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Digital blood flow response to body warming, cooling, and rewarming in patients with Raynaud's phenomenon.

Although the diagnosis of Raynaud's phenomenon (RP) is usually made easily from a careful history, the assessment of RP severity is difficult, for the vasopastic attacks are not easily induced under experimental conditions. In this study, the laser Doppler flowmetry (LDF) technique was used to quantify digital blood flow, which was standardized by body cooling and warming in patients with RP. Twenty-one healthy subjects and 56 RP patients were studied: 7 had primary RP, 22 had suspected secondary Raynaud's syndrome (susp RS), and 27 had systemic sclerosis (SSc)-associated secondary Raynaud's syndrome (SSc RS). The inherent variability in the acral cutaneous circulation was minimized by whole-body warming and cooling. Digital blood flow values at environmental temperatures of 40 degrees C, 12 degrees C, and after rewarming, to 40 degrees C were recorded, as was the time taken for blood flow to reach 25%, 50%, and 75% of the full effects of whole-body cooling and rewarming. Patients with primary RP and susp RS had normal blood flow values at ambient temperatures of 40 degrees C, 12 degrees C, and after rewarming to 40 degrees C when compared with controls, but they had significantly faster vasoconstrictor responses to whole-body cooling, suggesting a heightened sympathetic activity. Additionally, they had slower vasodilator responses with longer 25%max response time to whole-body rewarming. Patients with SSc RS had significantly lower blood flow values at 40 degrees C after initial warming and following subsequent rewarming, and despite a normal vasoconstrictor response to cooling, it took longer for them to vasodilate during rewarming, suggesting that poor digital blood flow in these patients may be more related to digital vasculature abnormalities and not an increase in sympathetic activity. In conclusion, our assessment technique can be used to quantify digital blood flow in patients with RP and may be potentially useful in the investigation of the etiologic role of the sympathetic nervous system in RP.

Adult↗

Rates of rewarming, heart and respiratory rates and their significance for oxygen transport during arousal from torpor in the smallest mammal, the Etruscan shrew Suncus etruscus.

We investigated the process of rewarming from torpor with respect to respiratory and circulatory oxygen transport properties in the smallest mammal, the Etruscan shrew Suncus etruscus. In seven adult Etruscan shrews with a mean body mass of 2.4g, torpor was induced by deprivation of food and a cold environment. During arousal from torpor at an ambient temperature of 22 degrees C, the shrews actively rewarmed from the lowest mean (+/- S.D.) body temperature (Tb) of 12.1 +/- 1.2 degrees C to 20 degrees C at a rate of 0.43 +/- 0.14 degree C min-1, from 20 to 24 degrees C at a rate of 0.8 degree C min-1, and from 24 to 36 degrees C at a rate of 1.1 +/- 0.1 degrees C min-1. The mean rate from 12 degrees C to normothermia amounted to 0.83 degree C min-1, which is among the highest values recorded in mammals. During rewarming, the heart rate increased exponentially (Q10 = 2.2) from 100 to 800-1200 min-1, whereas the respiratory rate increased linearly from 50 to 600-800 min-1. These rates are higher than the heart and respiratory rates reported for other small mammals at the same Tb. The fraction of brown adipose tissue (BAT) was 9.2 +/- 1.6% of body mass, which is higher than in any other mammal. Up to a body temperature of approximately 17 degrees C, the heat for rewarming was mainly produced in the BAT; above this value, considerable activity of the skeletal muscles enhanced thermogenesis. Estimation of the mixed venous oxygen partial pressure showed that, at the tissue level, the rewarming process corresponds to heavy work conditions. The ventilatory system is adapted such that during rewarming, in addition to the appropriate oxygen transport capacity, there is also a capacity for hyperventilation.

Animals↗

Cardiovascular changes during mild therapeutic hypothermia and rewarming in infants with hypoxic-ischemic encephalopathy.

