Comparative Whole-Transcriptome Profiling of Liver Tissue from Wistar Rats Fed with Diets Containing Different Amounts of Fat, Fructose, and Cholesterol.

Differential expression of 30,003 genes was studied in the liver of female Wistar rats fed with isocaloric diets with the excess of fat, fructose, or cholesterol, or their combinations for 62 days using the method of whole-transcriptome profiling on a microchip. Relative mRNA expression levels of the Asah2, Crot, Crtc2, Fmo3, GSTA2, LOC1009122026, LOC102551184, NpY, NqO1, Prom1, Retsat, RGD1305464, Tmem104, and Whsc1 genes were also determined by RT-qPCR. All the tested diets affected differently the key metabolic pathways (KEGGs). Significant changes in the expression of steroid metabolism gene were observed in the liver of animals fed with the tested diets (except the high-fat high fructose diet). Both high-fat and high-fructose diets caused a significant decrease in the expression of squalene synthase (FDFT1 gene) responsible for the initial stage of cholesterol synthesis. On the contrary, in animals fed with the high-cholesterol diet (0.5% cholesterol), expression of the FDFT1 gene did not differ from the control group; however, these animals were characterized by changes in the expression of glucose and glycogen synthesis genes, which could lead to the suppression of glycogen synthesis and gluconeogenesis. At the same time, this group demonstrated different liver tissue morphology in comparison with the animals fed with the high-fructose high-fat diet, manifested as the presence of lipid vacuoles of a smaller size in hepatocytes. The high-fructose and high-fructose high-fat diets affected the metabolic pathways associated with intracellular protein catabolism (endocytosis, phagocytosis, proteasomal degradation, protein processing in the endoplasmic reticulum), tight junctions and intercellular contacts, adhesion molecules, and intracellular RNA transport. Rats fed with the high-fructose high-fat or high-cholesterol diets demonstrated consistent changes in the expression of the Crot, Prom1, and RGD1305464 genes, which reflected a coordinated shift in the regulation of lipid and carbohydrate metabolisms.

[Role of contrast enhancement in ultrasound examination of the aortic abdominal portion]. / Rol' kontrastnogo usileniia pri ul'trazvukovom issledovanii briushnogo otdela aorty.

Analysed herein are the data of scientific literature over several last years concerning the use of contrast-enhanced ultrasonography (CEUS) of the abdominal portion of the aorta. Based on the results of Russian and foreign studies, the authors analyse feasibility of using CEUS in various pathological conditions of the aorta, as well as sensitivity, specificity, and prognostic value of the method. Also discussed are advantages and limitations of CEUS compared with other imaging techniques. This is followed by determining appropriate approaches to using the methodology of CEUS in examination of patients after endoprosthetic repair of abdominal aortic aneurysms. Finally, described is own experience of using CEUS in examination of a patient with chronic renal insufficiency after aortic endoprosthetic repair.

Transcription profile analysis of the endometrium revealed molecular markers of the personalized 'window of implantation' during in vitro fertilization.

To determine the most informative markers for assessing the functional state of endometrium during the 'window of implantation' and creating a model for assessment of the readiness of endometrium for embryo implantation. Forty-seven women with tubal infertility and a successful IVF pregnancy participated in the study. Pipelle endometrial sample was performed during the supposed 'window of implantation' in natural cycle with subsequent histological study, and transcriptional profile of genes GPX3, PAEP, DPP4, TAGLN, HABP2, IMPA2, AQP3, HLA-DOB, MSX1, POSTN determined by real-time quantitative polymerase chain reaction (qRT-PCR). Differences in the level of mRNA expression of all the studied genes in the receptive endometrium were found in comparison to the prereceptive one, which allowed us to classify two functional states of the endometrium. The results of histological examination responded to the stage of maturation of the endometrium in 78.7% of cases. Receptive endometrial status can be determined based on the integral evaluation of mRNA expression level of 4 PAEP, DPP4, MSX1, and HLA-DOB genes. The model for determining a personalized `window implantation' is offered for practical application in ART.

