A comparison of complete blood replacement with varying hematocrit levels on neurological recovery in a porcine model of profound hypothermic (<5 degrees C) circulatory arrest.

Profound hypothermia (<5 degrees C) may afford better neurological protection after circulatory arrest; however, there are theoretical concerns related to microcirculatory sludging of blood components at these ultra-low temperatures. We hypothesized that at temperatures <5 degrees C, complete blood replacement results in superior neurological outcome. Twelve Yorkshire pigs (30 kg) underwent thoracotomy, cardiopulmonary bypass (CPB), and were randomly assigned to one of three target hematocrits during circulatory arrest: 0%, 5%, 15%. Hextend (6% hetastarch in a balanced electrolyte vehicle) was used for the CPB prime and as an exchange fluid. Animals were cooled to a temperature <5 degrees C, underwent 1-h circulatory arrest, and were warmed to 35 degrees C with administration of blood to increase the hematocrit to >25% before separation from CPB. The primary outcome, peak postoperative neurobehavioral score, was compared between groups. The 0% group (mean +/- SD) had significantly (P: < 0.02) better neurobehavioral scores than the 5% and 15% groups (6.0 +/- 2.9 vs 1.3 +/- 1.0 and 1.5 +/- 0.6) respectively. Other variables (e.g., intracranial pressure) were similar between groups. In a porcine model of profound hypothermia (<5 degrees C) and circulatory arrest, complete blood replacement resulted in superior neurological outcome. This finding suggests that at ultralow temperatures, the presence of some blood component (e.g., erythrocytes, leukocytes) may be deleterious.

A new solution for life without blood. Asanguineous low-flow perfusion of a whole-body perfusate during 3 hours of cardiac arrest and profound hypothermia.

BACKGROUND: The benefits of hypothermia for preventing ischemic injury are well known, but its application in surgery to protect the whole body during procedures requiring circulatory arrest is currently limited to < 1 hour at 15 degrees C using 50% hemodilution. In a significant departure from previous methods, we have developed a technique of asanguineous blood substitution with low-flow perfusion and cardiac arrest at < 10 degrees C in a canine model. Our approach has been to design a hypothermic blood substitute that would protect the brain and visceral organs during several hours of bloodless perfusion. Two different solutions have been designed to fulfill separate requirements in the procedure. METHODS AND RESULTS: With the use of extracorporeal cardiac bypass, 14 adult dogs were exsanguinated during cooling; 11 dogs were blood substituted using in combination the "purge" and "maintenance" solutions (group 1), and 3 dogs were perfused throughout with the "purge" solution alone as controls (group 2). After cardiac arrest, the solutions were continuously circulated for 3 1/2 hours by the extracorporeal pump (flow rate, 40 to 85 mL.kg-1.min-1; mean arterial blood pressure, 25 to 40 mm Hg). The temperature was maintained at < 10 degrees C (nadir, 6.6 +/- 0.1 degrees C) for 3 hours, and the hematocrit was kept at < 1% before controlled rewarming and autotransfusion. In the experimental group, the heart always started spontaneously in the temperature range of 11 degrees C to 27 degrees C, and 8 animals have survived long-term (current range, 14 to 110 weeks) without any detectable neurological deficit. In contrast, two control animals survived after extensive and aggressive cardiac resuscitation efforts; after surgery they exhibited transient motor and sensory deficits for approximately 1 week. Evaluation of biochemical and hematological parameters showed only a transient and inconsequential elevation in enzymes (eg, brain, liver, cardiac) in group 1 compared with the markedly greater elevations in group 2. For example, immediate postoperative values (mean +/- SEM) for lactate dehydrogenase were 114 +/- 10 for group 1 versus 490 +/- 210 for group 2 (P < .03); for SGOT, values were 93 +/- 18 for group 1 versus 734 +/- 540 for group 2 (P < .05). On day 1 for creatine kinase (CK), the group 1 value was 7841 +/- 2307 versus 71,550 +/- 2658 for group 2 (P = .03), and for CK-BB, the group 1 value was 108 +/- 22 versus 617 +/- 154 for group 2 (P = .03). Neurological evaluation using deficit scores (NDS) was based on a modification of the Glasgow Coma Scale score: 0, normal; 1, minimal abnormality; 2, weakness; 3, paralysis; 4, coma; and 5, death. At days 1 and 2 after surgery, NDS (mean +/- SEM) were 0 +/- 0 for the experimental group versus 1.5 +/- 0.5 for the control group. At days 3 and 7 after surgery, NDS were 0 +/- 0 for group 1 versus 1.0 +/- 1.0 for group 2. CONCLUSIONS: The faster neurological recovery of dogs treated with the "intracellular-type" maintenance solution supports the biochemical data showing the benefits of this type of blood substitute for extending the safe limits of hypothermic cardiac arrest procedures to > 3 hours.

