The effects of normovolemic hemodilution on protein synthesis recovery following postischemic reperfusion in the rat brain

Normovolemic hemodilution is a possible way to improve the brain recovery after ischemia and reperfusion. Therefore we have decided to examine how this process may affect the post-ischemic protein synthesis machinery. We analysed rat brains after 4-vessel-occlusion and different time intervals of reperfusion using normovolemic hemodilution. We achieved an important increase of [4,5-3H]leucine incorporation into polypeptides in vitro in the rat brain neocortex 30 minutes after ischemia, but concurrently there was no significant change in the hippocampus and striatum. By extending the time course of reperfusion we did not observe any important deviation of in vitro [4,5-3H]leucine incorporation in the studied brain areas. Thus, although hemodilution increased protein synthesis in selective vulnerable regions after ischemia, this improvement is not of significant importance.     

Role and function of macrophages in the metabolic syndrome

Macrophages are key innate immune effector cells best known for their role as professional phagocytes, which also include neutrophils and dendritic cells. Recent evidence indicates that macrophages are also key players in metabolic homoeostasis. Macrophages can be found in many tissues, where they respond to metabolic cues and produce pro- and/or anti-inflammatory mediators to modulate metabolite programmes. Certain metabolites, such as fatty acids, ceramides and cholesterol crystals, elicit inflammatory responses through pathogen-sensing signalling pathways, implicating a maladaptation of macrophages and the innate immune system to elevated metabolic stress associated with overnutrition in modern societies. The outcome of this maladaptation is a feedforward inflammatory response leading to a state of unresolved inflammation and a collection of metabolic pathologies, including insulin resistance, fatty liver, atherosclerosis and dyslipidaemia. The present review summarizes what is known about the contributions of macrophages to metabolic diseases and the signalling pathways that are involved in metabolic stress-induced macrophage activation. Understanding the role of macrophages in these processes will help us to develop therapies against detrimental effects of the metabolic syndrome.     

Pentylenetetrazole decreases metabolic glutamate turnover in rat brain

Seizures were induced in rats by intraperitoneal injection of pentylenetetrazole (PTZ, 70 mg/kg), followed, 30 min later, by injection of [1-13C]glucose and [1,2-13C]acetate. Analyses of extracts from cortex, subcortex and cerebellum were performed using 13C magnetic resonance spectroscopy and HPLC. It could be shown that PTZ affected different brain regions differently. The total amounts of glutamate, glutamine, GABA, aspartate and taurine were decreased in the cerebellum and unchanged in the other brain regions. GABAergic neurones in the cortex and subcortex were not affected, whereas those in the cerebellum showed a pronounced decrease of GABA synthesis. However, glutamatergic neurones in all brain regions showed a decrease in glutamate labelling and in addition a decreased turnover in cerebellum. It could be shown that this decrease was in the metabolic pool of glutamate whereas release of glutamate was unaffected since glutamine labelling from glutamate was unchanged. Aspartate turnover was also decreased in all brain regions. Changes in astrocyte metabolism were not detected, indicating that PTZ had no effect on astrocyte metabolism in the early postictal stage.     

Acute severe carbon monoxide exposure in the rat: effects of hyperglycemia and hypoglycemia on mortality, recovery, and neurologic deficit.

