[The HRM-new molecular technique of detection of pharmaceutical resistance of micobacteria of tuberculosis].

The early detection of isolates of mycobacterium of tuberculosis resistant to anti-tuberculosis pharmaceuticals is ultimately needed for optimization of treatment scheme and prevention of propagation of resistant strains. The literature review presents analysis of data concerning the new, fast and inexpensive technique of detection of mutations in genes of mycobacterium of tuberculosis competent for multiple medicinal resistance. The molecular genetic technique HRM (high resolution melting curve analysis) is based on implementation of amplification of DNA fragments with subsequent analysis of their curves of melting with high resolution. The technique demonstrated its high sensitivity and specificity. In contrast with other molecular genetic techniques of detection of resistance of mycobacterium of tuberculosis, the makes it possible to detect all mutations in interested fragments of genes and to distinguish DNA of mutant and wild types in their mixture. This technique has no contamination danger. The HRM-analysis can become a basis for development of diagnostic kits with purpose to detect tuberculosis with multiple medicinal resistance.

[Quality assessment of HCV RNA detection by PCR method in Federal System of External Quality Assessment in 2005 - 2007].

Part of the Federal System of External Quality Assessment of Clinical Laboratory Tests named "PCR-identification of hepatitis C virus (HCV)" and aimed at detection of HCV RNA by polymerase chain reaction (PCR) assay is functioning from 2000. Ninety, 98, and 112 laboratories from more than 50 regions of Russian Federation participated in it in 2000, 2006, and 2007 respectively. Analysis of results of control samples tests, which were performed by participated laboratories, showed increasing proportion of method of amplification in the presence of fluorescent-marked probes both in real-time and in end-point regimens. In these circumstances, significant increase of specificity and sensitivity of tests was observed. In 2007, proportion of correct results obtained by the participants for tests with negative control samples was 96 - 97%, whereas during detection of HCV RNA in concentration 10(3) IU/ml the proportion of correct results was 80 - 83%. Significant proportion of laboratories, which did not detect HCV RNA in concentration 10(3) IU/ml, points on the necessity to improve sensitivity of this important method of laboratory tests.

[Results of assessing the quality of Chlamydia trachomatis, Mycoplasma hominis, and Ureaplasma urealyticum DNA by polymerase chain reaction in 2006-2007)].

Analyzing the results of detection of C. trachomatis, M. hominis, and U. urealyticum DNA by the participants of the Federal External Quality Assessment System in 2006-2007 indicated that the share of the fluorescent-probe amplification technique both in the real-time mode and by the end-point tended to increase. The above tendency coincided with the other--the higher proportion of laboratories detecting M. hominis and U. urealyticum at low concentrations (as low as 10(3) copies/ml) although no direct caused-and-effect relation could be established between these two tendencies. In 2006-2007, the section participants demonstrated a sufficiently high specificity (94-98%) in the detection of C. trachomatis, M. hominis, and U. urealyticum DNA while the sensitivity of studies left much to be desired--the proportion of correct results at the concentration of DNA of the above microorganisms of 10(3) copies/ml was as high as 73-82%. The considerable share of laboratories that fail to detect microorganisms at low concentrations suggests that it is necessary to continue work to enhance the sensitivity of this important type of laboratory studies.

[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).

[Quality of Neisseria gonorrhoeae DNA detection by polymerase chain reaction: the Federal External Quality Assessment System data].

An analysis of the data submitted in 2005 by 75 laboratories from 31 Russian Federation's regions, participated in the area of the Federal external quality assessment system (FEQAS) "Polymerase chain reaction detection of Neisseria gonorrhoeae" in 2005 indicated that for identification of Neisseria gonorrhoeae DNA, the FEQAS participants used mainly reagents made by Russian manufacturers; the proportion of correct results obtained in the detection of Neisseria gonorrhoeae DNA in the control samples with a DNA concentrations of 10(4) and 10(5) copies/ml was greater than 90% whereas it decreased to 65-75% when the concentration of DNA was 10' copies/ml; there were no significant differences in the proportion of correct results obtained when two most common sets were used to isolate DNA; the proportion of correct results for control samples with a DNA concentration of 10(3) copies/ml was distinctly higher when hybridization with fluorescence detection was employed as a method for detecting the products of Neisseria gonorrhoeae DNA amplification than electrophoresis was applied.

[PCR studies in laboratory diagnosis: problems of quality assurance].

