[Influence of the DAT gene knockout on exchange of essential and toxic trace elements in rats].

The maintenance of energy homeostasis of the body according to modern data is carried out with the active participation of dopaminergic neurons of the central nervous system. The synthesis and metabolism of dopamine (DA) occurs both in the brain and in peripheral tissues. Violation of the synthesis and metabolism of DA is considered as a link in the vicious cycle which it formed during the development of diet-induced obesity. According to modern data, a number of essential and toxic trace elements, such as Cd, Al, As, Mn, Fe, Cu, Zn, are actively involved in the exchange of DA in the brain and peripheral organs and tissues. One way to assess this relationship is to compare changes in the microelement status of the organism when consuming hypercaloric diets in animals with normal and impaired DA transport. The latter can be animals with a knockout of the DAT transporter gene, which performs DA reabsorption with subsequent storage in the composition of secretory granules. The aim is a comparative study of the content of a number of essential and toxic elements in the brain, liver, and kidneys of rats that differ in the allelic variants of the DAT gene fed balanced diet and the diet with an excess of energy value. Material and methods. The study was carried out on 30 male rats of the DAT-KO knockout line (homozygotes DAT-/- and heterozygotes DAT+/-), 8-10 weeks old, and 13 males rats of the outbred Wistar line (DAT+/+) of the same age. For 62 days the animals (6 groups) received a semi-synthetic diet containing essential elements in the salt mixture or a similar high-fat-highcarbohydrate diet (HFCD) with 30% fat and 20% fructose solution instead of drinking water. The content of 16 trace elements (Fe, Mg, Cu, Mn, Co, Se, Zn, Cr, V, Cs, Ag, Al, Cd, As, Pb, Ni) were determined by inductively coupled plasma mass spectrometry in the liver, kidneys, and brain of rats with a knockout of the dopamine DAT transporter gene: homozygotes (DAT -/-) and heterozygotes (DAT+/-), as well as wild-type rats (DAT+/+) of the Wistar strain. Results and discussion. In the liver, DAT knockout led to an increase in the content of As, Cd, Co, and Cs and a decrease in Fe; in the kidneys - to an increase in the levels of Pb, As, Cd and Se, in the brain - an increase in the content of most of the studied trace elements, including Pb, As, Cs, Al and Cu. Conclusion. Against the background of consumption of HFCD, the effect of DAT knockout on the content of a number of elements was more pronounced compared with the consumption of the control diet. The revealed changes in the trace element content in DAT knockout rats are considered in terms of the effect of DA metabolism in the central nervous system and in peripheral tissues on the status of trace elements.

Putative Trace-Amine Associated Receptor 5 (TAAR5) Agonist α-NETA Increases Electrocorticogram Gamma-Rhythm in Freely Moving Rats.

Cortical gamma rhythm is involved in transmission of information (communication) between brain areas that are believed to be involved in the pathogenesis of cognitive dysfunctions. Trace amines represent a group of endogenous biogenic amines that are known to be involved in modulation of function of classical monoamines, such as dopamine. To evaluate potential modulatory influence of a specific receptor for trace amines Trace Amine-Associated Receptor 5 (TAAR5) on the dopamine system, we used HPLC measurements of dopamine and its metabolites in the mouse striatum following administration of the putative TAAR5 agonist α-NETA. Administration of α-NETA caused significant modulation of dopaminergic system as evidenced by an altered dopamine turnover rate in the striatum. Then, to evaluate potential modulatory influence of TAAR5 on the rat brain gamma rhythm, we investigated the changes of electrocorticogram (ECoG) spectral power in the gamma-frequency range (40-50 Hz) following administration of the putative TAAR5 agonist α-NETA. In addition, we analyzed the changes of spatial synchronization of gamma oscillations of rat ECoG by multichannel recording. Significant complex changes were observed in the ECoG spectrum, including an increase in the spectral power in the ranges of delta (1 Hz), theta (7 Hz), and gamma rhythms (40-50 Hz) after the introduction of α-NETA. Furthermore, a decrease in the spatial synchronization of gamma oscillations of 40-50 Hz and its increase for theta oscillations of 7 Hz were detected after the introduction of α-NETA. In conclusion, putative TAAR5 agonist α-NETA can modulate striatal dopamine transmission and cause significant alterations of gamma rhythm of brain activity in a manner consistent with schizophrenia-related deficits described in humans and experimental animals. These observations suggest a role of TAAR5 in the modulation of cognitive functions affected in brain pathologies.

[Acute myocardial infarction in elderly and senile patients: epidemiology study according to the who program "registry of acute myocardial infarction].

