Ketamine modulates hippocampal neurochemistry and functional connectivity: a combined magnetic resonance spectroscopy and resting-state fMRI study in healthy volunteers.

A growing body of evidence suggests glutamate excess in schizophrenia and that N-methyl-d-aspartate receptor (NMDAR) hypofunction on γ-aminobutyric acid (GABA) interneurons disinhibiting pyramidal cells may be relevant to this hyperglutamatergic state. To better understand how NMDAR hypofunction affects the brain, we used magnetic resonance spectroscopy and resting-state functional magnetic resonance imaging (MRI) to study the effects of ketamine on hippocampal neurometabolite levels and functional connectivity in 15 healthy human subjects. We observed a ketamine-induced increase in hippocampal Glx (glutamate+glutamine; F=3.76; P=0.04), a decrease in fronto-temporal (t=4.92, PFDR<0.05, kE=2198, x=-30, y=52, z=14) and temporo-parietal functional connectivity (t=5.07, PFDR<0.05, kE=6094, x=-28, y=-36, z=-2), and a possible link between connectivity changes and elevated Glx. Our data empirically support that hippocampal glutamatergic elevation and resting-state network alterations may arise from NMDAR hypofunction and establish a proof of principle whereby experimental modelling of a disorder can help mechanistically integrate distinct neuroimaging abnormalities in schizophrenia.

Tumor-associated mutant p53 promotes cancer cell survival upon glutamine deprivation through p21 induction.

Cancer cells depend on glutamine to sustain their increased proliferation and manage oxidative stress, yet glutamine is often depleted at tumor sites owing to excessive cellular consumption and poor vascularization. We have previously reported that p53 protein, although a well-known tumor suppressor, can contribute to cancer cell survival and adaptation to low-glutamine conditions. However, the TP53 gene is frequently mutated in tumors, and the role of mutant p53 (mutp53) in response to metabolic stress remains unclear. Here, we demonstrate that tumor-associated mutp53 promotes cancer cell survival upon glutamine deprivation both in vitro and in vivo. Interestingly, cancer cells expressing mutp53 proteins are more resistant to glutamine deprivation than cells with wild-type p53. Depletion of endogenous mutp53 protein in human lymphoma cells leads to cell sensitivity to glutamine withdrawal, whereas expression of mutp53 in p53 null cells results in resistance to glutamine deprivation. Furthermore, we found that mutp53 proteins hyper-transactivate p53-target gene CDKN1A upon glutamine deprivation, thus triggering cell cycle arrest and promoting cell survival. Together, our results reveal an unidentified mechanism by which mutp53 confers oncogenic functions by promoting cancer cell adaptation to metabolic stress.

Acrylamide neuropathy. I. Spatiotemporal characteristics of nerve cell damage in rat cerebellum.

Based on evidence from morphometric studies of PNS, we suggested that acrylamide (ACR)-induced distal axon degeneration was a secondary effect related to duration of exposure [Toxicol. Appl. Pharmacol. 151 (1998) 211]. To test this hypothesis in CNS, the cupric-silver stain method of de Olmos was used to define spatiotemporal characteristics of nerve somal, dendritic, axonal and terminal degeneration in rat cerebellum. Rats were exposed to ACR at either 50 mg/kg per day (i.p.) or 21 mg/kg per day (p.o.) and at selected times (i.p. = 5, 8 and 11 days; p.o. = 7, 14, 21, 28 and 38 days) brains were removed and processed for silver staining. Results demonstrate that intoxication at the higher ACR dose-rate produced early (day 5) and progressive degeneration of Purkinje cell dendrites in cerebellar cortex. Nerve terminal degeneration occurred concurrently with somatodendritic argyrophilia in cerebellar and brainstem nuclei that receive afferent input from Purkinje neurons. Relatively delayed (day 8), abundant axon degeneration was present in cerebellar white matter but not in cortical layers or in tracts carrying afferent fibers (cerebellar peduncles) from other brain nuclei. Axon argyrophilia coincided with the appearance of perikaryal degeneration, which was selective for Purkinje cells since silver impregnation of other cerebellar neurons was not evident in the different cortical layers or cerebellar nuclei. Intoxication at the lower ACR dose-rate produced simultaneous (day 14) dendrite, axon and nerve terminal argyrophilia and no somatic Purkinje cell degeneration. The spatiotemporal pattern of dendrite, axon and nerve terminal loss induced by both ACR dose-rates is consistent with Purkinje cell injury. Injured neurons are likely to be incapable of maintaining distal processes and, therefore, axon degeneration in the cerebellum is a component of a "dying-back" process of neuronal injury. Because cerebellar coordination of somatomotor activity is mediated solely through efferent projections of the Purkinje cell, injury to this neuron might contribute significantly to gait abnormalities that characterize ACR neurotoxicity.