Method of Selection of Bacteria Antibiotic Resistance Genes Based on Clustering of Similar Nucleotide Sequences.

A new method for selection of bacterium antibiotic resistance genes is proposed and tested for solving the problems related to selection of primers for PCR assay. The method implies clustering of similar nucleotide sequences and selection of group primers for all genes of each cluster. Clustering of resistance genes for six groups of antibiotics (aminoglycosides, ß-lactams, fluoroquinolones, glycopeptides, macrolides and lincosamides, and fusidic acid) was performed. The method was tested for 81 strains of bacteria of different genera isolated from patients (K. pneumoniae, Staphylococcus spp., S. agalactiae, E. faecalis, E. coli, and G. vaginalis). The results obtained by us are comparable to those in the selection of individual genes; this allows reducing the number of primers necessary for maximum coverage of the known antibiotic resistance genes during PCR analysis.

SOD2 gene polymorphism and muscle damage markers in elite athletes.

Exercise-induced oxidative stress is a state that primarily occurs in athletes involved in high-intensity sports when pro-oxidants overwhelm the antioxidant defense system to oxidize proteins, lipids, and nucleic acids. During exercise, oxidative stress is linked to muscle metabolism and muscle damage, because exercise increases free radical production. The T allele of the Ala16Val (rs4880 C/T) polymorphism in the mitochondrial superoxide dismutase 2 (SOD2) gene has been reported to reduce SOD2 efficiency against oxidative stress. In the present study we tested the hypothesis that the SOD2 TT genotype would be underrepresented in elite athletes involved in high-intensity sports and associated with increased values of muscle and liver damage biomarkers. The study involved 2664 Caucasian (2262 Russian and 402 Polish) athletes. SOD2 genotype and allele frequencies were compared to 917 controls. Muscle and liver damage markers [creatine kinase (CK), creatinine, alanine transaminase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP)] were examined in serum from 1444 Russian athletes. The frequency of the SOD2 TT genotype (18.6%) was significantly lower in power/strength athletes (n = 524) compared to controls (25.0%, p = 0.0076) or athletes involved in low-intensity sports (n = 180; 33.9%, p < 0.0001). Furthermore, the SOD2 T allele was significantly associated with increased activity of CK (females: p = 0.0144) and creatinine level (females: p = 0.0276; males: p = 0.0135) in athletes. Our data show that the SOD2 TT genotype might be unfavorable for high-intensity athletic events.

Associations of single nucleotide polymorphisms with malignant and borderline bone tumors.

Bone neoplasms - are a rare group of diseases, which ethiology and pathogenesis are not fully understood. We have studied 6 single nucleotide polymorphisms rs792/(GHI), rs7956547(IGFI), rs3761243(GNRH2), rs11737764(FGF2), rs6599400(FGFR3), and rs1690916(MDM2) associations with bone tumors. In our work we've detected significant associations with some single nucleotide polymorphisms: IGFl.rs7956547, GNRH2.rs3761243 and FGFR3.rs6599400 in patients with malignant and borderline bone tumors.

Association of FGFR3 and MDM2 gene nucleotide polymorphisms with bone tumors.

Association study of 6 candidate single-nucleotide polymorphisms (rs7921, rs7956547, rs3761243, rs11737764, rs6599400, rs1690916) was carried out in a group of patients with bone tumors of different histological structure (n=68) and control group of normal subjects (n=96). Significant associations of rs6599400 and rs1690916 polymorphisms with disease risk were detected (odds ratio 2.15 [1.06-4.24] and 0.39 [0.19-0.78], respectively). These polymorphisms were located in untranslated genome regions: polymorphism rs6599400 in the 5' region of fibroblast growth factor-3 receptor gene (FGFR3), rs1690916 in the 3' region of mouse MDM2 p53-binding protein homolog (MDM2). These data indicated a possible role of hereditary genetic factors in the formation of predisposition to bone sarcomas and confirmed previous findings according to which these genes should be regarded among the most probable factors involved in tumor development, including tumors of the bone and cartilage tissues.

