Dysfunction of mitochondria induced by ischemia is considered to be a key event triggering neuronal cell death after brain ischemia. Here we report the effect of ischemia-reperfusion on mitochondrial protein synthesis and activity of cytochrome c oxidase (EC 1.9.3.1, COX). By performing 4-vessel occlusion model of global brain ischemia, we have observed that 15 min of global ischemia led to the inhibition of COX subunit I (COXI) synthesis to 56 % of control. After 1, 3 and 24 h of reperfusion, COXI synthesis was inhibited to 46, 50 and 72 % of control, respectively. Depressed synthesis of COXI was not a result of either diminished transcription of COXI gene or increased proteolytic degradation of COXI, since both Northern hybridization and Western blotting did not show significant changes in COXI mRNA and protein level. Thus, ischemia-reperfusion affects directly mitochondrial translation machinery. In addition, ischemia in duration of 15 min and consequent 1, 3 and 24 h of reperfusion led to the inhibition of COX activity to 90.3, 80.3, 81.9 and 83.5 % of control, respectively. Based on our data, we suggest that inhibition of COX activity is rather caused by ischemia-induced modification of COX polypeptides than by inhibition of mitochondrial translation., P. Racay ... [et al.]., and Obsahuje seznam literatury
K+-p-nitrophenylphosphatase (K+pNPPase) is the enzyme, which is considered to be involved in K+-dependent hydrolysis of the phosphoenzyme in the reaction cycle of Na+, K+-ATPase. The aim of our present study was to characterize some features of K+pNPPase in homogenates of the rat brain and liver. We determined p-nitrophenylphosphatase (pNPPase) activity in the presence of various ion combinations (Mg 2++K+, Mg2+, K+). We found a higher total pNPPase activity in the brain (0.8±0.079 nkat/mg protein) than in the liver (0.08±0.01 nkat/mg protein). Contrary to the liver, the main part of the total brain activity was K+-dependent. The activity of K+pNPPase was significantly higher in cerebral cortex homogenates (0.86±0.073 nkat/mg protein) in comparison to those of the whole brain (0.57±0.075 nkat/mg protein). The specific K+pNPPase activity was two times higher in the isolated pellet fraction (0.911±0.07 nkat/mg protein), rich in synaptosomes, compared to the whole brain homogenate (0.57±0.075 nkat/mg protein). Our results demonstrate the high activity of K+pNPPase in the brain tissue and its distribution mainly into the pellet fraction, what might indicate a possible role of K+pNPPase in specific structures of the brain, e.g. in synaptosomes., M. Ďurfinová, M. Brechtlová, B. Líška, Ž. Barošková., and Obsahuje seznam literatury
The aim of our in vitro studies was to understand the role of leptin in controlling proliferation, apoptosis, and protein kinase A (PKA) in human ovarian cells. We analyzed the in vitro effects of leptin (0, 1, 10 or 100 ng/ml) on the accumulation of proliferation-related peptides (PCNA, cyclin B1), apoptosis-associated peptide (Bax) and the intracellular signaling molecule PKA in cultured human granulosa cells using immunocytochemistry and Western immunoblotting. It was observed that leptin stimulated in a dose-dependent manner the accumulation of PCNA (at doses 1-100 ng/ml), cyclin B1 (at doses 10 or 100 ng/ml), Bax (at doses 10 or 100 ng/ml) and PKA (at doses 1-100 ng/ml) in cultured human ovarian cells. These observations suggest the ability of leptin to control directly human ovarian cell functions: proliferation, apoptosis, and intracellular messenger PKA., A. V. Sirotkin, M. Mlynček, A. V. Makarevich, I. Florkovičová, L. Hetényi., and Obsahuje bibliografii a bibliografické odkazy