BACKGROUND: Clinical trials of mild cooling to 35 degrees C or below in infants with early hypoxic-ischemic encephalopathy are under way. The objective of this study was to systematically document cardiovascular changes associated with mild therapeutic hypothermia and rewarming in such infants. PATIENTS AND METHODS: Nine infants with gestational ages of 36 to 42 weeks, with 10-minute Apgar scores of 5 or less, clinical encephalopathy, and an abnormal electroencephalogram before 6 hours were cooled by surface cooling the trunk (n = 3) or by applying a cap perfused with cooled water (n = 6) for a median of 72 hours. The target core temperature was 34.0 degrees C to 35.0 degrees C for head-cooled infants and 33.0 degrees C to 34.0 degrees C for surface-cooled infants. Maintenance heating and rewarming were provided by an overhead heater. RESULTS: Mean arterial blood pressure increased by a median of 10 mm Hg during cooling and fell by a median of 8 mm Hg on rewarming. Heart rate decreased by a median of 34 beats/minute on cooling and increased by a median of 32 beats/minute on rewarming. A large increase in the output of the overhead heater decreased mean arterial blood pressure in 5 infants. Anticonvulsant drugs, sedatives, or intercurrent hypoxemia also produced falls in temperature. The inspired oxygen fraction had to be increased by a median of.14 to maintain oxygenation during cooling with 2 infants requiring 100% oxygen, an effect probably attributable to pulmonary hypertension, which was reversible with rewarming. CONCLUSIONS: Therapeutic cooling produces changes in heart rate and blood pressure that are not hazardous, but the combination of inadvertent overcooling and inappropriately rapid rewarming, together with sedative drugs that can impair normal thermoregulatory vasoconstriction, can cause hypotension in posthypoxic newborn infants. Infants who already require 50% oxygen should be cooled cautiously because pulmonary hypertension may develop. Knowledge of these cardiovascular changes, careful monitoring, anticipation, and correction should help to avoid potential adverse effects in the upcoming clinical trials.

Anticonvulsants↗

[Successful rewarming technique of isolated rat hearts following a long term preservation--calcium increase in a step-wise fashion].

This study was designed to evaluate the successful rewarming technique of isolated rat hearts which was preserved for 24 hour. Isolated rat hearts preserved with hydroxy ethyl starch (3g/dl) solution at 15 degrees C, was rewarmed and calcium concentration of the perfusate was increased in a step-wise fashion from 0.25 to 2.5mM every 5 minutes. Other hearts were also rewarmed and perfused with normal calcium (2.5mM) solution, throughout the rewarming period. The hearts perfused with normal calcium solution contracted a few times, and then severely contractured. The heart lost its color and acquired a pale and mottled appearance. On the other hand, the hearts perfused low calcium solution started beating spontaneously, and their heart rate and left ventricular pressure was increased gradually according to the calcium increase. In addition, re-introduction of normal calcium concentration did not change the cardiac parameters. This results indicated that step-wise increase of calcium concentration of the perfusate prevented heart contracture after rewarming and was a useful rewarming technique.

Animals↗

Physiologic requirements during rewarming: suppression of the shivering response.

Intraoperative hypothermia has become a common occurrence. Postoperative rewarming often is accompanied by shivering and results in increased metabolic and circulatory demands. We examined the metabolic, hemodynamic, and biochemical variables in 2 groups of hypothermic (greater than 35.8 degrees C) patients requiring mechanical ventilation after a major operation. One was observed during routine medical management whereas the other group received 40 mg of metocurine iodide and then observed during routine medical management. All patients were allowed to rewarm passively. O2 consumption (VO2, ml/min, STPD), CO2 production (VCO2, ml/min, STPD) and respiratory quotient (RQ) measurements were made every 15 min using a Beckman Metabolic Measurement Cart. Esophageal temperature, arterial blood pressure, heart rate (HR), rate pressure product, CVP, arterial blood gases, serum lactate concentration, and duration of shivering also were recorded. Suppression of the shivering by metocurine increased rewarming time significantly and decreased VCO2, VO2, HR, rate pressure product, mean arterial pressure (MAP), and the O2 cost of rewarming. Thus, the elimination of shivering during postoperative rewarming is associated with a decrease in caloric, metabolic demands and myocardial work (as assessed by the rate pressure product) while rewarming time is prolonged. In the postoperative, hypothermic, critically ill patient, suppression of the shivering response in selected patients may be indicated.

Adult↗

Severe accidental hypothermia: rewarming strategy using a veno-venous bypass system and a convective air warmer.