Asanguineous whole body perfusion with a new intracellular acellular solution and ultraprofound hypothermia provides cellular protection during 3.5 hours of cardiac arrest in a canine model.

Temporary cessation of blood flow is a necessary aid for certain complex neurosurgical and cardiovascular procedures, and hypothermia is often used to help protect against the deleterious effects of ischemia and anoxia. In an attempt to protect cellular integrity during ultraprofound hypothermia (< 10 degrees C) and complete blood substitution, two new crystalloid-colloid blood substitutes (Hypothermosol-maintenance [HTS-M] and Hypothermosol-purge [HTS-P]) have been evaluated. Using extracorporeal bypass, 14 dogs were totally exsanguinated during cooling using the HTS-P solution, then perfused (40-85 ml/kg/min; mean arterial blood pressure = 25-40 mmHg) with either TS-M (Group I, n = 11), or with HTS-P as controls (Group II, n = 3) for 3 hr at 7 degrees C. During warming, the dogs were autotransfused and observed neurologically and biochemically during recovery. All dogs in Group I recovered and eight have survived long term (12-80 weeks) without apparent neurologic deficits. In contrast, dogs in Group II were more difficult to revive (cardiac resuscitation); two survived long term with delayed neurologic recovery. Evaluation of biochemical parameters showed only a transient and inconsequential elevation in enzymes (e.g., brain, liver, and heart) in Group I compared with the markedly greater elevations in Group II. The faster neurologic recovery of dogs treated with the "intracellular" maintenance solution supports the biochemical data showing the benefits of this type of blood substitute for extending the safe limits of hypothermic cardiac arrest to beyond 3 hr.

Rat model of delirium: atropine dose-response relationships.

The authors report dose-response relationships for atropine in their rat model of human delirium. In this model, anticholinergic mechanisms are an important cause and may be a neurochemical final common pathway for diverse etiologies of delirium. Four intravenous (i.v.) bolus doses of atropine (27.5, 13.5, 6.875, and 3.44 mg/kg) were studied along with the originally reported dose of 55 mg/kg and saline control. Subsequent iv infusions were also proportionately reduced. EEG frequency and amplitude, maze performance, and subjective behavioral descriptions were taken over 320-minute study periods. Repeated-measures analysis of variance compared data between groups, and effect sizes were calculated. Dosages to be used in future studies are discussed.

Rewarming, ultraprofound hypothermia and cardiopulmonary bypass.

Rewarming, a key event in resuscitation from accidental, experimental and clinical hypothermia, is sometimes followed by neurologic, cardiac, and respiratory sequelae and may lead to death. The rate of rewarming has been implicated but not quantified as etiologic in these sequelae. Under anesthesia fifteen dogs were cannulated and connected to an extracorporeal circuit for oxygenation, core cooling and rewarming. They were subjected to ultra-profound hypothermia with a core (esophageal) temperature as low as 1.3 degrees C, cardiac arrest, blood substitution, and continuous low flow perfusion. After 2-3 hours of cardiac arrest, rewarming began. Mechanical activity of the heart was seen between 10 degrees and 28 degrees C and respiration resumed at 29 degrees C. The rewarming rates of the 15 dogs were retrospectively studied. They were placed into three categories (G) based on the outcome. G-I (N=2):no neurological complications, G-II (N=8):transient neurological problems, and G-III (N=5):death, mainly from cardiovascular and respiratory complications confirmed at death by autopsy. Heat gain by each animal was recorded as a function of time for all experiments. The time it took each dog to reach 35 degrees C was determined and a mean was calculated (rewarming rate). Normal body temperature for a dog is 37.8 degrees C. Statistical analysis (ANOVA) was performed ex post facto to determine the relationship between rewarming rate and outcome. Our data contradicts the notion that slow core rewarming from nadir to normal temperature offers better outcome.(ABSTRACT TRUNCATED AT 250 WORDS)

Complete blood substitution during profound hypothermic cardiac arrest in dogs.

