Prolonged agonist stimulation results in specific transfer of activated Gα subunits of Gqα/G11α family from particulate membrane fraction to soluble (cytosol) cell fraction isolated as 250 000 x g supernatant. In this study, we have used 2D electrophoresis for more defined resolution of Gα subunits of Gqα/G11α family and followed the time course of solubilization effect. The small signal of soluble G proteins was already detected in control, hormone-unexposed cells. Hormone stimulation resulted in a slow but continuous increase of both intensity and number of immunoreactive signals/spots of these G proteins (10, 30, 60, 120 and 240 min). At longer times of agonist exposure (>2 hours), a marked increase of Gqα/G11α proteins was detected. The maximal level of soluble Gqα/G11α proteins was reached after 16 hours of continuous agonist exposure. At this time interval, eight individual immunoreactive signals of Gqα/G1 α proteins could be resolved. The relative proportion among these spots was 15:42:10:11:7:7:2:5. Solubilization of this class of Gα proteins was thus observed after prolonged agonist stimulation only, induced by ultra high concentration of hormone and in cells expressing a large number of GPCRs. Our data therefore rather indicate tight/persisting binding of Gqα/G11α proteins to the membrane., D. Durchánková, J. Novotný, P. Svoboda., and Obsahuje bibliografii a bibliografické odkazy
The aim of this study was to obtain a detailed analysis of the relationship between the finger arterial compliance C [ml/mm Hg] and the arterial transmural pressure Pt [mm Hg]. We constructed a dynamic plethysmograph enabling us to set up a constant pressure Pcss [mm Hg] and a superimposed fast pressure vibration in the finger cuff (equipped with a source of infra-red light and a photoelectric sensor for the measurement of arterial volume). Pcss could be set on the required time interval in steps ranging between 30 and 170 mm Hg, and on sinusoidal pressure oscillation with an amplitude Pca (2 mm Hg) and a frequency f (20, 25, 30, 35, 40 Hz). At the same time continuous blood pressure BP was measured on the adjacent finger (Portapres). We described the volume dependence of a unitary arterial length on the time-varying transmural pressure acting on the arterial wall (externally Pcss+Pca.sin(2πf), internally BP) by a second-order differential equation for volume. This equation was linearized within a small range of selected BP. In the next step, a Fourier transform was applied to obtain the frequency characteristic in analytic form of a complex linear combination of frequency functions. While series of oscillations [Pca, f] were applied for each Pcss, the corresponding response of the plethysmogram was measured. Amplitude spectra were obtained to estimate coefficients of the frequency characteristic by regression analysis. We determined the absolute value: elastance E, and its inverse value: compliance (C=1/E). Then, C=C(Pt) was acquired by applying sequences of oscillations for different Pcss (and thus Pt) by the above-described procedure. This methodology will be used for the study of finger arterial compliance in different physiological and pathological conditions., J. Moudr, J. Svačinová, E. Závodná, N. Honzíková., and Obsahuje bibliografii
To propose a test to evaluate endothelial function, based on VO 2 on-transition kinetics in sub -anaerobic threshold (AT) constant load exercise, we tested healthy subjects and patients with ischemic -hypertensive cardiopathy by two cardiopulmonary te sts on a cycle ergometer endowed with an electric motor to overcome initial inertia: a pre-test and, after at least 24 h, one 6 min constant load exercise at 90 % AT. We measured net phase 3 VO2-on kinetics and, by phase 2 time constant (τ), valued endothe lial dysfunction. We found shorter τ in repeated tests, shorter time between first and second test, by persisting endothelium -dependent arteriolar vasodilatation and/or several other mechanisms. Reducing load to 80 % and 90 % AT did not produce significant changes in τ of healthy volunteers, while in heart patients an AT load of 70 %, compared to 80 % AT, shortened τ(∆=4.38±1.65s, p=0.013). In heart patients, no correlation was found between NYHA class, ejection fraction (EF), and the two variables derived from incremental cycle cardio - pulmonary exercise, as well as between EF and τ; while NYHA class groups were well correlated w ith τ duration (r=0.92, p=0.0001). Doxazosin and tadalafil also significantly reduced τ. In conclusion, the O2 consumption kinetics during the on transition of constant load exercise below the anaerobic threshold are highly sensitive to endothelial functi on in muscular microcirculation, and constitute a marker for the evaluation of endothelial dysfunction. and D. Maione, A. F. G. Cicero, S. Bacchelli, E. R. Cosentino, D. Degli Esposti, D. N. Manners, E. R. Rinaldi, M. Rosticci, R. Senaldi, E. Ambrosioni, C. Borghi.