[Inflammation and atherosclerosis: the role of Renin-Angiotensin system and its inhibition].

The last studies show the renin angiotensin aldosterone system plays an important role in atherosclerotic processes. Vasoconstriction and sodium and water retention, due to angiotensin II and aldosterone promote atherosclerotic processes. Theres recent evidence from animal and clinical studies to consider angiotensin II as a mediator of inflammation, which induces atherosclerotic plaque development and heart remodeling. Also angiotensin II causes endothelial dysfunction by inducing cytokine and chemokine secretion. The pharmacological blockade of the renin angiotensin system has positive influence on clinical outcomes of patients with cardiovascular diseases independently of the blood pressure lowering effect. This review summarizes studies about the role of angiotensin converting enzyme (ACE) inhibitors, angiotensin II receptor antagonists (ARA) and renin inhibitors in vascular inflammatory processes in atherosclerosis.

[Effect of myocardial revascularization on dynamics of factors of angiogenesis in patients with ischemic heart disease].

Aim of the study was to assess effect of myocardial revascularization on levels of factors of angiogenesis in early and remote period after intervention. Main group comprised 228 patients with ischemic heart disease (n=228, 194 men, 34 women, mean age 57+/-8.7 years). Coronary bypass surgery was carried out in 29 patients while other 199 were subjected to percutaneous coronary intervention (PCI). Analysis of data was performed in the group as a whole and in 2 subgroups distinguished in dependence on type of invasive treatment. Levels of factors of angiogenesis - vascular endothelial growth factor (VEGF), transforming growth factor beta (TGFbeta), and endostatin - were measured before, in 6 days, and 6 months after invasive treatment. Compared with healthy persons patients with IHD had significantly higher level of VEGF and significantly lower levels of TGFbeta and endostatin. On day 6 after revascularization in the group as a whole level of VEGF insignificantly rose while level of TGFbeta insignificantly decreased. In 6 months after invasive treatment significant lowering of VEGF level and significant increase of TGFbeta was noted. Endostatin level was measured at baseline and in 6 months after invasive treatment. Significant elevation of endostatin level took place after 6 months. Thus PCI and coronary bypass surgery lead to lowering of VEGF level and elevation of levels of TGFbeta in 6 days after intervention. In remote period reverse dynamics was observed: of VEGF level rose and those of TGFbeta and endostatin increased.

[Disturbances of primary hemostasis in patients with dilation cardiomyopathy].

We studied morphological characteristics of platelets and parameters of platelet aggregation in patients with dilation cardiomyopathy. Augmented aggregatory activity of platelets was found in 76% of patients. In blood of majority of patients we found circulating leukocyte-platelet aggregates. This evidenced for development of inflammatory process and could be related to disturbances of blood rheology. In 2 patients examined by virusological method we revealed presence of a virus inside platelets. This phenomenon might serve as one of possible pathological pathways of disease progression at the account of spread of viral infection along vascular bed during thrombus formation.

[The role of atherosclerosis risk factors in development of ischemic heart disease in young men].

AIM: To determine distribution of different atherosclerosis risk factors and their correlation with severity of coronary arteries (CA) affection in young men. MATERIAL AND METHODS: A total of 235 males aged 28-45 years were examined with coronarography which detected more than 50% narrowing in one or several major CA in 200 patients (the study group with coronary artery disease), 35 males without coronary stenosis served control. Atherosclerosis risk factors, serum lipids, lipoprotein(a), fibrinogen, C-reactive protein (CRP) were assessed. RESULTS: Smoking and hyperlipidemia (HLE) were registered in the study group more frequently. The level of lipoprotein(a) above 30 mg/dl was recorded in 98 (49%) and 7 (20%) patients of the study and control group, respectively (p = 0.001). As shown by a correlation analysis, the number of affected CA was associated with smoking (r = 0.2; p < 0.001), HLE (r = 0.23; p < 0.001), concentration of total cholesterol (r = 0.23; p < 0.0.001), lipoprotein (a) (r = 0.26; p < 0.001) and CRP (r = 0.26; p < 0.05). The level of lipoprotein(a) in patients with myocardial infarction and occluded artery was significantly higher than in patients without myocardial infarction and non-occlusive CA affection. CONCLUSION: The presence and severity of coronary atherosclerosis in young men with coronary artery disease are associated with smoking, HLE and high concentration of lipoprotein (a).