This study compared the effects of 3 hours of cardiac arrest performed at 2 different levels of profound hypothermia in totally exsanguinated, blood substituted dogs. Dogs (N = 10) were anesthetized and esophageal temperature was lowered to 24 degrees at which time exsanguination began. Once exsanguination was complete and the heart had arrested, continuous whole body perfusion of an oxygenated blood substitute solution was performed for 3 hours. Core temperature during this period remained below 10 degrees C and reached nadirs of 1.3 degrees C in Group 1 (N = 5) versus 7.3 degrees C in Group II (N = 5). Once the dog was allowed to rewarm to 10 degrees C replacement of the perfusate with blood was begun. All dogs survived the procedure but 2 dogs from each group died by 4 days following the experiment. The remaining 6 dogs were observed for between 10 and 85 days. The group perfused with blood substitute at the warmer nadir had a faster recovery of motor behavior and showed smaller changes from normal in several biochemical and hematological parameters. These findings indicate that profound hypothermia and total blood substitution can be successfully achieved at either temperature nadir, but the warmer level appears to be associated with a faster recovery.

Comparison of the effects of total blood substitution during two different levels of hypothermic cardiac arrest.

The effect of total blood replacement with a solution containing neither hemoglobin nor fluorocarbon was studied under two different levels of hypothermia. Ten dogs were anesthetized and esophageal temperature lowered to about 24 degrees at which time exsanguination began. Upon cardiac arrest and the completion of exsanguination, continuous whole body closed chest extracorporeal circulation of an oxygenated blood substitute was begun. Hematocrit was 1% while core temperature remained less than 10 degrees C for 3 hours of perfusion during which nadirs of 1.3 degrees C (Group I, N = 5) and 7.3 degrees (Group II, N = 5) were achieved. Replacement of the perfusate with whole blood began once the dog was rewarmed to approximately 10 degrees C. All dogs survived the procedure. Two dogs from each group died by the fourth post-operative day but the others survived long term. Group II showed a faster return to normal based on motor behavioral, biochemical and hematological changes. Thus the combination of profound hypothermia and complete blood substitution with a solution lacking any special oxygen carrying molecule, can be tolerated for 3 hours using both levels of hypothermia, however, the warmer one appears to be associated with faster recovery.

Blood substitution: an experimental study.

Priming fluids for cardiopulmonary bypass have been extremely varied, with resultant hemodilution. Furthermore, major surgeries utilizing cardiopulmonary bypass require multiple postoperative transfusions of blood and blood products. The appeal of having a readily available blood substitute for major cardiovascular and neurosurgical operations could prove to be a life saver, while also eliminating the risk of diseases transmitted by transfusion. Blood substitutes could also lessen the reported complications resulting from blood damage due to prolonged circulation of the blood by the extracorporeal pump. A technique was examined in 15 dogs using hypothermia for maximum metabolic suppression, incorporating an aqueous blood substitute (Cryomedical Sciences, Inc., Rockville, MD). The anesthetized animals were cannulated for extracorporeal pump oxygenation. As temperature was lowered the dogs were exsanguinated and volume replaced with blood substitute to lower the hematocrit to less than 1%. After 3 hours of cardiac arrest and continuous perfusion at a core temperature less than 10 degrees C, rewarming began. When temperature reached greater than or equal to 10 degrees C, the blood substitute was drained and the animals were autotransfused. The heart was started at 15 degrees C and spontaneous respiration resumed at 29 degrees C. Using the first generation blood substitute the survival rate was maximal (100%) at 2.5 hrs under 10 degrees C and 3 hours of cardiac arrest. Research is underway on a new blood substitute, which is to serve as a universal hypothermic preservation solution (in situ organ preservation).(ABSTRACT TRUNCATED AT 250 WORDS)

Cutaneous immunopathology of androgenetic alopecia.

Male pattern baldness is assumed to result from a combination of normal serum concentrations of androgen and an appropriate genetic background. To study whether inflammation contributes to the development of androgenetic alopecia, direct immunofluorescence and dermatopathologic studies were performed on biopsy specimens from bald scalp of patients, with specimens from uninvolved scalp of these patients or from scalp of volunteers who were not bald serving as controls. Granular deposits of Immunoglobulin M or C3 (or both) were found at the basement membrane in 25 (96%) of 26 study patients and 1 (12%) of 8 control subjects. Granular C3 was also deposited on eccrine myoepithelial cells in 8 (31%) of 26 study patients, but no control subjects. Porphyrins were found in the pilosebaceous canal in 15 (58%) of 26 study subjects and in 1 (12%) of 8 control subjects. These results support an inflammatory pathogenesis of androgenetic alopecia. Propionibacterium acnes is known to produce porphyrins. Ultraviolet radiation may excite microbiologic porphyrins that could activate C3 and, subsequently, the complement cascade producing inflammatory mediators.

Ultraprofound hypothermia with complete blood substitution in a canine model.

