Phosphatidylethanol in rat organs after ethanol exposure

BACKGROUND: Phosphatidylethanol (PEth) is an abnormal phospholipid formed in mammalian cells that have been exposed to ethanol. It has been suggested that PEth mediates some of the damaging effects of ethanol on cells. This study was performed to investigate the level of PEth in organs of rats after in vivo alcohol exposure. METHODS: Three exposure models were studied: (1) acute, intraperitoneal injection of ethanol (n = 3 x 3); (2) chronic, forced ethanol drinking (n = 6); and (3) chronic, free choice of ethanol (n = 20). PEth was analyzed by high-performance liquid chromatography after lipid extraction of the organs. RESULTS: One acute injection gave detectable PEth levels in most organs analyzed, with maximal levels reached after 2 hr. The highest levels were reached in intestines, stomach, and lung. No PEth was detected in skeletal muscle, pancreas, or testis. The two exposure models for oral intake of ethanol also gave detectable PEth levels in most organs. The highest levels were reached in stomach, lung, and spleen. PEth was detected in muscle only in animals with heavy total alcohol intake. CONCLUSIONS: PEth is formed in most organs of rats exposed to ethanol acutely or chronically. Variations in PEth level and rates of PEth formation and PEth degradation are organ specific.     

Non-frozen cold storage is favorable for islet function and morphology

Freezing has been shown to damage pancreatic islets and to disrupt their insulin release, probably because of intracellular ice formation. We compared frozen islets with fresh ones and with others stored at temperatures above freezing from a standpoint of insulin release response to glucose and transplantation. Group A islets, isolated from rats and immersed in 10% dimethyl sulfoxide in RPMI 1640, were stored at -2 degrees C, and group B islets at -196 degrees C, for 7 days. As for group B, the islets were cooled at 1 degree C/min from room temperature to -40 degrees C, subsequently at 3 degrees C/min to -80 degrees C and then put into liquid nitrogen to be rapidly frozen to -196 degrees C. The control islets were fresh. In vitro, basal release at 3.3 mM glucose was similar in group A to that in the controls, but was higher in group B than group A. Stimulated release against 16.7 mM glucose was lower in group A than group B. However, insulin responsiveness, i.e., the ratio of insulin release at 16.7 mM glucose to that at 3.3 mM glucose, was lost in group B. Freezing also caused damage to the group B cells visible under the light and electron microscopes, while group A islets were largely intact. In vivo, after 600 islets were transplanted into streptozotocin-induced diabetic rats, group A was better able to lower fasting blood glucose than was group B, and remained so for 4 weeks. Above sub-zero preservation in the non-frozen state thus seems adequate for the short-term storage for 7 days.     

Cryopreservation of microfilariae and third-stage larvae of Brugia malayi and subsequent development in the hosts

Methods are studied for the cryopreservation of microfilariae (mf) and third-stage larvae (L3) of periodic Brugia malayi. RPMI-1640 tissue culture medium containing 6% dimethyl sulfoxide (DMSO) and 15% newborn calf serum were used as cryoprotectant. The larvae survived best when specimens were frozen at the rate of -0.5 degrees C to -1.0 degrees C per minute using the vapor phase of liquid nitrogen, when the temperature reached -70 degrees C to -90 degrees C the specimens were placed directly into the liquid nitrogen (-196 degrees C). After the thawing of the mf which had been stored for 6,212 and 375 days in cryogen, 96.2% of the mf were shown to be viable and developed in Aedes togoi. It was also shown that the survival rate of L3 cryopreserved for 28-321 days was also 96.2% and that, when 107 L3 frozen for 321 days were inoculated after being thawed into one jird, one live female adult was recovered at autopsy 71 days after inoculation, its morphology being the same as the unfrozen specimens. There was no correlation between the time of cryopreservation and the survival rate of the larvae.     

