Increased generation of reactive oxygen species results in the formation of fluorescent end-products of lipid peroxidation – lipofuscin-like pigments (LFP). LFP increased up to six-fold from the fetal value in the rat heart immediately after birth. In the experimental design of this study the fetuses were sampled 1 day before birth, and then the samples were collected on postnatal days 1, 4, 7, 10, 15, 30, and 60. Males and females were compared on day 30 and 60 when the difference between right and left ventricle was studied as well. Four LFP fluorophores were analyzed: F355/440, F310/470, F350/450, F315/450 (excitation/emission, nm). All fluorophores decreased on day 4 relative to day 1, subsequent transient increases ended in a significant decrease on day 60. However, the LFP levels on day 60 are still about threefold higher than those in fetuses. Differences between male and female hearts were observed on day 30. The corresponding male ventricles contained by one third higher concentration of LFP than the female counterparts. The increase in LFP concentration in male ventricles on day 30 was only transient, no difference between corresponding male and female ventricles was found on day 60. The most distinguished feature in the male heart was a sharp LFP decrease in the right ventricle on day 60., J. Wilhelm, I. Ošťádalová., and Obsahuje seznam literatury
Our present focus on the hypoxic immature heart is driven by clinical urgency: cyanotic congenital cardiac malformations remain the single largest cause of mortality from congenital defects and ischemic heart disease is no more the disease of the fifth and older decades but its origin as well as risk factors are present already during early ontogeny. Moreover, the number of adult patients operated for cyanotic congenital heart disease during infancy steadily increases. This group approaches the age of the rising risk of serious cardiovascular diseases, particularly ischemic heart disease. Experimental results have clearly shown that the immature heart is significantly more tolerant to oxygen deficiency than the adult myocardium. However, the mechanisms of this difference have not yet been satisfactorily clarified; they are likely the result of developmental changes in cardiac energy metabolism, including mitochondrial function. The high resistance of the newborn heart cannot be further increased by ischemic preconditioning or adaptation to chronic hypoxia; these protective mechanisms appear only with decreasing tolerance during development. Resistance of the adult myocardium to acute oxygen deprivation may be significantly influenced by perinatal hypoxia. These results suggest that the developmental approach offers new possibilities in the studies of pathogenesis, prevention and therapy of critical cardiovascular diseases., B. Ošťádal ... [et al.]., and Obsahuje seznam literatury
a1_The purpose of the present study was to compare the ontogenetic development of the activity of myocardial energy-supplying enzymes in two mammalian species, differing significantly in their level of maturation at birth. The animals were investigated during the late prenatal period and 2, 7, 14, 21, 25, 30, 63, 120 and 730 days after birth in the rat and 2, 21, 84 and 175 days in the guinea-pig. The following enzymes were assayed in the right and left ventricular myocardium: lactate dehydrogenase (LDH, lactate uptake and/or formation), triose phosphate dehydrogenase (TPDH, carbohydrate metabolism), glycerol phosphate dehydrogenase (GPDH, glycerol-P shuttle)), hexokinase (HK, glucose phosphorylation), malate dehydrogenase (MDH, tricarboxylic cycle), citrate synthase (CS, tricarboxylic cycle) and hydroxyacyl-CoA dehydrogenase (HOADH, fatty acid breakdown). The rat heart, highly immature at birth, exhibits three different developmental patterns of energy-supplying enzymes, identical in both ventricles: (i) two mitochondrial enzymes of aerobic metabolism (CS, HOADH) and GPDH have a relatively low activity at the end of prenatal life; thereafter their activity steadily increases, approaching the adult levels between the 3rd and 4th postnatal weeks. A significant decrease was observed between the 4th and 24th months. (ii) MDH and LDH: prenatal values were significantly higher as compared with the 2nd postnatal day; after this period the activities increased up to adulthood (4 months) and decreased during senescence. (iii) The activities of HK and TPDH are characterized by only moderate changes during development. HK differs from all other enzymes by the highest prenatal values, which exceed even adult values. In contradiction to the rat heart, the developmental differences in more mature guinea-pig heart were significantly less pronounced., a2_The only ontogenetic differences observed were the lower activities of enzymes connected with aerobic metabolism at the end of the prenatal period. Our results point to possible differences in the development of adaptive metabolic pathways in animals with different levels of maturation at birth., A. Bass, M. Stejskalová, A. Stieglerová, B. Ošťádal, M. Šamánek., and Obsahuje bibliografii
