Effects of interactions between cooling and rewarming conditions on survival of cells.
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The relevant dielectric properties of a practical perfusate formula have been measured at radiofrequencies from 27 to 2954 MHz and temperatures from -75 to +25 degrees C. Both dimethyl sulfoxide and 2,3-butanediol were investigated as the cryoprotectant component at concentrations from 15 to 50%. The dielectric properties are considered in terms of new indices which characterize uniformity of heating, the power delivered and the efficiency of heating, and the avoidance of localized thermal runaway during very rapid heating. It becomes clear that a compromise must be accepted between nonuniformity of heating at the higher frequencies and inconveniently low power delivery at the lower frequencies. The choice of CPA and the concentration are found to be not very significant in these respects.
Investigations were carried out on the respiratory function of isolated rat hepatocytes after cold storage alone for periods up to 48 h in either sucrose-based solution (SBS) or University of Wisconsin (UW) solution and after subsequent normothermic preincubation. In both SBS and UW, cold storage for 24 h depressed respiratory function (to 21 +/- 3 and 23 +/- 3 nmol O(2)/min/10(6) cells, respectively) compared to control cell values (31 +/- 3 and 33 +/- 5 nmol O(2)/min/10(6) cells; P < 0.01 in each case). However, normothermic preincubation for 60 min returned respiratory activity to control values (for SBS and UW storage: 41 +/- 6 and 40 +/- 5 nmol O(2)/min/10(6) cells; for control cells: 43 +/- 5 and 46 +/- 6 nmol O(2)/min/10(6) cells). Storage for 48 h in both SBS and UW allowed further depression of respiratory activity, with no recovery after preincubation. Stimulation of respiration by succinate in hepatocytes stored for longer periods was suggestive of increased membrane permeability. Both SBS and UW are effective storage solutions for isolated hepatocytes for up to 24 h as judged by aerobic metabolism, but significant damage was expressed in both solutions when preservation was extended.
The effects of electrical pacing during the early reperfusion following hypothermic global ischemia (60 min, at 25 degrees C) was studied in the isolated working rat heart model. The hearts were divided into three groups. Hearts in Group I (n = 8) were control without hypothermia, ischemia or pacing. Hearts in Group II (n = 16) were paced with ventricular rate at 300 beats/min with 1 mVolt for 10 min during the Langendorff mode after an initial 5 min of reperfusion. Hearts in Group III (n = 14) were not paced. The recovery of aortic flow (both absolute and percent) was significantly better in Group II than in Group III, but was significantly lower in both groups than in control. No significant differences were noted, however, in heart rate, aortic pressure or coronary flow between Group II and III. In contrast, the tissue concentration of adenosine triphosphate (ATP) in Groups II and III decreased significantly by the end of reperfusion relative to Group I, but no difference in ATP existed between Group II and III. Myocardial ATP concentrations did not correlate with percent recovery of aortic flow. The myocardial concentration of calcium in Groups II and III increased by the end of reperfusion as compared with Group I, but no difference in calcium existed between Group II and III. The myocardial concentration of calcium demonstrated a significant correlation with percent recovery of aortic flow (r = 0.71, n = 30, p < 0.005). Our results indicate that an electrical pacing during early reperfusion in the myocardium improves functional recovery of aortic flow.
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Oxygen consumption (VO2), carbon dioxide production (VCO2), end-tidal carbon dioxide partial pressure (PETCO2), mixed venous oxygen saturation (SvO2) and haemodynamic variables were recorded every 30 min for four hours in 15 patients recovering from hypothermic cardiopulmonary bypass (CPB). All patients had been anaesthetised with fentanyl 40 micrograms.kg-1, supplemented with isoflurane, and pancuronium 0.15 mg.kg-1 for muscle relaxation. Three of the 15 patients (20 per cent) shivered, defined as intermittent or continuous, vigorous movements of chest or limb muscles. Patients who shivered had a VO2 of 159 +/- 16.4 ml.min-1.m-2 on arrival in the ICU which rose to a maximum value of 254 +/- 28.3 ml.min-1.m-2 by 150 min post-CPB. In contrast, patients who did not shiver had a significantly lower VO2 of 93.1 +/- 6.9 ml.min-1.m-2 on arrival in the ICU which rose to a maximal value of only 168 +/- 11.5 ml.min-1.m-2 by 180 min post-CPB. Maximal VO2 in both groups was reached when the nasopharyngeal temperature (NPT) was approaching normal. VCO2 paralleled the increase in VO2 in both groups. By four hours there was no significant difference between the two groups; however, the VO2 in both groups (160.5 +/- 21.3 ml.min-1.m-2 and 173.9 +/- 12.3 ml.min-1.m-2 respectively) was approximately twice values commonly measured in anaesthetized patients. Patients who shivered had a significantly higher heart rate and cardiac index and significantly lower SvO2. We conclude that the high VO2 and VCO2 associated with shivering causing increased myocardial work may be detrimental to patients who have impaired cardiac function post-coronary artery surgery (CAS).
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BACKGROUND/PURPOSE: Increasing levels of glycerol in extracellular fluid indicate cell membrane disintegration, and can be continuously monitored with microdialysis. The aim of this study was to monitor liver cell membrane damage during liver transplantation in a pig model. METHODS: Thirteen donor and recipient pairs were divided into two groups; group I, with a liver ischemia time of 5 h (n = 6), and group II, with 15 h of ischemia (n = 7). Microdialysis samples from the liver graft were collected at 20-min intervals during donor operation, cold preservation, implantation, and reperfusion in the recipient. Glycerol concentrations were analyzed. RESULTS: During cold preservation, a continuous increase in glycerol levels was observed. This increase correlated with the time of cold ischemia (r2 = 0.88). During implantation, an accelerated increase of glycerol was observed, with a higher acceleration in group II (P < 0.005). During the first hour after reperfusion, a higher release of glycerol was observed in group II (P = 0.0005). CONCLUSIONS: Our data show a constant glycerol release during cold storage, indicating a continuous injury to the cell membranes over time. Improvement in cell membrane protection during cold preservation could lead to improvement in liver transplantation outcome and acceptance of an extended period of cold ischemia.
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The possibility that anterograde to retrograde reversal of axonal transport might take place in mid axon at a site distant from any nerve termination was investigated in sciatic nerve preparations from Xenopus laevis. The nerve, containing a pulse of anterogradely transported protein labeled with [35S]methionine, was kept in a two-compartment temperature controlled chamber. One compartment containing the proximal nerve was maintained at room temperature throughout the duration of an experiment while the second compartment containing the distal nerve, and separated from the first by a thermal barrier, was initially cooled to 3-4 degrees C and later warmed to room temperature. Transport of labeled proteins in the nerve was detected with a position-sensitive detector of ionizing radiation. With the distal portion of the nerve cold, the pulse of labeled protein transported up to the thermal barrier and stopped. When the distal part of the nerve was warmed to room temperature, retrograde and anterograde pulses of label propagated away from the thermal barrier with no time delay. The retrograde pulse could be collected on the distal side of a proximally placed tie and could be eliminated by treatment of the proximal nerve with vinblastine or dinitrophenol. Functional and structural evidence indicated that the cold block and thermal barrier were not destructive to the axons. Electron microscopy showed that the numerical density of axonal microtubules distal to the cold block was decreased about seven fold during the cold treatment and that this decrease could be prevented by 10 mumol/l taxol.(ABSTRACT TRUNCATED AT 250 WORDS)
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