The article presents results of the epidemiology study of acute myocardial infarction (AMI) among permanent Tomsk residents older than 60 years. The study was conducted in 2008-2009 according to the WHO program «Registry of Acute Myocardial Infarction¼. Data demonstrated that morbidity and mortality from AMI was higher in the above mentioned age group than in younger population. Rates of morbidity and mortality in men exceeded those in women in all age groups except population older than 80 years. The study showed high in-hospital lethality caused by high death rates in patients 60 years of age and older who were treated in non-specialized hospitals. Authors conclude that the severity of epidemiology situation with AMI is determined by the frequency of this pathology in population older than 60 years suggesting the necessity of change in the strategy of system development of medical care for AMI patients with focus on the older age groups.

[Factors determining application activity for and satisfaction with medical care among able-bodied population of closed industrial city].

According to a poll results, nearly 16.5% of questioned able-bodied inhabitants of industrial city mentioned unsatisfactory health state of self, with main reasons of negative environmental influence and low availability of quality medical care. Most respondents (81.1%) are characterized by poor medical activity. Over a quarter of the questioned (25.7%) are dissatisfied with provided medical care. The main factors determining medical activity and satisfaction with medical care are age, sex, employment place and income level.

[ACE and AGTR1 genes polymorphisms in left ventricular hypertrophy pathogenesis in humans]. / Polimorfizm genov ACE i AGTR1 v patogeneze gipertorofii levogo zheludochka serdtsa u cheloveka.

The role of A2350G polymorphism in exon 17 of the ACE gene and A1166C - in 3'-UTR of the AGTR1 in the pathogenesis of left ventricular hypertrophy was studied in patients with essential hypertension (EH) and arterial hypertension combined with diabetes mellitus type 2 (AH + DM2). Patients with EH and AH + DM2 did not differ from the control sample of healthy individuals by allele or genotype frequencies. However, an association of both polymorphisms with LVH was detected in EH patients. The frequency of 1166C allele was higher in patients with LVH (33.6% vs 20.7% without LVH). A1166C polymorphism determined the magnitude of left ventricular mass index (LVMI) in EH patients as well (p = 0.007). 2350G allele frequency of the ACE gene was in 1.5, and GG genotype--in 3.5-fold higher in EH patients with LVH, as compared without LVH. LVMI was significantly higher in patients with GG genotype as compared with heterozygotes and AA homozygotes (p = 0.002). Thus the presence of 1166C allele of AGTR1 and 2350G allele of ACE can be considered as predisposing factors for LVH development in EH. In contrast, association of studied polymorphisms with presence or LVH degree was not detected in patients with arterial hypertension combined with DM2. This may indicate another structure of genetic component of predisposition to LVH in different causes.

[Coronary flow vasodilator reserve in patients with type 2 diabetes mellitus-associated arterial hypertension]. / Vazodilatatsionnyi rezerv koronarnogo krovotoka u bol'nykh arterial'noi gipertoniei, assotsiirovannoi s sakharnym diabetom tipa 2.

The coronary flow vasodilator reserve (CFVR) in the proximal segment of the anterior descending coronary artery was studied in 50 patients with diabetes mellitus (DM), by Doppler study via transesophageal approach. Group 1 included 39 patients with DM concurrent with Stages 1-2 arterial hypertension (AH), of them 14 patients were documented as having coronary heart disease (CHD) in the presence of coronary atherosclerosis (Subgroup 1A) and CHD was excluded in the remaining 25 patients (Subgroup 1B). Group comprised 11 patients with normal blood pressure (BP). For comparison, 6 healthy individuals were examined. CFVR was calculated as a ratio of the peak diastolic coronary flow (CF) velocity during infusion of dipyridamole (0.56 mg/kg) to the baseline CF. CDVR was significantly decreased as compared with the controls (2.07 +/- 0.73 in Subgroup 1A, 2.15 +/- 0.67 in subgroup 1B, 1.78 +/- 0.33 in Group 2, and 3.68 +/- 0.26 in the controls), this decrease being due to low CF velocities in hyperemia in the majority of patients in Subgroup 1A and Group 2 and to higher baseline CF velocity in most patients from Subgroup 1B. In Group 1 patients, CFVR was not linear with age, the duration of the disease, BP and HbA1 levels, but it was related to the carotid distensibility coefficient (rho = 0.60, p = 0.004) and to the blood level of total cholesterol (rho = -0.43, p = 0.0107). In Group 2 patients, the least CF velocities in the presence of vasodilatation were detectable in older patients and in patients with hypercholesterolemia. An all the patients with left ventricular hypertrophy (LVH) had decreased CFVR whose values with the myocardial mass index above 130 g/m2 were significantly less than those in the absence of LVH. Thus, the limited reserve of coronary vasodilatation was detectable in patients with DM irrespective of BP levels and the status of epicardial arteries and it was most pronounced in LVH and hypercholesterolemia. The impaired elastic properties of peripheral arteries in the presence of cholesterolemia may be regarded as a marker of the low reserve of coronary vasodilatation in patients with DM concurrent with AH.

