Diagnostic accuracy of serum B-type natriuretic peptide for myocardial ischemia detection during exercise testing with spect perfusion imaging.

OBJECTIVE: To determine whether serum B-type natriuretic peptide measured at rest and peak exercise and DeltaBNP contribute to the predictive value and diagnostic accuracy of exercise test in the diagnosis of myocardial ischemia. BACKGROUND: Ventricular myocytes release BNP in response to increased wall stress that occurs in acute ischemia. During exercise testing, transient myocardial ischemia could also cause acute myocardial stress and changes in circulating BNP. METHODS: BNP was measured before and immediately after exercise testing with radionuclide imaging in 203 consecutive subjects referred for chest pain evaluation. Tested subjects were classified as ischemic and non-ischemic based on exercise results, and no ischemia, mild-moderate, and severe ischemia according to perfusion scan results. A logistic regression model, constructed of an ROC and an AUC (area under the curve), was used. RESULTS: Ischemic ECG changes (> or =1 mm, horizontal S-T shift) were detected in the treadmill exercise test in 127 subjects (62.6%), and 76 (37.4%) had neither ST segment shift nor chest pain. Baseline BNP was higher in the ischemic group compared to the non-ischemic group (p=0.044); peak BNP was also higher in the ischemic group (p=0.025), as was DeltaBNP (p=0.0126). Of these 127 subjects, 106 (52% of all) had abnormal perfusion scan results. In the ischemic group, the median baseline, peak exercise BNP, and DeltaBNP values from baseline to peak were higher than in the non-ischemic group. In the severe ischemic group these variables were approximately three-fold higher than in the mild-moderate ischemic group (p<0.0001 for baseline; p<0.0001 for peak; and p<0.0001 for DeltaBNP). Rest, peak exercise, and DeltaBNP values were significantly higher in patients with previous myocardial infarction (p<0.001) and in patients treated with beta blockers; peak exercise BNP was higher in hypertensives and diabetics (p<0.05). The ROC convergence model showed that the AUC for peak-exercise BNP was best able to discriminate and predict severe ischemia and no ischemia, while DeltaBNP from rest to peak exercise discriminated best between mild-moderate and severe ischemia. CONCLUSIONS: Peak exercise BNP and DeltaBNP improved the sensitivity, specificity, positive likelihood ratio, predictive value, and diagnostic accuracy of severe ischemia detection during an exercise test. The contribution of BNP determination during exercise was, however, less impressive than previously reported by others.

Changes in creatine phosphokinase (CK) concentrations after minor and major surgeries in children.

BACKGROUND: During surgery, damage occurs to muscles in the area of the operation. The few studies that have examined creatine phosphokinase (CK) values after surgery have been in adults. The only study in children was after cardiac surgery. Understanding the normal enzyme pattern of change may help to differentiate malignant hyperthermia, anaesthesia-induced rhabdomyolysis and elevated CK values resulting from inherited muscle disease in cases in which these are suspected. The aim of this study was to delineate the normal rise of CK after minor and major surgery in children. METHODS: A total of 71 patients aged 1 month-17 yr were studied. From the cohort of 71 patients, 46 underwent elective surgery (14 major, 32 minor) and in 25 the surgery was designated as an emergency surgery (21 major, 4 minor). The anaesthesia protocol was similar for both groups with halothane induction and isoflurane maintenance. Owing to its possible effect on CK, succinylcholine was avoided during the study. RESULTS: The mean values of CK concentration before and after surgery were 63.1 iu litre(-1) and 151.5 iu litre(-1), respectively. The median CK elevation (range) for the major and minor surgery groups was 43 iu litre(-1) (4-647) and 10 iu litre(-1) (-28 to 122), respectively (P<0.0001). CONCLUSIONS: CK concentrations in the major surgery group were significantly higher than the minor surgery group. This profile can contribute to the evaluation of patients who present with the possibility of malignant hyperthermia, anaesthesia-induced rhabdomyolysis and underlying muscle disease. Any rise of CK concentration above what is expected should prompt further investigation.

Cardiac troponin I serum concentrations in newborns: a study and review of the literature.

