Phosphorylation of extrajunctional Cx43 in ischemic-preconditioned rat hearts.

Ischemic preconditioning (IP) has been found to intensify ischemia-induced spreading of connexin 43 (Cx43) into the extrajunctional plasma membrane of the rat heart. Since ischemia is known to induce connexin dephosphorylation and plasmalemmal permeabilization, we considered whether IP might delay dephosphorylation of also extrajunctional connexin and thus impede myocyte permeabilization. Regional myocardial ischemia of both 15 and 45 min duration with and without IP, respectively, was induced in anesthetized rats. Sections of the hearts were immunostained for phosphorylated and dephosphorylated Cx43. The ratios of immunofluorescence in gap junctions and extrajunctional membranes, respectively, versus myocyte interiors were determined as relative fluorescence units (RFU). IP, however, was not found to reduce gap junctional dephosphorylated Cx43 (normal perfusion: 1.23 +/- 0.06 RFU; 15 min ischemia without and with IP: 1.80 +/- 0.09 and 2.10 +/- 0.24 RFU; 45 min ischemia without and with IP: 3.12 +/- 0.55 and 2.57 +/- 0.33 RFU, respectively). Extrajunctionally, only dephosphorylated Cx43 was found. Its occurrence was not prevented by IP, it was rather intensified by IP (normal perfusion: 1.02 +/- 0.01 RFU, 15 min ischemia without and with IP: 1.06 +/- 0.03 and 1.15 +/- 0.05 RFU (P < 0.02), and 45 min ischemia without and with IP: 1.30 +/- 0.07 and 1.27 +/- 0.07 RFU, respectively). Although under ischemia, junctional and extrajunctional Cx43 predominated in the dephosphorylated state, only a small fraction of myocytes were found permeabilized to propidium iodide. In conclusion, IP did not prevent ischemia-induced Cx43 dephosphorylation in gap junctions and extrajunctional plasma membranes. It is thus more likely that changes in non-channel functions or other localizations of Cx43 are involved in IP-induced myocardial protection.

Effect of lower body negative pressure and gravity on regional lung ventilation determined by EIT.

The aim of our study was to check the effect of varying blood volume in the chest and gravity on the distribution of ventilation and aeration in the lungs. The change in intrathoracic blood volume was elicited by application of lower body negative pressure (LBNP) of -50 cmH2O. The variation of gravity in terms of hypogravity (approximately 0g) and hypergravity (approximately 2g) was induced by changes in vertical acceleration achieved during parabolic flights. Local ventilation magnitude and end-expiratory lung volume were determined in eight human subjects in the ventral and dorsal lung regions within a transverse cross-section of the lower chest by electrical impedance tomography. The subjects were studied in a 20 degrees head-down tilted supine body position during tidal breathing and full forced expirations. During tidal breathing, a significant effect of gravity on local magnitude of ventilation and end-expiratory lung volume was detected in the dorsal lung regions both with and without LBNP. In the ventral regions, this gravity dependency was only observed during LBNP. During forced expiration, LBNP had almost no effect on local ventilation and end-expiratory lung volume in either lung region. Gravity significantly influenced the end-expiratory lung volumes in dorsal lung regions. The results indicate that exposure to LBNP exerts a less appreciable effect on regional lung ventilation than the acute changes in gravity.

Contribution of myoglobin-induced increases in vascular resistance to shock decompensation in experimental Crush-syndrome in anesthetized rats.

Myoglobin is known to become nephrotoxic when released in greater amounts from skeletal muscle into the general circulation during shock. The present study deals with the question as to whether a myoglobin-induced increase in vascular tone additionally contributes to the detrimental role of this protein in hypovolemic shock. Anesthetized rats were subjected to 250 mg kg x h(-1) myoglobin infused i.v. during hemorrhagic hypotension of 50 mmHg. Shock survival time was measured, as were blood flow and vascular resistance in kidney, intestine, brain, and heart, using the microsphere method. Rats subjected to only myoglobin or hemorrhage survived a period of >120 min; in contrast, rats, exposed to both myoglobin and hemorrhage died at 68 +/- 9 min. When the animals subjected to only hemorrhage and to myoglobin/ hemorrhage were compared, significantly lower values were found in the latter group with respect to blood flow in the kidney (1.7 +/- 0.1 vs. 0.2 +/- 0.05 ml x min(-1) x g(-1)), small intestine (1.0 +/- 0.1 vs. 0.5 +/- 0.1 ml x min(-1) x g(-1)), cardiac output (112 +/- 5 vs. 62 +/- 10 ml(-1) x min(-1) x kg(-1)), and significantly higher values of total peripheral vascular resistance (0.45 +/- 0.02 vs. 0.81 +/- 0.12 mmHg x min x ml(-1) x kg) at 50 min of hypotension. It is assumed that these effects of myoglobin are induced by its ability to scavenge endogenous nitric oxide, because a modified, non-nitrosylable myoglobin was unable to induce such effects. The results support the view that a pathological release of myoglobin into the general circulation causes increases in vascular resistance of vital organs that may contribute to decompensation of tissue supply when occurring in hypovolemic shock.