OBJECTIVE: To study a rewarming strategy for patients with severe accidental hypothermia using a simple veno-venous bypass in combination with a convective air warmer. SETTING: Eighteen beds in a university hospital intensive care unit. PATIENTS: Four adults admitted with a core temperature less than 30 degrees C. Hypothermia was caused by alcoholic intoxication in three patients and by drug overdose in one patient. MEASUREMENTS AND MAIN RESULTS: All patients were rewarmed by a venovenous bypass and in three cases a convective air warmer was also used. At a bypass flow rate of 100-300 ml/min the mean increase in core temperature was 1.15 degrees C/h (Range: 1.1-1.2 degrees C/h). One patient died 2 days after rewarming as a consequence of a reactivated pancreatitis. The other three patients survived without neurological sequelae. CONCLUSION: This rewarming technique seems safe and effective and allowed the controlled rewarming of our patients who suffered from severe accidental hypothermia

Adult↗

Cardiopulmonary bypass, rewarming, and central nervous system dysfunction.

BACKGROUND: During cardiopulmonary bypass a nasopharyngeal temperature greater than 38 degrees C at the end of rewarming may indicate cerebral hyperthermia. This could exacerbate an ischemic brain injury incurred during cardiopulmonary bypass. METHODS: In a cohort of 150 aortocoronary bypass patients neuropsychologic test scores of 66 patients whose rewarming temperature exceeded 38 degrees C were compared with those who did not. There were no differences between groups with respect to demographic and intraoperative variables. RESULTS: A trend was seen for hyperthermic patients to do worse on all neuropsychologic tests in the early postoperative period but not at 3-month follow-up. By analysis of covariance hyperthermic patients did worse on the visual reproduction subtest of the Weschler memory scale at 3 months (p = 0.02), but this difference was not found by linear regression (p = 0.10). CONCLUSIONS: We were unable to demonstrate any significant deterioration in patients rewarmed to greater than 38 degrees C in the early postoperative period. The poorer performance in the visual reproduction subtest of the Wechsler memory scale at 3 months in the group rewarmed to more than 38 degrees C is interesting but far from conclusive. Caution with rewarming is still advised pending more in-depth study of this issue.

Body Temperature↗

Glucose and fatty acid oxidation by the in situ dog heart during experimental cooling and rewarming.

BACKGROUND: Reduced myocardial function after hypothermia may be metabolic in origin, but the relationship between myocardial metabolism and the various components of hypothermia-mediated dysfunction has not been thoroughly investigated. METHODS: In the present study we measured myocardial uptake and oxidation of glucose and oleate in mongrel dogs undergoing cooling to 25 degrees C followed by rewarming to 37 degrees C, using radiolabeled substrates. RESULTS: Segment work index declined from 39.3 +/- 5.1 to 15.1 +/- 2.4 mm Hg in response to cooling from 37 degrees to 25 degrees C and did not recover completely on rewarming (27.2 +/- 4.2 mm Hg, p < 0.05). Oleate uptake declined from 3,251 +/- 619 to 1,043 +/- 356 nmol.min-1.100 g-1 (p < 0.05) when the dogs were cooled from 37 degrees to 25 degrees C. Simultaneously, oxidation rate fell from 1,089 +/- 158 to 354 +/- 83 nmol.min-1.100 g-1 (p < 0.05). On rewarming, oleate uptake was restored to prehypothermic values, whereas its rate of oxidation remained depressed (480 +/- 129 nmol.min-1.100 g-1; p < 0.05). Uptake and oxidation of glucose also declined significantly during cooling. However, both uptake and oxidation of glucose recovered fully on rewarming. CONCLUSIONS: The results of the present study demonstrate a reduced capacity to oxidize fatty acids by the myocardium during rewarming after hypothermia.

Animals↗

Postbypass effects of delayed rewarming on cerebral blood flow velocities in infants after total circulatory arrest.

Cerebral perfusion is reduced after prolonged periods of total circulatory arrest in infants. Methods of rewarming after arrest may modify the flow pattern of recovery, and a single report has suggested that using cold reperfusion to delay rewarming could mitigate abnormalities in cerebral blood flow. Cerebral perfusion was evaluated by transcranial Doppler sonography in 16 infants who required periods of total circulatory arrest of 35 minutes or more. In group A (n = 9) rewarming was begun immediately on reperfusion, whereas in group B (n = 7) a 10-minute period of cold reperfusion was instituted before rewarming was begun. The mean and end-diastolic flow velocities were measured before incision (baseline) and at 20, 45, and 90 minutes after conclusion of cardiopulmonary bypass. Mean arterial pressure, hematocrit value, and arterial carbon dioxide tension were controlled, with no significant differences between the two groups (p > 0.05). In group A, the mean cerebral blood flow velocity was below the baseline level at all three postbypass measurements (p < 0.001). In group B, however, mean velocity did not differ significantly from the baseline value (p > 0.05). Twenty minutes after bypass, 89% of the patients in group A had no diastolic Doppler signal, indicating absence of perfusion during diastole, compared with only 28% in group B (p = 0.02). These preliminary results suggest that a delay in rewarming on reperfusion may be beneficial in infants after circulatory arrest.

