Role of autophagy in oncolytic herpes simplex virus type 1-induced cell death in squamous cell carcinoma cells.

Herpes simplex virus type 1 (HSV-1) is one of the most widely studied viruses for oncolytic virotherapy. In squamous cell carcinoma (SCC) cells, the role of autophagy induced by neurovirulence gene-deficient HSV-1s in programmed cell death has not yet been elucidated. The oncolytic HSV-1 strain RH2, which lacks the γ34.5 gene and induces the fusion of human SCC cells, was used. RH2 replicated and induced cell death in SCC cells. RH2 infection was accompanied by the aggregation of microtubule-associated protein 1 light chain 3 (LC3) in the cytoplasm, the conversion of LC3-I to LC3-II and the formation of double-membrane vacuoles containing cell contents. No significant changes were observed in the expression of Bcl-2 or Bax, while a slight decrease was observed in that of Beclin 1. The autophagy inhibitors, 3-methyladenine (3-MA) and bafilomycin A1, did not affect viral replication, but significantly inhibited the cytotoxicity of RH2. The caspase-3 inhibitor z-DEVD-fmk and caspase-1 inhibitor z-YVAD-fmk also reduced the cytotoxicity of RH2. These results demonstrated that γ34.5 gene-deficient HSV-1 RH2 induced autophagic cell death in SCC cells as well as pyroptosis and apoptosis.

Immunogenic cell death by oncolytic herpes simplex virus type 1 in squamous cell carcinoma cells.

Molecules essential for the induction of immunogenic cell death (ICD) are called damage-associated molecular patterns (DAMPs). The effects of oncolytic herpes simplex virus type 1 (HSV-1) on the production of DAMPs were examined in squamous cell carcinoma (SCC) cells. The cytopathic effects of HSV-1 RH2 were observed in mouse SCCVII cells infected at a high multiplicity of infection (MOI), and the amounts of viable cells were decreased. After being infected with RH2, ATP and high mobility group box 1 (HMGB1) were released extracellulary, while calreticulin (CRT) translocated to the cell membrane. A flow-cytometric analysis revealed an increase in the number of annexin-V and propidium iodide (PI)-stained cells; and the amount of cleaved poly (ADP-ribose) polymerase (PARP) was increased. The killing effect of RH2 was reduced by pan-caspase inhibitor z-VAD-fmk and the caspase-1 inhibitor z-YVAD-fmk, suggesting the involvement of apoptosis and pyroptosis. In C3H mice bearing synergic SCCVII tumors, the growth of tumors injected with the supernatant of RH2-infected cells was less than that of tumors injected with phosphate-buffered saline (PBS). These results indicate that oncolytic HSV-1 RH2 produces DAMPs from SCC cells to induce cell death. This may contribute to the enhancement of tumor immunity by oncolytic HSV-1.

Ultrasound as a method to enhance antitumor ability of oncolytic herpes simplex virus for head and neck cancer.

Low-intensity ultrasound is a useful method to enhance the delivery of drugs to target cells via a range of mechanisms including the transient formation of micropores in the cell membrane, a process known as sonoporation. The effect of ultrasound on oncolytic herpes simplex virus type-1 (HSV-1) infection in oral squamous cell carcinoma (SCC) was examined. Human SCC cell line SAS and oncolytic HSV-1 RH2, which was deficient in the neurovirulent γ134.5 gene and exhibited cell fusion actions, were used. Cells grown in multi-well plates were infected with HSV-1 and exposed to ultrasound in the presence or absence of microbubbles after an adsorption period. The number of plaques was significantly greater than that of the untreated control. SAS cells were inoculated subcutaneously into nude mice and tumors were produced. Tumors were injected with HSV-1 RH2 with or without microbubbles and then exposed to ultrasound through the covering skin. The amount of the virus in tumor tissues 3 days after the injection was higher in tumors treated with HSV-1 RH2 and ultrasound than in tumors treated with RH2 only. The expression of the HSV-1 antigen was also increased by ultrasound and microbubbles. Tumor growth was suppressed with HSV-1 RH2 in combination with ultrasound, especially with microbubbles. These results indicated that ultrasound increased the efficiency of the HSV-1 infection in SAS cells and nude mouse tumors. This method can potentially be useful to enhance the antitumor effects of oncolytic HSV-1 on head and neck cancer treatment.

