Vol 96, No 5 (2015)

Readings named after Zubairov: novel in coagulation science. Medical Biochemistry: achievements and prospects
Development of D.M. Zubairov ideas by the Chita school of hemostasis
Kuznik B.I.
Abstract

Review is dedicated to the influence D.M. Zubairov ideas on the further research of hemostasis system in Chita scientific school. Even in 1957, D.M. Zubairov showed that the state of coagulation largely depends upon the reflex reactions from chemo- and baroreceptors of the carotid sinus and aortic arch. Building on this theory, we established that reflex reactions of hemostatic system can arise not only from known receptor zones, but also originate from the vessels of kidney, liver, lung and other organs. D.M. Zubairov et al. in 1963 dispelled the myth of the non-wetting vascular endothelium, ostensibly to retain the liquid state of the blood. Later we have shown that the increase in the potential difference between the intima and adventitia increased regardless of primary or secondary hypercoagulability. Our data indicate that when the crash syndrome lymph fibrinolysis is 23 times faster compared to the blood, which indicates a better adaptability against thrombosis. We have established the proof for the role of hemostatic system as a cellular regulator of the functional activity and survival. Pathological conditions associated with significant increase in the coagulation process, may not only lead to disseminated intravascular coagulation but also blood, extracellular fluid and cytoplasm of cells coagulation - a syndrome of intra- and extravasal coagulation. To date we can not tell the threshold determining when the hemostasis system is a cell regulator and when «executioner». Development of D.M. Zubairov ideas have led to new concepts of the «continuous liquid coagulation», «common transport system of the body», which helped to produce effective prophylactic and therapeutic methods for a significant umber of life threatening conditions.

Kazan medical journal. 2015;96(5):709-715
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Theoretical and clinical medicine
Features of coagulation hemostasis and anticoagulation system in patients with chronic heart failure due to ischemic heart disease
Mirsaeva G.K., Mironchuk N.N.
Abstract

Aim. To study the features of coagulant and anticoagulant systems in patients with varoious functional classes of congestive heart failure secondary to coronary heart disease (according to New York Heart Association criteria) and to determine the most informative method for diagnosing the hemostasis disorders.

Methods. 82 patients with congestive heart failure, who had previously survived myocardial infarction with Q-wave formation, and left ventricle ejection fraction (by Simpson) less than 50%, were included. Average age was 54.0 [50.0-64.0] years. Control group of 30 healthy people was formed. All patients were allocated to two groups. 1st group included patients with I-II functional class of congestive heart failure (n=57), 2nd group - patients with III-IV functional class of congestive heart failure (n=25). Activated partial thromboplastin time, prothrombin and fibrinogen levels, international normalized ratio, heparin cofactor activity of antithrombin, D-dimer level were measured in blood serum.

Results. Only D-dimer level and heparin cofactor activity of antithrombin were changed in the main group compared to the controls. D-dimer level in patients of the general group was 0.2 [0.2-0.4] mg/L versus 0.2 [0.2-0.2] mg/L in control group (p=0.000064). There was no obvious deficit of heparin cofactor activity of antithrombin in patients with congestive heart failure, but the values were lower in the main group - 90.5 [82,7-100,4]% compared to the control group - 102 [90,0-104,0]% (p=0.0068). In subgroups, there were indicators (prothrombin, international normalized ratio, D-dimer) that changed according to the functional class. The lowest values of prothrombin were found in patients with III-IV functional class, the highest - in patients with I-II functional class. Indicators (prothrombin, INR, D-dimer) which changing with the growth of CHF FC were identified by separating the patients into subgroups. FC, the largest - in patients with FC I-II. Serum D-dimer level increased in accordance with congestive heart failure functional class (р1-2=0.0065).

Conclusion. Congestive heart failure contributes to prothrombotic changes in hemostasis. The severity increases with increasing congestive heart failure functional class. D-dimer is the most sensitive marker of hemostasis activation at congestive heart failure. This parameter should be included in the routine hemostasis testing in patients with congestive heart failure.

Kazan medical journal. 2015;96(5):716-722
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Thrombocyte hemostasis disorders in patients with arterial hypertension at different terms after survived hemorrhagic stroke
Mirsaeva G.K., Khakimova R.A., Timershina I.R.
Abstract

Aim. To analyze the indices of platelet link of hemostasis in patients with arterial hypertension at different periods after surviving hemorrhagic stroke.

Methods. 82 patients with arterial hypertension stage 3 were followed up after surviving the hemorrhagic stroke. The following indicators of platelet hemostasis were studied: platelet count, spontaneous aggregation of thrombocytes; adenosine diphosphate aggregation of thrombocytes induced by adenosine diphosphate, collagen and ristomycin. The laboratory test were performed at admission before the standard treatment was initiated, and at discharge. Statistical data were processed using the integrated Statistica 8.0 software package for Windows (StatSoft).

Results. All examined patients had signs of thrombocyte hemostasis activation. In patients with arterial hypertension, reduced platelet count and increased functional activity of thrombocytes was revealed in early rehabilitation period after hemorrhagic stroke, characterized by non-significant increase of spontaneous and induced platelet aggregation. In patients with arterial hypertension in late rehabilitative period and in residual period after surviving the hemorrhagic stroke, a study of primary stage of platelet hemostasis showed statistically significant deviations of platelet aggregation, seen as the increased spontaneous and induced platelet aggregation, indicating chronic hypercoagulation.

Conclusion. Presented data of platelet link of hemostasis alterations may indicate a higher risk of thrombosis in patients with arterial hypertension over time after surviving the hemorrhagic stroke which may lead to recurring vascular complications.

Kazan medical journal. 2015;96(5):722-727
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Hemostatic system state and cytokine profile in patients with metabolic syndrome
Sumerkina V.A., Chulkov V.S., Chulkov V.S., Ozhigina E.V.
Abstract

Aim. Examine the hemostasis state and cytokine profile in patients with metabolic syndrome.

Methods. The study included 71 patients aged 20 to 44 years. Patients were allocated to four groups matched on age: the first group - females with the metabolic syndrome (n=15), the second - healthy females (control group, n=20), the third - males with the metabolic syndrome (n=19), the fourth group - healthy males (control group, n=17). Laboratory parameters of lipid, carbohydrate metabolism, hemostatic parameters, the levels of cytokines were measured in peripheral blood.

Results. Changes in plasma, fibrinolytic and anticoagulant links were revealed in metabolic syndrome (increased concentration of fibrinogen, soluble fibrin monomer complex, D-dimer - in patients of both gender, increased level of tissue factor pathway inhibitor and plasminogen activator inhibitor type 1 - in females). Metabolic syndrome was associated with an increased concentration of interleukin-6 (males and females), interleukin-10 (females) and decreased level of interleukin-8 (males) in peripheral blood. In females, the interrelation between fibrinolytic system activity and pro-inflammatory cytokines serum levels was revealed. In males with metabolic syndrome, a correlation of interleukin-6 pro-inflammatory cytokine level and D-dimer, interferon γ, insulin and insulin resistance index (HOMA) levels was discovered. The level of anti-inflammatory cytokine interleukin-10 correlated with serum triglycerides level.

Conclusion. Metabolic syndrome in patients of younger age is accompanied by changes in plasma fibrinolytic links of hemostasis, regardless of gender. In patients with metabolic syndrome, there are no gender differences in levels of peripheral blood cytokines inherent in healthy people.

Kazan medical journal. 2015;96(5):728-734
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Clinical and biochemical aspects of coagulation control in patients with atrial fibrillation and excessive alcohol consumption
Novikov D.G., Indutnyy A.V., Trofimovich N.A., Sidenko N.I., Borzenok G.A., Gorbunova L.V.
Abstract

Aim. Reveal the influence of excessive alcohol consumption on blood coagulation system, levels of blood proteins in patients with atrial fibrillation treated with warfarin.

Methods. The study included 80 patients with a diagnosis of atrial fibrillation, treated with vitamin K antagonists. The study group included 34 patients with excessive alcohol consumption (according to the questionnaire), the comparison group - 46 patients with no excessive alcohol consumption. Transferrin, hemopexin, aspartate transaminase, alanine transaminase, gamma-glutamyl transferase, carbohydrate-deficient transferrin levels were measured in venous blood serum, serum protein electrophoresis was performed. Parameters of plasma hemostasis tests and D-dimer level were also analyzed.

