alpha-Keto acid omega-amidase from rat liver.
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Amidohydrolases↗
Biomedical subjects
T E Duffy
Publications and source records attributed to T E Duffy.
Regional acetylcholine metabolism in brain during acute hypoglycemia and recovery.
Insulin-induced hypoglycemia in normothermic rats caused progressive neurological depression and differentially altered regional cerebral acetylcholine metabolism. Reductions of plasma glucose from 7.7 mM (control) to 2.5-1.7 mM (moderate hypoglycemia associated with decreased motor activity) or 1.5 mM (severe hypoglycemia with lethargy progressing to stupor) decreased glucose concentrations in the cerebral cortex, striatum, and hippocampus to less than 10% of control. Moderate hypoglycemia diminished acetylcholine concentrations in cortex and striatum (21% and 45%, respectively) and reduced [1-2H2, 2-2H2]choline incorporation into acetylcholine (62% and 41%, respectively). Severe hypoglycemia did not reduce the acetylcholine concentration or synthesis in cortex and striatum further. The concentrations of choline rose in the cortex (+53%) and striatum (+130%) of animals that became stuporous but a similar rise in [1-2H2, 2-2H2]choline left the specific activities of choline in these structures unchanged. Even severe hypoglycemia did not alter the hippocampal cholinergic system. In rats that developed hypoglycemic stupor and were then treated with glucose, the animals recovered apparently normal behavior, and the concentrations of acetylcholine and the incorporation of [1-2H2, 2-2H2]-choline into acetylcholine returned to control values in the striatum but not in the cerebral cortex. Thus, impaired acetylcholine metabolism in selected regions of the brain may contribute to the early symptoms of neurological dysfunction in hypoglycemia.
Acetylcholine↗
Local cerebral glucose metabolism in rats with chronic portacaval shunts.
Local cerebral glucose utilization was measured with the autoradiographic [14C]deoxyglucose method in rats at 2 days and 1, 4, 8, and 12 weeks after the construction of a portacaval shunt, and in weight-matched controls. Local glucose utilization in brain was altered in shunted animals, but the magnitude and direction of the changes differed among 36 neuroanatomical structures, depending upon the length of time that the animals had been shunted. In rats shunted for 4 weeks or less, glucose utilization did not differ from control (30 of 36 structures) or was decreased (6 structures). The largest decreases of glucose utilization, noted at 1 week, occurred in the parietal (-25%) and frontal cortices (-28%) and subcortical white matter (-50%. In rats shunted for 8 weeks, however, glucose utilization was higher than control in many brain structures (13 of 36), and after 12 weeks it was higher than control in most structures (25 of 36). Only the parietal cortex did not follow this trend; it exhibited a decreased rate of glucose utilization in rats shunted for 8 weeks (-21%) that normalized in animals shunted for 12 weeks. Portal-systemic shunting of blood increased arterial blood ammonia concentrations to twice the control value of 85 +/- 3 microM in animals shunted for 1 week, and to approximately 2.5 times control in animals shunted for 4-12 weeks. Kety-Schmidt measurements of cerebral blood flow and arterial-venous (torcular) differences for ammonia across the brains of control rats and rats with portacaval shunts for 8 weeks revealed an increased cerebral uptake of ammonia in the shunted animals. The late-developing morphological changes known to occur in astrocytes and the delayed increases in local glucose consumption in rats with portacaval shunts may be related, and represent an adaptive response to sustained hyperammonemia.
Ammonia↗
Use of beta-methylene-D,L-aspartate to assess the role of aspartate aminotransferase in cerebral oxidative metabolism.
