Specific Features of Interactions between Megakaryocytic and Granulocytic Hematopoiesis Lineages and Myelofibrosis during the Acute Phase of Chronic Myeloid Leukemia, Chronic Lymphocytic Leukemia, and Multiple Myeloma.

The specific features of interactions between megakaryocytic and granulocytic hematopoiesis lineages and myelofibrosis were studied in patients with active phase (before treatment) of chronic myeloid leukemia, chronic lymphocytic leukemia, and multiple myeloma. In patients with chronic myeloid leukemia, a direct correlation was found between the severity of both early and advanced myelofibrosis and the number of megakaryocytes in the bone marrow irrespectively of the type of the bone marrow tumor. The parameters of granulocytic hematopoiesis lineage were higher in myelofibrosis. In patients with chronic lymphocytic leukemia and multiple myeloma, negative correlations between the severity of early and advanced myelofibrosis and the number of megakaryocytes in the bone marrow and platelets in the peripheral blood were found. In chronic lymphocytic leukemia, negative correlations between the severity of early and advanced myelofibrosis and the number of neutrophils in the bone marrow and peripheral blood were detected. In patients with multiple myeloma, negative correlations between the severity of advanced myelofibrosis and number of neutrophils in the bone marrow and peripheral blood were detected.

Ultrastructure of the Liver in Response to Cyclophosphamide and Triterpenoids.

Ultrastructural reorganization of liver cells after isolated injections of cyclophosphamide, betulonic acid or its ß-alanylamide, and combined treatment with the cytostatic and each of the triterpenoids is studied. Cyclophosphamide causes significant ultrastructural changes in all intracellular compartments of hepatocytes. Both triterpenoids cause moderate cytotoxic and stimulatory effects on the liver cell populations (hepatocytes, sinusoidal endotheliocytes, and Kupffer cells), when used alone. The cytotoxic effect of betulonic acid manifests in modification of the fine structure of hepatocyte mitochondria, sequestration of glycogen, intensification of autophagic processes, emergence of necrobiotic changes in hepatocytes and endotheliocytes; betulonic acid amide actively modifies the mitochondrial fine structure (hypertrophic organelles, matrix rarefaction, uneven dilatation of cristae). The effects of combinations of cyclophosphamide with betulonic acid or its amide on liver are polytarget: the cytotoxic activity of the cytostatic is potentiated towards some cells, while in other cells the regeneratory reactions are stimulated. The common cytological cytoprotective effects of betulonic acid and its amide used alone and in combination with cytostatics include stimulation of the endocytotic (pinocytotic) activities of the cells and stimulation of intracellular regeneration processes in them.

Quantitative Analysis of Red Bone Marrow Microenvironment Cells in Patients with Chronic Myeloid Leukemia, Multiple Myeloma, and Chronic Lymphocytic Leukemia in the Dynamics of Chemotherapy.

We performed comparative analysis of quantitative changes in the populations of bone marrow microenvironment cells in patients with chronic myeloid leukemia, multiple myeloma, and chronic lymphocytic leukemia in the debut, during response to chemotherapy, and during relapse/progression/loss of response. It was shown that in the active phase of hemoblastoses, the number of reticular cells and fibroblasts in trephine biopsy specimens was higher than in the phase of response to chemotherapy and than in the control group. In patients with relapse of multiple myeloma and loss of response in chronic myeloid leukemia, the percentage ratio of adipocytes in the bone marrow significantly (by 9-13-fold) increased. In addition, endotheliocytes appear in the active phase of all hemoblastoses in trephine biopsy specimens, while in the phase of response to chemotherapy and in the control group, these cells were absent. The revealed quantitative changes in bone marrow stromal cells can be taken into account during assessing the phase of hemoblastosis and effectiveness of chemotherapy.

Efficacy and safety of the use of rebamipide in the scheme of triple eradication therapy of Helicobacter pylori infection: a prospective randomized comparative study.