Metabolic damage to human saphenous vein during preparation for coronary artery bypass grafting

Segments of saphenous vein from patients undergoing coronary artery by-pass graft surgery were frozen in liquid nitrogen immediately on dissection (control), after stripping of the adventitia and side branch ligation (manipulation), after distention with blood (distention), or at completion of the last proximal anastomosis (prepared vein). Vein was stored during the operation in patient's heparinised arterial blood at room temperature. Frozen vein was extracted with perchloric acid. ATP, ADP, and AMP, adenosine, inosine and hypoxanthine concentrations were measured by high pressure liquid chromatography. Prepared vein had ca 50% lower ATP concentrations and ATP/ADP ratio than control vein, higher concentrations of inosine and hypoxanthine and lower concentrations of AMP and adenosine. ATP concentration and ATP/ADP ratio did not correlate with the time elapsed between dissection and freezing of the prepared vein. The characteristic changes seen in prepared vein were not seen when control vein was simply stored in arterial blood at 23 degrees C, in normal saline at 23 degrees C or 4 degrees C, in Krebs-Ringer bicarbonate buffer at 37 degrees C or at St Thomas's Hospital cardioplegic solution at 4 degrees C. Distention with unlimited pressure did not distension at less than 300 mmHg gave rise to the same changes in ATP concentration and ATP/ADP ratio as in the prepared vein. These results show that vein suffered metabolic changes during preparation for bypass grafting and suggest that uncontrolled distention may contribute to these changes. Such biochemical measurements provide a quantitative estimate of tissue damage and allow objective comparison of different preparative techniques.     

Cryopreservation of Human Platelets Using 6% Dimethyl Sulfoxide and Storage at -80°

Platelet studies were done in healthy male volunteers and in thrombocytopenic patients. Some of the platelets used in the study were isolated by mechanical apheresis using either the Haemonetics blood processor 30, the IBM blood processor 2997 or the Fenwal CS-3000 blood processor before freezing. Other platelets were isolated from individual units of whole blood and pooled before freezing. The platelets were frozen with a 6% cryoprotectant (DMSO) in a polyvinylchloride (PVC) plastic bag or a polyolefin plastic bag at -80 degrees C in a mechanical freezer and stored for as long as 3 years. Some of the frozen platelets were transported in dry ice in polystyrene foam containers to determine whether they would be adversely affected by such treatment. Platelet recovery after freezing, thawing and washing was about 75%. In the healthy male volunteers, in vivo recovery of autologous platelets 1-2 h after transfusion was about 33%, and the life span was about 8 days. In the thrombocytopenic patients, in vivo recovery values were 50% of those from fresh platelets. The transfusion of previously frozen washed platelets reduced clinical bleeding in the thrombocytopenic patients with bleeding. There was no evidence of quality deterioration in platelets after storage at -80 degrees C for at least 2 years, as determined from in vivo recovery and in vivo survival values, nor was there any adverse effect as a result of shipment of the frozen platelets in dry ice in polystyrene foam containers from one facility to another.     

Spermatozoa and spermatids retrieved from frozen reproductive organs or frozen whole bodies of male mice can produce normal offspring

Cryopreservation of male germ cells is a strategy to conserve animal species and strains of animals valuable to biomedical research. We tested whether mouse male germ cells could be cryopreserved without cryoprotection by simply freezing epididymides, testes, or whole bodies. The reproductive organs were isolated from killed mice and frozen for 1 week to 1 year at -80 degrees C before spermatozoa and spermatids were collected and injected into mature oocytes. Normal pups were born irrespective of strains tested (ICR and C57BL/6). Epididymides and testes frozen and transported internationally to another laboratory by air could produce pups of inbred C57BL/6 mice. Testicular spermatozoa retrieved from the bodies of male mice (BALB/c nude and C3H/He strains) that had been kept frozen (-20 degrees C) for 15 years could also produce normal offspring by microinsemination. Thus, freezing of either male reproductive organs or whole bodies is the simplest way to preserve male germ cells. Restoration of extinct species could be possible if male individuals are found in permafrost.     

Plasma atrial natriuretic peptide is unstable under most storage conditions.

BACKGROUND. Atrial natriuretic peptide (ANP) is a hormonal regulator of cardiovascular fluid volume. More than 1,000 scientific articles were written about ANP between 1987 and 1991. Because some articles hinted at problems with storing ANP, this study examined the effect of numerous techniques for storing and processing human ANP samples. METHODS AND RESULTS. Samples were obtained repeatedly from three patients, treated, and stored under a variety of conditions. Experiment 1 evaluated the effects of different preservatives at 35, 21, 14, 10, and 7 days before assay. Experiment 2 evaluated nonspecific binding of ANP to different storage tubes during 28 days of storage. Experiment 3 evaluated the effect of storage at -20 degrees C, -80 degrees C, and -196 degrees C for 1 month. ANP was very unstable, degrading as much as 30% after 3 days of storage and by more than 50% in 1 month even when stored at -80 degrees C. Only storage at -196 degrees C (in liquid nitrogen) kept ANP stable for 1 month. Extraction and lyophilization of the samples before freezing and assay within 7 days of freezing only partially minimized the amount of degradation. All other processing techniques had little effect on slowing the degradation of ANP. CONCLUSIONS. These findings raise disturbing questions about the interpretation of the substantial literature on ANP.     