Results. Gamma-glutamyl transferase activity in patients of study group was 1.5 times higher compared to the comparison group (p=0.021). There was no statically significant differences in aspartate transaminase and alanine transaminase activity, but their levels were increased in alcohol abusers with gamma-glutamyl transferase above normal values (55 U/l). None of the patients had increased level of carbohydrate-deficient transferrin. Absolute and relative serum β1-globulin concentrations were significantly reduced in study group patients, while hemopexin and transferrin levels were at the same level as in the control group. No differences of coagulation parameters were between the groups. In the study group, in patients with β1-globulin levels below the median values, international normalized ratio was significantly closer to the target values, but the same patients had higher D-dimer levels, which apparently indicates low effect of anticoagulants.

Conclusion. The effect of anticoagulant was low in patients with excessive alcohol consumption and atrial fibrillation, as evidenced by elevated D-dimer levels at international normalized ratio close to the target values. Patients with atrial fibrillation and excessive alcohol consumption with elevated D-dimer levels had serum β1-globulin levels reduced compared to median values.

Kazan medical journal. 2015;96(5):734-741
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Platelet aggregation in hypothyroidism and graves’ disease
Chepis M.V., Ral’chenko E.S., Klimenova A.A., Ral’chenko I.V.
Abstract

Aim. To study platelet aggregation in subclinical and symptomatic hypothyroidism and Graves’ disease.

Methods. The results of clinical and laboratory study of 50 patients with hypothyroidism and 20 patients with Graves’ disease are described. To access platelets aggregation function aggregatogramms were decoded by setting the maximum aggregation value; maximum aggregation rate and platelet aggregates maximum size were evaluated according to the results of the dynamic measurements on aggregometer. A control group consisted of 40 healthy donors.

Results. Patients with subclinical and symptomatic hypothyroidism had prolonged activated partial thromboplastin time, reflecting the propensity for hypocoagulation, reduced total platelets number, spontaneous and ADP-induced platelet aggregation, aggregates formation rate and their maximum size. Changes were more pronounced in patients with Graves’ disease: weakening of platelet component of hemostasis, reduction of the platelets total number and the maximum size of platelet aggregates, significant reduction of the rate to reach the maximum size of platelet aggregates and platelet aggregation were observed.

Conclusion. Thyroid dysfunction is accompanied by dysfunction of hemocoagulation processes, signs of chronic disseminated intravascular coagulation syndrome; these changes are clinically more severe in patients with Graves’ disease.

Kazan medical journal. 2015;96(5):742-744
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Influence of concomitant diseases on health resort treatment in patients with coronary heart disease
Agasiev A.R.
Abstract

Aim. To study the influence of concomitant diseases on health resort treatment of patients with coronary heart disease in recreation period.

Methods. The study performed in «Bilgah» cardiologic resort, which applies the standards of health resort treatment. General sample included 3421 patients with confirmed diagnosis of «coronary heart disease». Patients were grouped according to the number of concomitant diseases (0, 1, 2, 3 or more). In each group, the frequency of prescribing the certain treatment methods was determined.

Results. The vast majority (94.5±0.4%) of patients with coronary heart disease had one or more associated chronic diseases. The proportion of patients with 1, 2, 3, 4, 5, 6 or more chronic diseases was 60.3±0.8; 13.7±0.6; 6.0±0.4; 5.5±0.4; 5.1±0.4 and 0.3±3.9% respectively. In addition to the main disease, a patient had 1.77 compensated chronic diseases in average. There were 56.4±0.85 cases of arterial hypertension, 29.7±0.78 cases of respiratory diseases, 26.8±0.76 cases of gastrointestinal diseases (excluding dental and other diseases of the oral cavity), 23.7±0.73 cases of genitourinary diseases, 18.6±0.66 cases of musculoskeletal system diseases, 9.2±0.49 cases of diabetes on average in 100 patients. In 100 patients with coronary heart disease and 1, 2, 3-4, 5 or more chronic comorbidities were administered 474.8, 524.5, 542.4 and 617.7 treatment methods, respectively.

Conclusion. In patients with coronary heart disease, comorbidities are often identified, requiring administration of additional treatment methods, indicating that polymorbidity is an objective reason for the increased volume of medical services at health resort treatment.

Kazan medical journal. 2015;96(5):744-748
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Inhibin B and activin A in the pathogenesis of idiopathic male infertility
Galimova E.F., Akhmadullina G.K., Bulygin K.V., Mochalov K.S., Galimov S.N.
Abstract

Aim. To assess the role of serum and ejaculate inhibin-activin status disorders in the development of reproductive disorders at idiopathic male infertility.

Methods. The research includes data of laboratory and clinical examination of 82 infertile and 60 fertile men. Serum and semen levels of reproduction regulatory peptides inhibin B and activin A were determined using standard commercially available kits for enzyme-linked immunosorbent assay. Other parameters of ejaculate were also investigated.

Results. In men with infertility of the unknown cause, sperm cell concentration was significantly decreased, and the proportion of sex cells with morphological anomalies was increased. The main features of inhibin-activin profile of biological fluids in healthy males with proven fertility were revealed, which were the significant gradient of inhibin and activin intertissue concentrations and domination of these molecular fertility factors in seminal plasma, corresponding with the views of their key role in sperm fertilizing capacity. In patients with pathospermia, the significant decrease of inhibin B concentration in the ejaculate but not in serum, associated with increased activin A levels was revealed, accompanied by characteristic shift of inhibin-activin ratio associated with the deterioration of quality and quantity parameters of semen.

Conclusion. Revealed changes in the inhibin-activin sperm plasma mirror may be a pathogenetic basis for the idiopathic infertility in couples. Inhibin-activin coefficient in blood serum and seminal fluid may be used as a promising diagnostic and prognostic marker of reproductive dysfunction risk.

Kazan medical journal. 2015;96(5):749-752
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The level of vitamin D and parameters of calcium and phosphorus metabolism in primary school children living in the south of Bashkortostan
Bikmetova E.R., Golovatskikh I.V., Kuznetsova E.V., Kozlov V.N., Ivanov V.G.
Abstract

Aim. Estimation of vitamin D level and calcium-phosphorus metabolism in urban and rural school children of 8-9 years old, living in the south of Bashkortostan.

Methods. The study included two groups of 2-3 class pupils: 87 children living in Meleuz and 93 children of Meleuz Region (Zirgan village). The average age was 8.3±0.66 years. The following parameters were analyzed: serum levels of 25-hydroxyvitamin D, phosphorus, total calcium level, urine calcium level, and daily calcium intake.

Results. 72.4% of children from Meleuz and 30.1% of rural children had vitamin D deficiency, subclinical vitamin D-deficiency was detected in 27.6 and 63% of primary school children, respectively. The average serum calcium level in schoolchildren living in town was somewhat lower than in rural residents (p=0.0012), and was close to the lower limit of the reference values. Serum phosphorus levels and urine calcium levels were within the physiological range in the examined children no matter of where they lived. The leading causes of calcium-phosphorus metabolism alterations are reduced calcium intake and vitamin D deficiency.

Conclusion. The vast majority of primary school children on the south of Bashkortostan had vitamin D-deficiency or subclinical vitamin D-deficiency. Dietary calcium intake is 60 to 70% of the age norm, which is characterized as a moderate deficit.

Kazan medical journal. 2015;96(5):752-756
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Features of free radical lipid peroxidation and serum proteins at dental implants and peri-implantitis
Plyukhin D.V., Tseylikman V.E., Tseylikman O.B., Sinitskiy A.I.
Abstract

Aim. Determine the most informative indicators of free radical peroxidation in blood plasma for predicting the complications of dental implantation.

Methods. The study included 84 patients of both with dental implants, including 31 patients without any complications and 53 patients with peri-implantitis. The levels of lipid peroxidation products and protein carbonylation were measured in peripheral blood.

Results. Moderate intensification of lipid peroxidation and protein carbonylation was discovered in blood serum of patients with dental implants. Increased free radical oxidation in the blood was not limited to lipid peroxidation and resulted in increased serum levels of carbonylated proteins. Facilitation of oxidative degradation of proteins in the blood was associated with decreased resistance to «carbonyl stress», manifested as increased level of Fe+2/H2O2-induced carbonylation. Development of peri-implantitis resulted in a greater enhancement of free radical oxidation of blood. Isopropanol-soluble and heptane-soluble Schiff bases level was increased in blood serum as well. Similar to patients with dental implants, increased levels of carbonylated proteins was observed in blood serum at peri-implantitis. However, free radical oxidation was more intense in patients with peri-implantitis compared to patients with dental implants. Indeed, the level of heptane-soluble Schiff bases at the group with peri-implantitis was 2 times higher as compared to dental implants group, and the level of isopropanol-soluble lipid peroxidation products in inflammatory complications after implantation has reached fourfold increase compared with patients who underwent operation without complications.