Several inhibitors of aspartate aminotransferase, a key enzyme of the malate-aspartate shuttle, were investigated for their effects on cerebral oxidative metabolism in vitro. beta-Methylene-D,L-aspartate (2 mM), aminooxyacetate (0.1 mM), and D,L-vinylglycine (20 mM) all significantly reduced the activity of aspartate aminotransferase and the rate of oxygen consumption of rat cerebral cortex slices respiring on glucose. In the presence of beta-methyleneaspartate, a one-to-one correlation was found between the degree of inhibition of tissue respiration and the degree of inhibition of transaminase activity. Slices of rat liver incubated in the presence of glucose and beta-methyleneaspartate showed a similar one-to-one relationship between inhibition of oxygen comsumption and inhibition of aspartate aminotransferase activity, whereas with rat kidney cortex slices, the inhibition of aspartate aminotransferase activity was greater than the inhibition of oxygen consumption. Structural analogs of beta-methyleneaspartate (D,L-beta-methyl-D,L-aspartate, gamma-methyl-D,L-glutamate, and alpha-methyl-D,L-didehydroglutamate) that did not inhibit the activity of aspartate aminotransferase similarly did not inhibit the rate of oxygen consumption by cerebral cortex slices. In the presence of beta-methyleneaspartate, pyruvate oxidation by cerebral cortex slices was inhibited to almost the same extent as was glucose oxidation, and the oxidation of succinate was decreased by approximately 20%. The artificial electron acceptor phenazine methosulfate (0.1 mM) only partially overcame the beta-methyleneaspartate-mediated inhibition of respiration with glucose as substrate. The content of ATP and phosphocreatine declined steadily in slices incubated with glucose and beta-methyleneaspartate. At 1 h the concentration of lactate and the lactate/pyruvate ratio, an indicator of the cytoplasmic redox state, increased threefold, whereas the concentrations of malate, citrate, and aspartate decreased. The findings are interpreted in the context of the hypothesis that enzymes common to the malate-aspartate shuttle and the tricarboxylic acid cycle are physically complexed in brain, so that inhibition of aspartate aminotransferase, a component of the complex, impedes the flow of carbon through both metabolic pathways.(ABSTRACT TRUNCATED AT 400 WORDS)
Aminooxyacetic Acid↗
Regional energy balance in rat brain after transient forebrain ischemia.
Phosphocreatine, ATP, and glucose were severely depleted, and the lactate levels were increased in the paramedian neocortex, dorsal-lateral striatum, and CA1 zone of hippocampus of rats exposed to 30 min of forebrain ischemia. Upon recirculation of the brain, phosphocreatine, ATP, and lactate concentrations recovered to control values in the paramedian neocortex and CA1 zone of hippocampus and to near-control values in the striatum. The phosphocreatine and ATP concentrations then fell and the lactate levels rose in the striatum after 6-24 h, and in the CA1 zone of hippocampus after 24-72 h. The initial recovery and subsequent delayed changes in the phosphocreatine, ATP, and lactate concentrations in the striatum and hippocampus coincided with the onset and progression of morphological injury in these brain regions. The results suggest that cells in these regions regain normal or near-normal mitochondrial function and are viable, in terms of energy production, for many hours before unknown mechanisms cause irreversible neuronal before unknown mechanisms cause irreversible neuronal injury.
Adenosine Triphosphate↗
Acute hyperammonemia in the young primate: physiologic and neuropathologic correlates.
Infusion-induced acute (less than or equal to 24 h) hyperammonemia to concentrations up to five times normal (0.19 +/- 0.03 versus 0.90 +/- 0.08 mM) was studied in eleven 6-9-month-old Macaca mulatta. The young primates developed a progressive reduction of consciousness that correlated in severity directly with the elevation of blood ammonia concentration. Hyperventilation, electroencephalographic slowing, occasional seizure activity, and, eventually, apneustic breathing also occurred. Intracranial pressure rose from 76 +/- 7 to 167 +/- 12 mmH2O. Arterial oxygen and blood pressure remained within normal limits. Neuropathologic examination showed early astrocytic changes, consisting primarily of swollen perikaryal cytoplasm and processes, and membranous whorls. The absence of neuronal pathology suggests that the acute, limited insult, as occurs in many of the childhood hyperammonemic syndromes, is fully reversible.
Acetates↗
Local cerebral glucose metabolism in newborn dogs: effects of hypoxia and halothane anesthesia.