Survival and infectivity of Brugia malayi microfilariae after cryopreservation

Methods were studied for the cryopreservation of microfilariae of periodic Brugia malayi. RPMI-1640 tissue culture medium containing 6% dimethyl sulfoxide (DMSO) and 15% newborn calf serum was used as cryoprotectant. Samples were frozen slowly in the vapor phase of liquid nitrogen prior to emersion in liquid nitrogen (-196 degrees C). The freezing rate was -0.5 to -1.0 degrees C per minute, microfilariae remained viable for as long as, 212 and 375 days, survival rates were 94 to 98% and they were infective to Aedes togoi mosquitos. The infective larvae (L3) were obtained for 10-11 days after feeding at 28 degrees C room-temperature and the infection rate of L3 in test mosquitos was 22.4-30.6%. All DMSO should be removed from the freezing medium to restore microfilariae activity after freezing.     

Retention of coagulation factors in plasma frozen after extended holding at 1-6 degrees C

BACKGROUND AND OBJECTIVES: The ability to use plasma, isolated from units of whole blood and frozen within 24 h of phlebotomy, as a substitute for plasma frozen within 8 h of phlebotomy would have several advantages for blood centers. It should provide increased flexibility pertaining to the freezing of plasma for clinical use. We have conducted studies to assess the influence of an extended holding time for separated plasma, prior to freezing, on the retention of coagulation factor activity. STUDY DESIGN AND METHODS: Freshly harvested plasma from each of 10 units of CPD-whole blood was divided into four equal aliquots. These aliquots were held in plastic packs at 1-6 degrees C for a total of 0, 8, 15 and 24 h. Subsequently, the plasma aliquots were frozen rapidly and stored at -20 degrees C for 4 months. The thawed plasma was tested for coagulant factors V and IX, factor VIII coagulant activity (factor VIII:C), von Willebrand factor antigen (vWF:Ag) and ristocetin cofactor of von Willebrand factor. RESULTS: The levels of factor V, factor vWF:Ag, factor IX and ristocetin cofactor were not influenced by holding the plasma for up to 24 h prior to freezing. Factor VIII:C activity was reduced with extended holding at 1-6 degrees C; the percentage at time zero activity was 75.9+/-2.4% for samples frozen immediately after a 24-hour period. CONCLUSIONS: The data indicate that coagulation factor properties of harvested plasma are retained except for factor VIII for at least 24 h prior to freezing.     

Hatchling turtles survive freezing during winter hibernation.

Hatchlings of the painted turtle (Chrysemys picta marginata) are unique as the only reptile and highest vertebrate life form known to tolerate the natural freezing of extracellular body fluids during winter hibernation. Turtles survived frequent exposures to temperatures as low as -6 degrees C to -8 degrees C in their shallow terrestrial nests over the 1987-1988 winter. Hatchlings collected in April 1988 had a mean supercooling point of -3.28 +/- 0.24 degrees C and survived 24 hr of freezing at -4 degrees C with 53.4% +/- 1.98% of total body water as ice. Recovery appeared complete after 20 hr of thawing at 3 degrees C. However, freezing at -10.9 degrees C, resulting in 67% ice, was lethal. A survey of possible cryoprotectants revealed a 2- to 3-fold increase in glucose content of liver and blood and a 3-fold increase in blood glycerol in response to freezing. Although quantitatively low, these responses by spring turtles strongly indicate that these may be the winter-active cryoprotectants. The total amino acid pool of blood also increased 2.25-fold in freezing-exposed turtles, and taurine accounted for 52% of the increase. Most organs accumulated high concentrations of lactate during freezing, a response to the ischemic state imposed by extracellular freezing. Changes in glycogen phosphorylase activity and levels of glucose 6-phosphate and fructose 2,6-bisphosphate were also consistent with a dependence on anaerobic glycolysis during freezing. Studies of the molecular mechanisms of natural freeze tolerance in these turtles may identify protective strategies that can be used in mammalian organ cryopreservation technology.     