Conclusion. Investigated indicators of free radical oxidation may be used as markers of peri-implantitis. Among them, the most informative indicators are Schiff bases and metal-catalyzed protein oxidation.

Kazan medical journal. 2015;96(5):756-759
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Periodontal tissue status and «lipid peroxidation - antioxidant protection» system in saliva and gingival fluid in petrochemical industry workers
Sabitova R.I., Bulyakov R.T., Shakirov D.F., Kamilov F.K.
Abstract

Aim. To study periodontal tissue status and «lipid peroxidation - antioxidant protection» system in saliva and gingival fluid in petrochemical industry workers.

Methods. 185 employees of JSC «Experimental Plant Neftechim» were included into the study. Besides the evaluation of dental status saliva and gingival fluid were analyzed for lipid peroxidation products - diene and triene conjugates, malondialdehyde, hydroperoxides and Schiff bases.

Results. The rate of chronic generalized periodontitis in petrochemical industry workers was 100%, including moderate (47.03%), severe (35.13%) and mild (17.84%) degree. Enhancement of lipid peroxidation and antioxidant defense system activity were detected (in mild and moderate chronic generalized periodontitis) with increasing inflammatory and destructive processes in periodontal tissues. In severe chronic generalized periodontitis reduction in the antioxidant protection activity was observed along with the increasing of lipid peroxidation. Correlation analysis revealed a direct relationship between the periodontal tissue status and «lipid peroxidation - antioxidant protection» system.

Conclusion. The prophylactic work should be organized among the petrochemical industry employees aiming to decrease risks of diseases; using the obtained data can be used in clinical practice to identify high-risk groups of patients with periodontal pathology.

Kazan medical journal. 2015;96(5):759-763
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The role of membrane lipids in forming the electrokinetic properties of red blood cells in solid tumors and psoriasis
Obukhova L.M., Erlykina E.I., Kopytova T.V., Makarova M.N., Shchelchkova N.A., Beschastnova E.S.
Abstract

Aim. Analysis of the electrophoretic mobility of red blood cells in patients with the diseases caused by increased cell proliferation activity, and its relationship with the lipid spectrum of erythrocyte membranes.

Methods. The blood of 42 patients with malignant tumors of epithelial tissues (colon, kidney, prostate, urine bladder and cervical cancer) and 16 patients with skin diseases was studied. The control group consisted of 20 healthy individuals. Electrophoretic mobility of red blood cells was assessed by microelectrophoresis. Lipid fractions levels in erythrocytes was determined by one-dimensional thin layer chromatography.

Results. Over than two-fold decrease of electrophoretic mobility of erythrocytes was observed in patients with malignant tumors of epithelial tissues and skin diseases associated with cell proliferation processes (psoriasis) - by 59 and 60%, respectively In inflammatory skin diseases (microbial eczema), the decrease was less pronounced (30%). The contents of charged phospholipid in erythrocyte membranes in case of cancer and psoriasis has changed in a similar way: a significant reduction in sphingomyelin level in the absence of significant differences in phosphatidylethanolamine and phosphatidylcholine levels. Multidirectional statistically significant changes in the contents of lysophosphatidylcholine were revealed depending on the stage of cancer, their level was decreased two-fold in case of psoriasis. The changes of cholesterol, diacylglycerols, free fatty acids levels in the membranes of red blood cells in case of cancer and psoriasis were divergent.

Conclusion. In diseases caused by cell proliferation activation, electrophoretic mobility of erythrocytes is reduced by more than 50%. Kinetic characteristic of erythrocytes may be used for detecting the conditions of pathologic cell proliferation, including malignant tumors, since they alter in cases of phospholipid membranes lipid spectrum disorders.

Kazan medical journal. 2015;96(5):764-768
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The study of albumin dialysis efficiency in acute liver failure using the «Bioartificial liver» device
Polevshchikova E.E.
Abstract

Aim. Clinical testing of albumin dialysis method in patients with acute liver failure of different etiologies on the experimental sample of the «bioartificial liver» device.

Methods. Clinical assessment of treatment using the «bioartificial liver» device was conducted on 20 patients aged 22-68 years old who had acute liver failure of different etiology. The «bioartificial liver» device consisted of a perfusion unit, the dialyzer, dialysis circuit with albumin solution and carbon haemosorbent mode. A 10% donor human albumin solution was used as the dialysis fluid in recirculation mode. For restoration of the accepting ability albumin was exposed to perfusion through the carbonic hemosorbent. Duration of the treatment was 3 hours, control of liver damage biochemical markers was performed before and after the session.

Results. There was a statistically significant reduction in biochemical markers of cytolysis and cholestasis: transaminases, alkaline phosphatase and bilirubin levels. We also observed a statistically significant reduction of ammonia level, which is one of the main clinical and laboratory signs characterizing the severity of encephalopathy. Improvement of liver synthetic function reflected a statistically significant increase in glucose level. Clinical stabilization as well as reduction of encephalopathy clinical signs were observed after the procedure. No side effects were notices.

Conclusion. Implementation of albumin dialysis in patients with acute liver failure using the «bioartificial liver» device was accompanied by an improvement of their functional state. This gives reason to use this method for the treatment of acute liver failure in intensive care units.

Kazan medical journal. 2015;96(5):768-771
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Peculiarities of immunogramm shifts and free radical oxidation markers of blood plasma in HIV-infected heroin addicts
Shatirko M.A., Reshetniov I.V., Golodnii S.V., Mingazov A.C., Kozochkin D.A., Izarovsky B.V., Tseilikman V.E.
Abstract

Aim. The contribution of heroin addiction in particular immunogram and oxidative stress in HIV-infected individuals.

Methods. The study was performed on 104 patients of working age (23-40 years). The first group consisted of 17 HIV-infected patients with stage III disease, intravenous drug users; the second group - 30 HIV-infected patients not taking the drugs; the third group - 25 injecting drug users not infected with HIV; the fourth group - 32 apparently healthy person. Determination of leukocyte subpopulation composition performed on a flow cytometer «Beckman CoulterFC 500». The blood was determined by the content of lipid peroxidation products and carbonylated proteins.

Results. In HIV-infected heroin addicts compounded was the deficit of T-helper cells. Furthermore, there was increase in CD16+-monocytes. In the group of HIV not-infected drug users also showed a reduction in the number of T-helper cells. Other parameters of the immunogram did not differ significantly from control values. In the group of HIV-infected drug addicts found was more distinct decrease in T-helper cells. The number of T-helper cells decreased both in relation to a group of HIV-infected persons who do not inject drugs and in relation to a group of non-infected drug users. Oxidative stress was manifested in a further strengthening of the carbonylation of proteins. In HIV-infected drug users carbonylated protein content was 4.5 times higher than the control level, and was significantly higher than non-infected drug addicts and HIV-infected persons who do not inject drugs.

Conclusion. Heroin addiction exacerbates characteristic for HIV-infected individual decrease in CD4+-lymphocytes along with increased free radical oxidation.

Kazan medical journal. 2015;96(5):772-775
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Postmortem diagnosis of shock conditions
Akimov P.A., Terekhina N.A.
Abstract

Aim. Develop a method for postmortem diagnosis of shock conditions.

Methods. Brood serum obtained from 72 dead bodies of people who died from different causes such as anaphylaxis, toxic shock, cardiovascular diseases, traumatic injury and thermal burns was examined. The level of fibrinogen fraction was performed by sulfite-alkaline method. Plasma soluble fibrin monomer complexes were identified by paracoagulation tests (ethanol and protamine sulfate tests).