BACKGROUND: Serum cardiac troponin I (cTnI) is a specific marker of cardiac injury. The use of cTnI in neonates, especially in relation to perinatal asphyxia has not been extensively examined. We defined the range of normal values of cTnI in newborns, and study factors that may influence these concentrations. METHODS: Serum cTnI concentrations were measured on the third day of life in 179 normal newborns: 157 were term (after 37 weeks, mean: 39.7+/-1.1, range: 37-42) and 22 were premature infants (mean: 32.6+/-2.9, range: 27-36 weeks). RESULTS: Mean cTnI for the term infants was 0.63+/-0.58 ng/ml (median: 0.50, range: 0.00-4.30). The concentration of 1.80 ng/ml, can serve as the upper limit of normal values. There was a borderline significant trend for higher cTnI in preterm infants. The number of newborns with cTnI>1.80 ng/ml was significantly higher after delivery by caesarean section, compared to vaginal delivery (14.6% vs. 2.9%, p<0.02). No other significant associations were found between cTnI and perinatal or neonatal parameters. CONCLUSIONS: Normal reference values for cTnI in healthy term newborns were defined, but need to be addressed with caution due to the wide range of normal values.

Value of laboratory studies in assessment of dehydration in children.

BACKGROUND: Due to the lack of a reliable way of clinically measuring dehydration, laboratory tests are usually used to improve the accuracy of clinical assessment of dehydration in children. The purpose of this study was to compare the relationship between clinical and laboratory parameters in the assessment of dehydration and to evaluate the improvement of those parameters over time. METHODS: We conducted a retrospective study to assess the relationship between clinical assessment of dehydration and laboratory findings. RESULTS: Three hundred children were eligible for the study. Twenty-six per cent of those with mild dehydration had serum urea concentrations greater than 14.3 mmol/L, compared with 38% and 5% of those with moderate or no dehydration, respectively. Urea concentration showed a good specificity, 95%. Creatinine concentrations and mean pH were similar whether or not dehydration was present. Bicarbonate and base excess concentrations decreased with the increasing severity of dehydration and were significantly greater in subjects with moderate dehydration than in those without. The sensitivity (71%) and specificity (74%) of both tests were rather poor. All groups had an abnormal anion gap, which was significantly greater in those with mild or moderate dehydration. CONCLUSION: This study confirms that there is a discrepancy between clinical assessment and laboratory parameters of dehydration. Urea showed good specificity, and anion gap was the most sensitive laboratory parameter for assessment of dehydration. These findings need further validation.

The role of high-dose oral iron supplementation during erythropoietin therapy for anemia of prematurity.

OBJECTIVE: To assess whether a high intake of oral iron would increase the effect of recombinant human erythropoietin (rHuEPO) on hemoglobin synthesis. METHODS: We studied 30 preterm infants (gestational age 29+/-1.8 weeks, birth weight 1161+/-200 g, at age of 28+/-10 days) who were randomly assigned to receive either 8 mg/kg per day (n=15) or 16 mg/kg per day of oral iron during a course of rHuEPO therapy (900 microg/kg per week) for a duration of 4 weeks. Both groups were comparable in regard to clinical and laboratory data at the time of enrollment. RESULTS: rHuEPO caused a significant increase in reticulocyte count in the low- and high-dose iron groups, 17.1+/-5.3 to 34.7+/-9.2 and 16.3+/-3.3 to 42.5+/-5.6 (10(9)/l), respectively (p<0.05). However, in both groups, hematocrit values remained stable at the end of the study as compared to baseline (0.35+/-0.03% vs. 0.30+/-0.03%, 0.35+/-0.05% vs. 0.30+/-0.03%, NS) and in both groups there was a comparable and significant decrease in ferritin level (259+/-109 to 101+/-40 and 168+/-54 to 69+/-38 microg/l, respectively; p<0.01). The rates of bloody stools without any evidence of necrotizing enterocolitis were not significantly different between the two treatment groups (1/15 vs. 4/15, NS). CONCLUSION: We conclude that a higher dose (16 mg/kg per day) of oral iron is not more beneficial when compared to a lower dose (8 mg/kg per day) during rHuEPO therapy for anemia of prematurity. Further studies will define the optimal dosage and route of administration of iron supplementation during rHuEPO therapy.

Cancer antigen 125: a sensitive marker of ascites in patients with liver cirrhosis.