The aim of our study was to explore the effects of regular aerobic exercise on anthropome tric, biochemical and hormonal parameters and mRNA expression of selected factors involved in metabolic regulations in subcutaneous adipose tissue of patients with obesity. Fifteen obese wome n with arterial hypertension underwent a three-month exercise program consisting of 30 min of aerobic exercise 3 times a we ek. Fifteen healthy lean women with no intervention served as a control group. Obese group underwent anthropometric measurements, blood sampling, subcutaneous adipose tissue (SCAT) biopsy and 24-h blood pressure monitoring at baseline and after three months of exercise, while control group was examined only once. At baseline, obese group had increased SCAT expression of proinflammatory cytokines and adipokines relative to control group. Three months of regular exercise improved anthropometric parameters, decreased CRP, blood glucose and HOMA-IR, while having no significant effect on lipid profile and blood pressure. Gene expressions in SCAT were not affected by physical activity with the exce ption of increased aquaporin-3 mRNA expression. We conclude that three months of regular exercise decrease systemic subc linical inflammation with only minor influence on the blood pressure and the endocrine function of subcutaneous fat., P. Trachta, J. Drápalová, P. Kaválková, V. Toušková, A. Cinkajzlová, Z. Lacinová, M. Matoulek, T. Zelinka, J. Widimský Jr., M. Mráz, M. Haluzík., and Obsahuje bibliografii
Patch clamp recordings carried out in the inside-out configuration revealed activity of three kinds of channels: nonselective cation channels, small-conductance K+ channels, and large-conductance anion channels. The nonselective cation channels did not distinguish between Na+ and K+. The unitary conductance of these channels reached 28 pS in a symmetrical concentration of 200 mM NaCl. A lower value of this parameter was recorded for the small-conductance K+ channels and in a 50-fold gradient of K+ (200 mM/4 mM) it reached 8 pS. The high selectivity of these channels to potassium was confirmed by the reversal potential (-97 mV), whose value was close to the equilibrium potential for potassium (-100 mV). One of the features of the large - conductance anion channels was high conductance amounting to 493 pS in a symmetrical concentration of 200 mM NaCl. The channels exhibited three subconductance levels. Moreover, an increase in the open probability of the channels at voltages close to zero was observed. The anion selectivity of the channels was low, because the channels were permeable to both Cl - and gluconate - a large anion. Research on the calcium dependence revealed t hat internal calcium activates nonselective cation channels and small-conductance K+ channels, but not large - conductance anion channels., M. Koselski, A. Olszewska, A. Hordyjewska, T. Małecka-Massalska, K. Trebacz., and Obsahuje bibliografii
Three-dimensional electrogram was used for analysis of ischemia manifestation in isolated hearts. Three parameters based on spherical coordination system were used in this study - amplitude of electrical heart vector, its azimuth and elevation. The parameters were presented as a trend. This approach reflected ischemic changes in a manner which can be easily observed and evaluated. Ischemia was analysed in seven isolated hearts of New Zealand white rabbits. It was found that (a) ischemia changes heart electric vector, (b) ischemic preconditioning has a protective effect, and (c) both of these findings can be clearly observed by the proposed method., O. Janoušek ... [et al.]., and Obsahuje bibliografii a bibliografické odkazy