Distribution of lung ventilation in spontaneously breathing neonates lying in different body positions.

OBJECTIVE: The aim of our study was to determine the effect of the irregular spontaneous breathing pattern and posture on the spatial distribution of ventilation in neonates free from respiratory disease by the non-invasive imaging method of electrical impedance tomography (EIT). Scanning of spontaneously breathing neonates is the prerequisite for later routine application of EIT in babies with lung pathology undergoing ventilator therapy. DESIGN: Prospective study. SETTING: Neonatal intensive care unit at a university hospital. PATIENTS: Twelve pre-term and term neonates (mean age: 23 days; mean body weight: 2,465 g; mean gestational age: 34 weeks; mean birth weight: 2,040 g). INTERVENTIONS: Change in body position in the sequence: supine, right lateral, prone, supine. MEASUREMENTS AND RESULTS: EIT measurements were performed using the Göttingen GoeMF I system. EIT scans of regional lung ventilation showing the distribution of respired air in the chest cross-section were generated during phases of rapid tidal breathing and deep breaths. During tidal breathing, 54.5+/-8.3%, 55.2+/-10.5%, 59.9+/-8.4% and 54.2+/-8.5% of inspired air (mean values +/- SD) were directed into the right lung in the supine, right lateral, prone and repeated supine postures respectively. During deep inspirations, the right lung ventilation accounted for 52.6+/-7.9%, 68.5+/-8.5%, 55.4+/-8.2% and 50.5+/-6.6% of total ventilation respectively. CONCLUSION: The study identified the significant effect of breathing pattern and posture on the spatial distribution of lung ventilation in spontaneously breathing neonates. The results demonstrate that changes in regional ventilation can easily be determined by EIT and bode well for the future use of this method in paediatric intensive care.

Detection of local lung air content by electrical impedance tomography compared with electron beam CT.

The aim of the study was to validate the ability of electrical impedance tomography (EIT) to detect local changes in air content, resulting from modified ventilator settings, by comparing EIT findings with electron beam computed tomography (EBCT) scans obtained under identical steady-state conditions. The experiments were carried out on six anesthetized supine pigs ventilated with five tidal volumes (VT) at three positive end-expiratory pressure (PEEP) levels. The lung air content changes were determined both by EIT (Goe-MF1 system) and EBCT (Imatron C-150XP scanner) in six regions of interest, located in the ventral, middle, and dorsal areas of each lung, with respect to the reference air content at the lowest VT and PEEP, as a change in either local electrical impedance or lung tissue density. An increase in local air content with VT and PEEP was identified by both methods at all regions studied. A good correlation between the changes in lung air content determined by EIT and EBCT was revealed. Mean correlation coefficients in the ventral, middle, and dorsal regions were 0.81, 0.87, and 0.93, respectively. The study confirms that EIT is a suitable, noninvasive method for detecting regional changes in air content and monitoring local effects of artificial ventilation.

Regional lung perfusion as determined by electrical impedance tomography in comparison with electron beam CT imaging.

