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Using continuous two wavelength near-infrared technology to detect the variation in the consistency of oxygen hemoglobin in the muscle and the sports heart rate wireless real time collection technology, we devised the real time muscle tissue oxygenation and instantaneous heart rate experiment scheme and implemented it for the process of the 100 m run with two parameters given simultaneously. The experiment shows that the concentration of the oxygen hemoglobin in the muscle tissue continues decreasing after the end of the 100 m run, and the time interval between the moment when the concentration of the oxygen hemoglobin attains the minimum value and the moment when the athletes finish the 100 m run is (6.65 +/- 1.10) sec; while the heart rate continues increasing after the end of the 100 m run, and the time interval between the moment when the heart rate attains the maximum value and the moment when the athletes finish the 100 m run is (8.00 +/- 1.57) sec. The results show that the two wavelength near-infrared tissue oxygenation detection technology and the sports heart rate real time collection equipment can accurately measure the sports tissue oxygenation and the heart rate in the extreme intensity sport, and reveal the process of muscle oxygen transportation and consumption and its dynamic character with the heart rate in the extreme intensity sport.

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To avoid cerebral hypoxia caused by the imbalance between cerebral oxygen supply and consumption, regional cerebral oxygenation of patients need to be monitored at real time during cardiopulmonary bypass (CPB) surgery, and the physiological parameters can be regulated and emergent treatment can be used according to it. Using the near infrared (NIR) instrument developed by our group, cerebral oxygenation of the patients under cardiac surgery was monitored. The instrument consists of a two-wavelength near infrared light source and two near infrared detectors. Hemoglobin concentration changes of regional cerebral tissue were calculated, and by steady-state spatially resolved spectroscopy (SRS) algorithm, regional cerebral oxygen saturation (rSO2) was also calculated. Physiological parameters of patients, such as mixed venous oxygen saturation (SvO2), were measured by another monitor during CPB. Hemoglobin concentration changes were easily disturbed, but the anti-disturbance ability of rSO2 was good. The value of rSO2 could be detected all over the surgeries, but SvO2 could be detected only during CPB. There were positive correlations between rSO2 and SvO2 in most of the patients, but the correlation coefficients were not very high. This was because SvO2 reflects the saturation of the main venous, but rSO2 reflects regional cerebral oxygenation. So the physiological meaning of rSO2 and SvO2 is different. The results indicate that cerebral oxygenation of patients can be reflected by rSO2 during CPB, while only monitoring SvO2 is not enough.

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This study was designed to evaluate the role of bcl-2 transcriptional regulation induced by calmodulin I (CaM I) in pressure overload rat hypertrophic hearts. The model of hypertensive Sprague-Dawley rat was established by abdominal aortic constriction. The hearts were collected four weeks after abdominal aortic constriction. Velocity and isopyknic gradient centrifugation was employed to fractionate rat myocardial nuclei. Western blot analysis revealed a marked increase in phosphorylated cAMP response-element binding protein (pCREB) of cardiac hypertrophy group compared with that in control group (P<0.05), while the protein level of cAMP response-element binding protein (CREB) was constant (P>0.05). Immunohistochemistry results showed a significant increase of CaM I protein in cardiac hypertrophy group relative to the control group (P<0.05). Nuclear run off transcription assay displayed a significant increase in bcl-2 mRNA treated with trifluoperazne compared with non-drug treatment (P<0.05). The results obtained suggest that the transcription of bcl-2 is possibly regulated by CaM I hypertrophic rat hearts, and CREB phosphorylation seems to be a minor factor in bcl-2 transcriptional regulation.

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AIM: The effects of angiotensin II on the changes of Ca2+ signal in cultured rat neonatal myocytes were investigated in order to reveal the localization and distribution of elementary Ca2+ signaling units. METHODS: The cultured neonate rat myocytes were treated with angiotensin II, and calcium signal was detected using confocal laser scanning microscopy and fluo-4/AM calcium probe. RESULTS: The propagation of Ca2+ waves was observed in rest and angiotensin II stimulated cardiac myocytes. Calcium fluorescent intensity oscillated slightly in myocytes and the average intensity was much higher in the nucleus than in the cytosol, all of which could be magnified significantly by AngII (10(-6) mol/L). Ca2+ oscillation induced by Ang II was completely blocked by NO donor sodium nitroprusside. AngII evoked Ca2+ sparks close to the cell surface membrane, and couldn't be abolished by sodium nitroprusside. CONCLUSION: There are spatiotemporal dynamics of Ca2+ signaling patterns such as Ca2+ wave, Ca2+ spikes, Ca2+ oscillation and the whole cell Ca2+ transients induced by angiotensin II, which might play very important roles in cellular cardiac function.

