The CD8+ natural killer (NK) subpopulation has recently been identified as a fast and reliable biodosimetric indicator within human peripheral blood mononuclear cells (PBMC) in vitro. In irradiated and subsequently cultivated PBMC, a decrease of the relative number of intact CD3-CD8+ lymphocytes 16 and 48 h after treatment has allowed for estimating the received dose in the range of 0 - 10 Gy and lethal/sublethal dose discrimination, respectively. Here we show that suitable biodosimeters can also be found in the peripheral blood B-cell compartment. Multiparameter flow cytometric analysis of irradiated and subsequently cultivated human PBMC revealed that both the CD27+ and CD21- B-cell subpopulations can be used as biodosimeters and the CD19+CD27+ lymphocytes have proved useful for retrospective determination of the received dose in the range of 0 - 6 Gy. In addition, several CD19+ lymphocyte subsets characterized by co-expression of CD21, CD27 and CD38 have been shown to bear biodosimetric potential, too. However, when important parameters like the original size within the CD19+ compartment, its radiation-induced changes and data variation had been taken into account, the CD27+ subpopulation proved superior to the other B-cell subpopulations and subsets. It appears that, in the dose range of 0 - 6 Gy, the relative decrease of CD27+ B lymphocytes provides more sensitive and reliable data than that of CD8+ NK-cells due mainly to lower data variation. In contrast to CD27+ B-cells, the proportions of CD27+ subpopulations of T-cells were not affected by irradiation. We have also proposed a simple experimental protocol based on full blood cultivation and three-color CD27/CD3/CD19 immuno-phenotyping as a time-saving and inexpensive approach for practical biodosimetric evaluations on simple, three-to-four color flow cytometers., Z. Řeháková, J. Šinkora, M. Vlková, D. Vokurková, J. Österreicher, J. Vávrová, D. Driák., and Obsahuje bibliografii a bibliografické odkazy
In the current study, we tested a hypothesis that CD36 fatty acid (FA) transporter might affect insulin sensitivity by indirect effects on FA composition of adipose tissue. We examined the effects of CD36 downregulation by RNA interference in 3T3-L1 adipocytes on FA transport and composition and on sensitivity to insulin action. Transfected 3T3-L1 adipocytes, without detectable CD36 protein, showed reduced neutral lipid levels and significant differences in FA composition when levels of essential FA and their metabolites were lower or could not be detected including gamma linolenic (C18:3 n6), eicosadienic (C20:2 n6), dihomo-gamma linolenic (C20:3 n6), eicosapentaenoic (EPA) (C20:5 n3), docosapentaenoic (DPA) (C22:5 n3), and docosahexaenoic (DHA) (C22:6 n3) FA. Transfected 3T3-L1 adipocytes exhibited a significantly higher n6/n3 FA ratio, reduced Δ5-desaturase and higher Δ9-desaturase activities. These lipid profiles were associated with a significantly reduced insulin-stimulated glucose uptake (4.02±0.1 vs. 8.42±0.26 pmol.10-3 cells, P=0.001). These findings provide evidence that CD36 regulates FA composition thereby affecting sensitivity to insulin action in 3T3-L1 adipocytes., K. Kontrová, J. Zídková, B. Bartoš, V. Skop, J. Sajdok, L. Kazdová, K. Mikulík, P. Mlejnek, V. Zídek, M. Pravenec., and Obsahuje bibliografii a bibliografické odkazy
The aim of this study was to compare the central and peripheral components of cardiorespiratory fitness during incremental to maximal exercise between older men who were either recreational athletes (RA) or leisurely active (LA) men, i.e., those who fall between trained and untrained. This was a crosssectional study in which all subjects completed an exercise test on a cycle ergometer. Maximal oxygen consumption (VO2max) and ventilatory threshold (VT) were assessed using gas analysis, and central components of VO2max were assessed using a non-invasive thoracic bio-impedance device. VO2max (RA: 45.1±4.8 ml/kg/min; LA: 32.2±4.6 ml/kg/min, p≤0.001) and SV at maximal exercise (RA: 133.5±24.96 ml/beat; LA: 107.9±17.6 ml/beat, p=0.005) were higher in the RA group compared to the LA group. A plateau in SV occurred between 30-45 % of maximal exercise capacity in the RA group. No differences in SV were observed across workloads in the LA group. No differences in the calculated arterio-venous oxygen difference ((a-v)O2diff) were observed between groups. In conclusions, training volume appears to influence central components of cardiorespiratory fitness among a matched sample of older men who are neither trained nor untrained. This builds a case for increasing the volume of training to preserve cardiorespiratory fitness among older men., C. D. O'neill, D. S. Kimmerly, S. Dogra., and Obsahuje bibliografii