Enhancement of systemic tumor immunity for squamous cell carcinoma cells by an oncolytic herpes simplex virus.

RH2 is a neurovirulent γ134.5 gene-deficient herpes simplex virus type 1 (HSV-1) with a lytic ability in human squamous cell carcinoma (SCC) cells; it is related to spontaneously occurring HSV-1 mutant HF10. The effect of RH2 on SCC was examined using a syngeneic C3H mouse model. After infection of mouse SCCVII cells with RH2, cell viability was decreased at first, but recovered by prolonged culture, indicating the limited replication of RH2. The antitumor ability of RH2 was examined using a bilateral SCCVII tumor model. The growth of the RH2-injected tumors was suppressed compared with that of phosphate-buffered saline-injected tumors. Moreover, the growth of contralateral tumor of RH2-treated mice was also suppressed significantly. The splenocytes of C3H mice treated with RH2 lysed more SCCVII cells than NFSaY83 cells and YAC-1 cells. The cytotoxicity of the splenocytes on SCCVII cells was significantly greater than that of splenocytes from tumor-bearing mice. Removal of CD8(+) T cells from splenocytes decreased their cell killing activity remarkably. The antitumor effect of RH2 on SCCVII xenografts in nude mice was not demonstrated. These results indicate that RH2 exhibited a suppressive effect on mouse SCC, even if the replication of RH2 was limited. This is ascribed to the ability of RH2 to enhance existing tumor-specific cytotoxic T lymphocyte activity.

Deposits on the intraluminal surface and bacterial growth in central venous catheters.

Central venous catheter (CVC) tip and blood cultures are generally used to diagnose a catheter-related infection. Such methodology does not confirm the presence of bacterial colonisation on parts of CVCs other than the CVC tip. In order to assess the extent of bacterial colonisation, 10 catheters were examined in detail from patients admitted to intensive care unit. Swabs from the lumen at several sites (hub, indwelling and non-indwelling) were cultured and the intraluminal surface of the device subjected to scanning electron microscopy (SEM). Bacteria were detected on five out of 10 catheters (50%), and bacterial contamination of CVCs was common in the hub area of the device. Deposits (crystallisation) that differed from bacterial colonisation or biofilm were observed on the intraluminal surface of used CVCs. SEM showed bacteria firmly anchored to the deposits. Experimental flow studies demonstrated that deposits were more likely to appear after exposure to solutions such as total parenteral nutrition rather than distilled water. These deposits facilitated bacterial colonisation 30 times more than CVCs free from deposits.

Hyperthermia: is it an ominous sign after cardiac arrest?

OBJECTIVE: To clarify the clinical characteristics of hyperthermia at an early stage after resuscitation from cardiac arrest (CA). MATERIALS AND METHODS: We reviewed the medical records of 43 adult patients with non-traumatic out-of-hospital CA, who survived for longer than 24 h after admission to our intensive care unit (ICU) between January, 1995, and December, 1998. The patients were divided into two groups: a clinical brain death (CBD) group (n=23) and a non-CBD group (n=20), and various factors relating to hyperthermia were compared between the two groups. RESULTS: The mean value of peak axillary temperatures within 72 h of admission was 39.8+/-0.9 degrees C for the CBD group, which was significantly greater than 38.3+/-0.6 degrees C for the non-CBD group (P<0.0001). The time of occurrence of the peak axillary temperature was at 19+/-16 h of admission in the CBD group and 20+/-18 h in the non-CBD group (not significantly different). There were no significant differences in risk factors relating to the occurrence of hyperthermia between the two groups, except for the number of patients who received epinephrine at ICU. In 23 patients with a peak axillary temperature of > or =39 degrees C during the first 72 h of hospitalization, brain death was diagnosed in 20 patients, whereas only 3 of 20 patients having a peak axillary temperature of <39 degrees C developed brain death (odds ratio, 37.8; 95% confidence interval, 6.72-212.2). CONCLUSION: Hyperthermia at an early stage after resuscitation from CA may be associated with the outcome of brain death.