[Effect of therapy with atorvastatin on parameters of postprandial lipemia and factors of inflammation in patients with ischemic heart disease].

Reaction of parameters of lipid transport system to standard fat load (J.Patsch method), C-reactive protein (CRP) and fibrinogen levels were studied in 30 patients with ischemic heart disease before and after 3 months of therapy with atorvastatin (10 mg/day). Atorvastatin therapy resulted in 25, 34, 17 and 16% lowering of concentrations of total cholesterol (CH), low density lipoprotein CH, triglycerides (TG), and apolipoprotein (apo) B, respectively, 6 and 9% elevation of high density lipoprotein CH and apo A-1 levels, respectively. Moreover atorvastatin improved although not completely normalized parameters of postprandial lipemia: significant lowering of baseline (-17%), 3 (-19%) and 6 hour (-14%) post load TG levels was noted. However relative TG elevation from baseline level to 3 and 6 hours after fat load did not change (+100 and 148% before, +95 and 156% after treatment, respectively). Changes of CRP (-22%) and fibrinogen (-8%) were not significant (p>0.05).

[Effect of atorvastatin therapy on the level of secretory phospholipase A2 group IIA and on the modification of low density lipoproteins in patients with ischemic heart disease].

We studied effect of atorvastatin on secretory phospholipase A2 group IIA (sPLA2-IIA) in blood serum of patients with ischemic heart disease (IHD), lipid composition of low density lipoproteins (LDL) and process of modification of LDL induced by sPLA2-IIA in 20 patients taking 20 mg/day of atorvastatin for 3 months. In patients with initially high level of sPLA2-IIA ( > 8 mcg/l) its concentration significantly decreased. Amount of total cholesterol, triglyceride, lecithin, and lysolecithin remained unchanged, however in equimolar relations there occurred decrease of amount of total cholesterol and increase of cholesterol esters. At incubation of LDL, extracted from patient s plasma before initiation of the study, with human sPLA2-IIA from cardiac myxoma, 3.5 nmol of lysolecithin per 1 mg of LDL protein was formed while at incubation of LDL of same patients, extracted after 3 months of atorvastatin administration, amount of lysolecithin was 1.54 nmol/mg LDL protein. Thus atorvastatin therapy causes lowering of sPLA2-IIA in patients with initially high blood level of the enzyme and to a great extent precludes sPLA2-IIA induced LDL modification.

[Cardiac natriuretic peptides in patients with hypertrophic cardiomyopathy].

AIM: To study interrelation of main clinico-hemodynamic parameters and levels of natriuretic peptides (NUP) in patients with hypertrophic cardiomyopathy (HCMP). MATERIAL AND METHODS: We measured concentrations of N-terminal fragments of precursors of plasma brain and atrial natriuretic peptides (NT-proBNP and NT-proANP) and conducted complex echocardiographical examination in 94 patients (mean age 42.3 +/- 133.8 years) with obstructive (n=43) and nonobstructive (n=51) HCMP. RESULTS: Content of NUP did not depend on patients age, contractile function and dimensions of the left ventricle (LV), moderate and significant correlation between content of NT-proBNP, NT-proANP and type of ventricular filling (r=0.45 and 0.46, respectively), heart failure functional class (r=0.41 and 0.44, respectively), severity of LV hypertrophy (r=0.42 and 0.34, respectively), left atrial dimension (r=0.26 and 0.35, respectively), magnitude of pressure gradient in LV outflow tract (r=0.35 and 0.26, respectively). Degree of mitral regurgitation correlated significantly only with NT -proBNP level (r=0.34). The patients who eventually died (n=6) were characterized by higher plasma content of NUP. CONCLUSIONS: In patients with HCMP elevated plasma content of N-terminal precursors of brain and atrial natriuretic peptides are associated with severity of LV diastolic dysfunction and hypertrophy of LV as well as of hemodynamic and functional disturbances.

