This work evaluates the myocardial protective potential of potassium cardioplegia on ischaemically arrested and reperfused hearts by two cardioplegic solutions: the University of Wisconsin solution (UW) and the standard crystalloid solution of St. Thomas’ Hospital (ST). Evaluation of myocardial preservation was based on creatine kinase and lactate releases and on high-energy phosphate preservation of isolated rabbit hearts after 4 hours’ hypothermic ischaemia. A morphometric ultrastructural evaluation of mitochondria in cardiomyocytes was also performed. The hearts of 24 rabbits were normothermally perfused with oxygenated Krebs-Henseleit solution for 30 min (Langendorff preparation), and the baseline contractile performance and biochemical parameters were evaluated. The hearts were then arrested and stored in the cardioplegic solutions (12 UW and 12 ST) at 4 °C for 4 hours. The hearts were then rewarmed and reperfused with oxygenated Krebs-Henseleit solution for further 30 min. At the end of reperfusion, creatine phosphate and high energy phosphates were higher with UW (p<0.05); creatine kinase release during reperfusion was significantly lower with UW both at 15 min (p<0.01) and at 30 min (p<0.05). Lactate release during the first 15 min of reperfusion was about doubled (p<0.05) with respect to controls in both groups; at 30 min this increase had almost vanished ( + 8 %) with UW but not with ST ( + 30 %). Ultrastructural morphometry did not show any significant difference at the level of mitochondria between the two treatments. The results indicate, for UW, an improved myocardial preservation associated with relative retention of high-energy phosphates and higher recovery of mechanical function, accelerated metabolic recovery and reduced stress of cell membranes.
The aim of this study was to evaluate myofibrillar creatine kinase (EC 2.7.3.2) activity on the background of the effect of substrate channeling by myosin ATPase and to compare it with creatine kinase (CK) activity of whole skinned fibers. In order to assess CK activity, skinned fibers were prepared from the rat psoas major muscles defined by light microscopy. The activity in permeabilized fibers after treatment with saponin, Triton X-100 and Ca2+-free medium reached 2.80, 6.97 and 3.32 m mol ATP min-1 mg-1 protein, respectively, when a coupled enzyme assay system with external hexokinase and glucose-6-phosphate dehydrogenase was used. Transmission electron microscopy (TEM) revealed a possible interference among activities of sarcolemmal, sarcoplasmic, myofibrillar and mitochondrial CK from persisting structures. For evaluation of the myofibrillar CK itself, a pure myofibrillar fraction was prepared. Fraction purity was confirmed by TEM and by enzymatic assays for marker enzymes. Two procedures, i.e. the coupled enzyme assay and the evaluation of phosphocreatine (Pcr) concentration before and after the CK reaction, were used for measurement of CK activity in this fraction. The procedures resulted in 3.2 nmol ATP min-1 mg-1 protein and 7.6 nmol PCr min-1 mg-1 protein, respectively. These alternative approaches revealed a discrepancy between the reacting portions of Pcr by more than 50 % , which provides information about the size of the effect, generally described as substrate channeling., M. Gregor, J. Mejsnar, A. Janovská, J. Žurmanová, O. Benada, B. Mejsnarová., and Obsahuje bibliografii
Our aim was to investigate whether hyperthermia before exercise protects against exercise-induced skeletal muscle damage. Two hyperthermia protocols were evaluated. In the first, male ICR mice were exposed to 30 min of whole-body heat in an environmental chamber at an ambient temperature of 42 °C. Heat-exposed and non-heat-exposed mice subsequently completed 60 min of downhill running on a treadmill, 24 h after exposure. Heat exposure significantly increased HSP70 and HSP25 content in the soleus muscle compared to controls. Plasma creatine kinase, muscle β-glucuronidase, and histochemical (hematoxylin and eosin stain) analysis demonstrated that muscle damage was lower in the heatexposed mice than in the non-heat-exposed mice. In the second, the effect of regional heating of the legs, by microwave diathermy, on the prevention of exercise-induced muscle damage was evaluated in male Wistar rats. Microwave-treated and nonmicrowave-treated rats again completed the running protocol 24 h after exposure. Microwave diathermy increased the muscle temperature to 40 °C, significantly increased HSP70 and HSP25 content in the soleus muscle, and significantly attenuated exercise-induced muscle damage. Therefore, hyperthermia before exercise increases skeletal muscle HSPs and attenuates the risk of exercise-induced muscle injury.
Troponin T (TnT) is recently being considered to be an important diagnostic marker of myocardial damage in adults, but this marker has not yet been used in neonates. The present study was designed to determine the normal level of cardiac TnT in the cord blood of healthy term neonates. Cardiac troponin T concentration in cord blood was measured in 15 healthy term neonates using commercial kit (Enzymun-Test System, Boehringer, Mannheim). TnT serum concentration was 0.05±0.04 /ig/1 in 10 of 15 babies whereas in the remaining 5 haemolysed samples its concentration was elevated (mean 0.19±0.07 /vg/1). It is important to consider that incidental haemolysis of blood samples can mimic pathological elevation of TnT by interfering with the assay.