A characteristic of mast cells is the degranulation in response to various stimuli. Here we have investigated the effects of various physical stimuli in the human mast-cell line HMC-1. We have shown that HMC-1 express the transient receptor potential channels TRPV1, TRPV2 and TRPV4. In the whole-cell patchclamp configuration, increasing mechanical stress applied to the mast cell by hydrostatic pressure (–30 to –90 cm H2 O applied via the patch pipette) induced a current that could be inhibited by 10 µM of ruthenium red. This current was also inhibited by 20 µM SKF96365, an inhibitor that is among TRPV channels specific for the TRPV2. A characteristic of TRPV2 is its activation by high noxious temperature; temperatures exceeding 50 °C induced a similar ruthenium-red-sensitive current. As another physical stimulus, we applied laser light of 640 nm. Here we have shown for the first time that the application of light (at 48 mW for 20 min) induced an SKF96365-sensitive current. All three physical stimuli that led to activation of SKF96365-sensitive current also induced pronounced degranulation in the mast cells, which could be blocked by ruthenium red or SKF96365. The results suggest that TRPV2 is activated by the three different types of physical stimuli. Activation of TRPV2 allows Ca2+ ions to enter the cell, which in turn will induce degranulation. We, therefore, suggest that TRPV2 plays a key role in mast-cell degranulation in response to mechanical, heat and red laser-light stimulation., D. Zhang ... [et al.]., and Obsahuje seznam literatury
The biochemical model of excitation-contraction coupling in cardiomyocyte is presented and the validity of simulations of both physiological and pathological processes is discussed. The model of regulatory and actomyosin subsystems, even if it is rather simple in its regulatory subunit, gives results well consistent with experimental data. Specifically, intracellular free calcium levels ([Ca2+]i) were computed under various states of sarcoendoplasmic reticular Ca2+-ATPase (SERCA2) and compared to experimental findings. Computed results reproduced well both the increase in resting [Ca2+]i level and the attenuation of [Ca2+]i decline commonly observed in heart failure. Thus the computational simulations could help to identify core relations in studied systems by comparing results obtained using similar models of various complexities., M. Mlček, J. Neumann, O. Kittnar, V. Novák., and Obsahuje bibliografii
It is well known that the consumption of moderate doses of alcohol leads to the increase of HDL-cholesterol (HDL-C). Atheroprotectivity of HDL particles is based primarily on their role in reverse cholesterol transport (RCT). In the study with a crossover design 13 male volunteers were studied in two different regimens: i) drinking of 36 g alcohol daily and ii) drinking only non-alcoholic beverages, to test whether alcohol-induced increase of HDL cholesterol can affect cholesterol efflux (CHE) from cell culture of labeled human macrophages. Alcohol consumption induced significant (p<0.05) increases of HDL cholesterol from 1.25±0.32 to 1.34±0.38 mmol/l and Apo A1 from 1.34±0.16 to 1.44±0.19 g/l. These changes were combined with a slight increase of cholesterol efflux from 13.8±2.15 to 14.9±1.85 % (p=0.059). There were significant correlations between individual changes of HDL-C and Apo A1 concentrations and individual changes of CHE (0.51 and 0.60, respectively). In conclusion, moderate alcohol consumption changes the capacity of plasma to induce CHE only at a border line significance., I. Králová Lesná ... [et al.]., and Obsahuje bibliografii a bibliografické odkazy
We evaluate the mRNA expression of monocarboxylate transporters 1 and 4 (MCT1 and MCT4) in skeletal muscle (soleus, red and white gastrocnemius), heart and liver tissues in mice submitted to a single bout of swimming exercise at the maximal lactate steady state workload (MLSSw). After 72 h of MLSS test, the animals were submitted to a swimming exercise session for 25 min at individual MLSSw. Tissues and muscle samples were obtained at rest (control, n=5 ), immediately (n=5 ), 5 h (n=5 ) and 10 h (n=5 ) after exercise for determination of the MCT1 and MCT4 mRNA expression (RT-PCR). The MCT1 mRNA expression in liver increased after 10 h in relation to