AIM: To evaluate the effectiveness and safety of the use of rebamipide as part of the triple eradication therapy (ET) scheme of Helicobacter pylori infection. MATERIALS AND METHODS: A prospective, randomized comparative study included 94 patients with uncomplicated H. pylori-associated stomach / duodenal ulcer. In the process of randomization, patients are divided into three groups depending on the intended therapy. The first group (n=36) received a classical triple scheme of the first-line ET (omeprazole 20 mg twice a day, amoxicillin 1000 mg twice a day, clarithromycin 500 mg twice a day) for 10 days. Patients of the second group (n=33) were assigned a classical triple scheme of ET with the inclusion of rebamipide (omeprazole 20 mg twice a day, amoxicillin 1000 mg twice a day, clarithromycin 500 mg twice a day, rebamipide 100 mg 3 times a day day) for 10 days. Patients of the third group (n=25) were assigned a classical triple scheme of ET with the inclusion of rebamipide (omeprazole 20 mg twice a day, amoxicillin 1000 mg twice a day, clarithromycin 500 mg twice a day, rebamipide 100 mg 3 times a day) in for 10 days, with the prolongation of the administration of rebamipide for the next 20 days. The effectiveness of ET was determined by the respiratory test after 6 weeks after the end of treatment. Adverse events were recorded by patients in specially developed diaries. All patients with gastric ulcer at the 6th week underwent a histological examination of the biopsy specimens of the antrum and the body of the stomach, assessing the inflammatory activity of the process on a point system in accordance with the updated Sydney system. RESULTS: Efficiency of H. pylori eradication in the first group was 77.7% (ITT), 82.3% (PP), in the second group - 81.8% (ITT), 84.4% (PP), and in the third group - 84% (ITT), 87.5% (PP). The use of rebamipide in the triple ET regimen was associated with an increase in H. pylori eradication efficiency, both with simultaneous use with the scheme [odds ratio (OR) 1.16; 95% confidence interval (CI) 0.32-4.24], and with subsequent prolonged admission (OR 1.5, 95% CI 0.34-6.7). A somewhat more pronounced dynamics of the epithelization of erosive and ulcerative changes in the mucous membrane of the stomach and duodenum to the 21st and 28th days in the third group of patients was noted. The incidence of adverse events between the groups was comparable: 22.2% in the first group, 24.2% in the second group and 20% in the third group. In the pathomorphological evaluation of biopsy specimens of patients with gastric ulcer at the 6th week after the treatment, significant differences were revealed between the first and third groups in terms of the inflammatory activity in the antrum stomach (2±0.63 vs. 1.4±0.52; p=0,0399). CONCLUSION: The inclusion of rebamipide in the classical triple scheme of H. pylori ET increases the effectiveness of treatment and does not affect the safety profile. In the post-eradication period, it is advisable to continue the use of rebamipide to potentiate the repair of the gastric mucosa and regress the inflammatory processes.

Evaluation of the efficacy and safety of the hybrid scheme for eradication therapy of Helicobacter pylori infection.

AIM: The aim is to conduct a comprehensive comparative study of the efficacy and safety of the hybrid scheme of eradication therapy (ET) in patients with peptic ulcer of the stomach or duodenum associated with Helicobacter pylori. MATERIALS AND METHODS: Materials and methods. In a prospective, randomized comparative study, 180 patients were divided into three equal groups of 60 people, depending on the prescribed 10-day ET regimen. Group 1 - the standard triple scheme (omeprazole, amoxicillin and clarithromycin); group 2 - four-component therapy with preparations of bismuth (omeprazole, tetracycline, metronidazole, bismuth tricalium dicitrate); group 3 - hybrid scheme (first 5 days: omeprazole and amoxicillin, the next 5 days: omeprazole, amoxicillin, clarithromycin, metronidazole). The effectiveness of ET was determined with the help of a breath test a month after the end of therapy. Adverse events were recorded by patients in specially developed diaries. Pharmacoeconomic analysis was carried out using the "cost-effectiveness" method with calculation of the CER coefficient. RESULTS: Results and discussion. The effectiveness of standard triple therapy was 73.3% (ITT), 75.9% (PP); four-component therapy with bismuth preparations - 78.3% (ITT), 82.4% (PP); hybrid scheme - 85% (ITT), 91% (PP). Hybrid therapy proved to be significantly more effective than standard triple therapy with a odds ratio (OR) of 3.25; 95% confidence interval (CI) 1.08-9.73 (p=0.043, χ2=4.75, p-level=0.029298). The incidence of adverse events with the use of triple, four-component and hybrid ET regimens was 15; 18.3 and 28.3% respectively. The OR of at least one adverse event in patients receiving a hybrid ET regimen compared with triple therapy was 2.24 (95% CI 0.91-5.53, p=0.0823, χ2=3.14, p-level=0.076394), and compared with the four-component therapy - 1.76 (95% CI 0.74-4.17, p=0.2804, χ2=1.68, p-level=0.194924). According to the results of the pharmacoeconomic analysis, the most profitable from an economic point of view was a hybrid ET scheme with a CER of 20.1. CONCLUSION: The conclusion. Hybrid therapy showed the greatest effectiveness in comparison with the triple and four-component ET regimens, however, the incidence of side effects in patients receiving the hybrid ET scheme was higher, although it remained within the acceptable level for use in clinical practice. Pharmacoeconomic analysis also showed the advisability of designating a hybrid ET scheme. The obtained data allow to draw a conclusion about the necessity of further study of the efficiency and safety of the hybrid ET scheme.