OBJECTIVE: Cancer antigen 125 (CA 125) is a high molecular mass glycoprotein, usually used for monitoring the course of epithelial ovarian cancer. Recently it has been shown that liver cirrhosis is associated with increased levels of CA 125, particularly in the presence of ascites. The aim of this study was to evaluate CA 125 as a marker for the detection of ascites in patients with chronic liver disease. METHODS: A total of 170 patients were studied. All had ultrasound scanning for detection of ascites. Group I consisted of 123 patients with chronic liver disease without ascites; whereas group II consisted of 47 patients with chronic liver disease with ascites. CA 125 levels were measured in all patients and also simultaneously in the ascitic fluid of 31 patients from group II. RESULTS: Of 47 patients, 46 (97.8%) of group II had elevated serum levels of CA 125 (mean 321 +/- 283 U/ml) as compared with only nine of 123 (7.3%) patients of group I [mean 13 +/- 15 U/ml]), p < 0.001. The mean CA 125 concentration in the ascitic fluid of 31 cirrhotic patients (group II) was 624 +/- 397 U/ml and was always higher than corresponding serum levels (p < 0.01). Serum CA 125 levels correlated with the amount of ascitic fluid (r = 0.78). A profound decrease in serum CA 125 concentration was noted 2-3 and 10 days after large volume paracentesis. CA 125 was more sensitive and preceded ultrasonography in detection of ascites in few cirrhotic patients. CONCLUSIONS: CA 125 is a highly sensitive marker to detect ascites in patients with liver cirrhosis. This marker may be useful to detect small to moderate amounts of ascitic fluid in cirrhotic patients when physical examination is difficult or equivocal for ascites.

Decreased ferritin levels, despite iron supplementation, during erythropoietin therapy in anaemia of prematurity.

Erythropoietin (rHuEPO) therapy has been shown to be beneficial in preventing and treating anaemia of prematurity and to decrease the need for blood transfusions. There is, however, only scanty data on the effect of rHuEPO therapy on iron metabolism. We studied 29 preterm infants (age 34 +/- 14 days) who were randomly assigned to receive either rHuEPO 900 U kg-1 week-1 with 6 mg kg-1 day-1 of iron for 4 weeks (n = 15) or no therapy. The following parameters were evaluated and compared between and within groups at the beginning, during and at the end of the study: Haematocrit (SI), reticulocytes (10(9) micrograms l-1), serum ferritin (microgram 1-1) and iron (mumol l-1). The results were as follows. At the baseline, erythropoietin levels were similar in both groups: 7.2 +/- 5.6 versus 6.2 +/- 3.2 mU ml-1 (NS). In the treated infants the haematocrit remained stable during the study and was significantly higher than in the control group by the end of the study: 0.34 +/- 0.03 versus 0.28 +/- 0.05 (p = 0.001). rHuEPO therapy increased the reticulocyte count from 130 +/- 70 to 430 +/- 200 (p = 0.0002). However, rHuEPO therapy depleted both serum ferritin and iron levels from 321 +/- 191 to 76 +/- 58 micrograms l-1 (p = 0.04) and from 18 +/- 5 to 13 +/- 4 mumol l-1 (p = 0.03), respectively. We conclude that rHuEPO therapy prevented anaemia and its sequelae; however, serum ferritin and iron levels were depleted. We suggest that the effect of rHuEPO may be further increased by higher iron supplementation.

Alcohol-induced changes in urinary aminolevulinic acid and porphyrins: unrelated to liver disease.

Urinary porphyrins and their metabolites aminolevulinic acid (ALA) and porphobilinogen (PBG) were determined in 15 normal volunteers and in 45 alcoholics, subdivided into three groups according to their liver function tests and histology: alcoholics exhibiting no evidence of hepatocellular damage; alcoholics with fatty liver and impaired function of liver enzymes; and alcoholics with proven liver cirrhosis. The dominant trend observed in those alcoholics devoid of any evidence of liver disease was increased ALA, PBG, and uroporphyrin. Coproporphyrinuria was shared by the patients exhibiting liver damage. The data shown enabled us to differentiate between the direct, primary effect of alcohol on the heme biosynthetic pathway and the secondary indirect effect, which is probably related to liver damage that follows alcohol consumption. Evaluation of the results led to the suggestion that urinary ALA could possibly serve as a marker of alcoholism. The specificity and sensitivity of the test were found to be 87% and 80%, respectively.

Neonatal urinary uric acid/creatinine [correction of ceratinine] ratio as an additional marker of perinatal asphyxia.