the control, immediate and 5 h groups, but the MCT4 remained unchanged. The MCT1 mRNA expression in heart increased by 31 % after 10 h when compared to immediate, but no differences were observed in relation to the control group. No significant differences were observed for red gastrocnemius in MCT1 and MCT4 mRNA expression. However, white gastrocnemius increased MCT1 mRNA expression immediately when compared to rest, 5 and 10 h test groups. In soleus muscle, the MCT1 mRNA expression increased immediately, 5 and 10 h after exercise when compared to the control. In relation to MCT4 mRNA expression, the soleus increased immediately and 10 h after acute exercise when compared to the control group. The soleus, liver and heart were the main tissues that showed improved the MCT1 mRNA expression, indicating its important role in controlling MLSS concentration in mice., G. G. de Araujo, C. A. Gobatto, F. de Barros Manchado-Gobatto, L. F. M. Teixeira, I. G. M. dos Reis, L. C. Caperuto, M. Papoti, S. Bordin, C. R: Cavaglieri, R. Verlengia., and Obsahuje bibliografii
Glucose-dependent insulinotropic peptide (GIP) contributes to incretin effect of insulin secretion which is impaired in Type 2 diabetes mellitus. The aim of this study was to introduce a simple meal test for evaluation of GIP secretion and action and to examine GIP changes in Type 2 diabetic patients. Seventeen Type 2 diabetic patients, 10 obese non-diabetic and 17 nonobese control persons have been examined before and after 30, 60 and 90 min stimulation by meal test. Serum concentrations of insulin, C-peptide and GIP were estimated during the test. Impaired GIP secretion was found in Type 2 diabetic patients as compared with obese non-diabetic and non-obese control persons. The AUCGIP during 90 min of the meal stimulation was significantly lower in diabetic patients than in other two groups (p<0.03). Insulin concentration at 30 min was lower in diabetic than in non-diabetic persons and the GIP action was delayed. The ΔIRI/ΔGIP ratio increased during the test in diabetic patients, whereas it progressively decreased in obese and nonobese control persons. Simple meal test could demonstrate impaired GIP secretion and delayed insulin secretion in Type 2 diabetic patients as compared to obese non-diabetic and nonobese healthy control individuals., J. Škrha., and Obsahuje bibliografii a bibliografické odkazy
Patch clamp method developed more than 30 years ago is widely used for investigation of cellular excitability manifested as transmembrane ionic current and/or generation of action potentials. This technique could be applied to measurement of ionic currents flowing through individual (single) ion channels or through the whole assembly of ion channels expressed in the whole cell. Whole cell configuration is more common for measurement of ion currents and the only one enabling measurement of action potentials. This method allows detailed analysis of mechanisms and structural determinants of voltagedependent gating of ion channels as well as regulation of channel activity by intracellular signaling pathways and pharmacological agents., M. Karmažínová, L'. Lacinová., and Obsahuje bibliografii a bibliografické odkazy
The multi-functional proteins, insulin-like growth factor-I (IGF-I) and leptin were present in seminal plasma from different species. Concentrations of IGF-I in equine and porcine semen were 20 and 17.5 ng/ml, respectively. Seminal plasma concentrations of leptin were 1 ng/ml in human and 11 ng/ml in porcine samples., B. R. Lackey, S. L. Gray, D. M. Henricks., and Obsahuje bibliografii
The Vasotrac monitor provides non-invasive near-continuous blood pressure monitoring and is designed to be an alternative to direct intra-arterial blood pressure (BP) measurement. As compared to radial artery invasive BP and upper arm noninvasive BP, Vasotrac readings have been found to have a good agreement with them. However, discrepancies have been reported when rapid changes in BP exist. In the present study we compared BP measured by the Vasotrac monitor on the radial artery with that recorded on the finger arteries by the differential oscillometric device allowing measurement on the beat-to-beat basis. Comparisons were performed on the mean arterial pressure (MAP) level. Special attention was paid to the