The determination of DNA and RNA of causative agents by the nucleic acid amplification techniques (NAT) presented in the Russian laboratory diagnosis of infections solely by polymerase chain reaction (PCR) have currently found the widest application in world practice and, in some cases, is a basic procedure for detecting an infectious agent, particularly in the diagnosis of infections caused by human immunodeficiency virus, hepatitis C virus, hepatitis B virus, cytomegalovirus, Chlamydia trachomatis, Neisseria gonorrhoeae, and others. The paper presents the data available in the literature on the main reasons for false positive and false negative results in PCR studies, international and national programs for the outside quality control (OQC) of detection of causative agents for various diseases, by applying NAT. It is emphasized that the regular participation in OQC programs is likely to be useful in improving the quality of PCR studies.

[Combination therapy with fluvastatin and fenofibrate in ischemic heart disease patients with combined hyperlipidemia and type 2 diabetes]. / Kombinirovannaia terapiia fluvastatinom i fenofibratom bol'nykh ishemicheskoi bolezn'iu serdtsa i sakharnym diabetom 2-go tipa so smeshannoi giperlipidemiei.

AIM: To assess efficacy of combination therapy with fluvastatin and fenofibrate in ischemic heart disease (IHD) patients with combined hyperlipidemia and type 2 diabetes. MATERIAL: Patients with IHD and combined hyperlipidemia with (n=56)) or without type 2 diabetes (n=30). METHODS: After 8-week diet period the patients were randomized to 4 weeks monotherapy with either fluvastatin (40 mg/day) or micronized fenofibrate (200 mg/day). In patients whose low-density lipoprotein cholesterol (LDL CH) remained > 2,6 mmol/1 and triglycerides (TG) > 2.3 mmol/1 combination of fluvastatin 40 mg/day and fenofibrate 200 mg/day was used for the next 12 weeks. RESULTS: Target levels of LDL CH and TG were achieved in 75 and 88%, respectively, of diabetics, and in 73 and 88%, respectively, of non-diabetics. CONCLUSION: The use of combination of fluvastatin and fenofibrate was more effective then monotherapy for correction of lipid abnormalities in combined hyperlipidemia both in diabetics and non-diabetics with IHD.

Macrophage-granulocyte colony-stimulating factor and fibroblast growth factor in the blood of patients with hyperlipoproteinemia.

Solid-phase ELISA revealed increased content of macrophage-granulocyte colony-stimulating factor and basic fibroblast growth factor in the serum of hyperlipoproteinemic patients.

[The content of double bonds and lipids in blood serum of healthy people and patients with hyperlipidemia]. / Soderzhanie dvoinykh sviazei i lipidov syvorotki krovi u zdorovykh liudei i patsientov s giperlipidemiei.

The number of dual bonds (DB) was determined by the ozonization method in the lipid fraction of blood serm (BS) in healthy subjects and in patients with hyperlipidemia (HLP). Single-direction changes were detected in the DB quantity and in the lipid level in case of HLP; besides, a highly reliable positive correlation was established between the DB concentration and the basic BS lipid classes. The results suggest that an increasing DB quantity in HLP is preconditioned by a growing lipid contents and show their tense relation with components of the lipid transport system (LTS). The determination of the DB concentration, as of an index of the total unsaturation of the BS lipid fraction, provides for obtaining extra data useful in describing the LTS in different-type impairments of lipid metabolism.

[The use of insulin-modified intravenous glucose tolerance test in the diagnosis of insulin resistance in patients with metabolic syndrome]. / Ispol'zovanie insulinmodifitsirovannogo vnutrivennogo gliukosotolerantnogo testa dlia diagnostiki insulinrezistentnosti u patsientov s metabolicheskim sindromom.

Fifty-five patients (mean age 42.3) with clinical signs of metabolic syndrome were investigated. The peroral glucose-tolerance and insulin-modified intravenous glucose-tolerance tests were used in the diagnosis of insulin-resistance. According to the SI index, a lower tissue sensitivity to insulin was detected in 48 (87%) of patients; it is noteworthy, that a low SI index was registered in some of them, when indirect signs of insulin-resistance, like hyperglycemia, hyperinsulinemia, were absent.

[The lipid composition of high-density lipoproteins in patients with psoriasis]. / Lipidnyi sostav lipoproteinov vysokoi plotnosti u bol'nykh psoriazom.