Blood Phosphatidylethanol as a Marker of Alcohol Abuse: Levels in Alcoholic Males during Withdrawal

Phosphatidylethanol (PEth) is formed only in the presence of ethanol, via the action of phospholipase D. We studied PEth in blood as a possible marker of alcohol abuse in 15 male alcoholics admitted for detoxification. Blood was drawn on the first day after admission and up to 28 days thereafter. PEth in whole blood was 13.2 ± 2.2 pmol liter⁻¹ (mean ± SE) at first sampling and remained detectable up to 14 days after admission. Blood ethanol was 0 on the morning after admission. The time courses of PEth disappearance varied among individuals. No PEth could be found in blood of control persons who had abstained from ethanol for 4 days. Levels of PEth and carbohydrate-deficient transferrin or γ-glutamyltranspeptidase did not correlate. Its high specificity and prolonged detectability suggest PEth in blood as a marker of recent alcohol abuse.     

Post-thaw storage at 4 degrees C of previously frozen red cells with retention of 2,3-DPG

Fresh human blood was collected in CPD, frozen by either the Meryman or the Valeri high glycerol technique, and stored at -80 degrees C. Later the red cells were thawed, deglycerolized by the appropriate technique and resuspended in either saline-glucose wash solution or an additive solution containing ascorbate-2-phosphate, adenine, glucose (dextrose), mannitol and sodium phosphate. The cells were stored at 4-6 degrees C for 21 days and assayed weekly for ATP, 2,3-DPG, pH, P50, glucose utilization and lysis. The additive solution maintained red cell 2,3-DPG at fresh blood levels for 3 weeks and maintained ATP levels sufficiently well to suggest good red cell viability for 21 days. There was no difference in results between the Meryman or the Valeri freezing methods if sodium phosphate was used with the saline-glucose wash solution in the Valeri method. If this additive solution is coupled with sterile deglycerolization techniques, 3 weeks of post-thaw red cell preservation would be practical. Using this additive solution would make frozen blood a reasonable source of red cells for emergency needs in both military and civilian blood banking.     

Effects of storage treatment on fecal steroid hormone concentrations of a rodent, the Cape ground squirrel (Xerus inauris)

Fecal steroid analysis is an increasingly common non-invasive technique used in both captive and field studies to measure an animal's approximate hormonal levels and corresponding physiological state. Fecal collection in the field necessitates storage and transportation methods that will prevent the degradation of hormonal metabolites by fecal bacteria. To determine the most stable and therefore preferred method of storage, 48 fecal samples were collected from six captive female Cape ground squirrels (Xerus inauris). Each sample was randomly divided into three sub-samples to be processed for storage through freezing, drying, or preservation in ethanol. Frozen samples were stored at -20 degrees C, dried-treated samples were desiccated in a conventional oven at 40 degrees C for 4 h, and alcohol-treated samples were preserved in 3 ml of 95% ethanol. Samples were stored for 330 days followed by enzyme immunoassay analysis (EIA) to determine their progestogen and estrone conjugate (E(1)C) concentrations. Validations were performed to establish that the progestogen and E(1)C assays accurately measure fecal progestogen and estrone conjugate concentrations and were sensitive enough to detect biologically meaningful differences in these steroid metabolite concentrations in female X. inauris. Validation results showed a significant difference in progestogen concentrations of gravid females compared to sub-adults and non-gravid females. There was also a significant difference in estrone conjugates between sub-adult and adult females. Duration of storage time did not affect progestogen or estrone metabolite concentrations after being frozen for 3 months. Storage treatment results showed no significant difference between frozen and dried samples, but a significant difference was found between frozen and ethanol samples in both progestogen and estrone conjugate concentrations demonstrating that drying feces provides a reliable method for long-term preservation of fecal steroid concentrations and is the better alternative when freezing is not a viable option.     

An experiment with glycerol-frozen red blood cells stored at -80 degrees C for up to 37 years

BACKGROUND AND OBJECTIVES: Red cells frozen using 40% W/V glycerol are currently FDA approved for frozen storage at -80 degrees C for up to 10 years. MATERIALS AND METHODS: Red cells frozen with 40% W/V glycerol and stored at -80 degrees C for up to 37 years were thawed, deglycerolized, and stored at 4 degrees C for 24 h. RESULTS: Red cells frozen for up to 37 years had mean freeze-thaw-wash recovery values of 75%, less than 1% hemolysis, and normal ATP, 2,3-DPG and P50 levels, and 60% of normal RBC K(+) levels. CONCLUSIONS: Red cells frozen with 40% W/V glycerol can be stored at -80 degrees C for up to 37 years with acceptable in vitro results.     