Involvement of the p38 MAPK signaling cascade in stress response of RAW 264.7 macrophages.

The role of the p38 MAPK signaling cascade was studied in stress response of RAW 264.7 macrophages to extremely low-intensity centimeter microwaves. Irradiation stimulated production of a number of cytokines (IL-1, IL-6, TNF-α, INF-γ and IL-10), as well as induced activation of the signaling cascades NF- κB and p38 MAPK, and enhanced expression of Hsp72 heat shock protein. In the presence of the cascade p38 MAPK inhibitor (p38 MAP kinase inhibitor XI), the stimulating effects of electromagnetic waves were abrogated either completely (for NF-κB and Hsp72) or partially (for p38 MAPK and cytokines). The results obtained are indicative of a high sensitivity of the signaling cascade p38 MAPK to the effect of low-intensity physical fields.

Experimental Study of Yeast RNA Preparation as a Possible Radioprotective Agent for Radiotherapy of Malignant Tumors.

We studied radioprotective effects of a preparation based on yeast RNA and its influence on therapeutic efficiency of ionizing radiation against transplanted tumors. Parenteral administration of yeast RNA preparation to mice in a dose of 10 mg 1 h prior to exposure to ionizing γ-radiation (137Cs) in a lethal dose (LD80/30) increased 30-day survival by 66%; by day 80, 80% of animals survived (vs. 2.5% in the control). Whole-body exposure to ionizing γ-radiation in a dose of 7 Gy significantly increased the mean lifespan of mice with experimental lung metastases or intraperitoneally transplanted leukemia L-1210 by 42 and 20.8%, respectively. RNA preparation injected to the mice with tumors 1 h before irradiation did not affect the therapeutic efficiency of ionizing radiation or significantly potentiated it (in mice with transplanted leukemia L-1210). These results suggest that yeast RNA preparation protects healthy tissues during radiotherapy of malignant tumors.

Incidence of Myelofibrosis in Chronic Myeloid Leukemia, Multiple Myeloma, and Chronic Lymphoid Leukemia during Various Phases of Diseases.

Pathomorphological study of trephinobiopsy specimens from 129 patients with lymphoproliferative and myeloproliferative diseases was carried out over the course of chemotherapy. Combinations of initial and manifest myelofibrosis (loose network of reticulin fibers and extensive network of reticulin and collagen fibers, respectively) predominated at the debut of chronic myeloid leukemia, chronic lymphoid leukemia, and multiple myeloma. Manifest myelofibrosis was detected in patients with chronic myeloid leukemia without hematological response (failure of normalization of hematological values) and in patients with progressing and relapsing multiple myeloma. Combinations of foci of initial and manifest myelofibrosis were most incident in patients with progressing and relapsing chronic lymphoid leukemia. The incidence of myelofibrosis was higher in patients with multiple myeloma and chronic lymphoid leukemia progression and relapses and in patients with chronic myeloid leukemia without hematological response than at the disease debut and in case of response to chemotherapy. The response to chemotherapy in patients with chronic myeloid leukemia and chronic lymphoid leukemia was associated with a decrease in the incidence of myelofibrosis. In patients with multiple myeloma responding to chemotherapy, the incidence of myelofibrosis did not change in comparison with the disease debut.

[The efficiency and safety of anti-Helicobacter pylori therapy in patients with concomitant chronic hepatitis C]. / Effektivnost' i bezopasnost' antikhelikobakternoi terapii u patsientov s soputstvuyushchim khronicheskim gepatitom S.

