The skin matrix metalloproteinase 3, tissue inhibitors of matrix metalloproteinase 2 and collagen III content changes in type 1 diabetes and insulin resistance treated with insulin a nd metformin were studied. Healthy adult male Wistar rats were obtained from experimental animal house, Department of Experimental Pharmacology, Medical University in Bialystok. The rats were divided randomly into five groups of 8 rats each. Control rats were injected intraperitoneally by NaCl. Type IDDM was induced by a single injection of Streptozocin. Insulin resistance was induced by a high -fat diet. The chosen groups of rats were also treated with insulin or metformin. ELISA K its (USCN Life Science, China) were used to measure content of matrix metallo - proteinase 3 (ELISA Kit for Matrix Metalloproteinase 3 - MMP3), tissue inhibitor of matrix metalloproteinase 2 (ELISA Kit for Tissue Inhibitors of Metalloproteinase 2 - TIMP2) and content of collagen type 3 (ELISA Kit for Collagen Type III - COL3). The results were reported as a median. The statistical significance was defined as p<0.05. Type 1 diabetes and insulin resistance have significantly reduced the quality of the skin, shown by the increase in cont ent of matrix metalloproteinase 3 and the decrease in content of tissue inhibitors of matrix metalloproteinase 2. Type 1 diabetes and insulin resistance have reduced the quality of the skin expressed by type III collagen content decrease but for future studies it is recommend to determine rat interstitial collagenase, MMP -13, as well. Insulin and metformin treatment improved the quality of the diabetic skin, demonstrated by the type III collagen content increase., M. Knaś, K. Wołosik, A. Zalewska, A. Mikucka-Niczyporuk, I. Kasacka, M. Niczyporuk., and Obsahuje bibliografii
The mood and behavior of individuals result from an orchestra of many factors. Among them steroids play an important role; however, only several common hormones have been investigated in this respect. It has been demonstrated that some steroid metabolites long considered merely the products of steroid hormone metabolism in fact possess considerable activity in the CNS. For this reason we studied the steroid metabolome including 50 analytes in 20 men with depression, 20 men with anxiety and 30 healthy controls. Significant differences were found not only between controls and men with either depression or anxiety, but also between men with depression and anxiety. Particularly striking were those steroids until now not generally associated with depression or anxiety, namely conjugated steroid forms, especially sulfates., M. Dušková, M. hill, M. Bičíková, M. Šrámková, D. Řípová, P. Mohr, L. Stárka., and Obsahuje bibliografii
Addiction to tobacco results in an imbalance of endocrine homeostasis in both sexes. This can also have impacts on fertility problems. The male reproductive system is less susceptible than that of females, with a worsening spermiogram in smokers, the most cited effect in the literature. However, the literature is inconsistent as to the effects of smoking on steroid hormone levels in men, and there is very little data on the effects of quitting smoking in men. In this study we followed 76 men before quitting smoking, and then after 6, 12, and 24 weeks and 1 year of abstinence. We measured basic anthropomorphic data and steroid hormone levels along with steroid neuroactive metabolites using GC-MS. We demonstrate lower androgen levels in men who smoke, and these changes worsened after quitting smoking. There was a drop in SHBG already in the first week of non-smoking, and levels continued to remain low. Male smokers have lower androgen levels compared to non-smokers. The lower the initial level of androgen, the lower the likelihood of success in quitting smoking. Changes in steroid hormones proved to be a promising marker for the prediction of success in quitting smoking., H. Jandikova, M. Duskova, K. Simunkova, B. Racz, M. Hill, E. Kralikova, K. Vondra, L. Starka., and Obsahuje bibliografii
The transformed C6 glial cells in cultures were treated with sodium mercaptoborate (Na2B12H11SH, BSH), a carrier of atomic targets (10B) of thermal neutrons for the neutron capture therapy of brain tumors. As shown by light microscopy, the therapeutic dose of BSH (100 µg/ml) did not alter the gross morphology and growth of the population of cells within a 72 h treatment interval. Electron microscopic analysis of these cells revealed activation of nucleoli and, occasionally, enlarged and bifurcated mitochondria. After 200 µg BSH/ml and 72 h treatment, growth of the cell population was inhibited and ultrastructural changes became more profound. They included condensation of chromatin and its allocation to the nuclear envelope which formed deeper invaginations. Mitochondria further increased in size and were characterized by slim or angular cristae. Moreover, in circumscribed segments of some of the slightly swollen mitochondria their cristae disappeared or were reduced to fine pouch-like structures localized near the continuous outer membrane, suggestive for a non-destructive restructuring of the inner mitochondrial membrane. The smooth pinocytotic vesicles near the plasma membrane, lysosomes and heterogeneous dense bodies were more frequent. The revealed subcellular targets of BSH may initiate the development of pharmacological protocols aimed to further improve the tolerance to BSH by the healthy tissues of patients undergoing BNCT of brain tumors, e.g. by intervention into the oxidative stress triggered likely by the altered mitochondria., V. Mareš, D. Krajčí, V. Lisá., and Obsahuje bibliografii
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 solid-state NMR measurements play an indispensable role in studies of interactions between biological membranes and peptaibols, which are amphipathic oligopeptides with a high abundance of α-aminobutyric acid (Aib). The solid-state NMR investigations are important in establishing the molecular models of the pore forming and antimicrobial properties of peptaibols, but rely on certain simplifications. Some of the underlying assumptions concern the parameters describing the 15N NMR chemical shielding tensor (CST) of the amide nitrogens in Aib and in conventional amino acids. Here the density functional theory (DFT) based calculations were applied to the known crystal structure of one of peptaibols, Ampullosporin A, in order to explicitly describe the variation of the 15N NMR parameters within its backbone. Based on the DFT computational data it was possible to verify the validity of the assumptions previously made about the differences between Aib and other amino acids in the isotropic part of the CST. Also the trends in the magnitudes and orientations of the anisotropic components of the CST, as revealed by the DFT calculations of the full periodic structure of Ampullosporin A, were thoroughly analyzed, and may be employed in future studies of peptaibols., J. Czernek, J. Brus., and Obsahuje bibliografii
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