The aim of the experiments was to check the feasibility of pulmonary perfusion imaging by functional electrical impedance tomography (EIT) and to compare the EIT findings with electron beam computed tomography (EBCT) scans. In three pigs, a Swan-Ganz catheter was positioned in a pulmonary artery branch and hypertonic saline solution or a radiographic contrast agent were administered as boli through the distal or proximal openings of the catheter. During the administration through the proximal opening, the balloon at the tip of the catheter was either deflated or inflated. The latter case represented a perfusion defect. The series of EIT scans of the momentary distribution of electrical impedance within the chest were obtained during each saline bolus administration at a rate of 13/s. EBCT scans were acquired at a rate of 3.3/s during bolus administrations of the radiopaque contrast material under the same steady-state conditions. The EIT data were used to generate local time-impedance curves and functional EIT images showing the perfusion of a small lung region, both lungs with a perfusion defect and complete both lungs during bolus administration through the distal and proximal catheter opening with an inflated or deflated balloon, respectively. The results indicate that EIT imaging of lung perfusion is feasible when an electrical impedance contrast agent is used.

Distribution of ventilation in young and elderly adults determined by electrical impedance tomography.

To determine the effect of age and posture on regional lung ventilation, eight young (26 +/- 1 years, mean +/- S.D.) and eight old (73 +/- 5 years) healthy men were studied by electrical impedance tomography in four body positions (sitting, supine, right and left lateral). The distribution of gas into the right and left lung regions was determined in the chest cross-section during tidal breathing at the resting lung volume, near residual volume and total lung capacity, as well as forced and slow vital capacity maneuvers. In the young, significant posture-dependent changes in gas distribution occurred during resting tidal breathing whereas they were absent in the elderly. In the older subjects, the contribution of the right lung to global ventilation fell with the transition from sitting to supine posture during both full expiration maneuvers. During forced vital capacity, the high flow rate and early airway closure in the dependent lung, occurring at higher volumes in the elderly, minimized the posture-dependency in gas distribution which was present during the slow maneuver. Our study revealed the significant effect of age on posture-dependent changes in ventilation distribution.

Acoustic phenomena and valve dysfunction in cardiac prostheses: data acquisition and collection via the Internet.

BACKGROUND AND AIM OF THE STUDY: The study aim was to investigate whether: (i) by detection of changing acoustic sound phenomena, minimal changes in prosthetic valve function may be detected earlier than with echocardiography, invasive diagnosis or clinically; (ii) patients can record and pass on signals with a high level of reproducibility from any location via the Internet; and (iii) clinical data evaluation permits conclusions to be drawn on changes in the functional state of a prosthetic replacement valve. METHODS: Simulation studies were carried out using a mock circulation device. Aortic valve replacement (AVR) using extracorporeal circulation was performed in pigs, valve function was artificially disturbed, and valve sounds were recorded. Patients were equipped with briefcase-like devices to record their valve sounds after AVR and to transfer them via the Internet. RESULTS: Simulation studies produced a typical sound spectrum for each tested valve that remained constant under variable conditions. Experiments in animals proved that minimal changes in prosthetic valve function led to a significant change in the sound phenomena. The degree of sensitivity was significantly greater than that in echocardiographic control experiments. All patients in the clinical study regularly recorded and passed on their signals. Surveys revealed high acceptance and easy handling of the compiled devices. Valve dysfunctions were not detected. CONCLUSION: On-line registration of acoustic sound phenomena and ECG seems suited to detect minimal changes in prosthetic function. In particular, the registration of flow, acoustics and ECG envisaged at the next level opens up diverse potential applications for Internet-based, remote monitoring of patients following cardiac surgery or cardiologic treatment.

Efficacy of postoperative rewarming after cardiac surgery.

OBJECTIVE: To compare the efficacy of forced-air warmers and radiant heaters on rewarming after cardiac surgery in a prospective randomized study. METHODS: Fifty male patients who had undergone coronary artery bypass graft surgery were studied. The control group (Gr. C, n=10) was nursed under a standard hospital blanket. Two groups were treated with forced-air warmers: WarmTouch 5700 (Gr. WT, n=10) and Bair Hugger 500 (Gr. BH, n=10). Two other groups were treated by radiant heaters: the Aragona Thermal Ceilings CTC X radiant heater (Gr. TC, n=10) and a self assembled radiant heater of 4 Hydrosun 500 infrared lamps (Gr. HY, n=10). Changes of oesophageal temperature, mean skin temperature, mean body temperature and relative heat balance were calculated from oesophageal temperature, 4 skin temperatures and oxygen consumption (VO(2)). RESULTS: All actively treated groups with exception of the TC group showed significantly faster oesophageal warming than the control group. The mean body temperature increased 1.1 (0.7-1.7) degrees Ch(-1) in Gr. WT, 1.3 (0.7-1.5) degrees Ch(-1) in Gr. BH, 0.8 (0.5-1.4) degrees Ch(-1) in Gr. TC and 0.7 (0.4-1.0) degrees Ch(-1) in Gr. HY compared to Gr. C with 0.4 (0.2-0.7) degrees Ch(-1). The mean VO(2) and the maxima of the VO(2) during the study period did not differ significantly between the groups. CONCLUSION: In the current setting active warming, forced-air warming more than radiant warming, increased speed of rewarming two- to threefold in comparison to insulation with a blanket.