AIM: To evaluate the efficiency and safety of two eradication therapy (ET) regimens for Helicobacter pylori infection in patients with concomitant chronic hepatitis C (CHC) in relation to the stage of liver fibrosis (LF). MATERIALS AND METHODS: A prospective clinical trial was conducted in parallel groups. Group 1 included 50 HCV-negative patients with H. pylori-associated peptic ulcer of the stomach or duodenum; Group 2 consisted of 50 HCV-positive patients with H. pylori-associated peptic ulcer of the stomach or duodenum concurrent with CHC. Each group was divided in 2 subgroups according to the used triple ET (a proton pump inhibitor (PPI) in a standard dose + amoxicillin 1000 mg twice daily + clarithromycin 500 mg twice daily for 10 days) or sequential therapy (PPI in a standard dose + amoxicillin 1000 mg twice daily within the first 5 days and then PPI in a standard dose + clarithromycin 500 mg twice daily + metronidazole 500 mg twice daily for the next 5 days). LF was assessed using indirect elastometry. The efficiency of ET was evaluated by a breath test (after 4 weeks) and an analysis depending on intention-to-treat (ITT) and per-protocol (PP) treatments. A patients recorded adverse events in specially developed diaries. RESULTS: The efficiency of ET was 74% (ITT) and 80.4% (PP) in Group 1 and 76 (ITT) and 79.1% (PP) in Group 2. Both groups displayed a tendency towards an 11.9-12.4% increase in the efficiency of the sequential therapy versus the classical triple (PP) one. The rate of totally found side effects was 20% in Group 1 and 28% in Group 2. During sequential therapy, the rate of side effects was lower than that during the classical one. The efficiency of ET did not significantly depend on the stage of LF. Only the presence of concomitant type 2 diabetes mellitus and the use of macrolides (12 months before treatment) significantly lowered the efficiency of ET (OR 0,21; 95% CI 0,06-0,69, p=0,0102 and OR 0,27 95% CI 0,08-0,9, p=0,0342). LF regardless of its magnitude significantly determined the risk of adverse events during ET (OR 3,33 95% CI 1,19-9,31, p=0,0217). A group at the highest risk of adverse events included patients with liver cirrhosis (OR 4,87; 95% CI 1,01-23,5, p=0,0492). CONCLUSION: It is appropriate to prescribe a sequential ET regimen as more effective and safe for patients with concomitant CHC during therapy for H. pylori infection-associated diseases. LF increases the risk of adverse events during ET.

Structural Characteristics of Gastric Cell Populations in Chronic Gastritis and Chronic Hepatitis under Conditions of Helicobacter pylori Persistence.

Helicobacter pylori persistence in patients with chronic gastritis is associated with a complex of nonspecific structural reactions, the type of these reactions correlates with the severity of infection: catarrhal fibrotic changes in the gastric mucosa predominate in cases with manifest colonization, while the absence of H. pylori is associated with predominance of fibrotic process. Analysis of the incidence of some pathomorphological phenomena (degeneration, atrophy, metaplasia, and dysplasia of the surface epithelium) shows no relationship between the presence of H. pylori and colonization intensity. In all patients with chronic hepatitis, the gastric mucosa is involved in the pathological process; fibrosis (gastropathy) was the most common process. No appreciable correlations between the structural changes and hepatitis activity and the presence of H. pylori were detected.

[METHODOLOGICAL FEATURES OF THE pH-METRY IN PATIENTS WITH A COMBINATION OF GASTROESOPHAGEAL REFLUX DISEASE AND BRONCHIAL ASTHMA].

Conducting daily pH monitoring in patients with asthma has a number of features. Due to the fact that the introduction of the pH probe in such patients may provoke an asthma attack, necessary pre-treatment of the patients to the study. It is necessary to perform the procedure for achieving remission of asthma medication. Utility of the modified pH measuring techniques (prolonged, 48-hour) is the need to identify pathological gastroesophageal reflux in patients with asthma and evaluate the effectiveness of antisecretory drugs, allowing for improved asthma, increasing the period of remission, and to improve the quality of life of patients.

[Design of cell line stable expressing proteasomal subunit PSMD14 fused to the fluorescent protein EGFP and HTBH tag based on HEK293 cells].

A stable cell line based on HEK293 cells that expresses proteasome subunit PSMD14 fused to the fluorescent protein EGFP and HTBH tag has been selected. This chimera was shown to be incorporated completely into proteolitic active proteasomes. The created cell line can be used for further fluorescent studies of proteasomes localization in the cell.

Ultrastructure of hepatorenal cell populations in patients with HCV and HBV infection markers. Hepatorenal associations.

Hepatorenal cell populations were studied in patients with HCV and HBV infection markers and renal dysfunction. Pronounced mosaicism of ultrastructural changes in hepatocytes was associated with polymorphic cytopathic effects caused by RNA-genome hepatitis C virus and DNA-genome hepatitis B virus. The destructive component of the tubular compartment predominated in renal biopsy specimens from patients, with subsequent degeneration of the tubular epithelium associated with progressive interstitial fi brosis. Immunohistochemical studies detected HCV NS3Ag and HBcAg structural marker in the tubular epitheliocytes. An appreciable part of the structural and functional changes in the liver in patients with HCV and HBV infections was caused by the therapeutic complex, including programmed hemoperfusion.