[Polymorphic markers of GNB3 (C825T), AGTR1 (A1166C) and ACE (A2350G and I/D) genes in patients with arterial hypertension combined with diabetes mellitus type 2]. / Polimorfnye markery genov GNB3(C825T), AGTR1(A1166C), and ACE(A2350G and I/D) u bol'nykh arterial'noi gipertoniei, sochetaiushcheisia s sakharnym diabetom tipa 2.

AIM: To elicit correlations of polymorphic markers of GNB3 (C825T), AGTR1 (A1166C), ACE (A2350G and I/D) genes with arterial pressure, left ventricular hypertrophy (LVH) and blood concentrations of proinflammatory cytokines in hypertensive patients with diabetes mellitus type 2 (DM2). MATERIAL AND METHODS: Clinical parameters (24-h arterial pressure profile, echocardiographic findings, immunoenzymes level) were studied in 89 hypertensive patients with DM2. These patients had different genotypes by the studied allele variants of the genes determined by polymerase chain reaction. RESULTS: Polymorphism of A1166C gene of type 1 vascular receptor of angiotensin II (AGTR1) contributes to formation of arterial hypertension (AH) signs diversity in DM2 patients. GNB3, a gene C825T polymorphic marker, showed a correlation with diastolic arterial pressure but this variant of the gene locus is not associated with LVH. However, G-allele of ACE gene contributes much to appearance of this pathological sign. Mean values of IL-1beta and TNF-alpha as well as the presence of LVH depended on genotypes by ACE gene (polymorphism I/D). CONCLUSION: Polymorphic markers of ACE and GNB3 candidate genes influence clinical diversity of pathological signs in DM2 patients through modification of AH and LVH severity and the level of proinflammatory cytokines.

[Clinical Efficacy of Combination Therapy With Trimetazidine and Angiotensin Converting Enzyme Inhibitors in Patients With Hypertension and Ischemic Heart Disease Associated With Type II Diabetes]. / Klinicheskaia éffektivnost' kombinirovannoi terapii trimetazidinom i ingibitorami angiotenzinprevrashchaiushchego fermenta u bol'nykh arterial'noi gipertoniei i ishemicheskoi bolezn'iu serdtsa, assotsiirovannykh s sakharnym diabetom 2-go tipa.

AIM: To assess in a randomized open study effect of 12-15 week use of angiotensin converting enzyme inhibitors (ACEI) with and without trimetazidine on myocardial perfusion reserve in patients with ischemic heart disease (IHD) and/or hypertension associated with type II diabetes. MATERIAL: Patients (n=69) receiving long term ACEI therapy with transient myocardial perfusion defects during dipyridamole stress test. METHODS: Control patients (n=29, including 15 with IHD) continued to receive an ACEI, while in trimetazidine group (n=40, including 21 IHD patients) trimetazidine (60 mg/day) was added to ACEI. Single photon emission computer tomography with (199)Thallium Chloride was used for measurement of myocardial perfusion reserve. Changes of physical working capacity, intracardiac hemodynamics and glycemia were studied only in trimetazidine group. RESULTS AND CONCLUSION: Significant 52% (mean) decrease (32.5% in IHD patients) of perfusion defects and acceleration on total clearance of Tl-199 were registered in trimetazidine group while no considerable changes of myocardial perfusion were revealed in control group. Most substantial changes of myocardial blood flow reserve occurred in patients with moderate alterations of left ventricular diastolic filling, and among IHD patients - in those without cardiac dilatation and pronounced diastolic left ventricular dysfunction. Significant increase (45.9 and 23.9% in patients with and without IHD, respectively) of total work performed during bicycle exercise was registered in trimetazidine treated patients. In IHD patients decline of initially elevated intramyocardial tension was also observed.

[Effect of long term therapy with captopril on dopplerographic parameters of intrarenal blood flow and renal function in patients with hypertension and diabetes]. / Vliianie dlitel'noi antigipertenzivnoi terapii kaptoprilom na dopplerograficheskie pokazateli vnutripochechnogo krovotoka i funktsiiu pochek u bol'nykh arterial'noi gipertoniei na fone sakharnogo diabeta.