Local cerebral glucose utilization (LCGU) was measured in 36 neuroanatomical structures of normal awake, halothane-anesthetized, and hypoxic newborn puppies by the autoradiographic 2-[14C]deoxyglucose method. In normal animals, LCGU was highest in the vestibular nucleus and in other gray matter nuclei of the brainstem and declined in a caudal-to-rostral progression through the neuraxis (i.e., LCGU of cerebellum greater than thalamus approximately equal to caudate-putamen greater than cerebral cortex). Lowest rates of glucose metabolism were detected in white matter structures. Halothane anesthesia (1.5% inspired) caused few changes in local glucose metabolism, the most notable being decreased LCGU among structures of the auditory system (cochlear nucleus, lateral lemniscus, inferior colliculus) and increased LCGU in the interpeduncular nucleus. Acute systemic hypoxia (arterial oxygen tension of approximately 12 mm Hg) produced markedly heterogeneous effects on local glucose metabolism: LCGU was increased in some gray matter structures, decreased in the thalamus, and substantially increased in the subcortical white matter and corpus callosum. In puppies whose brains were frozen in situ after 55 minutes of hypoxia, the concentration of lactate was increased ten- to elevenfold in cortical gray and subcortical white matter, but the concentrations of glucose, adenosine triphosphate, and phosphocreatine declined to a greater extent in the white matter. The results suggest that during hypoxia the high rate of glycolysis in white matter exceeded substrate supply so that glucose availability became the limiting factor for local energy production. Such a mechanism may contribute to the white matter injury that often develops following hypoxic-ischemic insults in the perinatal period.
Anesthesia, General↗
Regulation of local cerebral blood flow in normal and hypoxic newborn dogs.
Local cerebral blood flow (LCBF) was measured autoradiographically in newborn puppies by an indicator fractionation technique using 4-iodo-[14C]antipyrine as the diffusible indicator. Measurements were obtained in unanesthetized, normotensive animals, and the sensitivity of blood flow to hypercapnia and acute hypoxia was determined in 32 brain structures. LCBF in normal and hypoxic puppies was correlated with local cerebral glucose utilization (LCGU) obtained under the same experimental conditions (Duffy et al, 1982). In normocapnic (PaCO2 33 mm Hg) control animals, highest rates of blood flow were found in gray matter nuclei of the brainstem, in the medulla oblongata, and in the posterolateral nucleus of the thalamus (50 to 77 ml/100 gm/min); far lower flows were recorded among white matter structures (5 to 11 ml/100 gm/min). The vasodilatory response to both hypercapnia and hypoxia was greatest among brainstem gray matter structures, intermediate among cortical and diencephalic gray matter structures, and least in white matter. When LCBF was plotted as a function of LCGU for control animals, a positive linear correlation was obtained for all structures (p less than 0.001), implying that in newborns, as in adults, cerebral blood flow and metabolism are physiologically coupled. In hypoxic puppies, no consistent relationship between LCGU and LCBF could be demonstrated; however, there was suggestion that the two measurements correlated inversely, presumably reflecting enhanced anaerobic glycolysis in structures (e.g., hemispheric white matter) that were not adequately protected by compensatory hyperemia. White matter damage, a frequent complication of perinatal hypoxia-asphyxia, may be a consequence in part of the limited capacity of white matter to vasodilate in response to te chemical "signals" of hypercapnia and lactic acidosis.
Animals↗
Regional cerebral blood flow and glucose metabolism following transient forebrain ischemia.
Progressive brain damage after transient cerebral ischemia may be related to changes in postischemic cerebral blood flow and metabolism. Regional cerebral blood flow (rCBF) and cerebral glucose utilization (rCGU) were measured in adult rats prior to, during (only rCBF), and serially after transient forebrain ischemia. Animals were subjected to 30 minutes of forebrain ischemia by occluding both common carotid arteries 24 hours after cauterizing the vertebral arteries. Regional CBF was measured by the indicator-fractionation technique using 4-iodo-[14C]-antipyrine. Regional CGU was measured by the 2-[14C]deoxyglucose method. The results were correlated with the distribution and progression of ischemic neuronal damage in animals subjected to an identical ischemic insult. Cerebral blood flow to forebrain after 30 minutes of moderate to severe ischemia (less than 10% control CBF) was characterized by 5 to 15 minutes of hyperemia; rCBF then fell below normal and remained low for as long as 24 hours. Post-ischemic glucose utilization in the forebrain, except in the hippocampus, was depressed below control values at 1 hour and either remained low (neocortex, striatum) or gradually rose to normal (white matter) by 48 hours. In the hippocampus, glucose utilization equaled the control value at 1 hour and fell below control between 24 and 48 hours. The appearance of moderate to severe morphological damage in striatum and hippocampus coincided with a late rise of rCBF above normal and with a fall of rCGU; the late depression of rCGU was usually preceded by a period during which metabolism was increased relative to adjacent tissue. Further refinement of these studies may help identify salvageable brain after ischemia and define ways to manipulate CBF and metabolism in the treatment of stroke.