signal conditioning before comparison of pressures of different temporal resolution. Altogether 383 paired MAP measurements were made in 14 healthy subjects. Based on all 383 paired measurements, the MAP values measured at the radial artery at rest were 4.8±6.0 mm Hg higher than those measured on fingers. The observed difference between the Vasotrac and differential oscillometric device can be explained by different measurement sites. This result is consistent with previous investigations, and the Vasotrac monitor can be considered to adequately track relatively rapid MAP changes on the radial artery. Attention should be paid to a proper signal conditioning before comparison of results obtained by different devices., K. Jagomägi ... [et al.]., and Obsahuje bibliografii a bibliografické odkazy
A mitral allograft is us ed exceptionally in the mitral, as well as in the tricuspid position, mostly as an experimental surgical procedure. The authors decided to evaluate the possibility of inserting a cryopreserved mitral allograft into the tricuspid position in a sheep experimental model. Within the framework of this experimental project the mechanical properties of the cryopreserved mitral allograft were tested. A novel methodology studying the functional unit composed of mitral annulus, leaflet, chordae tendinaea, and papillary muscle is presented. A five-parameter Maxwell model was applied to characterize the viscoelastic behavior of sheep mitral valves. A control group of 39 fresh mitral specimens and a test group of 13 cryopreserved mitral allografts from tissue bank were tested. The testing protocol consisted of six loading cycles with 1 mm elongation every 5 min. There was no significant difference in the mean values of the determined parameters (p> 0.05) which confirms the main hypothesis that cryopreservation does not influence significantly material parameters characterizing the tissue mechanics. Slight discrepancy is observed in variances of viscous parameters suggesting that the values of the test group may be spread over larger interval due to the treatment., J. Hlubocký ... [et al.]., and Obsahuje bibliografii a bibliografické odkazy
a1_Mechanical properties of scaffolds seeded with mesenchymal stem cells used for cartilage repair seem to be one of the critical factors in possible joint resurfacing. In this paper, the effect of adding hyaluronic acid, hydroxyapatite nanoparticles or chitosan nanofibers into the cross-linked collagen I on the mechanical response of the lyophilized porous scaffold has been investigated in the dry state at 37 oC under tensile loading. Statistical significance of the results was evaluated using ANOVA analysis. The results showed that the addition of hyaluronic acid significantly (p<<0.05) reduced the tensile elastic modulus and enhanced the strength and deformation to failure of the modified cross-linked collagen I under the used test conditions. On the other hand, addition of hydroxyapatite nanoparticles and chitosan nanofibers, respectively, increased the elastic modulus of the modified collagen ten-fold and four-fold, respectively. Hydroxyapatite caused significant reduction in the ultimate deformation at break while chitosan nanofibers enhanced the ultimate deformation under tensile loading substantially (p<<0.05). The ultimate tensile deformation was significantly (p<<0.05) increased by addition of the chitosan nanofibers. The enhanced elastic modulus of the scaffold was translated into enhanced resistance of the porous scaffolds against mechanical load compared to scaffolds based on cross-linked neat collagen or collagen with hyaluronic acid with similar porosity. It can be concluded that enhancing the rigidity of the compact scaffold material by adding rigid chitosan nanofibers can improve the resistance of the porous scaffolds against compressive loading, which can provide more structural protection to the seeded mesenchymal stem cells when the construct is implanted into a lesion., a2_Moreover, scaffolds with chitosan nanofibers seemed to enhance cell growth compared to the neat collagen I when tested in vitro as well as the scaffold stability, extending its resorption to more than 10 weeks., J. Jančář, A. Slovíková, E. Amler, P. Krupa, H. Kecová, L. Plánka, P. Gál, A. Nečas., and Obsahuje bibliografii