Endothelial cell growth factor enhances musculocutaneous flap survival through the process of neovascularization.

The effect of an angiogenic growth factor-endothelial cell growth factor (ECGF)-was tested in the rat transverse rectus abdominis musculocutaneous (TRAM) flap model based on a single inferior vascular pedicle. The animals were divided into three groups (N = 8 per group) after flap elevation. In group A (control), each animal received both local and local intra-arterial injections of 1 ml saline. In group B (local), each received a 2-mg ECGF local injection and 1-ml saline local intra-arterial injection. In group C (local intra-arterial), each received a 1-ml saline local injection and a 2-mg ECGF local intra-arterial injection. All animals were evaluated on postoperative day 7. There was a significant increase in the percentage of the skin paddle survival area of the TRAM flap in both ECGF-treated groups when compared with the control group (group B vs. group A, p < 0.001; group C vs. group A, p < 0.001). This correlated with a significant increase in vascularity in both ECGF-treated groups compared with the control group (group B vs. group A, p = 0.007; group C vs. group A, p = 0.021). The results between groups B and C were not significant. ECGF, when administered via either local or local intra-arterial route, enhances musculocutaneous flap survival through the process of neovascularization.

The effect of prolonged clamping and vascular stasis on the patency of arterial and venous microanastomoses.

During free tissue transfer, an artery and one or two veins must be sutured. Either the artery or the vein can be repaired first, but the optimal order of vascular repair has not been established. Maintaining a clamp on a vessel is associated with vessel injury, and releasing the clamp may cause bleeding and vascular stasis. The purpose of this study was to determine if the order of vessel repair and the length of vascular clamping affects surgical outcome. Free flaps were created on Sprague-Dawley rats (400-450 g) as follows: Bilateral abdominal skin flaps (3 x 3 cm) based on the superficial inferior epigastric vessels were elevated. The femoral artery and vein were tied and divided distal to the branching of the inferior epigastric vessels. Proximal to the branching, the artery and vein were divided. The animals were then randomized into four groups as follows: In group I (N=16), the artery was repaired and then the clamps were released to revascularize the flap. Venous stasis occurred as the vein was being repaired because of the venous clamp. In group II (N=15), the artery was repaired, but the clamp was maintained to prevent blood from coming in contact with the fresh arterial anastomosis and to prevent venous stasis. The vein was then repaired. In group III (N=15), the vein was repaired first but the venous clamp was not released until the artery was repaired. In group IV (N=15), the vein was repaired first and the clamps were released, allowing venous blood to contact the fresh anastomosis while the artery was being repaired. After final clamp removal, all anastomoses were assessed immediately for evidence of thrombosis. Five days postoperatively the skin flap was evaluated for evidence of necrosis and the anastomosis was inspected for evidence of late thrombosis. The anastomoses were resected for histopathological evaluation. Flap success was compared between groups using chi-squared analysis. Eleven of 16 flaps failed (69%) in group I, and 3 of 15 flaps failed (20%) in each of groups II, III, and IV. Statistical significance was reached in comparing group I with the other three groups (p < 0.01). All failures in group I were caused by immediate venous thrombosis. The other failures were secondary to arterial thrombosis. Histopathological analysis failed to demonstrate any differences between the groups. According to these results, arterial repair followed by clamp release prior to the completion of the venous repair results in a low success rate, probably secondary to venous stasis within the draining vein while the venous anastomosis is being completed. If the arterial repair is performed first, then it is recommended that the arterial clamp be maintained until the venous repair is completed. If the vein is repaired first, then it can be clamped or unclamped with similar results.