Analysis of plasma nitrite/nitrate in human thermal injury.

The aim of this study is to examine the characteristics of nitrite/nitrate (NOx), the final metabolite of nitric oxides, in plasma after burn injury. A total of 83 blood samples were collected from 19 patients on arrival, day 1, day 3, and day 5 after suffering burn injuries and from 7 non-burned volunteers. We measured the NOx levels in plasma using the Griess method, and analyzed the relationships among plasma the NOx levels, the burn-magnitude, and the blood examination data using a stepwise multivariate regression analysis. The plasma NOx levels at hospital-arrival after burns significantly exceeded those of non-burned volunteers, and the NOx levels in the plasma returned to normal range after day 1. Based on the findings of a multivariate analysis, the plasma NOx levels at admission to the hospital were not found to be related to the total burn surface area, the burn index or inhalation injury, but they were significantly related to age. Furthermore, these plasma NOx levels were also related to the platelet count, neutrophil count and blood urea nitrogen. The increase in the plasma NOx level may therefore play an important role in the pathophysiology of elderly burned patients, while the nitric oxide levels in the plasma might also play a role in inhibiting the constriction of microvascular smooth muscle in extensively burned patients.

[11C]methionine positron emission tomography for the evaluation of pancreatic exocrine function in chronic pancreatitis.

Positron emission tomography (PET) can be used for the quantitative analysis of amino acid metabolism. The aim of this study was to investigate whether pancreatic exocrine function can be evaluated by [11C]methionine PET in chronic pancreatitis (CP) patients. Dynamic PET scan of the pancreas and liver was performed in eight healthy subjects and seven patients with CP after intravenous (i.v.) injection of [11C]methionine. Simultaneously, duodenal juice was collected with the background of continuous i.v. administration of secretin (125 ng/kg/h). The radioactivity ratio of the pancreas to that of the liver (PLR) was calculated by regions of interest (ROI) analysis. Amylase output and bicarbonate concentration were measured in the duodenal aspirates. The PLR of CP patients was significantly lower than that of healthy subjects at all time points after methionine injection. The PLRs at 4.5 minutes (PLR4.5) after methionine injection were positively correlated with the amylase output, mean bicarbonate concentration, and volume of duodenal aspirates (R = 0.74, 0.69, 0.46). It is concluded that [11C]methionine PET would be a noninvasive method for the evaluation of exocrine pancreatic function, which may represent total amino acids uptake of viable acinar cells in the pancreas.

[A case of disseminated intravascular coagulopathy (DIC) and multiple organ failure (MOF) after ingestion of hydrochloric acid].

Ingestion of caustic materials causes systemic damages and requires treatment in an intensive care unit. This report presents a case of disseminated intravascular coagulopathy (DIC) and multiple organ failure after ingestion of hydrochloric acid in an attempted suicide. The patient was admitted to the emergency ward within 1 hr after ingestion of 60 ml of 35% hydrochloric acid. Initial blood examination suggested hemolysis without anemia or thrombocytopenia. Arterial blood gas analysis exhibited evident metabolic acidosis with hypoxia. Two hours after ingestion, severe hemolysis, anemia, thrombocytopenia emerged with abnormal results from clotting and fiblinolysis tests, and a diagnosis of DIC was made. Subsequently, conjugate deviation and severe bleeding diathesis appeared. No evidence for gastrointestinal bleeding and brain hemorrhage was obtained in our abdominal echography and brain computed tomography, respectively. In spite of vigorous suppurative therapy including transfusion of blood, fresh frozen plasma, and catecholamines, the patient expired 29 hrs after ingestion. Autopsy was not carried out. Rapid progressive DIC as an acute complication of acid ingestion must be born in mind.

A refractory case of secondary erythermalgia successfully treated with lumbar sympathetic ganglion block.