As a consequence of enhanced production of oxygen free radicals, lipid peroxidation leads to the degradation of membrane lipids and disturbances of membrane permeability. Lipid peroxidation increases under stress conditions such as hypoxia, ischemia or acidosis as well as in metabolic diseases, e.g. diabetes mellitus. We have shown that subcomatous doses of insulin (6.0 IU/kg) significantly increase thiobarbituric acid reactive substances (TBARs), especially malondialdehyde (MDA) - the endproduct of lipid peroxidation, in the brain and heart of mice. In our model of insulin-induced hypoglycemia, mice were treated with the neuroprotective, peptide-containing drug Cerebrolysin (100 mg/kg b.w.). Animals were sacrificed by decapitation two or three hours after the injection of tested substance and samples were taken to determine several serum parameters (glucose, total protein, triglycerides and lactic acid) and TBARs in the brain and heart. Although Cerebrolysin was not able to affect serum parameters after subcomatous insulin injection, the drug significantly influenced lipid peroxidation. A single injection of Cerebrolysin already decreased TBARs levels in the brain and heart tissue. Presuming that an increase of TBARs reflects disturbances of the cell membrane, we have documented a promising effect of Cerebrolysin on cell integrity., J. Patočková, M. Kršiak, P. Marhol, E. Tůmová., and Obsahuje bibliografii
Short-term weight-reducing regimens were shown to influence fatty acid composition of serum lipids unfavorably. Adding long chain n-3 polyunsaturated fatty acids (n-3 LC PUFA) to a low-calorie diet (LCD) could avoid these changes. The aim of this study was to examine the effect of a short-term in-patient weight-reducing regimen including LCD with yogurt enriched by low doses of n-3 PUFA (n-3 LCD). The enriched yogurt contained 790 mg of fish oil, predominantly eicosapentaenoic (20:5n-3; EPA) and docosahexaenoic (22:6n-3; DHA). Forty obese women were randomly assigned to the group consuming LCD and joghurt either with or without n-3 enrichment. Following the 3-week diet in the n-3 LCD group a significantly higher increase in the proportion of n-3 LC PUFA (sum of n-3 FA, EPA and DHA) in serum lipids was confirmed. In phospholipids (PL) a significant difference in the sum of n-6 fatty acids was found, a decrease in the n-3 LCD group and an increase in LCD group. Significantly higher increase in the PL palmitate (16:0) was shown in the LCD group. The results suggest that low doses of n-3 fatty acid enrichment can help to avoid unfavorable changes in fatty acid composition in serum lipids after a short-term weight-reducing regimen., P. Hlavatý, M. Kunešová, M. Gojová, E. Tvrzická, M. Vecka, P. Roubal, M. Hill, K. Hlavatá, P. Kalousková, V. Hainer, A. Žák, J. Drbohlav., and Obsahuje bibliografii a bibliografické odkazy
Matrix metalloproteinases (MMPs) play an important role in the pathogenesis of heart failure (HF). Our aim was to determine the activities of circulating MMP-2 and MMP-9 in patients with HF in respect of gender, comorbidities and treatment (n=51). We did not reveal any differences in circulating pro-MMP-2 and pro-MMP-9 activities between the patients with HF and without it. However, there was a decrease in activity of pro-MMP-2 in treated hypertensive participants versus healthy ones. In contrast, we observed increased pro-MMP-2 activity in hypertensive participants with coexistent HF versus hypertensive participants without HF. In addition, a decrease in pro-MMP-2 activity was shown in women suffering from HF versus men suffering from HF. In conclusion, potential inhibitory effect of antihypertensive treatment on pro-MMP-2 activity was found. Coexistent HF with hypertension probably reduces the inhibitory effect of antihypertensive treatment on pro-MMP-2 activity. Our data also suggest the role of potential cardioprotective factors influencing the activity of pro-MMP-2 in women., E. Giannakos, E. Vardali, M. Bartekova, M. Fogarassyova, M. Barancik, J. Radosinska., and Obsahuje bibliografii
Acute phase proteins and markers of proteosynthetic activity reflect the clinical activity in Crohn´s disease (CD). The impact of anti-tumor necrosis factor antibody (anti-TNF) therapy on serum levels of acute phase proteins and proteosynthetic markers was studied. Fourteen patients with active CD were treated with 5 mg per kg of anti-TNF in intravenous infusion. Clinical activity (assessed by Crohn´s disease activity index - CDAI), α-1-acid glycoprotein, haptoglobin, cholinesterase and prealbumin were assessed before and in months 1 and 5 after treatment. A sustained decrease in CDAI was observed. This was accompanied by a significant decrease in α-1-acid glycoprotein and haptoglobin in month 1 (p=0.005 and p=0.01, respectively) while in month 5 the levels of both acute phase proteins rose significantly (p=0.003 for α-1-acid glycoprotein and p=0.02 for haptoglobin). Cholinesterase and prealbumin significantly increased in month 1 after the treatment (p=0.02 and p=0.0006, respectively), the increase was sustained in cholinesterase while prealbumin levels diminished in month 5. We conclude that the clinical improvement after anti-TNF therapy for CD is accompanied by changes of acute phase proteins and proteosynthetic markers. The assessment of these laboratory markers may be useful in the management of CD patients treated with anti-TNF., V. Kupčová, L. Turecký, Z. Detková, M. Príkazská, A. Keleov., and Obsahuje bibliografii