Maternal and neonatal outcome of pregnancies complicated with maternal HELLP syndrome.

UNLABELLED: Preeclampsia complicated by the HELLP syndrome is associated with poor maternal outcome; there is scant information on neonatal outcome. OBJECTIVE: To evaluate the outcome of infants born to mothers with HELLP syndrome. STUDY DESIGN: Chart review comparing perinatal variables and the clinical course of 23 infants born to mothers whose pregnancy was complicated with HELLP syndrome (H) with 23 infants of mothers with uncomplicated preeclampsia (P). RESULTS: Infants in the H group, when compared with those in the P group, had a higher incidence of low Apgar scores (52% vs 18%, p < 0.01), lower admission systolic blood pressure (45 +/- 7 vs 32 +/- 11, p < 0.01), and more frequent need for assisted ventilation (61% vs 30%, p < 0.05). There were no differences between the two groups of infants in hematocrit, leukocyte and platelet count, or duration of ventilation and hospitalization. CONCLUSIONS: These findings suggest that infants born to preeclamptic mothers who develop HELLP syndrome have an increased need for resuscitation at delivery and a higher incidence of postnatal cardiopulmonary instability. Thus mothers with HELLP syndrome should be identified promptly and delivered in level II or III centers with appropriate facilities for management of these newborn infants at risk for perinatal asphyxia and a potential for long-term neurologic sequelae.

Comparison of the respiratory and systemic kinetics of nitrous oxide in the sheep.

BACKGROUND: To determine whether discrepancies in views on the kinetics of nitrous oxide (N2O) may have a methodological basis, we compared its kinetics, simultaneously, in the respiratory system and systemic circulation. METHODS: Six merino ewes (40-50 Kg) were previously prepared with catheters in the pulmonary artery and aorta. The animals were anaesthetised with thiopentone then ventilated on a mixture of 70% N2O, 1% halothane in oxygen for 4 h. Simultaneous serial arterial and pulmonary arterial blood samples were assayed for N2O by gas chromatography and respiratory gases were monitored continuously by mass spectrometry. RESULTS: Marked differences were observed between the respiratory and systemic kinetics of N2O uptake. While the expired/inspired N2O concentration ratio rose within 30 min to a value close to unity, the pulmonary arterial/arterial blood N2O concentration ratio did not reach unity during the 4 h of each study, but approached a constant rate of uptake shown by the mean ratio of 0.94 (SD 0.01) from about 2 h onward. CONCLUSIONS: Discrepancies in fluid flow between respiratory gas and the cardiovascular systems, a concentration effect of N2O in the lungs, the relative solubility of N2O in blood and tissues, and ventilation/perfusion inequalities all may contribute to the observed differences. The ongoing uptake is consistent with persistent extrapulmonary losses. There remains a need for experimental data on the pharmacokinetics of N2O. Unequivocal studies on the disposition of N2O can be undertaken only by using direct measurement of fluxes of N2O across relevant organs or tissues.

Early programmes of high and low intensity exercise and quality of life after acute myocardial infarction.

OBJECTIVE: To determine whether a group programme of light exercise could improve quality of life in patients after acute myocardial infarction to the same extent as a high intensity exercise training programme. SETTING: Australian teaching hospital. PATIENTS: 224 men from a consecutive series of 339 men under 70 admitted to a coronary care unit with transmural acute myocardial infarction. INTERVENTION: Patients were randomly allocated in hospital to a group programme lasting eight weeks of either high intensity exercise training or light exercise. MAIN OUTCOME MEASURES: Physical working capacity based on metabolic equivalents achieved from treadmill exercise tests at entry, after 11 weeks, and after one year. Quality of life based on self report scores of anxiety, depression, denial, and wellbeing and interview assessments of activities and psychosocial adjustment at entry, after four months, and after one year. RESULTS: The two groups were well matched at entry. At 11 weeks the mean results of treadmill testing were 10.7 (95% confidence interval 10.20 to 11.20) metabolic equivalents for exercise training and 9.7 (9.26 to 10.14) for light exercise (t = 2.85, df = 181, p = 0.005). Apart from this small temporary benefit in mean physical working capacity, there were no significant differences between groups. Improvement in occupational adjustment score from baseline to four months was greater after exercise training than after light exercise, but at one year repeated measures analysis of variance showed no significant effects of treatment or interaction between treatment and time point. CONCLUSION: The effects on quality of life of a low cost programme of light exercise are similar to those obtained from a high intensity exercise training programme.