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AIM: To evaluate whether protein phosphorylation and dephosphorylation in nuclei play roles in the development of myocardial hypertrophy, distribution of protein kinases and phosphatases in cell fractions were determined. METHODS: The model of hypertensive rat was established by abdominal aortic constriction. Velocity and isopyknic gradient centrifugation was employed to fractionate rat myocardium to membrane, cytosol and nuclei. Enzymatic methods were employed to determine kinases and phosphatases. RESULTS: Compare with control group, the activity of CaMK increased by 101.1% (P < 0.01) and 40.16% (P < 0.01) respectively in nuclear and membranous fractions, changed without significance in cytosolic fraction; the activity of calcineurin in nuclei increased by 43.57%, (P < 0.05), lightly changed without significance in membranous and cytosolic fractions. CONCLUSION: Nuclear translocation of CaMK and calcineurin, might play important roles on overload-induced cardiac hypertrophy.

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To investigate the regulation of Ca2+ in the isolated cardiac nuclei from rats which may illuminated the mechanism of nuclear calcium transport system. Elocity and isopyknic gradient centrifugation were employed to fractionate rat cardiac nuclei. Then fluo-4 confocal microscopy techniques was used to verify the changes of nuclear Ca2+. There are calcium-dependent Ca2+ uptake in the cardiac nuclear obtained from normal rats. The accumulation Ca2+ of cardiac nuclei in vitro from the incubating medium were not consistent with free [Ca2+] in incubating medium. The nuclear envelope was initially loaded with Ca2+ (1 mmol/L ATP and approximately 100 nmol/L Ca2+), Adequate Ca2+ loading was next confirmed by imaging the nuclear envelope and nucleoplasm. Exposure of Ca2+ -loaded nuclei to IP3, ryanodine or ryanodine + thapsigargin, respectively, resulted in a rapid and transient elevation of nucleoplasmic Ca2+ free concentration, this effects were abolished by pretreatment of cardiac nuclei with Ca2+ -ATPase inhibitor thapsigargin. Thapsigargin and IP3 receptor antagonist heparin induced nucleoplasmic Ca2+ free concentration decrease. Fluorescence experiments indicated that both ryanodine receptors and Ca2+ -ATPase were distributed in the outer layer of nuclear envelope, and inositol 1,4,5-trisphosphate receptors mainly dispersively localized at inner layer of nuclear envelope. The present study demonstrates that nuclear calcium were regulated by free Ca2+, IP3 and ryanodine. The results suggested calcium transport system might be present in the myocardial nuclei, the myocardial nuclei might served as one of calcium pools in myocardial cell.

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AIM: The characteristics of ryanodine receptor in rat cardiac sarcoplasmic reticulum (SR) and nuclear envelope (NE) were studied. METHODS: Velocity and isopyknic gradient centrifugation was employed to fractionate rat SR and NE. Ryanodine receptor was assayed with [3H] ryanodine saturate binding to the preparations. RESULTS: The maximal binding (Bmax) and dissociating constant (Kd) of ryanodine receptor in rat cardiac NE were, 1.7% and 60% of those in SR respectively. Phosphorylation in vitro by PKA and PKC increased Bmax of the receptors in SR by 372% and 121%, and augmented those in NE by 221% and 306%, without any effects on Kd. CONCLUSION: Ryanodine receptors were present in rat myocardial NE, with lower density and higher affinity than those located in SR, which can be activated by PKA and PKC.

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The characteristics of nuclear calcium regulation were investigated in isolated rabbit myocardial nuclei. It was found that calcium concentration in myocardial nuclei was 2.6 fold more than that in myocardial homogenate (P<0.O1), and the nuclear calcium content was only l/6 of the total cellular calcium. Ca-ATPase of myocardial nuclei was [Ca(2+] and [ATP] dependent. [Ca2+] dependent K(a) and V(max) at 2.O mmol/L [ATP] were 226 nmol/L and 3 460 nmol/(h.mg) protein respectively. [ATP] dependent K(m) value and V(max) at 400 nmol/L [Ca2+] were 376.5 &mgr;mol/L and 2 445 nmol/(h.mg) protein respectively. A positive correlation between nuclear 45)Ca2+ transport and Ca-ATPase activity was observed (r=O.945, P<0.01). The above result suggests that myocardial nuclei are able to transport calcium actively. The pathophysiological role of myocardial nuclear calcium transport should be further determined.