Redistribution of connexin43 in regional acute ischemic myocardium: influence of ischemic preconditioning.

Connexins are known to play an essential role in the ischemic preconditioning (IP) of the heart; their functional role in this process, however, has not been clearly defined. For this reason, anesthetized rats were subjected to regional myocardial ischemia, with or without IP or reperfusion. In frozen sections of hearts, fluorescence immunohistochemical staining for connexin43 (Cx43) was performed. In contrast to undisturbed zones, tissue that had been subjected to ischemia revealed Cx43 immunostaining not only in the gap junctions but also in a conspicuous pattern in the free cellular membranes of the myocytes. In myocardium that was exposed to IP only, the ratio of immunofluorescence intensity in the free cellular membrane to that in the interior of the cell was 1.22 +/- 0.04 (ratio in non-ischemia-exposed area = 1.04 +/- 0.01). When 15 or 45 min of permanent ischemia followed IP, the effect became more evident (ratio = 1.31 +/- 0.03 and 1.46 +/- 0.03, respectively) and proved to be significantly greater than in the corresponding non-IP groups (ratio = 1.16 +/- 0.03 and 1.30 +/- 0.03, respectively, P < 0.01). Reperfusion led to an overall weakening of fluorescence intensities and a disappearance of the observed IP-specific differences. We conclude that IP initiates a redistribution of Cx43 from its natural position in the gap junctions toward the free plasma membrane, thereby improving the cell's chance of survival during the subsequent phase of prolonged ischemia by an unknown, supposedly gap junction-independent, mechanism.

Gene expression of the Na-Ca2+ exchanger, SERCA2a and calsequestrin after myocardial ischemia in the neonatal rabbit heart.

BACKGROUND: Neonatal hearts are less susceptible to developing myocardial dysfunction after hypoxia and/or ischemia than adult hearts. Differences in intracellular calcium homeostasis may be responsible for reduced calcium overload of the immature myocardium leading to the observed protection against ischemia. OBJECTIVE: To assess differences in baseline and post-ischemic gene expression of calcium handling proteins after ischemia in neonatal and adult rabbit hearts. METHODS: We used isolated antegrade perfused rabbit hearts (age 2 days, 28 days, n = 32), which were exposed to ischemia and hypothermia simulating myocardial stunning comparable to neonatal asphyxia. Gene and protein expression of the sodium-calcium exchanger (NCX), the sarco-endoplasmatic reticulum Ca2+-ATPase 2a (SERCA) and calsequestrin (CSQ) were measured using quantitative real-time PCR and Western blotting. RESULTS: After ischemia and reperfusion in neonatal and adult hearts, a significant decrease in myocardial performance was recorded. At the mRNA level, significant differences in the baseline expression of NCX, SERCA and CSQ between neonatal and adult hearts were observed. In neonatal post-ischemic hearts, NCX and CSQ expression were significantly higher at the mRNA level than in controls. In contrast, SERCA expression remained unchanged in neonatal hearts and decreased in adult hearts compared to the non-ischemic controls. CONCLUSION: These findings suggest that changes in gene expression of calcium handling proteins may be involved in the different susceptibility of neonatal compared to adult hearts to developing myocardial dysfunction after ischemia.

Na(+)-channel modulation, a new principle of inotropic intervention: effects on hemodynamic and myocardial energetics in the immature rabbit heart.