A 59-year-old Japanese man with myasthenia gravis, who had a 10-year history of temperature-sensitive pain in the lower extremities, i.e. improved by cooling and worsened by warming, consulted us because the pain had become intolerable during the previous 4 months. Bilateral erythema, swelling and large ulcers were noted on the calves, dorsal aspects of the feet, and soles. Laboratory data showed thrombocythaemia and a positive antibody to the acetylcholine receptor, but were negative for antinuclear and antiphospholipid antibodies. A diagnosis of secondary erythermalgia was made because of the clinical features, the laboratory data, and the lack of family history of this disease. Although steroid pulse therapy, oral aspirin and antiserotonin drugs were ineffective, bilateral lumbar sympathetic ganglion block succeeded in relieving the severe pain and curing the ulcers. The clinical course in our patient suggests that sympathetic ganglion block may be one of the most effective treatments for secondary erythermalgia. Although the mechanism of this effect is uncertain, microcirculation disturbance in secondary erythermalgia, if any, may be improved by this block.

Effects of increased oxygen breathing in a volume controlled hemorrhagic shock outcome model in rats.

It is believed that victims of traumatic hemorrhagic shock (HS) benefit from breathing 100% O(2). Supplying bottled O(2) for military and civilian first aid is difficult and expensive. We tested the hypothesis that increased FiO(2) both during severe volume-controlled HS and after resuscitation in rats would: (1) increase blood pressure; (2) mitigate visceral dysoxia and thereby prevent post-shock multiple organ failure; and (3) increase survival time and rate. Thirty rats, under light anesthesia with halothane (0. 5% throughout), with spontaneous breathing of air, underwent blood withdrawal of 3 ml/100 g over 15 min. After HS phase I of 60 min, resuscitation phase II of 180 min with normotensive intravenous fluid resuscitation (shed blood plus lactated Ringer's solution), was followed by an observation phase III to 72 h and necropsy. Rats were randomly divided into three groups of ten rats each: group 1 with FiO(2) 0.21 (air) throughout; group 2 with FiO(2) 0.5; and group 3 with FiO(2) 1.0, from HS 15 min to the end of phase II. Visceral dysoxia was monitored during phases I and II in terms of liver and gut surface PCO(2) increase. The main outcome variables were survival time and rate. PaO(2) values at the end of HS averaged 88 mmHg with FiO(2) 0.21; 217 with FiO(2) 0.5; and 348 with FiO(2) 1. 0 (P<0.001). During HS phase I, FiO(2) 0.5 increased mean arterial pressure (MAP) (NS) and kept arterial lactate lower (P<0.05), compared with FiO(2) 0.21 or 1.0. During phase II, FiO(2) 0.5 and 1. 0 increased MAP compared with FiO(2) 0.21 (P<0.01). Heart rate was transiently slower during phases I and II in oxygen groups 2 and 3, compared with air group 1 (P<0.05). During HS, FiO(2) 0.5 and 1.0 mitigated visceral dysoxia (tissue PCO(2) rise) transiently, compared with FiO(2) 0.21 (P<0.05). Survival time (by life table analysis) was longer after FiO(2) 0.5 than after FiO(2) 0.21 (P<0. 05) or 1.0 (NS), without a significant difference between FiO(2) 0. 21 and 1.0. Survival rate to 72 h was achieved by two of ten rats in FiO(2) 0.21 group 1, by four of ten rats in FiO(2) 0.5 group 2 (NS); and by four of ten rats of FiO(2) 1.0 group 3 (NS). In late deaths macroscopic necroses of the small intestine were less frequent in FiO(2) 0.5 group 2. We conclude that in rats, in the absence of hypoxemia, increasing FiO(2) from 0.21 to 0.5 or 1.0 does not increase the chance to achieve long-term survival. Breathing FiO(2) 0.5, however, might increase survival time in untreated HS, as it can mitigate hypotension, lactacidemia and visceral dysoxia.

Mild or moderate hypothermia, but not increased oxygen breathing, increases long-term survival after uncontrolled hemorrhagic shock in rats.