Blood Flow Velocity↗

Regional cerebral blood flow during rewarming of cardiopulmonary bypass correlates with posthypothermic regional glucose use.

BACKGROUND AND OBJECTIVES: Although global measurements of cerebral blood flow and metabolism during and after profoundly hypothermic cardiopulmonary bypass have been performed both in experimental animals and in human beings, little is known about their regional changes. The purpose of this study was to investigate the changes in regional cerebral blood flow during profoundly hypothermic cardiopulmonary bypass and regional cerebral glucose use after cardiopulmonary bypass. METHODS: We measured regional cerebral blood flow with positron emission tomography during both the cooling (n=5) and rewarming (n=5) of hypothermic cardiopulmonary bypass in anesthetized dogs by continuously infusing 15O-labeled water. We altered the core temperature between 20 degrees and 37 degrees C. To assess the integrity of brain metabolism, we measured the regional cerebral glucose use by bolus injections of 18F-labeled 2-fluoro-2-deoxy-D-glucose. RESULTS: Regional cerebral blood flow decreased homogeneously during cooling. The regional cerebral blood flow at 20 degrees C was about one fourth of that at 37 degrees C. In contrast, at 24 degrees, 28 degrees , and 32 degrees C during rewarming, there were significant interregional differences in the regional cerebral blood flow for given temperatures (p=0.0075, 0.034, and 0.048, respectively). These interregional differences disappeared after rewarming. Although the regional cerebral blood flow significantly correlated with the regional cerebral glucose use in the control condition at 37 degrees C without cardiopulmonary bypass (r=0.75; p=0.00012), this correlation disappeared after profoundly hypothermic cardiopulmonary bypass (r=0.204; p=0.388). Regional cerebral blood flow at 32 degrees C during rewarming positively correlated with the regional cerebral glucose use after cardiopulmonary bypass (r=0.655; p=0.0017). CONCLUSION: The altered regional cerebral blood flow during rewarming of profoundly hypothermic cardiopulmonary bypass might affect regional brain metabolism.

Animals↗

Heat production and body temperature during cooling and rewarming in overweight and lean men.

OBJECTIVE: To compare overweight and lean subjects with respect to thermogenesis and physiological insulation in response to mild cold and rewarming. RESEARCH METHODS AND PROCEDURES: Ten overweight men (mean BMI, 29.2 +/- 2.8 kg/m(2)) and 10 lean men (mean BMI, 21.1 +/- 2.0 kg/m(2)) were exposed to cold air for 1 hour, followed by 1 hour of rewarming. Body composition was determined by hydrodensitometry and deuterium dilution. Heat production and body temperatures were measured continuously by indirect calorimetry and thermistors, respectively. Muscle activity was recorded using electromyography. RESULTS: In both groups, heat production increased significantly during cooling (lean, p = 0.004; overweight, p = 0.006). The increase was larger in the lean group compared with the overweight group (p = 0.04). During rewarming, heat production returned to baseline in the overweight group and stayed higher compared with baseline in the lean group (p = 0.003). The difference in heat production between rewarming and baseline was larger in the lean (p = 0.01) than in the overweight subjects. Weighted body temperature of both groups decreased during cold exposure (lean, p = 0.002; overweight, p < 0.001) and did not return to baseline during rewarming. DISCUSSION: Overweight subjects showed a blunted mild cold-induced thermogenesis. The insulative cold response was not different among the groups. The energy-efficient response of the overweight subjects can have consequences for energy balance in the long term. The results support the concept of a dynamic heat regulation model instead of temperature regulation around a fixed set point.

Adult↗

The failure of negative pressure rewarming (Thermostat) to accelerate recovery from mild hypothermia in postoperative surgical patients.