Results. The method of shock conditions postmortem diagnosis was determined, regardless of shock etiology. The lowest levels of fibrinogen fraction in cases of anaphylactic and toxic shock were 19.3 and 11.9 g/l, respectively, which is significantly higher compared to other groups. In cases of death due to acute burn, fibrinogen level was almost tenfold higher (p <0.01) compared to patients with burn disease, indicating the presence of a shock at the time of death. In acute burn injury (burn shock), the lowest level of fibrinogen fraction was 10.1 g/l, while in burn disease the maximal content of fibrinogen fraction was 9.6 g/l. Disseminated intravascular coagulation syndrome was diagnosed in 5 (62.5%) of 8 cases of road traffic injury and in 20 (57.1%) of 35 cases of sudden death, indicating the traumatic and cardiogenic shock before the death. Paracoagulation tests were positive result in 6 (8.3%) of 72 cases. Ethanol test was positive in traumatic injury and cardiovascular diseases, protamine sulfate test was positive in acute burns and anaphylactic shock.

Conclusion. The postmortem diagnosis of shock conditions regardless of etiologic factor leading to death might be performed by detecting the signs of disseminated intravascular coagulation. The level of fibrinogen is significantly elevated at anaphylaxis, toxic shock and other types of shock. Positive results of paracoagulation tests indicate the stage of the process.

Kazan medical journal. 2015;96(5):775-779
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Structure, diagnosis, prevention and treatment of postoperative complications of childbirth perineal tears complicated by anal incontinence
Sultanova S.G.
Abstract

Aim. Improving the results of surgical treatment of complications of second and third degree perineal tears complicated by anal incontinence.

Methods. The study included 248 patients aged 16 to 50 years. The patients were allocated to three groups: the first group - 40 women who underwent traditional surgery and conservative treatment; the second group - 128 women with second degree perineal tear; group III - 80 women who underwent sphincteroplasty (I option) and sphincteroplasty combined with levatorplasty (II option) in our modification (sphincter-saving surgery with precision sutures). The main group included 111 patients who additionally received conservative treatment [1 mL of 1% enoxaparin sodium intravenously and 1 ml of 30% vitamin E (alfa-tocopherol acetate) by intramuscular injection]; control group - 97 patients who underwent standard treatment. Long-term results were assessed by a survey, physical examination in the clinic, telephone and Internet surveys in 35 patients of the main group and 31 women of the control group.

Results. In 91 (82%) patients of the main group, the pain intensity decreased after 7-10 days of treatment was antioxidants, 8 (7.2%) patients had pain in the perineum or anal canal, in 12 (10.8%) cases intense pain in the anal canal were still present. Long-term results were evaluated as good in 16 (51.6%) and 25 (69.4%) patients, as satisfactory - in 9 (29%) and 8 (22.2%) patients, as unsatisfactory - 6 (19.4 %), and 3 (8.3%) patients of the main and the control groups, respectively. The test group showed earlier formation of granulation tissue (3.1±0.3 days earlier compared to the control group). At sphincterometry on the 12th day, 6 patients of the control group had first degree anal incontinence, 3 - second degree anal incontinence compared with only 1 (1.8%) case in the study group (second degree sphincter incontinence).

Conclusion. A proposed diagnostic strategy in women with anal incontinence due to perineal tears of II-III degrees after the labor trauma, allowed choosing the optimal method of surgical treatment and improving treatment outcomes.

Kazan medical journal. 2015;96(5):779-783
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Influence of copper-zinc sulfide ores elements on bone tissue remodeling and regulative factors
Farshatova E.R., Ganeev T.I., Men’shikova I.A., Sarmeneeva L.V., Nurgaleev N.V., Kamilov F.K.
Abstract

Aim. Characterize the intensity of bone remodeling, balance of hormones and local cytokines regulating bone remodeling and bone metabolism, at chronic intake of copper-zinc sulfide ores elements.

Methods. A total of 101 miner, producing copper-zinc sulfide ore by underground mining, and 30 employees of ground services of OAO «Uchaly Mining and Processing Plant», were examined. Experimental studies were performed on 60 white adult male rats, distributed to control and experimental groups. The experimental animals of the study group got copper-zinc sulfide ore powder in a 2% starch solution daily for 3 months as a suspension at the dose of 60 mg per 100 g of body weight. The serum levels of testosterone, parathyroid hormone, total thyroxine and triiodothyronine, cortisol, 25-hydroxyvitamin D, soluble Receptor activator of nuclear factor kappa-B ligand, osteoprotegerin, sclerostin and C-terminal telopeptide of collagen type I, as well as bone alkaline phosphatase activity were determined.

Results. Miners who were diagnosed with decreased bone density had increased level of C-terminal telopeptide of collagen type I, with bone alkaline phosphatase activity similar to the control group. In miners with physiological level of bone density, there was no statistically significant decrease in blood testosterone level, in the groups with low and very low bone mineral density there was a statistically significant decrease in testosterone level and increased level of parathyroid hormone. Experimental animals exposed to sulfide ore had serum levels of testosterone, 25-hydroxyvitamin D, thyroxine and triiodothyronine decreased, and increased level of parathyroid hormone and cortisol. Together with that, blood concentration of sclerostin was increased, level of osteoprotegerin - decreased, and soluble Receptor activator of nuclear factor kappa-B ligand was not changed.

Conclusion. Long-term intake of copper-zinc sulfide ore leads to an imbalance of bone remodeling with a predominance of resorption. It is associated with the reduction of testosterone, calcidiol and thyroid hormones levels providing anabolic and anti-catabolic effect on bone metabolism, and overproduction of parathyroid hormone and cortisol, stimulating osteolysis. Receptor activator of nuclear factor kappa-B ligand / osteoprotegerin ratio and sclerostin level increases.

Kazan medical journal. 2015;96(5):783-787
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Hemostasis state, adipokine levels and markers of endothelial dysfunction in young patients with components of the metabolic syndrome
Chulkov V.S., Sumerkina V.A., Chulkov V.S., Sinitsin S.P., Vereina N.K.
Abstract

Aim. To carry out a comparative assessment of hemostasis, adipokines levels and markers of endothelial dysfunction in young patients with a variety of components of the metabolic syndrome.

Methods. The study included 154 patients aged 18-44 years who were divided into four groups matched for age and sex: the first group - 35 patients with metabolic syndrome, the second group - 25 patients with hypertension without abdominal obesity, the third group - 22 patients with abdominal obesity without hypertension, the fourth group - 72 healthy subjects (control group). We studied the vascular-platelet, coagulation and anticoagulation units of the hemostasis system, the fibrinolytic system, the levels of adipokines and markers of endothelial dysfunction.

Results. Patients with metabolic syndrome showed signs of activation of coagulation together with a slowdown functions of the fibrinolytic system and activation of the anticoagulation system. In young patients with hypertension without abdominal obesity increased were the levels of endothelin, plasminogen activator inhibitor type 1, angiotensin II inhibitor and extrinsic pathway of coagulation. Patients with abdominal obesity without arterial hypertension had no significant change in adipokin levels and markers of endothelial dysfunction compared to control group.

Conclusion. In young patients with a variety of components of the metabolic syndrome revealed were signs of intravascular activation of blood coagulation in conjunction with the imbalance of adipokines levels and endothelial dysfunction.

Kazan medical journal. 2015;96(5):787-791
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The effect of sorbents on the serum levels of copper, iron and their transporting proteins at alcohol intoxication
Terekhina N.A., Zhidko E.V., Terekhin G.A., Orbidans A.G.
Abstract

Aim. Evaluate the effect of sorbents on erythrocyte membrane permeability and serum levels of copper, iron and their transporting proteins at acute ethanol intoxication.

Methods. The study was performed on 94 rats. Acute alcohol intoxication was simulated on intact animals and in animals with prior artificial alcohol abuse. Acute ethanol intoxication was caused by intragastric administration of 40% ethanol at a dose of 0.5 of median lethal dose. Polysorb, Litovit, and Sapropel sorbents were administered at a dose of 3000 mg/kg 30 minutes after ethanol administration. permeability of erythrocyte membrane, serum levels of copper, iron, ceruloplasmin and transferrin were measured by spectrophotometry.

Results. Levels of copper and iron in rat serum and erythrocyte membrane permeability significantly dropped compared to the control level at acute ethanol intoxication, ceruloplasmin level raised by 1.5 times, transferrin level did not change significantly. At acute ethanol intoxication in animals with prior artificial alcohol abuse, copper and iron levels and erythrocyte membrane permeability remained low, ceruloplasmin level remained high, transferrin level was decreased for 2 times. All sorbents were able to compensate the serum levels of copper, iron and ceruloplasmin in animals with prior artificial alcohol abuse, and Litovit and Polysorb also influenced on transferrin level.