OBJECTIVE: To test the hypotheses that, for uncontrolled hemorrhagic shock (UHS) in rats, mild hypothermia, compared with normothermia, would increase long-term survival as well as moderate hypothermia, oxygen breathing would increase survival further, and hypothermia and oxygen would mitigate visceral ischemia (dysoxia) during UHS. DESIGN: Prospective, randomized study. SETTING: Animal research laboratory. SUBJECTS: A total of 54 male Sprague-Dawley rats. INTERVENTIONS: Under light anesthesia and spontaneous breathing, rats underwent UHS phase I of 75 mins, with initial withdrawal of 3 mL/100 g of blood over 15 mins, followed by UHS via tail amputation and limited fluid resuscitation to maintain mean arterial pressure at > or =40 mm Hg; resuscitation phase II of 60 mins (from 75 mins to 135 mins) with hemostasis and aggressive fluid resuscitation to normalize hemodynamics; and observation phase III to 72 hrs. Rats were randomly divided into nine groups (n = 6 each) with three rectal temperature levels (38 degrees C [normothermia] vs. 34 degrees C [mild hypothermia] vs. 30 degrees C [moderate hypothermia]) by surface cooling; each with 3 FIO2 levels (0.25 vs. 0.5 vs. 1.0). MEASUREMENTS AND MAIN RESULTS: Hypothermia increased blood pressure compared with normothermia. Increased FIO2 had no effect on blood pressure. Additional blood loss from the tail cut was small, with no differences among groups. Hypothermia and FIO2 of 0.5 decreased visceral hypoxia, as measured by the difference between visceral (liver and jejunum) surface Pco2 and PaCO2 during UHS. Compared with normothermia, mild hypothermia increased the survival time and rate as well as moderate hypothermia (p < .01 by life table), without a significant difference between mild and moderate hypothermia. Increased FIO2 had no effect on survival time or rate. CONCLUSIONS: After severe UHS and resuscitation in rats, mild hypothermia during UHS, compared with normothermia, increases blood pressure, survival time and 72-hr survival rate as well as moderate hypothermia. Mild hypothermia is clinically more feasible and safer than moderate hypothermia. Increased FIO2 seems to have no significant effect on outcome.

Surface cooling, which fails to reduce the core temperature rapidly, hastens death during severe hemorrhagic shock in pigs.

OBJECTIVE: To examine whether surface cooling (SC) would rapidly decrease the core temperatures and prolong the survival time during volume-controlled lethal hemorrhagic shock in pigs. METHOD: Twelve pigs were randomly assigned to the SC group (group 1, n = 6) or the no cooling control group (group 2, n = 6), after blood withdrawal of 30 mL/kg over 15 minutes, and maintained under spontaneous breathing by light anesthesia with 1.0% halothane. SC was performed by applying ethanol to the skin, blowing with an electric fan, and placing ice packs. Pigs were observed without fluid resuscitation until their death (apnea and no pulse). RESULTS: SC did not lower the rectal temperature (Tr) to 35 degrees C at any time point until death, except one pig; in that animal, Tr was decreased to 34 degrees C after 135 minutes from the start of SC. The survival time was 108 +/- 43 minutes in group 1 and 175 +/- 55 minutes in group 2 (p < 0.05, life table analysis). CONCLUSION: In lightly anesthetized pigs during hemorrhagic shock, SC without resuscitation did not rapidly reduce the core temperature and rather hastened death for reasons that remain to be explored.

Adenosine by aortic flush fails to augment the brain preservation effect of mild hypothermia during exsanguination cardiac arrest in dogs - an exploratory study.