Human and animal studies suggest a poorer outcome in the presence of abnormal blood glucose concentration during cerebral hypoxia-ischemia. It is unknown whether this is also the case in acute severe carbon monoxide poisoning. Using Levine-prepared rats, three groups were established and exposed to CO to answer this question: (1) hyperglycemics resulting from the administration of a 50% glucose solution, (2) hypoglycemics resulting from the administration of normal saline, and (3) untreated controls. The rats inhaled 2400 ppm CO for 90 min in the absence of anesthesia. Blood glucose was raised to a mean value of 402 mg/dL just prior to CO exposure in group 1. This resulted in an increased mortality rate (i.e., 54%), and during 4 h of room air recovery an impaired ability to regain body temperature, an increased plasma lactate dehydrogenase activity, and an increased neurologic deficit as compared with group 3. Hypoglycemia, which developed during CO exposure in group 2 (mean minimum glucose after 90 min, 44 mg/dL), resulted in an increased mortality rate (i.e., 46%), and during 4 h of room air recovery an impaired ability to regain body temperature and an increased neurologic deficit as compared with group 3. Blood glucose concentration in the rats in groups 2 and 3 that died during or shortly after CO exposure was significantly depressed relative to the survivors of those groups. Plasma insulin activity was elevated during CO exposure in group 1 as compared with group 3, but fell during recovery; insulin remained low throughout CO exposure and recovery in group 2. The results demonstrate the deleterious effects of both a very high and a very low blood glucose concentration during acute CO exposure.     

Role of glucose metabolism in the recovery of postischemic LV mechanical function: effects of insulin and other metabolic modulators

The role of proton (H+) production from glucose metabolism in the recovery of myocardial function during postischemic reperfusion and its alteration by insulin and other metabolic modulators were examined. Rat hearts were perfused in vitro with Krebs-Henseleit solution containing palmitate (1.2 mmol/l) and glucose (11 mmol/l) under nonischemic conditions or during reperfusion following no-flow ischemia. Perfusate contained normal insulin (n-Ins, 50 mU/l), zero insulin (0-Ins), or supplemental insulin (s-Ins, 1,000 mU/l) or other metabolic modulators [dichloroacetate (DCA) at 3 mmol/l, oxfenicine at 1 mmol/l, and N6-cyclohexyladenosine (CHA) at 0.5 micromol/l]. Relative to n-Ins, 0-Ins depressed rates of glycolysis and glucose oxidation in nonischemic hearts and impaired recovery of postischemic function. Relative to n-Ins, s-Ins did not affect aerobic glucose metabolism and did not improve recovery when present during reperfusion. When present during ischemia and reperfusion, s-Ins impaired recovery. Combinations of metabolic modulators with s-Ins stimulated glucose oxidation approximately 2.5-fold in nonischemic hearts and reduced H+ production. DCA and CHA, in combination with s-Ins, improved recovery of function, but addition of oxfenicine to this combination provided no further benefit. Although DCA and CHA were each partially protective in hearts perfused with n-Ins, optimal protection was achieved with DCA + CHA; recovery of function was inversely proportional to H+ production during reperfusion. Although supplemental insulin is not beneficial, elimination of H+ production from glucose metabolism by simultaneous inhibition of glycolysis and stimulation of glucose oxidation optimizes recovery of postischemic mechanical function.     

The recovery of sensory function following skin flaps in humans

Two cross-sectional studies were made of the recovery of tactile and pain sensitivity in subjects having skin flaps in the region of the chest and neck as a result of tumor excision. In experiment 1, stimuli ranging from 2.46 to 17.10 gm of force were delivered by von Frey hairs to the flaps and comparable normal sites in 35 subjects at times ranging from 1 month to 10 years after surgery. No subjects perceived stimuli of less than 11.80 gm, thermal, or moving touch applied to flaps, whereas 21 percent perceived 11.80 gm or greater force (judged as painful applied to normal skin). The results of experiment 2 showed that these findings were not due to visual information available to subjects. Possible explanations for the fact that these results are radically different from those reported in the literature are discussed.     

Somatostatin concentration and binding in the rat hypothalamus and striatum during severe insulin-induced hypoglycemia

Hypoglycemia was induced by administration of insulin (40 I.U./kg) to 24 h fasted rats. Somatostatinlike immunoreactivity (SLI) and¹²⁵I-Tyr¹¹-somatostatin binding were measured in the striatum and hypothalamus at the onset of hypoglycemic coma (5–10 min). No significant changes in SLI concentration were detected in either site although the total number of specific somatostatin receptors in the striatum membranes, but not in the hypothalamus, decreased in insulin-injected rats when compared with the control group.     