Ten lipid fractions were detected in patients with psoriasis by the method of unidimensional chromatography in a thin layer of silica gel with two solvent systems designed for separating a lipid extract of high-density lipoproteins (HDLP). Four phospholipids fractions were identified as follows: three main ones, i.e. phosphatidylcholine, lisophosphatidylcholine and sphyngomielin, and one minor fraction, i.e. phosphatidylethanolamine. The HDLP lipid comprises free cholesterol and its esters. Three glyceride fractions were detected: mono-, di- and triglicerides and free fatty acids. The ratio of the individual phospholipid fractions, on the one hand, and of free to ester-bound cholesterol was found in HDLP by computer-aided densitometry of chromatograms and coincided with the routinely obtained data. The described method designed to evaluate the HDLP lipid composition can be handy in investigations of HDLP in patients with psoriasis.

[Effect of hormone replacement therapy on blood serum lipids in postmenopausal women with type 2 diabetes]. / Dinamika pokazatelei lipidnogo obmena pod vliianiem zamestitel'noi gormonal'noi terapii u zhenshchin v postmenopauze s sakharnym diabetom 2-go tipa.

There are limited data concerning influence of hormone replacement therapy (HRT) on lipid profile in women with type 2 diabetes. Aim of the study was to compare changes of blood lipids during HRT in postmenopausal women with and without type 2 diabetes. Seventy seven women included in the study were assigned to 1 of 4 groups, basing on being diabetic or nondiabetic, and further subdivided into users of estrogen alone (ERT), and of estrogen plus progestin (EPRT). Effect of 6-month ERT (oral estradiol valerate 2 mg/day) and EPRT (oral estradiol valerate 2 mg/day sequentially combined with cyproterone acetate 1 mg/day) on total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), triglycerides and lipoprotein (a) [Lp(a)] was separately assessed. EPRT and ERT caused decrease in LDL-C by 15% and 12%, and increase in HDL-C by 12% and 13%, respectively, in patients with diabetes (p<0.05 in all cases). LDL-C decreased by 11% and 15%, respectively, in women without diabetes (p<0.05 in all cases). Lp(a) was also reduced 25% with EPRT (p<0.01) and ERT (p<0.05). HDL-C increased 10% (p<0.05) with ERT but remained unchanged with EPRT. In conclusion, changes in all lipid parameters except Lp(a) caused by ERT and EPRT were comparable in postmenopausal women with and without type 2 diabetes.

[Effect of atorvastatin on endothelial function in patients with familial hypercholesterolemia]. / Vliianie atorvastatina na funktsional'noe sostoianie éndoteliia u bol'nykh s nasledstvennoi giperkholesterinemiei

AIM: To study the influence of treatment with HMG-CoA reductase inhibitor atorvastatin on endothelial function in patients with familial hypercholesterolemia type IIa. MATERIALS AND METHODS: Sixteen patients (5m/11w, 51-/+3 years) with familial hypercholesterolemia were studied before and after 3 months of therapy with atorvastatin 20 mg/day. EDRF release test (D.Celermajer, 1992) was used to assess flow-mediated endothelium-dependent vasodilatation (FMD) of the brachial artery in response to reactive hyperemia. Plasma nitrite/nitrate (NOx) levels were measured as an indirect index of nitric oxide (NO) production in vivo using HPLC. RESULTS: Atorvastatin treatment resulted in a 32% reduction in total serum cholesterol (CH), 41% reduction in low density lipoprotein (LDL) CH, 16% reduction in triglycerides and a 21% increase in high density lipoprotein CH. Flow mediated dilatation (FMD) was impaired at baseline (5.8-/+0.9%) and significantly improved up to 9.5-/+0.9% after 3 month atorvastatin therapy (p<0.002). Change in FMD inversely correlated with baseline FMD (r = -0.58, p<0.05). There was no significant correlation between FMD and neither total serum CH nor LDL CH levels at baseline. During atorvastatin therapy significant reduction of plasma NOx levels occurred from 53.4-/+5.1 mcmol/l at baseline (range 42.6-86.2 mcmol/l) to 35.5-/+5.1 mcmol/l (18.4-46.0 mcmol/l) after treatment (p<0.02, n=7). CONCLUSION: In patients with familial hypercholesterolemia atorvastatin produced beneficial effect on endothelial function (increase in flow-mediated dilatation, decrease in NOx).

[Six-month xenical (orlistat) therapy of patients with stable angina pectoris concomitant with obesity and hyperlipidemia]. / Opyt 6-mesiachnogo primeneniia ksenikala (orlistata) u bol'nykh stabil'noi stenokardiei s ozhireniem i giperlipidemiei.