Changes in extracellular space (ECS) diffusion parameters, DC potentials and extracellular potassium concentration were studied during single and repeated cortical spreading depressions (SD) in 13-15 (P13-15), 21 (P21) and 90-day-old (adult) Wistar rats. The real-time iontophoretic method using tetramethylammonium (TMA+)-selective microelectrodes was employed to measure three ECS parameters in the somatosensory cortex: the ECS volume fraction α (α = ECS volume/total tissue volume), ECS tortuosity λ (increase in diffusion path length) and the nonspecific TMA+ uptake k’. SD was elicited by needle prick. SD was significantly longer at P13-15 than at P21 and in adults. During SD, α in all age groups decreased from 0.21-0.23 to 0.05-0.09; λ increased from 1.55-1.65 to 1.95-2.07. Ten minutes after SD, α (in adults) and λ (all age groups) increased compared to controls. This increase persisted even 1 hour after SD. When SD was repeated at 1 hour intervals, both α and λ showed a gradual cumulative increase with SD repetition. Our study also shows that cortical SD is, as early as P13, accompanied by severe ECS shrinkage and increased diffusion path length (tortuosity) with values similar to adults, followed by a long-lasting increase in ECS volume and tortuosity when compared to pre-SD values., T. Mazel, F. Richter, L. Vargová, E. Syková., and Obsahuje bibliografii
Patients with obesity and type 2 diabetes often display high levels of the anti-diabetic factor fibroblast growth factor-21 (FGF21), suggesting that the overproduction of FGF21 may result from increased adiposity in an attempt by white adipose tissue (WAT) to counteract insulin resistance. However, the production of FGF21 diabetes in the absence of WAT has not been examined. In this study, we investigated the effects of lipodystrophy in A-ZIP F-1 mice on FGF21 production in relation to diabetes. A-ZIP F-1 mice displayed high FGF21 plasma levels resulting from enhanced FGF21 mRNA expression in the liver. Concomitant enhancement of FGF21 receptor (FGFR1) and glucose transporter 1 (GLUT-1) mRNA expression was observed in the muscles of A-ZIP F-1 mice. Furthermore, the activation of hypothalamic NPY and AgRP mRNA expression positively correlated with plasma levels of FGF21 but not active ghrelin. Our study demonstrates that an increased FGF21 plasma level in lipodystrophic A-ZIP F-1 mice results mainly from up-regulated liver production but does not suffice to overcome the lipodystrophy-induced severe type 2-diabetes and insulin resistance in the liver linked to the augmented liver fat deposition., A. Špolcová, M. Holubová, B. Mikulášková, V. Nagelová, A. Štofková, Z. Lacinová, J. Jurčovičová, M. Haluzík, L. Maletínská, B. Železná., and Obsahuje bibliografii
The study was designed to prove the hypothesis that lipopolysaccharide (LPS)-induced fever elicits the changes in surfactant specific proteins, potentially related to thermal tachypnea. In adult rats fever was induced by intraperitoneal administration of LPS at a dose 100 μg/kg of body weight; control group received saline. Respiratory parameters, arterial blood gases and pH and colonic body temperature (BT) were recorded. Five hours later, surfactant proteins (SP) A, B, C and D were evaluated in bronchoalveolar lavage fluid (BALF) and lung tissue (LT). LPS evoked monophasic thermic response (at 300 min 38.7±0.2 vs. 36.4±0.3 °C, P<0.05) and an increase in minute ventilation due to changes in breathing rate and tidal volume. LPS-instilled animals had higher levels of SP-A and SP-D in LT (P<0.05 and 0.01), and higher SP-D in BALF (P<0.01) than controls. SP-B increased in LT and SP-C in BALF of animals with LPS (both P0.05 vs. controls). The changes in all surfactant specific proteins are present in LPS-induced fever. Alterations of proteins related to local immune mechanisms (SP-A, SP-D) are probably a part of general inflammatory response to pyrogen. Changes in proteins related to surface activity (SP-B and SP-C) might reflect the effort of the body to stabilize the lungs in thermal challenge., M. Kolomaznik, I. Zila, J. Kopincova, D. Mokra, A. Calkovska., and Obsahuje bibliografii