia by activating iron absorption. The curreni literature contains few data on the use of hepcidin as a diagnostic marker of anemia. The necessity oftreating anemia in patients with pneumonia under hospital conditions is a matter of discussion. Direct involvement of hepcidin in iron metabolism creates a prerequisite for the treatment of anemia. Medicamental suppression of its activity by stimulating erythtropoiesis can facilitate normalization of iron metabolism and restoration of hemoglobin level.

[The use of lithium electrophoresis combined with balneotherapy in hypertension depending on the type of central hemodynamics]. / Primenenie élektroforeza litiia v sochetanii s bal'neoterapiei pri gipertonicheskoi bolezni v zavisimosti ot tipa tsentral'noi gemodinamiki.

Central chest rheography was used to study the central hemodynamics in 62 patients with hypertensive disease (grades I-II) before and after treatment. 23 patients (hyperkinetic hemodynamics) were treated with oxygen baths and endonasal lithium chloride electrophoresis; 20 with en--and hypokinetic types of hemodynamics received CO2 baths and endonasal lithium chloride electrophoresis. It is concluded that lithium chloride electrophoresis in association with balneotherapy increases the therapeutic effect in hypertensive disease, especially in patients with the hyperkinetic type of blood circulation.