AIM: To evaluate the efficiency of 6-month therapy with xenical (gastrointestinal lipase inhibitor) in combination with diet in patients with stable angina pectoris associated with obesity and hyperlipemia. MATERIAL AND METHODS: An open comparative randomized study of the efficiency of xenical in combination with diet was carried out in patients with stable angina pectoris concomitant with obesity and hyperlipemia. Thirty coronary patients aged 45-65 years with stable angina of effort (functional class I-II) with body weight index 28.1-45.6 kg/m2 (mean 33.5 kg/m2) were examined. All patients presented with dyslipemia (low density lipoprotein (LDL) cholesterol more than 4.14 mmol/liter, triglycerides (TG) more than 2.2 mmol/liter). Controls (n = 15) were treated with diets alone for 6 months. In the main group diets were supplemented by xenical in a dose of 360 mg/day. RESULTS: Body weight index decreased in both groups (by 9.9% in the main group and by 4.2% in the control). Body weight stabilization during 6 months of treatment and the fact that it was slow and gradual were essential. In patients treated with xenical total cholesterol level decreased by 10.9% and of LDL cholesterol by 12.2% after 6 months (p < 0.05). Changes in the levels of high density lipoprotein cholesterol and TG were insignificant. The drug did not affect the incidence of angina attacks and improved exercise tolerance after 6-month therapy. Blood biochemistry (transaminases, alkaline phosphatase, glucose, and creatinine) changed negligibly. No side effects were observed; all patients received a complete 6-month course. CONCLUSION: The results confirm that xenical (orlistat) can be used for long therapy of patients with stable angina of effort concomitant with obesity and hyperlipemia.

[Diagnostic significance of albumin transporting characteristics and blood level of troponin T in myocardial infarction]. / Diagnosticheskoe znachenie transportnykh svoistv al'bumina i soderzhaniia v krovi troponina T pri infarkte miokarda.

Serum albumin binding capacity (ABC) was studied using K-35 hydrophobic fluorescent probe in order evaluate the passive transfer and absorption of saturated fatty acids (s-FA) by the cells. ABC decreases on day 3 of myocardial infarction and remains low till discharge and is paralleled by hyperTG. We believe that ABC decrease reflects activation of passive transfer and absorption of s-FA by the cells during simultaneous blocking (presumably by acute phase proteins) of active apoE/B-100 receptor endocytosis of s-FA in the form of TG as VLDL remnants. Decrease of ABC reflects a later than erythrocyte sedimentation rate and acute phase proteins normalization of metabolic consequences of inflammation. ABC helps indirectly evaluate the blocking of active (receptor) absorption of s-FA in the form of TG and activation of passive absorption of s-FA from associations with albumin by the cells.

[The content of double bonds in blood serum lipids from patients with myocardial infarction]. / Soderzhanie dvoinykh sviazei v lipidakh syvorotki krovi pri infarkte miokarda.

Hypocholesterolemia and hypotriglyceridemia during the first days of myocardial infarction (MI) is a result of enhanced absorption of saturated fatty acids in the form of triglycerides within very low density lipoproteins (VLDL) through apoE/B-100 receptors. This is followed by inhibition of this endocytosis, accumulation of saturated fatty acids VLDL in the blood, and hypertriglyceridemia. Permanent level of double bonds in MI (ozone titration) results not from MI but from blocking of cell absorption of polyenic fatty acids in the form of cholesterol polyesters in low-density lipoproteins (LDL) through apoB-100 receptors, which is characteristic of atherosclerosis. High molar ratio of double bonds/cholesterol in comparison with the double bonds/glycerol ratio indicates that the majority of polyenic fatty acids in the blood lipoproteins present as cholesterol polyesters. Lipoprotein transfer and cell absorption of saturated fatty acids alone are selectively impaired in MI. Selective impairment of cell absorption of saturated or polyenic fatty acids alone or a combination of both is observed in some diseases. Ozone titration of serum lipid double bonds reflects impaired absorption of essential polyenic fatty acids by cells.

[The determination of the double bounds in blood serum lipids by titration with ozone: the pathophysiology and diagnostic significance]. / Opredeleneie dvoinykh sviazei v lipidakh syvorotki krovi metodom titrovaniia ozonom: patofiziologiia i diagnosticheskoe znachenie.

In this article the method of definition of double binders in lipids of blood serum in patients with different diseases basically atherosclerosis and ischemic heart disease, with use titration by ozone is given. The received data testify to an opportunity of use of this method in the diagnostic purposes.