[Pilot experience in Russia with external contrapulsation in patients with coronary heart disease].

AIM: To examine effects of programmed external contrapulsation (ECP) on the clinico-functional status, quality of life (QOL) in patients with coronary heart disease (CHD), stable angina of FC II-IV resistant to medication. MATERIAL AND METHODS: Eighteen patients with CHD, stable angina of FC III-IV (2 females and 16 males, mean age 63.6 +/- 7.4 years) have completed a course of ECP including 35 one-hour procedures 5-6 times a week for 7 weeks. Ten patients had cardiac failure (NYHA FC II-III). Before and after ECP course the patients were examined using QOL assessment by Minnesota questionnaire, complex echo-cardiography, bicycle exercise test, perfusion myocardial scintigraphy with 99m-Tc-4,2-methoxy-isobutilisonitril, 24-h Holter ECG monitoring, enzyme immunoassay for plasma natriuretic propeptides. RESULTS: Significant subjective QOL improvement (p < 0.01) was noticed by all the patients. Anginal attacks and nitrates doses reduced at least 2-fold. Exercise tolerance rose significantly (p < 0.01), bicycle exercise test was positive in 5 patients, the rest stopped the test after achievement of submaximal heart rate (HR) and fatigue. Most of the patients exhibited improvement of myocardial perfusion. Patients with abnormal myocardial contractility showed a moderate trend (p < 0.03) to an increase in left ventricular ejection fraction. By 24-h ECG, mean HR diminished significantly (p < 0.02). CONCLUSION: ECP is highly effective and safe in combined therapy of CHD, stable angina resistant to drug therapy, in impossibility of myocardial revascularization, including patients with cardiac failure. This manifests in a significant abatement of angina, lower doses of nitrates, improvement of exercise tolerance, quality of life, myocardial perfusion and hemodynamic indices.

[Therapeutic plasmapheresis in patients with destructive pulmonary tuberculosis].

Therapeutic plasmapheresis was performed in 159 patients with pulmonary tuberculosis treated in the clinic in 2000 to 2005. Four hundred and seventy-seven manipulations were made. Among its clinical forms, there was prevalent fibrocavernous tuberculosis (in 136 patients); 9 had infiltrative tuberculosis; caseous pneumonia was present in 5 patients; 4 had tuberculoma; 3 and 2 patients had focal and disseminated tuberculosis, respectively. Out of the 159 patients, 42 (26.4%) underwent therapeutic plasmapheresis at the stages of surgical treatment. Plasmapheresis was made in 61.9% (26/42) before surgery and in 38.1% (n = 16). The study defined indications for and contraindications to therapeutic plasmapheresis in patients with destructive pulmonary tuberculosis, including adolescents and patients at the stages of surgical treatment. A procedure has been developed for discrete and membranous plasmapheresis for this category of patients. In patients with destructive patients, therapeutic plasmapheresis was shown: 1) to level the symptoms of antituberculous chemotherapy intolerance; 2) to correct homeostatic disorders; 3) to enhance the efficiency of basic drug therapy and to promote timely preparation of patients for surgery; 4) to improve the quality of life.

[Assessment of clinical course, prognosis and effectiveness of drug and nondrug treatment of patients with ischemic heart disease and occlusive coronary artery atherosclerosis (data of 5-year prospective follow-up)].

Data of 5-year prospective follow-up were used for assessment of clinical course, prognosis and effectiveness of drug and nondrug treatment of 202 patients with ischemic heart disease, occlusive coronary artery atherosclerosis and preserved left ventricular function. It was found that 5-year cardiovascular mortality and rate of nonfatal myocardial infarction did not differ significantly between groups of patients subjected to drug treatment only, transluminal balloon angioplasty, and coronary artery bypass grafting.