Animals↗
Cerebral oxidative metabolism and blood flow during acute hypoglycemia and recovery in unanesthetized rats.
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Adenosine Triphosphate↗
Comparison of radio-labeled butanol and iodoantipyrine as cerebral blood flow markers.
Cerebral blood flow (CBF) techniques based on the principle of indicator fractionation rely upon free diffusibility of the blood flow indicator into brain (i.e. complete cerebral extraction). Extraction of two commonly-used indicators, iodoantipyrine and n-butanol, was evaluated in rats by measuring torcular venous efflux after systemic injection of the indicator under conditions of normal and high CBF. The extraction of n-butanol was found to be virtually complete at all blood flows examined; iodoantipyrine, on the other hand, was completely extracted only at flows under 180 ml/100 g/min, despite the fact that the oil: water partition coefficient for iodoantipyrine exceeds that for n-butanol. Brain uptake indices for the two indicators were also measured: brain uptake of n-butanol was greater than that of iodoantipyrine, and the difference was more marked if the indicator entered brain mixed with blood than if it entered as a bloodless bolus. Blood components may thus interact with iodoantipyrine to retard its movement across the blood-brain barrier and thereby limit extraction of this lipid-soluble substance. Inasmuch as iodoantipyrine is diffusion-limited at blood flows above 180 ml/100 g/min, butanol is a more accurate CBF indicator above the normal flow range in the rat.
1-Butanol↗
Brain dysfunction in mild to moderate hypoxia.
Hypoxia is commonly invoked to explain alterations in mental function, particularly in patients with cardiac pulmonary failure. The effects of acute graded hypoxia or higher integrative functions are well documented experimentally in man. Hypoxia in experimental animal models demonstrates that the pathophysiology is complex. In mild to moderate hypoxia, in contrast to severe hypoxia and to ischemia, the supply of energy for the brain is not impaired; cerebral levels of adenosine triphosphate (ATP) and adenylate energy charge are normal. In contrast, the turnover of several neurotransmitters is altered by mild hypoxia. For example, acetylcholine synthesis is reduced proportionally to the reduction in carbohydrate oxidation. This relationship holds in vitro and with several in vivo models of hypoxia. Pharmacologic and physiologic studies in man and experimental animals are consistent with acetylcholine having an important role in mediating the cerebral effects of mild hypoxia. These observations raise the possibility that treatments directed to cholinergic or other central neurotransmitter systems may benefit patients with cerebral syndromes secondary to chronic hypoxia.
Acetylcholine↗
Simultaneous measurement of cerebral blood flow and unidirectional movement of substances across the blood-brain barrier: theory, method, and application to leucine.
The uptake of compounds by the brain depends upon cerebral blood flow. To determine the normal blood flow-cerebral extraction relationship, a method for rapid, simultaneous measurement of cerebral blood flow and brain extraction was developed and applied to blood-brain leucine transfer. Awake rats were injected intravenously with a mixture of n-[(14)C]butanol and [(3)H]leucine. The quantities of indicators accumulated over the following 5-12 s in brain and in a sample of arterial blood withdrawn at a know rate were used to determine the flux of butanol and leucine into brain. Butanol extraction was assessed independently by measuring arterial and cerebral venous concentrations of the indicator after a bolus injection. Cerebral blood flow was equal to the ratio of butanol flux into brain to butanol extraction by brain; leucine extraction was then calculated as the ratio of leucine influx to cerebral blood flow. Leucine extraction by brain and cerebral blood flow were shown to be related exponentially. The maximum velocity of active leucine transport was virtually the same at flows of 150 and 400 ml/100 g/min. The present method is theoretically applicable to the measurement of the extraction of any compound from blood by brain. By measuring the normal blood flow-extraction relationship, one can differentiate changes in extraction secondary to altered flow from changes intrinsic to pathologic conditions with inconstant cerebral blood flow.