Failure of the nitrous oxide tissue equilibration method for the determination of brain and myocardial blood flow under controlled conditions.

1. Two adult merino ewes were prepared with intravascular cannule for sampling aortic root blood, sagittal sinus blood and coronary sinus blood. 2. One week after preparation the animals were anaesthetized then ventilated with a gas mixture containing 10% nitrous oxide (N2O) for 60 min. Serial measurements of brain and myocardial blood flow were made using the N2O tissue equilibration method of Kety and Schmidt. 3. N2O failed to achieve matching arteriovenous blood concentration equality and saturation of the relevant tissues. Valid use of the Kety-Schmidt method, therefore, could not be confirmed. 4. Because of the failure of the arteriovenous equilibration, serially determined brain and myocardial blood flows were found to decrease with time. 5. The use of this method in circumstances where tissue saturation with the indicator gas cannot be ascertained is arbitrary.

Failure of the Kety-Schmidt nitrous oxide method for determination of myocardial blood flow.

1. The reliability of the Kety-Schmidt nitrous oxide (N2O) blood-tissue equilibration method was examined in 50 studies of myocardial blood flow in seven conscious, unrestrained sheep using a newly developed carefully validated gas chromatographic assay for N2O. 2. In 10 studies the arterial and coronary sinus N2O blood concentration-time curves converged as expected at the end of the 10 min sampling period. In 14 studies they crossed over, and in 26 studies, the curves failed to converge. 3. A survey of the literature revealed that such results have been encountered previously but have not been accorded particular significance. An ultimate matching equilibrium between arterial and venous blood N2O concentration-time curves is, however, fundamental to the validity of the method. 4. The results indicate that the use of the Kety-Schmidt method with N2O as the indicator gas is invalid as applied to the measurement of myocardial blood flow in this preparation.

Effect of early programmes of high and low intensity exercise on physical performance after transmural acute myocardial infarction.

Does a programme of light exercise training after acute myocardial infarction produce the same improvement in treadmill performance as aerobic exercise training? Three hundred and eight men from a consecutive series of 479 men with transmural (Q wave) acute myocardial infarction, admitted to a single coronary care unit, were randomly allocated to eight weeks of group aerobic exercise training or group light exercise. Groups were well matched for all characteristics other than site of infarction, which did not significantly affect results. Mean (SD) physical working capacity (metabolic equivalents) determined by treadmill testing at the start of the study (in the third week after infarction) was 6.8 (2.2) v 6.7 (2.5) METs, at the end (in the eleventh week after infarction) 10.8 (2.3) v 9.9 (2.4) METs, and at 12 month review 10.8 (2.4) v 10.7 (1.9) METs for the exercise training group and the light exercise group respectively. The difference of 0.9 METs at the end of the study was the only significant difference between groups. There were no significant intergroup differences at any stage in resting and maximal heart rate, resting and maximal systolic blood pressure, or rate-pressure product. Apart from a small temporarily greater physical working capacity, the physical benefits of aerobic exercise training were equally well achieved by group light exercise.

Patient-orientated gastroenterology.

Analysis of the first year's working of a combined gastroenterology clinic in a district hospital has shown that the major benefit was improved patient management. Hospital attendances were reduced, the diagnostic process accelerated, and unnecessary radiological investigations and surgical operations avoided. There were no obvious major disadvantages.