Most trauma cases with rapid exsanguination to cardiac arrest (CA) in the field, as well as many cases of normovolemic sudden cardiac death are 'unresuscitable' by standard cardiopulmonary-cerebral resuscitation (CPCR). We are presenting a dog model for exploring pharmacological strategies for the rapid induction by aortic arch flush of suspended animation (SA), i.e. preservation of cerebral viability for 15 min or longer. This can be extended by profound hypothermic circulatory arrest of at least 60 min, induced and reversed with (portable) cardiopulmonary bypass (CPB). SA is meant to buy time for transport and repair during pulselessness, to be followed by delayed resuscitation to survival without brain damage. This model with exsanguination over 5 min to CA of 15-min no-flow, is to evaluate rapid SA induction by aortic flush of normal saline solution (NSS) at room temperature (24 degrees C) at 2-min no-flow. This previously achieved normal functional recovery, but with histologic brain damage. We hypothesized that the addition of adenosine would achieve recovery with no histologic damage, because adenosine delays energy failure and helps repair brain injury. This dog model included reversal of 15-min no-flow with closed-chest CPB, controlled ventilation to 20 h, and intensive care to 72 h. Outcome was evaluated by overall performance, neurologic deficit, and brain histologic damage. At 2 min of CA, 500 ml of NSS at 24 degrees C was flushed (over 1 min) into the brain and heart via an aortic balloon catheter. Controls (n=5) received no drug. The adenosine group (n=5) received 2-chloro-adenosine (long acting adenosine analogue), 30 mg in the flush solution, and, after reperfusion, adenosine i.v. over 12 h (210 microg/kg per min for 3 h, 140 microg/kg per min for 9 h). The 24 degrees C flush reduced tympanic membrane temperature (T(ty)) within 2 min of CA from 37.5 to approximately 36.0 degrees C in both groups. At 72 h, final overall performance category (OPC) 1 (normal) was achieved by all ten dogs of the two groups. Final neurologic deficit scores (NDS; 0-10% normal, 100% brain death) were 5+/-3% in the control group versus 6+/-5% in the adenosine group (NS). Total brain histologic damage scores (HDS) at 72 h were 74+/-9 (64-80) in the control group versus 68+/-19 (40-88) in the adenosine group (NS). In both groups, ischemic neurons were as prevalent in the basal ganglia and neocortex as in the cerebellum and hippocampus. The mild hypothermic aortic flush protocol is feasible in dogs. The adenosine strategy used does not abolish the mild histologic brain damage.

Classification of 3D structural character of RNA by hydrogen bond and base stacking.

We are developing a computational system to classify RNA structures by its structural character. Here, an improved grouping algorithm was introduced to the system and the base-stacking pattern (BSP) is used as a criterion for the classification in addition to hydrogen-bond pattern (HBP). 279 conformers of 15 mer RNA hairpin were classified into 89 and 36 groups by HBP and BSP, respectively, suggesting that HBP represents conformational character better than BSP.

Hypothermic aortic arch flush for preservation during exsanguination cardiac arrest of 15 minutes in dogs.

BACKGROUND: Trauma victims rarely survive cardiac arrest from exsanguination. Survivors may suffer neurologic damage. Our hypothesis was that a hypothermic aortic arch flush of 500 mL of isotonic saline solution at 4 degrees C, compared with 24 degrees C (room temperature), administered at the start of prolonged exsanguination cardiac arrest (CA) would improve functional neurologic outcome in dogs. METHODS: Seventeen male hunting dogs were prepared under light N2O-halothane anesthesia. The animals were randomized into two groups: group I (n = 9) received 4 degrees C isotonic saline flush and group II (n = 6) received 24 degrees C flush. Two additional dogs received no flush. While spontaneously breathing, the dogs underwent normothermic (tympanic membrane temperature [Ttm] = 37.5 degrees C) exsanguination over 5 minutes to cardiac arrest, assured by electric induction of ventricular fibrillation. After 2 minutes of arrest, the flush was administered over 1 minute into the aortic arch by means of a 13 French balloon-tipped catheter inserted by means of the femoral artery. After 15 minutes of CA, resuscitation was with closed-chest cardiopulmonary bypass, return of shed blood, and defibrillation. For the first 12 hours after CA, core temperature was maintained at 34 degrees C. Mechanical ventilation was continued to 20 hours and intensive care to 72 hours, when final evaluation and perfusion-fixation killing for brain histologic damage scoring were performed. RESULTS: Three dogs in group I were excluded because of extracerebral complications. All 14 dogs that followed protocol survived. During CA, the Ttm decreased to 33.6 +/- 1.2 degrees C in group I and 35.9 +/- 0.4 degrees C in group II (p = 0.002). At 72 hours, in group I, all dogs achieved an overall performance category (OPC) of 1 (normal). In group II, 1 dog was OPC 2 (moderate disability), 3 dogs were OPC 3 (severe disability), and 2 dogs were OPC 4 (coma). Both dogs without flush were OPC 4. Neurologic deficit scores (NDS 0% = normal, 100% = brain death) were 1 +/- 1% in group I and 41 +/- 12% in group II (p < 0.05). The two dogs without flush achieved an NDS of 47% and 59%. Total brain histologic damage scores were 35 +/- 28 in group I and 82 +/- 17 in group II (p < 0.01); and 124 and 200 in the nonflushed dogs. CONCLUSION: At the start of 15 minutes of exsanguination cardiac arrest in dogs, hypothermic aortic arch flush allows resuscitation to survival with normal neurologic function and histologically almost clean brains.