UNLABELLED: The Thermostat device (Aquarius Medical Corp., Phoenix, AZ) is used in a new technique to accelerate recovery from hypothermia by mechanically distending blood vessels in the hand, thereby increasing transfer of exogenous heat to the body core. We evaluated the use of the Thermostat device in patients with mild postoperative hypothermia (< 36 degrees C). We studied adult patients undergoing elective surgery, general anesthesia, and neuromuscular blockade. Patients with an initial postoperative tympanic membrane temperature < 36 degrees C were randomized into two groups: 1) Thermostat, which consisted of a hypothermia warming mitt/seal and thermal exchange chamber for 60 min, and 2) conventional treatment, which consisted of warm blankets and/or radiant heat. Of the 191 patients enrolled, 60 (31%) developed hypothermia and were randomized to receive the Thermostat (n = 30) or conventional methods (n = 30). Fourteen patients in the Thermostat group and 17 patients in the conventional group rewarmed to 36 degrees C before discharge from the recovery room (P is not significant). There were no differences in vital signs, rewarming time, time to discharge from the recovery room, or postoperative temperature between groups. We conclude that patients with mild postoperative hypothermia rewarmed in a similar fashion, regardless of whether the Thermostat or conventional methods were used. IMPLICATIONS: We found that a commercially available negative pressure rewarming device (Thermostat; Aquarius Medical Corp., Phoenix, AZ) was not effective in accelerating rewarming in postoperative hypothermic surgical patients after general anesthesia.

Adult↗

Core rewarming via warm lavage liquid ventilation in a swine model of hypothermia-associated ventricular fibrillation.

OBJECTIVES: To determine whether warm lavage liquid ventilation (LV) would provide rapid cardiopulmonary rewarming in swine with severe hypothermia and ventricular fibrillation. METHODS: Intubated common swine (n = 3; mean +/- SEM weight 26+/-1.2 kg) were cooled to a mean aortic temperature of 26.4+/-0.9 degrees C. Ventricular fibrillation was induced by transthoracic electrical shock. Rewarming was initiated by continuous endotracheal instillation of warm (44 degrees C) pre-oxygenated, perfluorocarbon liquid at 5 mL/kg/min. Endotracheal instillation of perfluorocarbon occurred while standard gas ventilation continued. Manual chest compressions were performed throughout the 30-minute rewarming process. Outcome measures were the absolute and relative rates of change of all temperatures. RESULTS: After 30 minutes of warm lavage LV, the mean aortic and pulmonary artery temperatures increased by 6.6+/-0.6 degrees C, respectively. Esophageal, nasal, and rectal temperatures did not change significantly. In one animal, normal sinus rhythm spontaneously returned after 16 minutes of rewarming. CONCLUSIONS: During cardiac arrest, warm lavage liquid ventilation may produce rapid cardiopulmonary rewarming.

Analysis of Variance↗

Morphologic changes in tubular cells from in situ kidneys following experimental hypothermia and rewarming.

Although renal failure may occur following rewarming from deep accidental hypothermia, this subject has received little attention in experimental hypothermia and clinical case reports. In order to explore the integrity of hypothermic and posthypothermic renal morphology we used an experimental animal model of accidental hypothermia where the heart supports the circulation throughout cooling and rewarming without accompanying cardioplegia or ischemia. Ultrastructural changes in renal tubular cells from three groups of pentobarbital anesthetized Wistar rats: 1) controls (n=6) maintained at 37 degrees C for 4 h, 2) hypothermic rats (n=6) core-cooled and maintained at 15-13 degrees C for 4 h, and 3) rewarmed rats (n=10), were studied as a sensitive indicator of renal damage. Electron micrographs (EM) from hypothermic kidneys showed rounded up mitochondria with loss of contrast. These changes were observed in several though not all of the biopsies, but they were found in all kidneys. Areas exhibiting focal tubular necrosis were seen on most EM from three of these kidneys. EM from rewarmed kidneys showed alterations of mitochondrial ultrastructure with similarities to those observed after hypothermia, but in general the changes were more prominent. Extracellular edema, intracellular edema, swelling of mitochondria, margination of chromatin, necrosis of single tubular cells, and disrupting necrotic debris into tubular lumen could be found in micrographs from 7 of the 10 kidneys examined. Rewarming from experimental hypothermia, without episodes of ischemia or hypoxia, thus induces ultrastructural changes in renal tubular cells similar to changes observed in acute tubular necrosis, associated with renal failure.

Animals↗

Extracorporeal rewarming in a severely hypothermic patient using venovenous haemofiltration in the accident and emergency department.

Severe hypothermia is a medical emergency and requires active and occasionally rapid core rewarming to prevent cardiac arrhythmias and death. In the accident and emergency department rewarming is often limited to warmed intravenous fluids, heated blankets, gastric and bladder lavage. Extracorporeal methods, which rewarm core blood directly, for example haemodialysis and cardiopulmonary bypass, require expertise and equipment not always found in a district general hospital. Venovenous haemofiltration is now commonly found in district general hospitals around the country and can be used safely for core rewarming. A case is reported of a severely hypothermic elderly patient successfully rewarmed using venovenous haemofiltration, in an accident and emergency department, when other conventional methods had failed.