HDL cholesterol resp. apolipoprotein A1 concentrations are tools to estimate individual CVD risk, although only a part of HDL particles participate in reverse cholesterol transport (RCT). This discrepancy was analyzed in life style change based on increase of physical activity and dietary counseling. Efflux of cholesterol from pre-labeled macrophages to plasma acceptors of tested individuals was used as an RCT measure. Changes of lipoprotein parameters, glucose, fasting insulin concentrations and RCT were analyzed in 15 obese women after 9-week intervention consisted of 5 sessions of increased physical activity per week. Controlled increase in physical activity for 9 weeks induced a decrease of body weight averaging 9 kg (ranged from 2.3 to 15.5 kg). The intervention leads to significant decreases of triglycerides, apoprotein A1 and apoprotein B concentration, whereas total cholesterol, LDL cholesterol and HDL cholesterol did not change significantly. The increase of RCT was not significant, but there was highly significant negative correlation between individual decrease of body weight and an increase of RCT. Significant increase of RCT was found in 13 persons with a weight reduction more than 3.5 kg. Substantial weight loss is necessary to increase RCT., I. Králová Lesná ... [et al.]., and Obsahuje seznam literatury
The aim of the present study was to define the stress-induced pattern of cytosolic glucocorticoid receptor (GR) and Hsp70 protein in the liver of male Wistar rats exposed to different stress models: acute (2 h/day) immobilization or cold (4 °C); chronic (21 days) isolation, crowding, swimming or isolation plus swimming and combined (chronic plus acute stress). Changes in plasma levels of corticosterone were studied by radioimmunoassay (RIA). The results obtained by Western immunoblotting showed that both acute stressors led to a significant decrease in cytosolic GR and Hsp70 levels. Compared to acute stress effects, only a weak decrease in the levels of GR and Hsp70 was demonstrated in chronic stress models. Chronically stressed rats, which were subsequently exposed to novel acute stressors (immobilization or cold), showed a lower extent of GR down-regulation when compared to acute stress. The exception was swimming, which partially restores this down-regulation. The observed changes in the levels of these major stress-related cellular proteins in liver cytosol lead to the conclusion that chronic stressors compromise intracellular GR down-regulation in the liver., D. Filipović, L. Gavrilović, S. Dronjak, M. Demajo, M. B. Radojčić., and Obsahuje bibliografii a bibliografické odkazy
This study explores the biological validation of markers of acute stress in pigs subjected to transportation for slaughter. The stress markers selected for monitoring were neopterin and cortisol. Their levels in pig serum were measured for two porcine stress syndrome genotypes, NN and Nn, after a 30-min transport to a slaughterhouse. Blood samples were withdrawn before transport (control group) and immediately after the animals' arrival (experimental group). The values of neopterin and cortisol measured before the transport were 5.60±1.65 nm ol/l and 273.54±66.17 nmol/l respectively. After the transfer, the concentration of cortisol rose significantly compared to the control (355.69±85.13 nmol/l, p<0.01). Neopterin concentrations in the serum (8.25±1.60 nmol/l) were also significantly higher (p<0.01) after transportation. The elevated concentrations of both analytes were found to be independent of the genotype. These results document the stimulation of the endocrine system and the immune system that develops in animals undergoing transportation for slaughter., K. Breineková, M. Svoboda, M. Smutná, L. Vorlová., and Obsahuje bibliografii a bibliografické odkazy
The miniature excitatory postsynaptic currents (MEPCs) of the muscle cells of the earthworm Lumbricus terrestris were recorded by glass microelectrodes. In a single synaptic zone, three types of MEPC were recorded: a fast single-exponential type that decayed with τ=0.9 ms, a slow single-exponential with τ=9.2 ms and a two-exponential MEPC with τ = 1.3 and 8.5 ms, respectively. The muscle cells of earthworms contain populations of yet-unidentified ionic channels that might be different from the common nicotinic and muscarinic groups of acetylcholine receptors, since these MEPCs are not sensitive to d-tubocurarine, atropine, benzohexonium or proserine. Alternatively, besides ACh receptors, the membrane may contain receptors for an other yet-unidentified excitatory transmitter., E. M. Volkov, L. F. Nurullin, E. Nikolsky, F. Vyskočil., and Obsahuje bibliografii a bibliografické odkazy