Mild or moderate hypothermia but not increased oxygen breathing prolongs survival during lethal uncontrolled hemorrhagic shock in rats, with monitoring of visceral dysoxia.

OBJECTIVE: To test the hypotheses that during lethal uncontrolled hemorrhagic shock (UHS) in rats compared with normothermia and room air breathing: a) mild hypothermia would prolong survival time as well as moderate hypothermia; b) oxygen breathing would prolong survival further; and c) hypothermia and oxygen would mitigate visceral ischemia (dysoxia) during UHS. DESIGN: Prospective, randomized, controlled laboratory animal study. SETTING: Animal research facility. SUBJECTS: Male Sprague-Dawley rats. INTERVENTION: Fifty-four rats were lightly anesthetized with halothane during spontaneous breathing. UHS was induced by blood withdrawal of 3 mL/100 g over 15 mins, followed by 75% tail amputation with topical application of heparin. Five minutes after tail cut, rats were randomly divided into nine groups (6 rats each) with three rectal temperature levels (38 degrees C [100.4 degrees F; normothermia] vs. 34 degrees C [93.2 degrees F; mild hypothermia] vs. 30 degrees C [86 degrees F; moderate hypothermia]) by surface cooling; each with 3 FIO2 levels (0.25 vs. 0.5 vs. 1.0). Rats were observed without fluid resuscitation until death (apnea and pulselessness). Visceral ischemia was monitored by observing liver and gut surface PCO2. MEASUREMENTS AND MAIN RESULTS: Mean survival time, which was 51 mins in the control group with normothermia and FIO2 of 0.25, was more than doubled with hypothermia, to 119 mins in the combined mild hypothermia groups (p < .05) and to 132 mins in the combined moderate hypothermia groups (p < .05; NS for moderate vs. mild hypothermia). FIO2 had no statistically significant effect on survival time. Increases in visceral surface PCO2 correlated with hypotension (r2 = .22 for intestine and .40 for liver). Transiently, increased FIO2, not hypothermia, mitigated visceral ischemia. CONCLUSIONS: Both mild and moderate hypothermia prolonged survival time during untreated, lethal UHS in rats. Increased FIO2 had no effect on survival. The effects of hypothermia and increased FIO2 during UHS on viscera, the ability to be resuscitated, and outcome should be explored further.

11C-methionine uptake to the pancreas and its secretion: a positron emission tomography study in humans.

The aim of this study was to investigate the uptake of [11C-methyl]-L-methionine (11C-methionine) in the human pancreas by analyzing dynamic positron emission tomography (PET) images and the duodenal aspirate. A double-lumen tube was inserted in the duodenum and dynamic PET was performed in seven healthy volunteers for 110 min after intravenous (i.v.) injection of 11C-methionine during the continuous i.v. administration of secretin (125 ng/kg/h) and cerulein (30 ng/kg/h). For the calculation of the radioactivity in the pancreas, the regions of interest were set on the PET images. Radioactivity was measured in 10-min fractions of duodenal juice. After i.v. injection, 11C-methionine accumulated in the pancreas within a few minutes, and the radioactivity plateaued during the study. The radiolabeled proteins in the duodenal juice increased linearly 30 min after 11C-methionine injection, but the relative rates of radioactivity in the protein precipitate to the total count in the duodenal juice were 44-48%. From these findings, it was concluded that 11C-methionine accumulation in the pancreas is very rapid after the i.v. administration, and only a part of methionine uptake to the pancreas is incorporated into secretory proteins. The pancreatic 11C-methionine uptake detected by PET may represent a new aspect of exocrine pancreatic function that has not been expressed by the conventional intubation method.

Development of a system to classify 3D structural character of RNA.

We are developing a computational system to extract structural character of RNA. We made a program that classifies conformers by recognizing the hydrogen bonding patterns. The program was applied to a set of 279 conformers and they were classified into about 40 groups. The system is expected to be useful for searching structural motifs of RNA and classifying large number of generated conformers in structural modeling process.