UNLABELLED: The diagnosis and evaluation of perinatal asphyxia can be problematic and objective means of assessing its severity are lacking. To study the validity of urinary uric acid as a marker of the degree of perinatal asphyxia, the ratio of urinary uric acid to creatinine (UA/Cr) in urine specimens obtained after birth was measured in two groups of infants. Eighteen term infants with Apgar scores < or = 5 at 5 min and/or an umbilical cord blood pH < or = 7.2, and a base deficit > or = 12 meq/l were compared to 50 healthy controls. The severity of the perinatal asphyxia was determined by using an ASPHYXIA SCORE. The UA/Cr was higher in the asphyxiated group when compared to controls. (2.06 +/- 1.12, vs. 0.64 +/- 0.48; P < 0.001). Within the perinatal asphyxia group, a significant correlation was found between the UA/Cr ratio and the asphyxia score. (r = 0.86, P < 0.01). CONCLUSION: Infants with perinatal asphyxia have a significantly higher urinary UA/Cr ratio. This may be used as an indicator of the severity of perinatal asphyxia.

Aging and food restriction alter some indices of bone metabolism in male rhesus monkeys (Macaca mulatta).

Food restriction increases life span, reduces aging rate and affects a wide variety of biological functions. In rats, food restriction delays bone growth and reduces bone density and mineral content. We report the effects of aging and long-term (> 6.0 y) food restriction on several indices of bone growth and metabolism in rhesus monkeys (Macaca mulatta). Food allotments for controls approximated free access consumption, whereas food-restricted monkeys received 30% less food on a body weight basis. Cross-sectional and longitudinal age effects on serum alkaline phosphatase paralleled those reported for humans. Food restriction induced a significant delay in the developmental decline (to adult levels) in total alkaline phosphatase and significantly suppressed serum interleukin 6 concentrations, particularly in younger monkeys. Also, food restriction slowed skeletal growth, as reflected by shorter crown-rump length, and significantly reduced total body bone mineral content, but not bone mineral density, measured by dual energy X-ray absorptiometry. Analyses of serum parathyroid hormone, calcium, phosphate and osteocalcin concentrations suggested that the effects on skeletal growth were not related to alterations in calcium and phosphate homeostasis or a primary defect in bone formation. These findings suggest that long-term food restriction delays skeletal development in male rhesus monkeys while allowing the development of a reduced but otherwise normal skeleton.

Relationship of sweat electrolytes to apparent life-threatening events (ALTE): a case control study.

Twenty infants aged 25 days to 6 months who were consecutively investigated for apparent life-threatening events (ALTE) with negative results and 20 matched normal controls underwent an iontophoresis sweat test. A statistically significant elevated sweat potassium level (22.1 +/- 8.9 versus 12.4 +/- 6.5 mol/l) was noted in the ALTE patients compared with the control group (p < 0.001). No difference was found, however, between sweat sodium and chloride levels in the two groups. Na/K ratio in sweat was significantly different between the groups (p < 0.001). A between-groups discriminant analysis, using Na/K ratio as a discriminant variable, resulted in 80% accuracy in group assignment. A significant increase in sweat potassium concentration at night compared with day time was evident in ALTE patients. Elevated sweat potassium levels specifically characterized infants who experienced ALTE and may possibly indicate an underlying mechanism involving enhanced sympathetic activity.

Treatment of severe, resistant familial combined hyperlipidemia with a bezafibrate-lovastatin combination.

Familial combined hyperlipidemia (FCHL) is a common lipid disorder characterized by high levels of cholesterol, triglycerides, or both. The basic metabolic abnormality is overproduction of apolipoprotein B-100. High atherogenicity has been attributed to all forms of FCHL. We evaluated combined bezafibrate-lovastatin therapy in 10 patients (9 men and 1 woman) with FCHL and markedly high cholesterol and triglyceride levels who were at high risk of coronary artery disease and who had not responded to diet and bezafibrate treatment alone. Eight patients had coronary artery disease, 6 had hypertension, and 3 had noninsulin-dependent diabetes mellitus. Lovastatin 20 mg/day was added to the bezafibrate 600 mg/day regimen for 6 weeks; the lovastatin dosage was then doubled to 40 mg/day for an additional 6 weeks. The addition of 20 mg of lovastatin resulted in decreases of 15%, 20%, and 13% in total cholesterol, low-density lipoprotein (LDL) cholesterol, and triglyceride levels, respectively. Increasing the dose of lovastatin to 40 mg resulted in further moderate decreases of 4%, 3%, and 8% in total cholesterol, LDL cholesterol, and triglycerides, respectively, compared with the 20 mg/day dosage. Although previous reports have emphasized the potential side effects of combination treatment with lovastatin and fibric acid derivatives, our patients tolerated the regimen well, with no significant subjective complaints or laboratory abnormalities. The bezafibrate-lovastatin combination is a possible therapeutic option for severe, resistant FCHL, but close medical supervision is needed because of potential side effects.