Conclusion. Compensatory effect of Polysorb, Litovit, and Sapropel on the serum levels of ceruloplasmin, copper, iron and transferrin and on erythrocyte membrane permeability was discovered at acute ethanol intoxication in animals with prior artificial alcohol abuse.

Kazan medical journal. 2015;96(5):868-871
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Experimental medicine
Studying the blood clotting investigation in prescence of boron-dipyrrin fluorescent dyes
Aleksakhina E.L., Marfin Y.S., Merkushev D.A., Tomilova I.K., Rumyantsev E.V.
Abstract

Aim. To study the blood clotting in prescence of boron-dipyrrin fluorescent dyes.

Methods. Photophysical properties of several boron-dippiryn based fluorescent dyes were examined in presence of blood plasma biomolecules and in model system containing bovine serum albumin using electron absorption spectroscopy and fluorescence spectroscopy.

Results. The interaction between the investigated dyes and protein plasma components changes spectral characteristics of the dyes and leads to bathochromic and hypochromic shifts of absorption spectra accompanied by changing of fluorescence intensity. The mechanism of fluorescence changing was defined within the Stern-Folmer theory. It was shown that the static factor prevails due to dye-biopolymers molecular complex formation at plasma protein concentration up to 1 g/l, while the higher levels are characterized mainly by nonspecific interactions of fluorophores. The increase of fluorescent characteristics of phenyl-substituted BODIPY in the presence of proteins caused by resonance energy transfer and physicochemical features changes of the fluorophore molecular environment was shown.

Conclusion. The gained results demonstrate the possibility of using the BODIPY dye for determination of blood clotting activity. Specific interactions of dye molecules with studied biological objects allows to analyze the contents and dynamic processes in biological objects at blood serum clotting.

Kazan medical journal. 2015;96(5):792-798
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Association of blood and liver free radical oxidation indicators in rats with different resistance to hypoxia after survived anoxia
Bayburina G.A., Nurgaleeva E.A., Bashkatov S.A., Shibkova D.Z.
Abstract

Aim. To study the association of rat liver and blood free radical oxidation indicators with different resistance to hypoxia on the long run after ischemic damage caused by systemic circulation arrest.

Methods. Circulatory arrest lasting for 5 minutes was modeled at ether anesthesia on male white rats which were further allocated to 3 groups according to resistance to hypoxia. The observation period was 35 days. The levels of reduced glutathione, catalase, and products reacting with thiobarbituric acid were determined in liver homogenates, levels of erythrocyte catalase, total antioxidant activity, glutathione peroxidase, glutathione transferase, and products reacting with thiobarbituric acid - in blood. Statistical analysis was performed by parametric statistics, regression and factor analysis.

Results. Rats with different resistance to hypoxia initially differed significantly on the number of safety systems functioning indicators levels in liver tissue and blood cells. Total factor analysis of the entire body of empirical data revealed two factors, conventionally called «resistance to hypoxia» and «antioxidant activity of blood». Daily analysis of the figures recorded at 1, 3, 5, 7, 14, 21 and 35 day of the experiment revealed similarities in common and daily factor solutions, demonstrating the objectivity of the revealed patterns, which emphasizes the significant role of free radical oxidation in anoxic conditions pathogenesis.

Conclusion. The balance pro- and antioxidant systems activity in the liver and blood is an essential component of the body’s resistance to hypoxia affecting the survival of the animals after systemic circulation arrest. Their level predicts resistance to hypoxia at the recovery period.

Kazan medical journal. 2015;96(5):798-802
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Effect of chondroitin sulfate on the type I collagen metabolism in the compact bone in alloxan-induced rats
Vyatkin V.A., Butolin E.G., Ivanov V.G.
Abstract

Aim. To study the type I collagen metabolism in the compact bone in rats with alloxan-induced diabetes receiving sulfated glycosaminoglycans.

Methods. The study was performed on 67 white outbred male rats with body weight of 180-220 g. Mortality at diabetes reproduction was 44.8%. To clarify the role of exogenous glycosaminoglycans on bone collagen metabolism at diabetes mellitus, 16 animals with alloxan-induced diabetes received 1 mg/kg of chondroitin sulfate intramuscularly every second day. The second group (21 animals) with alloxan-induced diabetes did not received any chondroitin sulfate. Control group included 10 intact animals who were administered a single injection on 0.5% ml of normal saline. The levels of type I collagen metabolism markers (PINP - aminoterminal propeptide of type I procollagen, a marker of bone formation; β-CrossLaps - β-isomerized carboxy-terminal cross-linking region of type I collagen, a marker of bone resorption) and the amount of total collagen were determined in homogenates of femoral shaft.

Results. Administration of alloxan to the animals has induced the development of diabetes mellitus. The levels of PINP and β-CrossLaps was significantly higher in alloxan-induced rats which were administered chondroitin sulfate compared to rats with «isolated» alloxan-induced diabetes by 21 (p=0.001) and 28 (p=0.01) days of follow-up, the level of total collagen was higher at 70% at 28 day of the experiment (p=0.0004).

Conclusion. Effect of sulfated glycosaminoglycans on type I collagen metabolism of the compact bone in animals with «isolated» alloxan-induced diabetes is manifested by intensified catabolic and anabolic processes with a predominance of the latter over the control and alloxan-induced rats at 21 and 28 days of the experiment.

Kazan medical journal. 2015;96(5):802-806
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Levels of biogenic amines in the brain of rats at experimental post-traumatic stress disorder development
Deev R.V., Shatrova Y.M., Sinitskiy A.I., Molchanova N.S., Yunusova A.K., Tseylikman O.B., Kozochkin D.A., Lapshin M.S.
Abstract

Aim. To study the changes in levels of biogenic amines-neurotransmitters in the brain at experimental post-traumatic stress disorder development in rats.

Methods. Post-traumatic stress disorder was modeled by keeping 48 outbred male rats in under constant and inescapable strong unconditioned stimulus. The control group included 16 intact animals, not exposed to stress influences. The levels of 3,4-dihydroxyphenylalanine, dopamine, norepinephrine, epinephrine and gamma-aminobutyric acid were determined by fluorometric methods. Behavioral activity of animals was evaluated on the day 3, 7, 10 and 14 by «open field» and «elevated plus maze» actinographs.

Results. When comparing the concentrations of studied neurotransmitters in the brain of control animals with experimental groups, reflecting the development of post-traumatic stress disorder at the time, adrenaline and 3,4-dihydroxyphenylalanine levels were increased on the third day, level of norepinephrine was reduced on the seventh day, 3,4-dihydroxyphenylalanine, dopamine, norepinephrine levels were elevaled, gamma-aminobutyric acid level was reduced on the tenth day, gamma-aminobutyric acid level was increased on the fourteenth day after the stress.

Conclusion. According to the results of the correlation analysis, the largest contribution to the development of behavioral disorders are made by altered brain level of gamma-aminobutyric acid at the time of post-traumatic stress disorder formation (tenth and fourteenth day). At the earlier stages (third and seventh day), the relationship of rats behavioral activity and altered 3,4-dihydroxyphenylalanine and norepinephrine brain levels was shown.

Kazan medical journal. 2015;96(5):806-810
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Reperfusion endotoxin eye damage in experimental study
Drozdova G.A., Samigullina A.F., Nurgaleeva E.A., Sorokin A.A.
Abstract

Aim. To evaluate the level of endogenous intoxication in the blood and the status of the lipid peroxidation processes in eye tissues of experimental animals after acute circulatory arrest.

Methods. Chronic (5 weeks) pathophysiological experiments were performed in 72 nonlinear mature male rats, in which acute circulatory arrest was modeled by neurovascular heart bundle compression, followed by resuscitation. Endogenous intoxication intensity was assessed using endogenous intoxication integral index based on the detection of oligopeptides and low and medium molecular weight substances. Free radical oxidation in eye tissues was assessed by determining the level of 2’-thiobarbituric acid-reactive products, as well as by chemiluminescence. The condition of antioxidant system was investigated by catalase activity features and reduced glutathione level.

Results. There was a significant two-phased increase of endogenous intoxication index during the whole postoperative period. Fe2+-induced chemiluminescence with subsequent accumulation of lipid peroxidation secondary metabolites, as well as increased activity of catalase protective system both in early stages (days 1-3) and on the second week were revealed in eye tissues. A positive correlation of endogenous intoxication integral index and the level of 2’-thiobarbituric acid-reactive products on the first day after resuscitation was revealed.