Numerous abnormalities of thyroid hormones in end-stage renal disease (ESRD) have been described. Our aim was to analyze the impact of these abnormalities on survival. In 167 hemodialyzed ESRD patients, TSH and thyroid hormone levels (T4, fT4, T3, fT3, rT3) were determined. The patients were then prospectively followed up for up to 5 years and the possible impact of any observed abnormalities on their mortality was studied. Only 16.8 % patients had all six tests within the reference range. The pattern of nonthyroidal illness syndrome was found in 56.3 %. Low T3 was particularly common (44.3 %), and clearly associated with increased 6- and 12-month mortality and decreased overall survival (log rank test, P=0.007). Independent of T3 levels (Spearman correlation, NS), increased rT3 was more frequently observed (9.9 %) than expected from the literature, and was also related to increased mortality and decreased survival (log rank test, P=0.021). Increased rT3 may be more common in ESRD patients than previously described, and together with decreased T3 it may serve as an indicator of poor prognosis in subsequent months., J. Horáček ... [et al.]., and Obsahuje seznam literatury
Thyroid nodules are a very frequent pathology among common population. Despite the vast majority of them are of benign origin, the incidence of thyroid cancer is currently rather rising. Although there are several risk factors of thyroid cancer and several clinical, ultrasound, biochemical and molecular diagnostic markers, the exact mechanisms of thyroid oncogenesis and the linkage between thyroid nodule ultrasound appearance and its biological character remain unclear. While ionizing radiation is the only one well-known risk factor for thyroid cancer, the significance of some others remains unclear. The aim of our review was to discuss some not completely known pathophysiological mechanisms involved in thyroid oncogenesis as hypothyroidism, mutations of genes regulating cell proliferation, thyroid autoimmunity and pregnancy and to describe pathophysiological background of some ultrasound markers of thyroid cancer (size, echogenicity, vascularization, calcifications and stiffness). Better knowledge in this field is crucial for development of novel diagnostic techniques and therapeutic approaches. For exampl e, the analysis of BRAF, RAS and other mutations in cytological samples may help to distinction between follicular thyroid carcinoma and follicular thyroid adenoma and may signific antly decrease the number of unnecessary surgery among patients with thyroid nodules. Alternatively, the different malign cells growth, angiogenesis, destructions of thyroid follicles, reparative changes, growth retardation, fibrosis and increased interstitial fluid pressure implicate the typical ultrasound appearance of papillary thyroid cancer (hypoechogenicity, irregular vascularization, microcalcifications, stiffness) which is essential to catch the suspicious nodules on the basis of their ultrasound appearance among large amount of benign nodules., J. Krátký, H. Vítková, J. Bartáková, Z. Telička, M. Antošová, Z. Límanová, J. Jiskra., and Obsahuje bibliografii
The sequence of changes in circulating immune cells and in free radical production was studied during the small intestine reperfusion. Rat small intestine ischemia/reperfusion was induced by a 45 min superior mesenteric artery occlusion followed by a 4-hour reperfusion. Samples of peripheral blood were collected every 20 min during reperfusion. While the number of polymorphonuclear leukocytes increased significantly both in the sham-operated controls and the experimental group (about 400 % at the end of reperfusion), a decrease in lymphocyte counts to 60 % was observed in the experimental group only. Although there were no changes in the counts of total T lymphocytes, a significant reduction in B cell counts was observed. Flow-cytometrical measurements showed no changes in the Tc subpopulation, while the Th subpopulation increased in the experimental group only. Free radical generation in blood (luminometric measurements) increased gradually and reached an eight-fold level by the end of reperfusion in both groups. Thus, it has been shown that the increase in free radical production is mainly due to the increased number of polymorphonuclear leukocytes mobilized already at the initial stages of reperfusion. The reduction in B lymphocyte population is probably due to homing mechanisms., J. Hamar, I. Rácz, M. Číž, A. Lojek, É. Pállinger, J. Fűrész., and Obsahuje bibliografii