Our data indicate the significant intrinsic efficacy of GABABreceptors in rat brain cortex already at birth (PD1, PD2). Subsequently, baclofen- and SKF97541-stimulated G-protein activity, measured by agonist-stimulated, high-affinity [35S]GTPγS binding assay, was increased; the highest level of both baclofen and SKF97541-stimulated [35S]GTPγS binding was detected between PD10 and PD15. In older rats, baclofen- and SKF97541- stimulated [35S]GTPγS binding was continuously decreased so, that the level in adult, 90-days old animals, was not different from that in newborn animals. The potency of G-protein response to baclofen (characterized by EC50 values) was also high at birth but unchanged by further postnatal development. An individual variance among different agonists was observed in this respect as the potency of SKF97541 response was decreased between the birth and adulthood. Accordingly, the highest plasma membrane density of GABAB-R, determined by saturation binding assay with antagonist [3 H]CGP54626, was measured in 1-day old animals (2.27±0.08 pmol · mg-1). The further development was reflected in a decrease of [3 H]CGP54626 binding as the Bmax values of 1.38±0.05 and 0.93±0.04 pmol · mg-1 were determined in PM isolated from 13- and 90-days old rats, respectively., D. Kagan, ... [et al.]., and Obsahuje seznam literatury
Local application of four concentrations of bicuculline methiodide (a specific antagonist of GABAA receptors) was used to study a sensitivity of somatosensory cortex in four age groups of immature rats with implanted electrodes. Presence and latencies of two epileptic phenomena (focal discharges and seizures) were evaluated. Focal discharges exhibited moderate tendency to a decrease of sensitivity to bicuculline methiodide with maturation. Concentration-effect relation of incidence of focal discharges was observed only in 7- and 12-day-old but not in older animals. Results with incidence and latencies of seizures did not show relations to age or concentration of bicuculline. Neither of the epileptic phenomena can be used as a reliable index of cortical maturation., P. Mareš, K. Bernášková, H. Kubová., and Seznam literatury
An ontogenetic study of ecto-ATPase activity and the content of enzyme proteins was assessed in the caudate nucleus and hippocampal synaptic plasma membranes isolated from rats at various ages (15, 30, 90, 180 and 365 days). The ontogenetic profile revealed that the enzyme activities in both brain areas were the highest on day 30 and 365, while the ecto-ATPase protein abundance was the highest on day 15 after birth. Possible explanation for obtained ontogenetic profile and the discrepancy between activity and abundance may reside in the fact that ecto-ATPase during development could exert additional roles other than those related to metabolism of ATP. It is likely that ecto-ATPase, regulating the concentration of ATP and adenosine in synaptic cleft, has important role in the processes of brain development and aging., A. Banjac, N. Nedeljković, A. Horvat, D. Kanazir, G. Nizekić., and Obsahuje bibliografii
The article gives an overview of developmental aspects of the ontogeny of pain both in experimental models and in children. The whole article is devoted to the ontogenesis in pain perception and the possible influence on it. The role of endogenous opioids on the development of pain and other important substances such as serotonin, nerve growth factor (NGF) and nicotine are mentioned. There are also important differences of the ontogenesis of thermal and mechanical nociceptive stimulation. The physiological and pathophysiological findings are the backgrounds for principles of treatment, taking into account the special status of analgesics during ontogeny. In particular there are mentioned the special effects of endogenous opioids and especially morphine. It describes the role of vitamin D and erythropoietin during the development of pain perception. This article also mentioned the critical developmental periods in relation to the perception of pain. The attention is paid to stress and immunological changes during the ontogeny of pain. Another important role is played by microglia. The work is concluded by some statements about the use of physiological and pathophysiological findings during the treatment of pain in pediatric practice. Codein analgesia is also described because codein starts to be very modern drug with the dependence., R. Rokyta, J. Fricová., and Obsahuje seznam literatury