23Na- and 31P-NMR studies of perfused mouse liver during nitrogen hypoxia.

The effects of nitrogen hypoxia on isolated perfused mouse livers from fed mice were studied at 37 degrees C using 23Na and 31P nuclear magnetic resonance (NMR) spectroscopy. The paramagnetic shift reagent, dysprosium-triethylenetetraminehexaacetic acid, was used to distinguish intracellular from extracellular sodium. The area of the intracellular sodium resonance remained relatively constant over the first 30 min of hypoxia and then increased by a factor of approximately 2 relative to controls over the next 30 min. High-energy phosphate metabolites were measured using 31P-NMR. The beta-ATP resonance decreased to zero, and the intracellular pH decreased from 7.3 to 6.9 during 60 min of hypoxia. Liver enzyme activity in the effluent exiting the liver increased in direct proportion to the length of hypoxia up to 56 min. The sodium, ATP, and enzyme changes during hypoxia were correlated with histological and electron-microscopic findings. The morphology of liver specimens exposed to 30 min of hypoxia was close to normal, whereas extensive centrilobular and midlobular necrosis was seen in specimens subjected to 60 and 90 min of hypoxia. The effect of 30 min of reoxygenation after 15, 30, 45, 60, and 90 min of hypoxia was also studied. The level of beta-ATP recovery depended on the duration of hypoxia. For 60 min of hypoxia followed by reoxygenation, beta-ATP recovered to only 20% of control values. The morphology of hypoxic livers after 30 min of reoxygenation was similar to livers subjected to hypoxia alone.

Liver adenosine triphosphate and pH in fasted and well-fed mice after infusion of adenine nucleotide precursors.

Nutrition is a factor which may affect the liver energy charge. Experiments were performed to determine the effect of starvation and of ATP precursors, adenine and ribose on liver energy stores. The 31P NMR spectra of well-fed and starved mice livers were studied in a perfusion system using Krebs-Henseleit buffer (KHB). The ATP precursors, adenine (20 mmol/l) and ribose (80 mmol/l), were then added to determine their effect. Their effect on the ATP dynamics during ischemia and reperfusion were then evaluated. The effects of adenine alone and ribose alone were then determined. The 31P spectra of well-fed mice demonstrated high ATP content relative to Pi, phosphoesters and phospholipids. Animals starved for 24 h showed very low ATP, high Pi and little or no detectable phospholipids. In starved animals, ATP rose steadily to approximately 50% above the baseline level when precursors were added. Pi decreased to 30% of the baseline after 40 min. Little change was noted in well-fed animals. The rate of ATP decay did not change with the onset of ischemia, whether the livers were perfused with KHB alone or KHB with precursors. Upon reperfusion, precursors improved the recovery of ATP (81% vs 49% after 20 min ischemia, 44% vs 34% after 30 min ischemia). Addition of adenine alone produced similar results, but addition of ribose alone did not significantly alter ATP recovery. In conclusion, supplying starved or post-ischemic livers with adenine or ribose and adenine does improve ATP levels.

Hepatic transplantation survival: correlation with adenine nucleotide level in donor liver.

The hypothesis was tested that human donor livers with higher ATP content and energy charge achieve better results after hepatic transplantation. Biopsies were obtained from 25 donor livers immediately prior to implantation and analyzed for adenine nucleotides using high-performance liquid chromatography. The results were correlated with organ histology, transplant function and outcome. Significantly higher concentrations of ATP (4.22 +/- 2.87 vs. 0.71 +/- 0.69 nmoles per mg protein, p less than 0.01), ADP (8.75 +/- 2.96 vs. 4.49 +/- 1.95 nmoles per mg protein, p less than 0.01) and energy charge (0.43 +/- 0.15 vs. 0.21 +/- 0.04, p less than 0.02) were found in successful (n = 20) relative to failed (n = 5) livers. No significant differences were found in AMP, xanthine or hypoxanthine for the two groups, although the average values were higher in failed livers. Fifteen recipients with liver ATP concentration above 2 nmoles per mg protein and energy charge above 0.3 recovered well. Five other successful patients with lower ATP concentration (0.70 +/- 0.39 nmoles per mg protein) and energy charge (0.20 +/- 0.03) had postoperative courses complicated by infection or prolonged hyperbilirubinemia. In five patients whose livers failed, all had low ATP content and energy charge. Of these, three received a replacement liver and two died shortly after the transplantation. The study demonstrates a direct correlation between high ATP content and good posttransplant outcome.