Regional differences in the effects of insulin-induced hypoglycemia on phospholipid metabolism of rat brain

Insulin-induced hypoglycemia in rats may lead to stimulated brain activity and if severe enough, they may develop a stupor-coma condition. In this study, the effects of insulin-induced hypoglycemia on brain phospholipid metabolism were examined in rats which were prior injected with ³²Pi. Three hours after insulin injection (1 or 5 units/100 g body wt, i.p.), there was an increase (25%) in radioactivity of the lipid phase of cerebral cortex, but radioactivity in the cerebellum tended to decrease instead. Radioactivity in the aqueous phase of cortex was not altered after insulin injection, but that in the cerebellum was decreased by 30%. Differences were observed in labeling of individual phospholipids in response to the hypoglycemic treatment. A marked decrease in labelled phosphatidate was observed in the cerebellum from the hypoglycemic samples, but not in the cerebral cortex. In the cortex, hypoglycemic condition resulted in an increase in ³²Pi uptake into the phospholipids. However, the differences in the amount of label among individual phospholipids suggest that phosphatidylinositol and phosphatidylcholine are turning over more rapidly than other phospholipids. The hypoglycemic rats also showed a 3-fold increase in the brain free fatty acid level, but the level of diacylglycerol was not changed. Results thus suggested a correlation between the free fatty acid release and the increased turnover of phosphatidylinositol and phosphatidylcholine during brain stimulation due to insulin-induced hypoglycemia.     

Mechanisms for recovery of motor function following cortical damage

Recent studies of focal injury to the cerebral cortex have demonstrated that the remaining, intact tissue undergoes structural and functional changes that could play a substantial role in neurological recovery. New information regarding the molecular and cellular environment in the adjacent, intact tissue has suggested that waves of growth promotion and inhibition modulate the self-repair processes of the brain. Furthermore, recent studies have documented widespread neurophysiological and neuroanatomical changes in regions remote from a focal cortical injury, suggesting that entire cortical networks participate in the recovery process.     

Cellular mechanisms of brain hypoglycemia

Data on intracellular processes induced by a low glucose level in nerve tissue are presented. The involvement of glutamate and adenosine receptors, mitochondria, reactive oxygen species (ROS), and calcium ions in the development of hypoglycemia-induced damage of neurons is considered. Hypoglycemia-induced calcium overload of neuronal mitochondria is suggested to be responsible for the increased ROS production by mitochondria.     

Eicosanoids in human ventricular cerebrospinal fluid following severe brain injury

Recent evidence has shown that a variety of prostaglandins and leukotrienes can be produced in brain tissue after injury in animals. It has also been speculated that increases in brain prostaglandins occur in humans following injury. Ventricular cerebrospinal fluid (CSF) samples have been obtained from children with static lesions (controls) as well as children with acute brain injury and eicosanoids measured by immunologic techniques. Metabolites of prostacyclin (6-keto-PGF1 a) and thromboxane A2 (thromboxane B2) were the major eicosanoids found in CSF, and levels of these compounds were increased 3-10 times in acutely injured patients. Prostaglandin E2 was also found in lower amounts, although in one case its level was very high. Prostaglandin D2 was also present, but in low amounts. No leukotrienes were found in CSF samples that were purified by HPLC prior to immunoassay. Elevated levels of hydroxyeicosatetraenoic acids (HETEs) were observed in those samples stored frozen, but these metabolites were most probably due to autooxidation of arachidonic acid in CSF. Arachidonic acid concentration in CSF was typically found to be in the range of 10-200 ng/ml, but was found to be 5-10 fold higher in one severely injured patient. Thus, elevated free arachidonic acid and various oxygenated metabolites were observed in CSF following brain injury.     