Conclusion. Endogenous intoxication syndrome is formed after the acute circulatory arrest with maximal levels of endogenous toxins in the systemic circulation on 1-3rd and on 10-14th day after resuscitation, as well as with significant intensification of lipid peroxidation processes in eye tissues at the same period.

Kazan medical journal. 2015;96(5):811-814
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Studying the functional condition of rat epididymis mitochondria at nitric oxide (II) synthesis change
Zvyagina V.I., Bel’skikh E.S., Medvedev D.V., Golovach N.A.
Abstract

Aim. To study the levels of nitric oxide (II) metabolites, lactate, endogenic carnitine and mitochondrial oxidoreductase activity in epididymis tissue at nitric oxide low synthesis and, based on the obtained data, to assess the functional condition of rat epididymis mitochondria.

Methods. 16 Wistar rats were allocated to two equal groups: The first group was administered L-NG-nitroarginine methyl ester (L-NAME), which is non-selective NO-synthase inhibitor, at a dose of 25 mg/kg for 7 days. The second group (the control group) got 0.9% sodium chloride solution administered as intraperitoneal injection for 7 days. Homogenate was obtained from epididymis tissue (head and tail), and mitochondria were isolated from it by differential centrifugation. Activities of mitochondrial enzymes (lactate dehydrogenase, succinate dehydrogenase and superoxide dismutase) were measured, as well as concentrations of NO metabolites, lactate, total protein and endogenous carnitine.

Results. Activity of succinate dehydrogenase was significantly decreased by 55 and 68%, lactate dehydrogenase activity - by 78 and 92%, superoxide dismutase - by 16 and 43% in the mitochondria of epididymis head and tail tissue, respectively in the group receiving 25 mg/kg of L-NAME, compared to the control group. The concentrations of NO metabolites decreased by 18 and 30%, lactate levels increased by 43 and 35%, the share of bounded carnitine decreased by 25% in epididymis tail. Changes of carnitine concentrations in epididymis head were non-significant.

Conclusion. In L-NAME-induced deficiency of NO in epididymis tissues mitochondria, statistically significant decrease in the activity of mitochondrial oxidoreductases and lactate accumulation was noticed, indicating the development of secondary mitochondrial dysfunction.

Kazan medical journal. 2015;96(5):814-818
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Effect of L-arginine and carnitine on cathepsin L and H activity and lysosomal membranes permeability in myocardium in expressed hyperhomocysteinemia
Il’icheva A.S., Fomina M.A.
Abstract
Aim. To study the activity of lysosomal cysteine proteases (cathepsins L, H) and acid phosphatase, changing of permeability, stability of myocardial lysosomal membranes in rats in experimental expressed hyperhomocysteinemia model, and while administering L-arginine and carnitine. Methods. The study was performed on male Wistar rats kept on standard vivarium conditions divided into three control and three experimental groups of 8 animals each. Experimental samples were administered methionine, or combination of L-arginine and carnitine with methionine. The level of serum homocysteine was measured by ELISA. Cathepsin L and H activity was detected by spectrofluorimetric method. Acid phosphatase activity was recorded using the «end point» method. Results. In the model of expressed hyperhomocysteinemia the increase of cathepsin H total activity due to both lysosomal and nonlysosomal fractions was found. These changes were observed along with the general increase of lysosomal membranes permeability. When correcting hyperhomocysteinemia with L-arginine and carnitine a decrease of cathepsin L and H levels was noted as well as positive effect on the myocardial lysosomal membranes stability. Conclusion. Expressed hyperhomocysteinemia is accompanied by statistically significant increase of both lysosomal and cytoplasmic fractions of the cathepsin H activity, indicating the lysosomal membranes permeabilisation phenomenon; L-carnitine and arginine correct hyperhomocysteinaemia effects, leading to cathepsin L and H reduced activity and having a stabilizing effect on the lysosomal membranes of cardiomyocytes.
Kazan medical journal. 2015;96(5):819-824
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Preparation, purification and common characteristics of anticoagulant peptide from peloid
Kalinin E.P., Boyarintsev D.I., Galyan S.L.
Abstract
Aim. To obtain the peloid extract containing peptide anticoagulant, to separate it from co-extracted compounds, to confirm the chemical structure of the purified anticoagulant, to characterize the amino acid profile and to evaluate its effect on blood clotting activity in vitro and the effect on hemostasis in laboratory animals. Methods. The anticoagulant was derived from lake Taraskul (Tyumen region) peloid. The effector was prepared and purified by original method including extraction, sedimentation of humic acids by salting-out, and removal of fulvic acids and low molecular weight fractions of humic acids by ion exchange chromatography. Purity and homogeneity of gained anticoagulant was evaluated by high performance liquid chromatography and polyacrylamide gel electrophoresis. The chemical composition of the active substance was determined by acid hydrolysis and identification of phenylthiohydantoin amino acid derivatives by reversed-phase high performance liquid chromatography. Specific anticoagulant activity was evaluated by substance’s influence on effect of thrombin-dependent conversion of fibrinogen to fibrin and serum thrombin clotting time after being administered to laboratory animals. Results. Peptide anticoagulant was isolated of the peloid and purified. Its activity is realized at the stage of thrombin and fibrinogen interaction. The substance prolongs the fibrin clotting time both in a model test system and when administered intravenously to rats. The anticoagulant effect of the peptide when administered to animals lasts at least 60 min, and no significant toxic effects appear. Conclusion. An isolated active substance might be a basis for the drug development of anticoagulants.
Kazan medical journal. 2015;96(5):824-828
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Biochemical markers of bone and osteoclastic differons in plasma at subacute dichloroethane intoxication
Kamilov F.K., Farshatova E.R., Enikeev D.A., Ivanova G.V.
Abstract

Aim. To explore the plasma level of soluble receptor activator of nuclear factor kappa-B ligand (RANKL), osteoprotegerin and sclerostin in a model of subacute dichloroethane intoxication in rats.

Methods. Experiments were carried out on 20 adult male rats weighing 180-200 g. The rats of the experimental group were administered dichloroethane at the dose of 0.84 mg/kg, mixed with olive oil by a gastric tube daily for two months, which amounted to 0,1 LD50. Control animals received an adequate amount of olive oil. Levels of soluble RANKL, osteoprotegerin and sclerostin were determined in blood serum by ELISA using commercially available reagent kits (reagents «Free RANKL», «Osteoprotegerin» and «Sclerostin» by «Biomedica Medizinprodukte Gmb and CoKG» company. Statistical data processing was performed using the Statistica 6.0 (Stat Soft) software package.

Results. Exposure to dichloroethane resulted in increased levels of soluble RANKL, reduced levels of osteoprotegerin, reflecting the intense new bone formation and the functional state of osteoclasts. The level of sclerostin, which is the negative regulator for new bone formation, was elevated, indicating osteoblast precursors’ differentiation inhibition and decreased function of osteoblasts.

Conclusion. In subacute dichloroethane intoxication, the serum level of soluble receptor activator of nuclear factor kappa-B ligand (RANKL) increases, osteoprotegerin level reduces, sclerostin level increases.

Kazan medical journal. 2015;96(5):828-831
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The study of pituitary-thyroid hormones ratio based on regression analysis
Kamilov F.K., Kozlov V.N., Baymatov V.N., Mamtsev A.N., Smirnov D.Y.
Abstract

Aim. To work up a mathematical model for calculation of the levels of pituitary-thyroid hormones system by least squares method in rats with experimental hypothyroidism.

Methods. To study the relationship of hormones levels regression analysis was used. The search of coefficients was performed using the least squares method. Investigations were carried out on rats, which were divided into six groups of 12 rats each: the first group was control, in the animals of second, third, fourth, fifth and sixth groups hypothyroidism was induced by daily intragastric administration of tiamazol in following doses: 2.5; 20.0; 10.0; 5.0 and 1.0 mg per 100 g of rat body weight for 3 weeks.