Magnetic resonance imaging of gynecologic masses.

The pelvis of 21 women with various gynecological masses were imaged with magnetic resonance (MR) imagers at 2.0 and 0.5 T. Fifteen normal individuals were used for studying the normal appearance of the female genitalia using spin echo pulse sequences with various pulse repetition (TR) and spin echo (TE) time values. Images were compared with those of the ultrasonic images, intraoperative findings, and the histopathologic examinations. The masses included simple ovarian cyst, cystadenoma, serous cystadenocarcinoma, ovarian teratoma, dysgerminoma, and uterine myoma. MR imaging was useful in demonstrating the anatomy and pathology of the cases examined in this study. It detected the internal structure of some tumors which were sonographically homogeneous. The potential of MR in staging of malignancies was demonstrated.

Nuclear magnetic resonance assessment of adenosine triphosphate (ATP) dynamics in ischemic mouse livers perfused with adenine and ribose.

Hepatic energy stores are essential to liver viability. We used a mouse liver perfusion model and MR spectroscopy to study the effect of adding two precursors of ATP (adenine and ribose) on ATP dynamics during ischemia and reperfusion. Using Krebs-Henseleit buffer with or without added adenine and ribose made little difference in the ATP decay rate during ischemia, but the recovery of ATP during reperfusion was more complete when adenine and ribose were added to the buffer. These findings suggest that the addition of the precursors of ATP, adenine and ribose, to perfusate after ischemia can accelerate and enhance ATP recovery.

Liver adenine nucleotide metabolism during ischemia and reperfusion of mice livers studied by phosphorous-31 nuclear magnetic resonance.

Perfusion experiments were performed at 20 degrees C and 37 degrees C to study liver adenine nucleotide metabolism during ischemia and reperfusion of mouse livers using 31P NMR. Perfusing at 8 mL/min (Krebs-Henseleit buffer), ATP was shown to be stable for 6 hours. There was a progressive decrease in the phosphodiesters (glycerophosphorycholine and glycerophosphorylethanolamine) during the 6-hour period. Liver subjected to cold ischemia at 20 degrees C showed a slow decrease in the beta ATP peak during a 42 +/- 6-minute period with a rise in the inorganic phosphate accompanied by a shift of inorganic phosphate to the high field indicating intracellular acidosis. With reperfusion, the beta ATP returned to previous levels and the inorganic phosphate shifted to its original position. During warm ischemia (37 degrees C) the ATP peak disappeared within 5 +/- 1 minute and only returned to 34% of its original value after 30 minutes of ischemia, indicating damage to a certain percentage of liver cells. When the liver was subjected to multiple short periods of cold ischemia, there was complete recovery of the ATP after six cycles. Reperfusion after each period of cold ischemia resulted in an ATP recovery consistently greater than the initial amount, which gradually decreased to preischemic levels after a short period. This suggests that there is an as yet undelineated mechanism of ATP production during ischemia that attempts to protect the cell against ischemia.

Liver preservation for transplant. Evaluation of hepatic energy metabolism by 31P NMR.

31P NMR spectroscopy proved to be an excellent, dynamic, nondestructive method for assessing the liver during cold flush and pulsatile perfusion experiments. 31P NMR spectroscopy was used to measure ATP decay, inorganic phosphate appearance, and phosphate chemical shift in the excised mouse livers subjected to cold and warm ischemia. Cold flush followed by cold preservation in saline, Krebs-Henseleit buffer, or Collins' solution showed that Collins' solution resulted in the slowest ATP decay. In temperature-controlled experiments (5 degrees -37 degrees C), ATP decay was much slower with lower temperature. In separate pulsatile perfusion experiments with oxygenated Krebs-Henseleit buffer, hepatic ATP was unchanged for at least 6 hr at 20 degrees C. At 37 degrees C, the NMR spectrum showed changes in the diphosphoesters region, but the ATP remained stable during the 6-hr perfusion. These studies suggest that for long periods of liver preservation, an adequate perfusion method should be developed.