[Effects of activation and blockde of GABA(A)-receptors on activity of the nitrergic system of the N. accumbens].

In Sprague-Dawley rats by means of in vivo microdialysis it was shown that intra-accumbal infusion of a GABA(A) receptor antagonist bicuculline (20, 60, 120 µM) dose-dependently increased the local level of extracellular citrulline (a NO co-product). This increase was almost completely prevented by intra-accumbal infusion of 7-nitroindazol (0.5 mM), a neuronal NO synthase inhibitor. Intra-accumbal infusion ofa GABA(A) receptor agonist muscimol at concentrations of 10, 50 µM did not affect the citrulline extracellular level in this brain area, whereas at a concentration of 150 µM, muscimol decreased the accumbal citrulline extracellular level. Intra-accumbal infusion of NMDA (100 µM), an NMDA receptor agonist, increased the local level of extracellular citrulline, and intra-accumbal muscimol infusions (10, 50, 150 µM) dose-dependently decreased the NMDA-dependent citrulline rise. The data obtained indicate for the first time that in the n. accumbens, GABA(A)-receptors might regulate basal and NMDA-evoked activity of accumbal nitrergic system.

Cytochrome P450 4F2 polymorphism in patients with liver cirrhosis.

1297C/T polymorphism of CYP4F2 gene was studied in 108 patients with chronic liver diseases. No significant correlation with predisposition to rapid liver cirrhosis was revealed without consideration for cirrhosis etiology (OR=0.93, 95% CI=0.28-2.99, p=0.885). In patients with viral cirrhosis, a tendency to association of 1297T allele genotypes with rapid cirrhosis development was found (OR=6.0, 95% CI=0.28-382.64, p=0.222). At the same time, CYP4F2 1297T allele was associated with mild (Child-Pugh class A-B) cirrhosis (OR=2.9, p=0.044).

Role of lipid infiltration of hepatocytes in the morphogenesis of chronic hepatitis C.

Immunohistochemical and PCR analysis of the structure of lipid-containing hepatocytes and replication of chronic hepatitis C viral particles revealed no positive correlation between the presence of HCV RNA in blood specimens, number of infected hepatocytes (by expression of HCV NS3Ag) with clinical biochemical parameters and intensity of structural changes in biopsy specimens. Small-vesicular subcytolemmal lipid infiltration of hepatocytes was detected, presumably associated with hepatitis C virus replication phase. Characteristic subcytolemmal location of the lipid droplets can reflect their involvement in virus transport from the cell and indicate a virus-induced nature of lipid metabolism disorders.

Cytochrome P450 2D6 polymorphism is a molecular genetic marker of liver cirrhosis progression.

Patients with infectious viral or toxic cirrhosis of the liver participated in complex clinical pathomorphological and molecular-genetic study aimed at the search for markers of predisposition to accelerated liver fibrosis, in which the xenobiotic biotransformation system is involved. The results demonstrate association between CYP2D6 (1846G/A) genotype and rapid cirrhosis development and indicate the necessity of studying the mechanisms underlying this association.

Ultrastructural and stereological study of the liver in chronic mixed HCV+HBV infection.

The stereological values of the structural density of hepatocyte cytoplasmic organelles were similar in chronic mixed HCV+HBV infection irrespective of the form of HBV infection (seropositive or latent). High incidence of HBsAg- and HBeAg-negative forms of HBV infection determines the leading role of PCR diagnosis and studies of liver biopsy specimens with detection of HBcAg structural markers and HBV DNA in native liver tissue in the diagnosis of mixed HCV+HBV infection.

Structural modification of the gastrointestinal epithelium during immune-dependent granulomatosis.

We present results of pathomorphological study of the mucosa epithelium in various parts of the gastrointestinal tract in Crohn's disease. Particular attention was paid to structural transformation of epithelial cells. Barrier dysfunction of the gastrointestinal mucosa was shown to be one of the major pathogenetic factors for Crohn's disease.

Ultrastructural modification of the endothelium in placental insufficiency and microangiopathies.

Structural reorganization of endotheliocytes was studied on models of various pathological processes: placental dysfunction, glomerular pathology, vibration syndrome, antiphospholipid syndrome, and diffuse angiokeratoma, all of these characterized by endothelial insufficiency. Universal modification of endothelial associations was revealed. It included a chain of stereotypical reactions: from degeneration alternating with compensatory hypertrophy to subsequent atrophy and death of endotheliocytes. The time course of the process was confirmed by the results of light microscopy in combination with ultrastructural examination and by evaluation of the biosynthetic reactions by in vitro radioautography.