Effect of 9-12 month treatment with captopril on dopplerographic parameters of intrarenal blood flow and renal function was studied in 30 hypertensive diabetics without clinical signs of nephroangiopathy. There was an interrelationship between strict blood pressure (BP) control (average 24-hour BP below 135/83) and improvement of parameters of intrarenal hemodynamics. BP normalization and most pronounced positive changes of renal perfusion during therapy with captopril were achieved in patients with mild hypertension and initially high intrarenal resistance yet at the stage of normo- or microalbuminuria. In moderate hypertension with microalbuminuria treatment with captopril was associated with stabilization of parameters of renal blood flow and rate of 24-hour albumin excretion at their initial level despite less strict control of BP and unsatisfactory compensation of diabetes. BP response to captopril in patients with hypertension and diabetes was related to initial state of intrarenal hemodynamics. In patients with mild hypertension indexes of resistance of renal and intrarenal arteries could be used for prediction of sensitivity to antihypertensive action of captopril.

Contrasting effects of IG20 and its splice isoforms, MADD and DENN-SV, on tumor necrosis factor alpha-induced apoptosis and activation of caspase-8 and -3.

We identified a novel cDNA (IG20) that is homologous to cDNAs encoding a protein differentially expressed in normal and neoplastic cells (DENN-SV) and human MADD (MAPK-activating death domain-containing protein). Furthermore, we show that the above variants most likely result from alternative splicing of a single gene. Functional analyses of these variants in permanently transfected HeLa cells revealed that IG20 and DENN-SV render them more susceptible or resistant to tumor necrosis factor alpha (TNF-alpha)-induced apoptosis, respectively. All variants tested could interact with TNF receptor 1 and activate ERK and nuclear factor kappaB. However, relative to control cells, only cells expressing IG20 showed enhanced TNF-alpha-induced activation of caspase-8 and -3, whereas cells expressing DENN-SV showed either reduced or no caspase activation. Transfection of these cells with a cDNA encoding CrmA maximally inhibited apoptosis in HeLa-IG20 cells. Our results show that IG20 can promote TNF-alpha-induced apoptosis and activation of caspase-8 and -3 and suggest that it may play a novel role in the regulation of the pleiotropic effects of TNF-alpha through alternative splicing.

[The amplification and overexpression of mdr-family genes in ethidium bromide-resistant Chinese hamster CHO-K1 cells and in the hybrids of sensitive and resistant cells]. / Amplifikatsiia i sverkhékspressiia genov semeistva mdr v ustoichivykh k bromistomu étidiiu kletkakh kitaiskogo khomiachka CHO-K1 i v gibridakh chuvstvitel'nykh i ustoichivykh kletok.

Stable mutant cells Cebr-1 and Cebr-2, resistant to ethidium bromide (EB) in concentration of 2 micrograms/ml, have been isolated by a multistep selection in Chinese hamster ovary cells. It was shown that Cebr-1 and Cebr-2 cells acquired a cross-resistance to unrelated drugs. Stable changes in the structure of chromosomes 1, 2, 5 and 8 were revealed by karyological analysis. Overexpression and amplification of mdr genes were detected in Cebr-2 cells using Northern RNA and Southern DNA blot hybridization. Two independent hybrids Hybr-1 and Hybr-2 were obtained by fusion of Cebr-2 cells with Chinese hamster lung V-79 RJK cells, sensitive to EB. Hybr-2 cells were characterized by the same level of EB-resistance as Cebr-2 cells. Hybr-1 cells have a lower level of EB-resistance than Cebr-2 cells. Hybr-2 cells have demonstrated amplification and overexpression of mdr gene, the same as in Cebr-2 cells, whereas in Hybr-1 cells no mdr gene amplification was observed, but the level of mdr gene expression was higher than in sensitive cells. The data suggest that resistance of Chinese hamster cells to EB is mediated by amplification and overexpression of mdr genes.

[An increase in brain temperature in rats to the intranasal administration of thyroliberin]. / Povyshenie temperatury mozga krys pri intranazal'nom vvedenii tiroliberina.

The integral rat brain temperature was measured by radiothermoscope after intranasal instillation of thyroliberin in a dose 400 mkg. The recording increase of temperature lasted during two hours after instillation. This phenomenon is based both on the direct action of thyroliberin and on its capacity to stimulate the induction of other regulatory peptides.

[Comparison of the effects of thyroliberin and ACTH4-7 PGP on the learning of rats during the solving of spatial orientation tasks]. / Sravnenie vliianii tiroliberina i AKTG4-7 PGP na obuchenie krys pri reshenii zadach na prostranstvennuiu orientatsiiu.