b1_Large number of extracellular signals is received by plasma membrane receptors which, upon activation, transduce information into the target cell interior via trimeric G-proteins (GPCRs) and induce activation or inhibition of adenylyl cyclase enzyme activity (AC). Receptors for opioid drugs such as morphine ( μ-OR, δ-OR and κ-OR) belong to rhodopsin family of GPCRs. Our recent results indicated a specific up-regulation of AC I (8-fold) and AC II (2.5-fold) in plasma membranes (PM) isolated from rat brain cortex exposed to increasing doses of morphine (10-50 mg/kg) for 10 days. Increase of ACI and ACII represented the specific effect as the amount of ACIII-ACIX, prototypical PM marker Na, K-ATPase and trimeric G-protein α and β subunits was unchanged. The up-regulation of ACI and ACII faded away after 20 days since the last dose of morphine. Proteomic analysis of these PM indicated that the brain cortex of morphine-treated animals cannot be regarded as being adapted to this drug because significant up-regulation of proteins functionally related to oxidativ e stress and alteration of brain energy metabolism occurred. The number of δ-OR was increased 2-fold and their sensitivity to monovalent cations was altered. Characterization of δ-OR-G-protein coupling in model HEK293 cell line indicated high ability of lithium to support affinity of δ-OR response to agonist stimulation. Our studies of PM structure and function in context with desensitization of GPCRs action were extended by data indicating part icipation of cholesterol-enriched membrane domains in agonist-specific internalization of δ-OR. In HEK293 cells stably expressing δ-OR-G i 1 α fusion protein, depletion of PM cholesterol was associated with the decrease in affinity of G-protein response to agonist stimulation, whereas maximum response was unchanged., b2_drophobic interior of isolated PM became more “fluid”, chaotically organized and accessible to water molecules. Validity of this conclusion was supported by the analysis of an immediate PM environment of cholesterol molecules in living δ -OR-G i 1 α-HEK293 cells by fluorescent probes 22- and 25-NBD-cholesterol. The alteration of plasma membrane structure by cholesterol depletion made the membrane more hydrated. Unders tanding of the positive and negative feedback regulatory loops among different OR-initiated signaling cascades (μ-, δ -, and κ-OR) is crucial for understanding of the long-term mechanisms of drug addiction as the decrease in functional activity of μ-OR may be compensated by increase of δ-OR and/or κ-OR signaling., H. Ujčíková ... [et al.]., and Obsahuje bibliografii a bibliografické odkazy
Genetic factors may contribute to the differential response to opioids. The aim of this study was to evaluate the association between polymorphisms of μ1-opioid receptor gene OPRM1 (rs1799971), and P-glycoprotein transporter gene ABCB1 (rs1045642, rs2032582), and piritramide efficacy under postoperative patient-controlled analgesia (PCA). In 51 patients, OPRM1 variant was associated with decreased efficacy in early postoperative period evidenced by sum of pain intensity difference in the 0-6 h postoperative period (SPID0-6), (F=3.27, p=0.029). Mean (SD) SPID0-6 was observed in the 118AA genotype 22.9 (6.1) mm, which was significantly higher from the 118GG genotype 10.0 (4.4) mm, p=0.006. The lowest cumulative dose was recorded in 118AA genotype 19.1 (9.8) mg, which was significantly less than in the 118GG genotype group 36.6 (6.1) mg, p=0.017. Opioid-induced adverse effects were observed in 11, 30, and 100 % of patients in 118AA, 118AG, and 118GG genotype groups, respectively (p<0.05). Piritramide efficacy and safety was not significantly affected by ABCB1 (rs1045642, rs2032582) polymorphisms. Variant OPRM1 118G allele is associated with decreased acute postoperative pain relief after piritramide. Decreased efficacy leads to higher drug consumption under PCA settings, which however, does not fully compensate insufficient pain relief, but increases incidence of adverse effects., O. Bartošová, O. Polanecký, F. Perlík, S. Adámek, O. Slanař., and Obsahuje bibliografii
The determination of cytochrome c oxidase (COX) activity represents an important indicator for the evaluation of cell oxidative capacity. However, it has been shown repeatedly that different factors modify the rate of COX activity under various experimental conditions. The most important concern the ionic concentrations of the medium and the application of various detergents for the solubilization of mitochondrial membranes. We found the highest activity of COX in rat heart homogenates and mitochondria at 40-60 mM potassium phosphate. The rate of COX activity is dependent on the detergent/protein (P) ratio. Using n-dodecyl-b-D-maltoside (lauryl maltoside, LM) as the detergent, we obtained the highest activity at LM/P ratios of (50:100):1. By kinetic measurements of low-affinity binding sites in heart mitochondria, we found Vlim values of 4.3 and 22.2 mmol cytochrome c per min per mg P in the presence or absence of lauryl maltoside, respectively. The Km values were 16.7 mmol in the presence or absence of lauryl maltoside. Our results thus indicate that 1) the exact assessment of COX activity in heart homogenates and mitochondria requires the determination of optimum phosphate concentrations in the medium used, and 2) even small modifications of the experimental procedure may induce significant differences in the maximum values of COX activity., A. Stieglerová, Z. Drahota, B. Ošťádal, J. Houštěk., and Obsahuje bibliografii