Successful banking of pancreatic endocrine cells for transplantation

To determine optimal freezing and thawing conditions for rat pancreatic endocrine cells (PEC) and insulinoma cells, five different cryopreservation protocols were compared in this study. PEC and insulinoma cells were cooled at rates of between -0.3 degrees C/min and -5 degrees C/min to -70 degrees C in the presence of 10%, 15%, or 20% dimethylsulfoxide (DMSO) with a programmable temperature controller and then transferred to liquid nitrogen for storage. Frozen cells were thawed by either rapid (in 37 degrees C water bath) or slow (in air) thawing procedure. One hour after the thawing process, cellular viability was determined by trypan blue dye exclusion. The viability results for PEC and insulinoma cells were similar and showed that a slow cooling rate at -0.3 degrees C/min in combination with a rapid thawing in 37 degrees C water bath gave the best results, with up to 80% cellular viability. Cryoprotectant DMSO used at 10% concentration was the most effective among the three concentrations tested. Later, transplantation studies were performed with PEC cryopreserved with the best protocol, which is -5 degrees C/min to 4 degrees C, held for 3 minutes, -0.3 degrees C/min to -7 degrees C, held for 3 minutes, -0.3 degrees C/min to -40 degrees C, and -5 degrees C/min from -40 degrees C to -70 degrees C in 10% DMSO with a programmable temperature controller then transferred to liquid nitrogen for storage.(ABSTRACT TRUNCATED AT 250 WORDS)     

Survival of fetal rat pancreases frozen to -78 and -196 degrees.

Transplantation of pancreases may have clinical utility in the treatment of diabetes, for it has been shown that chemically induced diabetes in rats can be reversed by the transplantation of greater than or equal to four syngeneic fetal pancreases. Allogeneic transplants produce serious immunological problems, but the problems could be ameliorated if tissue-typed organs could be stored in the frozen state. Unfortunately, nearly all attempts to freeze organs have failed. Nevertheless, recent developments in the physical-chemical analysis of freezing injury and its successful application to the freezing of mammalian embryos encouraged us to attempt the freezing of 16 1/2- to 17 1/2-day intact fetal pancreases. The analysis indicated that to achieve success pancreases would have to be cooled less than 1 degree/min and diluted extremely slowly after thawing. Experiments with embryos and red cells indicated that high survivals might require high concentrations of protective solutes and slow warming. These predictions were accurate. After freezing to -78 or -196 degrees and thawing under optimal conditions, the fetal pancreases synthesized 80-100% as much protein as unfrozen controls and they yielded viable allografts. Optimal conditions included suspension in 2 M dimethylsulfoxide, freezing at 0.3 degrees/min, and slow dilution to preclude osmotic shock.     

Comparative Study on Ethanol Elimination and Blood Acetaldehyde between Alcoholics and Control Subjects

Factors influencing ethanol elimination rate and blood acetaldehyde concentration with regard to age, blood ethanol concentration, and degrees of alcoholic liver disorder were studied in alcoholic patients vis-à-vis healthy subjects. Ninety-four male alcoholic patients were divided into five groups according to age and into three groups according to blood ethanol concentration which ranged between 4.21 and 0.29 mg/ml. It was noted that as age increased, blood ethanol concentration decreased. Groups A, B, and C consisted of subjects whose ethanol levels were more than 2.5, between 1.0 and 2.5, and less than 1.0 mg/ml, respectively. Average values of ethanol elimination rate in Groups A, B, and C were 0.26, 0.23, and 0.18 mg/ml/hr, respectively. The rate increased as blood ethanol concentration elevated. The rate in Group B was significantly higher than that in the healthy control subjects who had blood ethanol levels between 1.0 and 2.5 mg/ml. The rate of ethanol elimination was independent of liver disorder judged by liver function test values. Blood acetaldehyde concentrations in the patients were less than 1.48 micrograms/ml, with an average value of 0.58 microgram/ml, and were significantly correlated with blood ethanol levels in both alcoholics and healthy subjects.     