The influence of TRH (100 micrograms/kg) and ACTH4-7 Pro-Gly-Pro (15 micrograms/kg and 25 micrograms/kg) involved an acceleration of the spatial learning on the 12-arm radial maze and on T-maze in rats; an increase in the number of correct choice after short (5-10 min) retention interval; and affected the working and reference spatial memory in the test of reinforced place performance. The effects of the TRH and ACTH4-7 Pro-Gly-Pro did not interfere with one another.

[Thyroliberin-induced motor asymmetry as an indicator of the individual behavioral strategy of rats in a radial labyrinth]. / Motornaia asimmetriia, vyzvannaia tiroliberinom, kak indikator individual'noi strategii povedeniia krys v radial'nom labirinte.

The individual peculiarities of functional changes in motor asymmetry determined by behavioral test in T-maze have been shown in experiments with rats (nonpedigree and line LATI). The application of thyroliberin (10 mkg/kg) causes and increases the left-side lateralization which introduces alteration in the individual strategy of the successive round of 12 rays of radial maze with food during solution of spatial orientation task.

[Early and late changes in potassium transport during the start of proliferation in CHO cell cultures. The action of serum, isoproterenol, propranolol and theophylline]. / Rannie i pozdnie izmeneniia transporta kaliia pri zapuske proliferatsii v kul'turakh kletok CHO. Deistvie syvorotki, izoproterenola, propranolola, teofillina.

The stimulation of DNA synthesis by serum is accompanied by early (30 minutes) and late (2-8 hours) increase in ouabain-sensitive rubidium (potassium) influx and the elevation of intracellular potassium content from 0.5-0.6 to 0.7-0.8 mmole per gram protein in CHO-K1 cells. Isoproterenol alone induces the transient increase both in potassium influx via Na,K-ATPase and in potassium efflux without any effect on intracellular potassium content and cell proliferation. Isoproterenol acts synergistically with serum in eliciting the early and late changes in potassium transport and in stimulating G1----S transition. The combination of serum and theophylline produces a rapid increase in potassium influx, however, it does not stimulate DNA synthesis and does not induce any later increase in intracellular potassium content. It is concluded that early and late activation of Na,K-ATPase by mitogens can be dissociated; the Na,K-ATPase activation is involved in mitogenic response when producing the sustained potassium influx and the elevation of intracellular potassium content during G1----S transition.

[Cation transport and content in serum-stimulated CHO-773 cells. III. The intracellular content of sodium, calcium and magnesium]. / Transport i soderzhanie kationov v kletkakh CHO-773, stimulirovannykh syvorotkoi. III. Vnutrikletochnoe soderzhanie natriia, kal'tsiia i magniia.

Intracellular sodium, calcium, and magnesium content as well as lithium influx have been examined in serum-stimulated CHO cultures using flame-emission technique. Intracellular sodium and lithium influx does not change during the G1----S transition, they increase by 1.3-2 times in the late S and in mitosis. In stationary cultures of CHO cells cellular magnesium is about 50-60 mumole/gr protein; its content increases in 2-3 hours after serum addition and remains constant during the G1----S transition. In stationary cultures of CHO cells cellular calcium is about 20 mumole/gr protein and it increases by 1.5-2 times in the late G1 and S phases. It is concluded that alterations of ion transport accompany not only the early cell response to mitogen but also the G1----S transition.

[Cation transport and content in serum-stimulated CHO-773 cells. I. Rapid changes in rubidium and lithium influxes and intracellular sodium content]. / Transport i soderzhanie kationov v kletkakh CHO-773, stimulirovannykh syvorotkoi. I. Bystrye izmeneniia vkhodnykh potokov rubidiia i litiia i vnutrikletochnoe soderzhanie natriia.

The cultures of Chinese hamster ovary cells (CHO-K1 clone 773) can be brought to the stationary state with most of cellular populations in G1 phase by growing continuously for 4 days up to the cultural density (10-12) X 10(4) cells/cm2. Upon introduction of fresh Eagle medium with 10% calf serum the cells progress from G1 to S phase for 7-9 hours. It is shown that within the first minutes of serum addition ouabain-sensitive rubidium influx increases, however, lithium influx, which serves a test for passive sodium pathways in the membrane, increases or does not change. No correlation was found between the rubidium influx and intracellular sodium changes, induced by serum. From comparative studies of ouabain-sensitive rubidium influx, lithium influx and intracellular sodium content it is concluded that the increase in intracellular sodium is not responsible for serum-induced Na,K-ATPase activation.