Na+-channel modulators exert their positive inotropic action without affecting the adenylate-cyclase pathway by an increase in the open probability of the sarcolemmal Na+ channels. Although inotropic effects in neonatal hearts are less pronounced compared with adult hearts, the Na+-channel modulator BDF 9148 increases contractility and relaxation velocity in immature myocardium. Effects on hemodynamics and myocardial energetics are not known. Therefore, we studied the Na+-channel modulator BDF 9148 in isolated antegrade perfused rabbit hearts of different ages (2-28 d) and compared the effects with isoproterenol, enoximone, and ouabain. ANOVA showed significant effects in the concentration response curves for heart rate, stroke volume, cardiac output, and oxygen consumption but not for myocardial efficiency (p = 0.06). Age-dependent differences were observed for heart rate and stroke volume. Administration of BDF 9148 resulted in a maximal increase in stroke volume and cardiac output up to 25% in neonatal and 40% to 60% in adult preparations. Heart rate decreased by 15% in adult hearts only. Myocardial oxygen consumption was increased in a concentration-dependent manner between 25% in neonatal and 50% in adult hearts. Myocardial efficiency was increased by 35% in adult and by 10% in neonatal preparations. Although positive hemodynamic and energetic effects were less pronounced in immature compared with adult hearts, neonatal hearts also profited from the administration of the Na+-channel modulator BDF 9148. Further studies are necessary to clarify the risk of arrhythmia during application of Na+-channel modulators such as BDF 9148.

Effects of different inotropic interventions on myocardial function in the developing rabbit heart.

The development of the mammalian heart is characterized by substantial changes in myocardial performance. We studied the ontogeny of myocardial function with and without various inotropic interventions in the developing isolated, antegrade-perfused rabbit heart (2d, 8d, 14d, 28d, n = 96). Myocardial function was related to the protein expression of the sarcolemmal Na(+)-Ca2+ exchanger and to the sarcoplasmic Ca(2+)-ATPase. In neonatal hearts an age-dependent increase in maximal developed pressure velocity (dP/dtmax) by 45% and peak negative pressure velocity (dP/ dtmin) by 75% within days 2 to 8 were observed. In response to inotropic intervention with isoproterenol, ouabain, calcium and the Na(+)-channel modulator BDF 9148, dP/dtmax and dP/dtmin increased in a concentration dependent manner. Significant differences between neonatal, juvenile and adult hearts could be demonstrated in a repeated measurement ANOVA model on the concentration-response curves for BDF 9148 (dP/dtmax and dP/dtmin), ouabain (dP/dtmin) and calcium (dP/dtmin), but not for isoproterenol. At the maximum isoproterenol concentration of 1 micromol/l, the increase in dP/dtmax and dP/dtmin was significantly higher in adult compared to neonatal hearts (t-test, p < 0.01). The significant decline of the Na(+)-Ca2+ exchanger protein expression from neonatal (1822 +/- 171) to adult hearts (411 +/- 96 S.E.M. [units per 20 microg protein], p < 0.01) was related to an increase in myocardial function (dP/dtmax r = 0.63, p < 0.01, dP/dtmin r = 0.62, p < 0.01). Contractility, relaxation and the observed positive inotropic effects were in general significantly lower in neonatal compared to adult hearts. In the individual heart an increase in contractility and relaxation was related to a decrease in Na(+)-Ca2+ exchanger expression.

Extent of damage in ischemic, nonreperfused, and reperfused myocardium of anesthetized rats.

To investigate the localization of the earliest damage in ischemic and ischemic-reperfused myocardium, anesthetized rats were subjected to coronary occlusion for 15, 30, 45, or 90 min. One-half of the animals in each group had no reperfusion, whereas the other half was reperfused for 14 min. With the use of histological methods, preferentially in the periphery of the area at risk, localized zones were detected that lacked the hypoxia-specific increase in NADH fluorescence. The extent of these areas displaying injured tissue was found to be significantly smaller in the ischemic-nonreperfused hearts than in the ischemic-reperfused organs (15-min ischemia: 0.22 +/- 0.12% vs. 43.0 +/- 5.0%; 30-min ischemia: 5.7 +/- 2.7% vs. 64.6 +/- 2.9%; 45-min ischemia: 5.6 +/- 1.2% vs. 66.0 +/- 7.5%; 90-min ischemia: 39.3 +/- 5.5% vs. 86.7 +/- 1.8% of the area at risk). The results point to a localized initiation of the damage close to the surrounding oxygen-supplied tissue during ischemia and an expansion of this injury by intercellular actions into yet-intact areas upon reperfusion.