Dipeptidyl peptidase-IV (DPP-IV, CD26) is a serine protease almost ubiquitously expressed on cell surface and present in body fluids. DPP-IV has been suggested to proteolytically modify a number of biologically active peptides including substance P (SP) and the chemokine stro mal cell derived factor-1α (SDF-1α, CXCL12). SP and SDF-1α have been implicated in the regulation of multiple biological processes and also induce responses that may be relevant for glioma progression. Both SP and SDF-1α are signaling through cell surface receptors and use intracellular calcium as a second messenger. The effect of DPP-IV on intracellular calcium mobilization mediated by SP and SDF- α was monitored in suspension of wild type U373 and DPP-IV transfected U373DPPIV glioma cells using indicator FURA-2. Nanomolar concentrations of SP triggered a transient dose dependent increase in intracellular calcium rendering the cells refractory to repeated stimulation, while SDF-1α had no measurable effect. SP signaling in DPP-IV overexpressing U373DPPIV cells was not substantially different from that in wild type cells. However, preincubation of SP with the DPP-IV overexpressing cells lead to the loss of its signaling potential, which could be prevented with DPP-IV inhibitors. Taken together, DPP-IV may proteolytically inactivate local mediators involved in gliomagenesis., P. Bušek, J, Stremeňová, E. Křepela, A. Šedo., and Obsahuje bibliografii a bibliografické odkazy
Mor je zoonóza, jejíž epidemie sužují lidstvo od starověku. Od objevu bakteriálního původce moru A. Yersinem a S. Kitasatem uplynulo 120 let, během kterých byla tato choroba velmi dobře popsána jak z epidemiologického, tak z molekulárně mikrobiologického a evolučního hlediska. Studiem DNA izolované z ostatků obětí moru byl původce moru přímo prokázán u epidemií starých až 650 let. Vysoká mortalita při moru je dána neefektivním přenosem mezi hostiteli pomocí blechy jako vektoru., Plague is a zoonotic disease, the epidemics of which have troubled mankind since ancient times. During the last 120 years that have passed since the discovery of the plague bacillus Y. pestis by A. Yersin and S. Kitasato this infectious disease was described in detail, including its epidemiology, molecular microbiology and evolution. Ancient DNA isolated from the remains of plague victims have enabled us to establish Y. pestis as the causative agent in epidemics more than 650 years old. The high mortality of the plague is caused by an ineffective transfer by its flea vector., and Ivo Konopásek.
Gastrointestinal form is the second stage of the Acute Radiation Syndrome (ARS) with a threshold dose of 8 Gy. It represents an absolutely lethal clinical-pathological unit, enteritis necro-hemorrhagica (duodenitis, jejunitis, ileitis, respectively) with unknown causal therapy. The purpose of our study has been to evaluate the morphological changes in a model of radiation-induced enteritis in rats and estimate the significance of changes in biodosimetry. Wistar rats were randomly divided into 21 groups, 10 animals per group. Samples of the jejunum were taken 24, 48, 72, and 96 h after the whole-body γ-irradiation with the doses of 1, 5, 10, 15, and 20 Gy, and routinely stained with hematoxylin and eosin. Five morphometric markers – intercryptal distance, enterocytal height on the top and base of villus, length of basal lamina of 10 enterocytes and enterocytal width – in irradiated rat jejunum were examined. The results were compared with sham-irradiat ed control group. After lethal doses of irradiation, all morphometric parameters of jejunum significantly changed. With the exception of intercryptal distance, they might be considered as suitable biodosimetric markers under these experimental conditions. Our morphometry results in radiation-induced jejunitis are in accordance with those in other studies. We were the first who quantified morphological post-irradiation changes in animal jejunum. Some of them might be used under experimental conditions. This experimental study is a predecessor of the clinical assessment of a specific marker. Under clinical practice, the sensitive biodosimetric parameter could serve as one of the guidance for evaluation of the absorbed dose in irradiated troops as well as rescue workers. This is in accordance with tasks and Standardization Agreement of the North Atlantic Treaty Organization., D. Driák, J. Österreicher, J. Vávrová, Z. Řeháková, Z. Vilasová., and Obsahuje bibliografii a bibliografické odkazy