Results. Regression analysis was carried out, the type of regression as well as parameters were chosen; statistical analysis of the relationship of hormones was conducted based on the obtained results. By comparing the calculation results of the laboratory analysis sufficiently high reliability of the developed model was set up. The deviation of the arithmetic mean value of the level of thyroid stimulating hormone, produced on the basis of experimental and calculated data, is 2.7%. The accuracy of thyroid stimulating hormone levels calculations increased with the decrease of thyrostatic medication dose. The relative error while calculating the free thyroxine levels in the same groups of rats did not exceed 2.15%, accounting for 1.64; 1.34; 0.36 and 2.15%, respectively, when administered daily 20.0; 10.0; 5.0 and 1.0 mg of antithyroid drug per 100 g body weight. One can argue about the reliability of the constructed model to reproduce the performance levels of the hormones of the pituitary-thyroid system.

Conclusion. In the absence of modern high sensitive immunochemiluminescent diagnostic methods the results can be used for thyroid-stimulating hormone levels calculation as one of the major markers of the thyroid gland functional state.

Kazan medical journal. 2015;96(5):831-837
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The study of cardiomyocytes mitochondrial dysfunction in hyperhomocysteinemia and nitric oxyde deficiency
Medvedev D.V., Zvyagina V.I.
Abstract

Aim. To study the influence of nitric oxide metabolism disturbance on the development of mitochondrial dysfunction in case of hyperhomocysteinemia.

Methods. The research was conducted on 32 Wistar male rats. Hyperhomocysteinemia was simulated by intragastric injection of methionine suspension prepared using starch and Tween-80 with addition of this amino acid into the drinking water. The nitric oxide deficiency was induced by intraperitoneal injection of L-Nω-nitroarginine methyl ester (L-NAME) solution.

Results. Hyperhomocysteinemia is accompanied by dysfunction of cardiac cells mitochondria, manifesting in growth of cytoplasmic lactate level and development of oxidative stress with increased mitochondrial proteins carbonylation. Oxidative stress is largely compensated by the activation of the antioxidant defense system (including superoxide dismutase), as evidenced by a slight decrease of succinate dehydrogenase and H+-ATPase activity, the absence of statistically significant changes of cytoplasmic lactate dehydrogenase activity. Tween-80 showed antioxidant properties, reducing the content of protein carbonyl derivatives and superoxide dismutase activity. Nitric oxide deficiency caused by the L-NAME injection was accompanied by an inhibition of aerobic oxidation processes in cardiomyocytes mitochondria, which was proved by a significant decrease in succinate dehydrogenase activity as well as slight reduction of lactate dehydrogenase activity and lactate accumulation in the cytoplasm, and an oxidative phosphorylation reduction which manifested with a decrease of H+-ATPase activity. One reason for these changes is increased carbonylation of proteins due to high production of reactive oxygen species, which is not sufficiently compensated by increased activity of superoxide dismutase.

Conclusion. Since hyperhomocysteinemia is associated with reduced concentrations of nitric oxide metabolites in cardiomyocytes mitochondria, and changes in these organelles after the administering of methionine have some similarities with those after injection of L-NAME, it can be argued that nitric oxide deficiency plays an important role in the pathogenesis of mitochondrial dysfunction of cardiomyocytes in case of hyperhomocysteinemia.

Kazan medical journal. 2015;96(5):837-843
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Influence of neurotransmitters on lipid peroxidation during immobilization stress exposure in rats of different ages
Meschaninov V.N., Scherbakov D.L.
Abstract

Aim. To identify age-specific changes of «peroxidation and antioxidant activity» system in rats during immobilization stress exposure and after correction with neurotransmitters.

Methods. The study was conducted on 410 male Wistar rats of mature and old age. Rats was immobilized in plastic canisters for 12 hours. Neurotransmitters - acetylcholine chloride, epinephrine (adrenaline hydrochloride), L-tryptophan, nicotinic acid solutions - were injected subcutaneously. After decapitation lipid peroxidation, antioxidant activity, routine biochemical and hematological parameters were studied in sacrified rats using standardized methods.

Results. Immobilization stress induced phase changes of lipid peroxidation and antioxidant activity in the blood of rats, which corresponds to the stages of stress reaction. Increasing age leads to earlier activation of lipid peroxidation which was observed in old rats. Weakening of the parasympathetic nervous system influence and enhantion of the sympathetic nervous system action was observed with age in the blood of rats, which can lead to age-related changes in the intensity of lipid peroxidation when exposed to stress. In mature and old rats exposed to stress L-tryptophan and nicotinic acid induced antioxidant, in old rats - hypolipidemic geroprophylactic effect.

Conclusion. Immobilization stress exposure causes adverse hyperlipidemic changes in peripheral blood of old age rats. However under the impact of L-tryptophan and nicotinic acid combination normalization of lipid and lipoprotein content of blood occurs showing geroprophylactic hypolipidemic qualities of non-antioxidant genesis.

Kazan medical journal. 2015;96(5):843-849
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Effect of erythropoietin on the content of lipid peroxidation products in lymphocytes in experimental thermal injury
Osikov M.V., Simonyan E.V., Saedgalina O.T.
Abstract

Aim. To investigate the effect of different concentrations of erythropoietin on the content of lipid peroxidation products in lymphocytes isolated from the blood of rats with thermal injury.

Methods. The study was performed on 22 white male rats. Thermal injury of IIIA degree on 4% of body surface area was simulated by immersion in water at a temperature of 98-99 °C. After 24 hours, blood lymphocytes were isolated and the content of the primary (diene conjugates), secondary (ketodienes and conjugated trienes) and final products (Schiff bases) of lipid peroxidation were determined spectrophotometrically. Erythropoietin was added to lymphocytes at concentrations of 0.01; 0.1 and 1 IU/ml.

Results. It was found that 24 hours after thermal injury there were the accumulation of primary, secondary and final products of lipid peroxidation in isopropanol fraction of lipid extracts of peripheral blood lymphocytes. Addition of erythropoietin to the rat lymphocytes resulted in a controversial change in the content of lipid peroxidation products: an increase in the heptane fraction, decrease - in the isopropanol fraction of lipid extract of lymphocytes. In the heptane fraction erythropoietin (at concentrations of 0.01, 0.1, and 1 IU/ml) increased the content of primary, end (at a concentration of 0.1 IU/ml) and secondary (at a concentration of 1 IU/ml) lipid peroxidation products. In isopropanol fraction erythropoietin reduced the content of primary (at concentrations of 0.01, 0.1, and 1 IU/ml), final (at concentrations of 0.01 and 0.1 IU/ml) and secondary (at concentrations of 0.01 and 1 IU/ml) products of lipid peroxidation.

Conclusion. It was found that there is an accumulation of lipid peroxidation products in the isopropanol fraction of lipid extract of lymphocytes isolated from peripheral blood of rats with thermal injury; erythropoietin application at concentrations of 0.01; 0.1 and 1 IU/ml increases the content of lipid peroxidation products in heptane fraction and decrease in the isopropanol fraction of lipid extract of lymphocytes.

Kazan medical journal. 2015;96(5):849-853
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Proteins of the oral mucosa in experimental herpetic stomatitis
Reuk S.E., Terekhina N.A.
Abstract

Aim. To conduct a comparative analysis of acute phase proteins levels and permeability proteins in the tissues of oral mucosa and blood plasma in case of experimental herpetic stomatitis.

Methods. Levels of total protein, C-reactive protein, orosomucoid, α1-antitrypsin, prealbumin, albumin and microalbumin were determined in the homogenates of oral mucosa and blood plasma of 30 rats with experimental herpetic stomatitis.

Results. Levels of C-reactive protein, orosomucoid and α1-antitrypsin in serum of rats with experimental herpetic stomatitis are increased. In the homogenates of the rat oral mucosa, levels of the studied proteins increased significantly, by 1.5-3.5 times compared to the control levels. Total protein, orosomucoid, prealbumin, and microalbumin levels increased not only in tissues affected by herpes, but also in the oral mucosa on the opposite side, indicating alterations of histochematic barrier transport of, most importantly, low molecular weight proteins, increased cell membrane permeability and changes in oral mucosa periodontal tissue structural integrity.

Conclusion. Herpetic infection of the oral cavity is accompanied by increased levels of acute phase reactants in rat oral mucosa both on the side of oral mucosa infected by herpes and on healthy side. The level of not only acute phase proteins, but also low-molecular transport proteins allows to evaluation the alterations of oral mucosa membrane permeability in herpetic stomatitis.