Aged↗

Heart rate variability and electrocardiogram waveform as predictors of morbidity during hypothermia and rewarming in rats.

This study examined electrocardiogram (ECG) waveform, heart rate (HR), mean blood pressure (BP), and HR variability as potential autonomic signatures of hypothermia and rewarming. Adult male Sprague-Dawley rats had telemetry transmitters surgically implanted, and 2 weeks were allowed for recovery prior to induction of hypothermia. Rats were lightly anesthetized (sodium pentobarbital, 35 mg/kg i.p.) and placed in a coil of copper tubing through which temperature-controlled water was circulated. Animals were cooled to a core temperature (Tc) of 20 degrees C, maintained there for 30 min, and then rewarmed. Data (Tc, BP, HR from ECG, and 10-s strips of ECG waveforms) were collected every 5 min throughout hypothermia and rewarming. Both HR and BP declined after initial increases with the drop in HR starting at a higher Tc than the drop in BP (29.6 +/- 2.4 degrees C vs. 27.1 +/- 3.3 degrees C, p < 0.05). Animals that were not successfully rewarmed exhibited a significant (p < 0.05) increase in the normalized standard deviation of interbeat intervals (IBI) throughout cooling compared with animals that were successfully rewarmed. The T wave of the ECG increased in amplitude and area with decreasing Tc. T-wave amplitude and IBI variability show potential as predictors of survival in hypothermic victims.

Animals↗

Significance of donor nutritional status for rewarming injury of the hepatic graft in rats.

We studied the significance of donor nutritional status for hepatic rewarming injury and the usefulness of the glucagon loading test for the assessment of donor nutritional status in rats. In experiment 1, the animals were either free fed or fasted for 6, 24 or 48 h. The livers were preserved in chilled lactated Ringer's solution for 4 h and were divided into eight groups according to the fasting and rewarming periods. The ammonia level in the graft effluent was increased in grafts preserved for 24 h or longer, which was augmented when rewarming time was increased from 15 to 30 min (p < 0.05). In experiment 2, the animals were divided into four groups (n = 3 each) according to the fasting period. The hepatic tissue glycogen content was measured after fasting, and the serum glucose was measured after the administration of 50 micrograms/kg of glucagon i.v. The hepatic tissue glycogen content correlated with the increase rate of serum glucose (p = 0.0001). We conclude that glycogen may protect the hepatic graft from rewarming injury by improving energy status, particularly in prolonged rewarming. The glucagon loading test seems to be useful for assessing the glycogen content of the hepatic graft.

Alanine Transaminase↗

Heat shock-induced protection of renal proximal tubular epithelial cells from cold storage and rewarming injury.

Cold storage and reperfusion injury to transplanted kidneys contributes to increased incidence of delayed graft function and may have a negative impact on graft survival. This study examined the mechanisms by which previous heat shock protects against cell death in an in vitro model of kidney storage. Cold storage is mimicked by incubating human renal proximal tubular epithelial (HK-2) cells in University of Wisconsin solution at 4 degrees C with and without subsequent rewarming. Heat shock was induced by incubation of cells at 42 degrees C for 1 h. Altered protein expression was measured by Western blot, and cell viability and apoptosis were measured by propidium iodide DNA staining using flow cytometry. The specific role of heat-shock protein 70 (HSP-70) was determined both by siRNA knockdown and by stable overexpression approaches. Cold storage and rewarming-induced cell death was associated with decreased expression of HSP-70, HSP-90, HSP-27, and Bcl-2. Previous heat shock significantly reduced HK-2 cell death after cold storage and rewarming and was associated with the maintenance of HSP-70, HSP-27, and Bcl-2 protein levels. Blocking heat stress-induced HSP-70 with siRNA did not significantly block the protective effect of heat stress against cold storage and rewarming cell death; however, overexpression of HSP-70 protected HK-2 cells from this stress. It is concluded that previous heat shock protects HK-2 cells from cold storage and rewarming injury. siRNA inhibition of HSP-70 induction did not block the protective effect of heat shock, indicating that HSP-70 is not essential to the heat stress-induced protective effect reported in this study.

Blotting, Western↗