[Effect of therapy with rosuvastatin on lipid spectrum, factors of inflammation and endothelial function in patients with ischemic heart disease].

Rosuvastatin (10 mg) was given for 3 months to 30 men (mean age 57+/-9 years) with total cholesterol (CH) above 5.2 mmol/l. Questioning, physical examination, registration of ECG, measurement of levels of total, low density lipoprotein (LDL), high density lipoprotein (HDL) CH, and triglycerides (TG), assessment of endothelium-dependent brachial artery dilation were carried out at baseline and in 3 months. It was noted that 3 months therapy with rosuvastatin exerted positive effect on blood lipid spectrum: lowering of concentration of total CH (-31%, baseline 6.52+/-0.92, after therapy 4.47+/-0.96 mmol/l, p<0.0001), TG (-39%, baseline 2.73+/-1.56, after therapy 1.67+/-0.71 mmol/l, p<0.001), LDLCH (-44%, baseline 4.11+/-0.85, after therapy 2.40+/-0.90 mmol/l, p<0.0001), elevation of HDLCH (+6%, baseline 1.15+/-0.27, after therapy 1.22+/-0.34 mmol/l, p=0.08). Target LDL CH level (<2.6 mmol/l) was achieved in 23 patients (77%). Significant lowering of concentration of C-reactive protein (CRP) (-56%) and interleukin 6 (-25%) was also established. Before beginning of therapy mean flow dependent dilation was 6.1+/-1.64%, after therapy -- 10.4+/-5.0% (p<0.05). Treatment of men with ischemic heart disease with rosuvastatin (10 mg for 3 months) led to achievement of target values of LDLCH in 77% of them, to significant lowering of concentrations of CRP and interleukin 6, and to improvement of endothelial function.

[Coronary calcium and statins therapy].

AIM: Assessment of the dynamics of coronary calcium score (CCS) evaluated with electron-beam tomography (EBT) under the influence of statin therapy in patients with coronary heart disease (CHD) and hyperlipidemia. MATERIAL AND METHODS: A total of 119 hyperlipidemic CHD patients (mean age 62.1 +/- 9.3 years, 75% males) were included in the study. 69 patients were treated with statins (the study group), 50 patients rejected statins (the control group). CCS was calculated according to Agatson. Initial study parameters in the groups were similar. Mean follow-up was 25.3 +/- 10.6 months. RESULTS: A mean increase of CCS in the study group was 62 +/- 84 units (27.1%) and 122 +/- 180 units (65.2%) in the control group (p = 0.044). Total plasma cholesterol (TC) in the study group decreased by 25.1% from 6.99 +/- 0.92 to 5.24 +/- 0.54 mmol/l), in the control group--by 0.5% (from 6.67 +/- 1.01 to 6.63 +/- 1.0 mmol/l). A weak but significant negative correlation was found between a fall in total cholesterol and CCS increment (r = -0.243, p = 0.008). CONCLUSION: Calcium scoring with EBT or multispiral computed tomography can be used both for pre-clinical diagnosis of coronary atherosclerosis and for monitoring of coronary atherosclerosis progression under influence of hyperlipidemic therapy.

[Disturbances of thyroid function after coronary angiography].

Frequency of thyroid gland functional disturbances after introduction of iodine-containing radiographic contrast agents during coronary angiography was studied in 146 patients, residents of the zone of borderline iodine deficit. Frequency of thyroid pathology was high at baseline (39%). Functional state of the thyroid gland was assessed before and 1, 3, 6, 12 months after investigation. During first month after coronary angiography there were several new cases of thyrotoxicosis and hypothyroidism, as well as deterioration of preexisting functional disturbances of the thyroid. Patients with baseline pathology and functional disturbances of thyroid gland before conduction of coronary angiography should be included into risk group of development of iodine induced states.