Transfer of phosphatidylethanol between lipoproteins

BACKGROUND: Phosphatidylethanol (PEth) is an abnormal phospholipid formed only in the presence of ethanol. It has been recently shown that lipoprotein-associated PEth may mediate the effects of ethanol on endothelial cells, and this may explain, at least in part, the beneficial effect of ethanol on atherosclerosis. This study was performed to investigate the transfer of PEth between lipoproteins and the effects of PEth on cholesteryl ester transfer protein (CETP) activity in plasma. METHODS: Lipoproteins were isolated from the plasma of healthy male volunteers (n = 16) and male alcoholics (n = 13). The transfer of cholesteryl esters and PEth was determined between labeled low-density lipoprotein (LDL) and unlabeled high-density lipoprotein particles in vitro. The electrophoretic mobility of PEth-modified LDL particles was determined by agarose gel electrophoresis. RESULTS: PEth was transferred from PEth-modified LDL to high-density lipoprotein at an initial rate of 25.9 nmol/ml/hr. Monoclonal antibody (TP2) against the putative lipid-binding domain of CETP inhibited the transfer rate of PEth by approximately 64%, whereas the cholesteryl ester transfer was inhibited by 86%. This indicates that most of PEth was transferred by transfer proteins other than CETP. CONCLUSIONS: The transfer of PEth between lipoproteins enables the redistribution of PEth from lipoprotein fractions with a slow turnover to those with a rapid clearance. Moreover, the PEth-induced change in the electrical charge of lipoproteins may affect the binding of lipoproteins to their receptors and binding proteins. This in turn may alter the metabolism of lipoproteins and lipid-mediated signaling pathways in the cells delineating the vascular wall.     

Use of ethanol for preserving steroid and indoleamine hormones in bird plasma

Endocrinological research on wild animals inhabiting remote areas has been hampered by the need to store plasma samples at subzero temperatures. In an attempt to remedy this logistical issue, we here investigate the use of ethanol as an alternative to freezing for the preservation of steroid and indoleamine hormones in avian plasma. Known quantities of the steroids 5alpha-dihydrotestosterone (DHT), testosterone, 17beta-estradiol, corticosterone, and the indoleamine melatonin were added to a stripped pool of chicken plasma. Samples were either immediately frozen at -40 degrees C, or treated with pure ethanol. Ethanol-treated samples were either immediately frozen, or-to simulate storage conditions at various field locations-left sitting at room temperature for one to two months, or incubated at 36 degrees C for one month before all treatment groups were frozen at -40 degrees C. All samples were then analyzed by radioimmunoassay. For DHT and estradiol there were no differences among treatment groups suggesting that ethanol-treatment is as effective as immediate freezing in preserving plasma steroid concentrations. For testosterone, corticosterone and melatonin ethanol-treated samples differed significantly from immediately frozen samples suggesting that caution is needed when comparing absolute concentrations of hormones between samples preserved in different ways. However, differences among ethanol-treated samples in general were small, demonstrating the feasibility of this preservation method in the field at remote locations.     

Cold preservation of rat pancreatic islets just above the freezing point using University of Wisconsin solution

AIMS: To confirm whether rat islets stored at a temperature just above the freezing point using University of Wisconsin (UW) solution would remain viable for the short term. METHODOLOGY: Rat islets were stored for 24 hours in UW solution, either at 4 degrees C or at -0.6 degrees C (just above the specific freezing point of the UW solution). After cold storage, the islets were assessed for in vitro viability by static incubation and for in vivo viability by a transplantation study. One thousand islets preserved under different conditions were injected intraportally into a streptozotocin-induced diabetic rat as an isograft. Four weeks after the transplantation, an intravenous glucose tolerance test was performed. RESULTS: Islets stored at -0.6 degrees C showed higher insulin secretion rates than those stored at 4 degrees C on a static challenge. The interval from transplantation to the achievement of normoglycemia was also shorter in the -0.6 degrees C group than in the 4 degrees C group. After islet transplantation, the daily nonfasting plasma glucose concentration was higher in the 4 degrees C group than in the -0.6 degrees C group. When compared with the 4 degrees C group, the -0.6 degrees C group showed lower blood glucose values during all investigational periods on an intravenous glucose tolerance test. CONCLUSION: Islet preservation at -0.6 degrees C using UW solution is more advantageous for short term.