Antipsychotic drugs differentially modulate apolipoprotein D in rat brain

Apolipoprotein-D (apoD), a member of the lipocalin family of proteins, binds to arachidonic acid and cholesterol among other hydrophobic molecules. Recently, elevated apoD levels have been reported in the post-mortem brains, as well as plasma, of schizophrenic patients and in rodent brains after chronic treatment with clozapine (CLOZ). These findings and the evidence for altered membrane lipid metabolism in schizophrenia suggest that apoD may have a role in the pathophysiology of illness, and also in the differential clinical outcome following treatment with typical and atypical antipsychotic drugs. Here, we compared the effects of these antipsychotics on the expression of apoD in rat brain. Chronic treatment with typical antipsychotic, haloperidol (HAL) reduced apoD expression in hippocampus, piriform cortex and caudate-putamen (p = 0.027-0.002), whereas atypical antipsychotics, risperidone (RISP) and olanzapine (OLZ) increased (p = 0.051 to < 0.001 and p = 0.048 to < 0.001, respectively) apoD expression. In hippocampus, HAL-induced changes were present in CA1, CA3 and dentate gyrus, however, apoD levels in motor cortex were unchanged. There were also very dramatic effects of HAL on the neuronal morphology, particularly, cellular shrinkage and disorganization with the loss of neuropil. Post-treatment, either with RISP or OLZ, was very effective in restoring the HAL-induced reduction of apoD, as well as cellular morphology. Similarly, pre-treatments were also effective, but slightly less than post-treatment, in preventing HAL-induced reduction of apoD. The increased expression of apoD by atypical antipsychotics may reflect a novel molecular mechanism underlying their favorable effects compared with HAL on cognition, negative symptoms and extra-pyramidal symptoms in schizophrenia.     

创伤性颅脑损伤脑脊液代谢谱改变及临床意义

目的:探讨创伤性颅脑外伤患者脑脊液中代谢谱的改变及其临床意义。方法:用高分辨率质子核磁共振代谢组学检验创伤性颅脑损伤对脑化学物质和代谢的影响。在重型创伤性颅脑损伤组(n=6),损伤后的脑脊液分析脑代谢的变化,并与轻,中型颅脑损伤组(n=6)相比较。结果:与轻型,中型颅脑损伤组相比,发现了乙酰乙酸,尿酸,3-硝基酪氨酸升高的证据。3组脑脊液中乙酰乙酸,尿酸,3-硝基酪氨酸含量有显著性差异(p<0.01)。结论:颅脑创伤后脑脊液中乙酰乙酸,尿酸,3-硝基酪氨酸值均有不同程度升高,且升高越明显则病情越严重。说明乙酰乙酸,尿酸,3-硝基酪氨酸可作为颅脑创伤病情的监测指标。     

Role of T-cell function in recovery from murine influenza infection.

Athymic (nude) mice and their normal littermates were intranasally inoculated with graded doses of A/WSN influenza virus. At a dose of 10³ EID₅₀, all mice survived the infection. In contrast, at a dose of 5 × 10⁴ EID₅₀, all mice died by 7 days. At intermediate doses of 5 × 10³ and 10⁴ EID₅₀, the nude mice were less resistant to the infection than their normal littermates, so that a higher proportion always died. Given a dose of 5 × 10³ EID₅₀, lung virus levels in both groups reached similar high levels by Day 5. Thereafter, virus levels in the normal mice rapidly fell so that no infectious virus could be detected by Day 18. In nude mice, the levels fell very slowly so that relatively high levels were still present at Day 18 in the surviving mice. At the height of the infection, high levels of cytotoxic T-cell activity was detected in the lungs of normal but not nude mice. Transfer to the nude mice of specific immune T cells raised from infected normal littermates enhanced survival of the nude mice and reduced the lung virus levels. Nude mice consistently showed a greater degree of lung consolidation than their normal littermates. Microscopically, the nude mouse lungs showed greater respiratory epithelial hyperplasia with minimal inflammatory cell infiltration in the foci of consolidation compared with their infected normal littermates. Under the conditions of these experiments, influenza-immune T cells seemed to inhibit rather than contribute to the generation of virus-mediated pulmonary pathology. The findings strongly suggest that T cells play an important positive role in the process of recovery from murine influenza infection.