Kazan medical journal. 2015;96(5):854-857
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Anti-aggregant activity of new xanthine derivative under conditions of hyper aggregation of platelets in vitro
Samorodov A.V., Kamilov F.K., Khaliullin F.A., Shabalina Y.V., Timirkhanova G.A., Muratayev D.Z.
Abstract

Aim. To study the anti-aggregant activity and the effectiveness of firstly synthesized cyclohexilammonium salt of 2-[1-ethil-3-methyl-7-(dioxotiethanyl-3)-xantinyl-8-thio]acetic acid as a potential antiplatelet agent under in vitro conditions.

Methods. Experimental work was carried out on the blood of healthy donors and 74 male patients diagnosed with thrombosis of various localization. Thromboelastography of citrate blood samples was performed in the presence of the test substances on the device TEG 5000. The analysis determined the overall trend of coagulation, functional activity of platelets and fibrinogen, fibrinolytic activity and physico-mechanical properties of clots formed. ADP-, collagen-, and epinephrine-induced platelet aggregations were registered. Agregatogramm analysis was performed using software AGGR. We evaluated the general nature of the aggregation, the maximum aggregate, maximum aggregation rate, the average size of platelet aggregates.

Results. Cyclohexilammonium salt of 2-[1-ethil-3-methyl-7-(dioxotiethanyl-3)-xantinyl-8-thio]acetic acid in in vitro conditions showed anti-aggregant activity greater than the reference substance, and effectively suppress hypercoagulation caused by an excess of thrombin and tissue factor.

Conclusion. The results of the study make it possible to establish in a potentially high ex vivo therapeutic effect of a cyclohexilammonium salt of 2-[1-ethil-3-methyl-7-(dioxotiethanyl-3)-xantinyl-8-thio]acetic acid under conditions involving platelet hyperaggregation.

Kazan medical journal. 2015;96(5):857-862
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Influence of zinc administration on its tissue concetration and activity of serum antioxidant enzymes in rats at physical activity
Skal’nyy A.A., Tin’kov A.A., Medvedeva Y.S., Alchinova I.B., Bonitenko E.Y., Karganov M.Y., Nikonorov A.A.
Abstract

Aim. To investigate the effect of zinc supplementation at physical exercise on the distribution of the metal in the tissues and the activity of serum antioxidant enzymes.

Methods. Physical activity was simulated using the treadmill. Laboratory animals were distributed to 6 even (n=12) groups. The first and fourth groups of animals received no zinc-containing additives and were imposed to low and high physical activity, respectively. Animals of the 2 and 3 as well as 5 and 6 groups received 5 and 15 mg/kg/day of zinc asparaginate intragastrically and were imposed to low and high physical activity, respectively. The zinc concentrantion in the organs and tissues was determined by inductively coupled plasma mass spectrometry. The activity of antioxidant enzymes was determined by spectrophotometry.

Results. Administration of zinc asparaginate to the laboratory animals with low physical activity resulted in a dose-dependent increase of the metal concentration in liver and kidney parenchyma and blood serum, as well as in increase of serum glutathione peroxidase activity. Intensive physical activity for 14 days was accompanied by a significant increase in serum and kidney tissue zinc level. At the 7-day exposure to zinc at physical activity, a dose-dependent increase in zinc concentration in the organs and increase of serum glutathione peroxidase activity was registered. Zinc administration together with physical activity for 14 days did not result in a significant change in the balance of metal in the body of animals. In contrast to the 7-day exposure, a combination of factors studied for 14 days was accompanied by increased activity of superoxide dismutase, but not glutathione peroxidase.

Conclusion. Physical activity of different duration has a significant effect on the zinc kinetics at oral administration, and the activity of serum antioxidant enzymes in laboratory animals.

Kazan medical journal. 2015;96(5):862-868
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Effect of plantago maxima extract on intensity of free radical oxidation in animals with alimentary obesity
Tinkov A.A., Gatiatulina E.R., Nemereshina O.N., Popova E.V., Nikonorov A.A.
Abstract

Aim. To study the effect of the Plantago maxima extract on the level of markers of free radical oxidation in the organism of laboratory animals with alimentary obesity.

Methods. 32 female Wistar rats were evenly divided into four groups. Animals of the first and second groups were kept in standard and high-fat diet and served as controls, respectively. Animals of the third and fourth groups received as drinking water extract of plantain Plantago maxima together with standard and high-fat diet, respectively. The total duration of the experiment was 3 months. After the experiment liver, adipose tissue as well as serum was extracted. Level of thiol compounds, carbonyl compounds, compounds that react with thiobarbituric acid, as well as diene and triene conjugates were determined spectrophotometrically.

Results. It was found that the alimentary obesity associated with increased oxidative stress markers in adipose tissue homogenate. The use of extracts of plantain in high-fat diet group prevented the development of obesity associated with oxidative stress in adipose tissue. At the same time keeping animals on a high-diet was not associated with increased levels of markers of free radical oxidation in the blood serum. Multivariate analysis of variance revealed a statistically significant impact on both the type of diet and the use of the extract on the studied parameters.

Conclusion. The results of the study indicate a protective effect of aqueous extract of plantain in relation to obesity associated with oxidative stress.

Kazan medical journal. 2015;96(5):872-876
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In vitro effects of L-arginine on lysosomal cysteine proteases activity in isolated experiment and in the state of oxidative stress
Fomina M.A., Kudlaeva A.M.
Abstract

Aim. Assessment of direct influence of arginine on lysosomal cysteine proteases activity in vitro, in isolation as well as the stimulation of oxidative stress.

Methods. The study was conducted on the 72 female conventional mature Wistar rats 280-320 g divided into 6 series of 12 rats each. Lysosome slurries were isolated from the liver of intact animals with a subsequent in vitro incubation in a sucrose solution, in the presence of L-arginine, as well as in the presence of L-arginine accompanied by the stimulation of oxidative stress. Samples of control groups were exposed in vitro with the addition of isolate and oxidant, respectively. Each batch was reproduced three times, incubation was performed at 37 °C in a water bath for 1, 2 and 4 hours. The activity of cathepsins B, L and H was studied using spectrofluorimetric method in two fractions - intra- and extralysosomal. Acid phosphatase activity was used as the main marker of membrane labialization.

Results. One hour Incubation with 5 mM arginine in vitro led to inhibition of the cathepsin H activity and lysosomal membrane damage, however, further increase in incubation time led to its stabilization. In vitro exposure to 5 mM H2O2 caused an increase in activity of cathepsines B and L and the drop in the cathepsin H activity without obvious changes in the distribution of enzymes between extra and intralysosomal fractions. In a state of oxidative stress 2-hour in vitro incubation with 5 mM arginine reduced the permeability of lysosomal membranes for cathepsines B, H and L; while 4-hour incubation led to the destabilization of lysosomal membranes.

Conclusion. The direct effect of arginine at a concentration of 5 mM within the 1,2 and 4-hour time intervals leads to a distinct change as a lysosomal cysteine protease activity and stability of lysosomal membranes.

Kazan medical journal. 2015;96(5):876-882
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History of medicine
V.P. Aksenov in Kazan medical institute
Kuznetsova E.G.
Abstract

Aim. To explore the archives, to analyze the biographical data and to uncover one of the life periods of the Russian writer Vasily Pavlovich Aksenov (1950-1954), when he studied at the medical-prophylactic faculty of Kazan State Medical Institute.

Methods. Biographical research methods (writer’s biographic data, the memories of relatives, friends, colleagues, and various records, interviews, etc.), local history (materials of Kazan period of the writer’s life), as well as historical and literary (all information about V.P. Aksenov fit into the context of country’s historical and literary situation) were used.

Results. The named period of writer’s life was studied in detail. Materials stored in the archives of the Kazan State Medical University, as well as biographical data of V.P. Aksenov, memories and text works that mention the Kazan period of writer’s student life were analyzed. Time of youth, open-mindedness, a desire to live are found side by side with mediocrity, fear of tomorrow and unknown. Harassment, shadowing, exile - all this «tempered and fostered» Aksenov. Friends, family, mainly the mother helped him to fight for his rights, to move on.

Conclusion. Time spent by V.P. Aksenov on the Volga bank, attending the Kazan Medical Institute largely determined his future career path, formed a worldview and sense of self as a person and a writer; Kazan story of student Aksenov is one of many examples of man’s destiny in the transitional period of life of the state.

Kazan medical journal. 2015;96(5):883-886
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Jubilees
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Kazan medical journal. 2015;96(5):887
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Obituary
Dotsent Tat'yana Pavlovna Shagivalieva
 
Kazan medical journal. 2015;96(5):888
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