The potential for hypothermia to prevent or ameliorate ischemic injury to the central nervous system is well known. To determine if a more prolonged period of metabolic suppression with blood substitution is possible, a method was developed to lower body temperature to near the freezing point. Eight adult mongrel dogs underwent closed-chest extracorporeal circulation with both external and internal body cooling. As they were cooled, progressive hemodilution was employed until complete exsanguination and blood substitution with an aqueous solution was accomplished. Continuous circulation and a core temperature at a mean of 1.7 degrees C were maintained from 2 1/2 to 3 hours. After rewarming to 20 degrees C, the animals were autotransfused and allowed to recover. Of the eight animals, two died due to technical factors related to cardiac defibrillation. Of the six surviving animals, five survived over a long period and one died on the 10th postoperative day with hepatorenal failure resulting from a presumed blood transfusion incompatability reaction. All six showed normal neurological function and kennel behavior, except one dog with mild weakness of a hindlimb. When the dogs were sacrificed 1 to 2 months postoperatively, all organs were histologically normal. Specifically, there was no gross or microscopic evidence of ischemic or hypoxic injury to any central nervous system structures. This pilot study demonstrates that it is possible to successfully achieve complete exsanguination, blood substitution, and ultraprofound body temperature, while continuous circulation of the blood substitute is maintained. With the capability of controlling and repeatedly performing washout of the extracellular environment and by reaching lower temperatures, it may be possible to attain greater cellular metabolic suppression. This perhaps will extend the allowable times for circulatory arrest procedures. In addition, "bloodless ischemia" may be beneficial in removing both blood substances and formed elements which may mediate organ ischemia. With replacement of blood at warm temperatures, coagulopathy is avoided. This preliminary evidence demonstrates potential in the combination of ultraprofound hypothermia and complete blood component substitution. However, further study is required to confirm the potential of achieving circulatory arrest of longer duration.

Effect of intraventricular nadolol infusion on shock-induced aggression in 6-hydroxydopamine-treated rats.

Increasingly, beta blockers are being utilized to treat patients with aggressive disorders secondary to brain lesions. To secure further data about the potential efficacy of beta blockers for this condition, a rat model was employed in which aggression was enhanced by 6-hydroxydopamine. Following lesioning of the brain, aggression in the rats increased from an average baseline of 10.9 +/- 2.1 to 42.7 +/- 2.0 fights per testing period. Thereafter, either pharmacologically inert, artificial CSF or nadolol was introduced through transcerebral, intraventricular perfusion with a significant reduction in aggression in the nadolol-treated animals when compared with those receiving artificial CSF.

Central cardiovascular activity of prostaglandin E2, prostaglandin F2 alpha and prostacyclin.

Prostacyclin (PGI2), prostaglandin E2 (PGE2) and prostaglandin F2 alpha (PGF2 alpha) were tested here in unanesthetized male Sprague-Dawley rats for their effects on the cardiovascular system as mediated by the central nervous system. Cannulae were chronically implanted into the third cerebral ventricle, femoral arteries and femoral veins of rats. Both PGE2 and PGF2 alpha induced increased arterial blood pressure and tachycardia by an action on the central nervous system. The changes seen with PGE2 were larger than those observed with PGF2 alpha. Only transient depressor effects were seen with PGI2 and these changes appeared to be due to the leakage of the substance into the peripheral vascular system.

Suppression of stimulated plasma renin by clonidine in the dog.

The mechanism by which clonidine suppresses plasma renin activity (PRA) was investigated in dogs anesthetized with pentobarbital. Injection of clonidine (1 micrograms/kg) into the cisterna magna decreased PRA from levels stimulated by prior hemorrhage into a blood reservoir to reduce mean blood pressure by 25% (21.7 ng/ml/hr +/- 6.6 SE leads to 11.1 ng/ml/hr +/- 2.4 SE; p less than 0.05). Clonidine reduced heart rate but mean arterial pressure remained constant due to fluid movement between the reservoir and the arterial circulation of the dog. These effects could not be attributed to leakage of clonidine from the cerebrospinal fluid since intravenous administration of the same dose had no effect on PRA. In animals bilaterally splanchnicotomized at the level of the diaphragm, elevated PRA was not reduced by intracisternal clonidine. When return of reservoir fluid was prevented, animals became hypotensive after central clonidine and renin tended to increase. These results indicate that clonidine reduces stimulated renin by a central mechanism that is dependent upon the integrity of the sympathetic innervation of the kidney. Other stimuli for renin release may override the inhibitory effect of central clonidine.