1-Butanol↗
Impaired synthesis of acetylcholine by mild hypoxic hypoxia or nitrous oxide.
The effect of mild hypoxic hypoxia on brain metabolism and acetylcholine synthesis was studied in awake, restrained rats. Since many studies of hypoxia are done with animals anesthetized with nitrous oxide (N2O), the effects of N2O were evaluated. N2O (70%) increased the cerebral cortical blood flow by 33% and the cortical metabolic rate of oxygen by 26%. In addition, the synthesis of acetylcholine in N2O-anesthetized animals, measured with [U-14C]glucose and [1-2H2,2-2H2]choline, decreased by 45 and 53%, respectively. Consequently, mild hypoxia was studied in unanesthetized rats. Control rats breathing 30% O2 (partial pressure of oxygen, PaO2 = 120 mm Hg) were compared with rats exposed to 15% O2 (PaO2 = 57 mm Hg) or 10% O2 (PaO2 = 42 mm Hg). The synthesis of acetylcholine, measured with [U-14C]glucose, was decreased by 35 and 54% with 15% O2 and 10% O2, respectively; acetylcholine synthesis, measured with [1-2H2,2-2H2]choline, was decreased by 50 and 68% with 15% O2 and 10% O2, respectively. Animals breathing either 15% or 10% O2 had normal cerebral metabolic rates of oxygen but had increased brain lactates and increased cortical blood flows compared with animals breathing 30% O2. These results show that even mild hypoxic hypoxia impairs acetylcholine synthesis, which in turn may account for the early symptoms of brain dysfunction associated with hypoxia.
Acetylcholine↗
Determination of 2-pyrrolidone-5-carboxylic and alpha-ketoglutaramic acids in human cerebrospinal fluid by gas chromatography.
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Amides↗
Regional protein synthesis in rat brain following acute hemispheric ischemia.
Regional protein synthesis was measured in rat brain at intervals up to 48 h following occlusion of the four major arteries to the brain for either 10 or 30 min. Four-vessel occlusions produces ischemia in the cerebral hemispheres and oligemia in the midbrain-diencephalon and brainstem. During the hour following 10 min of ischemia, protein synthesis, measured by incorporation of [14C]valine into protein, was inhibited in the cerebral cortex by 67%. Normal rates of protein synthesis were attained within 4 h of recirculation. In rats subjected to 30 min of ischemia, protein synthesis was inhibited by 83% during the first hour of recirculation in the cortex, caudate-putamen, and hippocampus. Recovery of protein synthesis in these regions was slow (25-48 h). The midbrain-diencephalon showed less inhibition, 67%, and faster recovery (by 12 h). Protein synthesis was unaffected in the brainstem. [14C]Autoradiography revealed that the pyramidal neurons of the hippocampus and areas of the caudate and cortex failed to recover normal rates of protein synthesis even after 48 h. The accumulation of TCA-soluble [14C]valine was enhanced (55-65%) in the cortex, caudate, and hippocampus after 30 min of ischemia; the increase persisted for 12 h. A smaller rise in [14C]valine content (30%) and more rapid normalization of valine accumulation (by 7 h) were observed in the midbrain-diencephalon; no changes were found in the brainstem. In the cortex, recovery was more rapid when the duration of ischemia was reduced. Thus, the degree of inhibition of protein synthesis, the accumulation of valine in the tissue, and the length of time required to reestablish normal values for these processes were dependent on both the severity and the duration of the ischemic insult. Restoration of normal rates of protein synthesis after ischemia was slow compared with the normalization of cerebral energy metabolites.
Animals↗
Glutathione and ascorbate during ischemia and postischemic reperfusion in rat brain.
Thirty minutes of total cerebral ischemia (decapitation) decreased total glutathione (GSH + GSSG) by 7% but had no detectable effect on the concentration of oxidized glutathione (GSSG), reduced ascorbate, or total ascorbate, In a model of reversible, bilateral hemispheric ischemia (four-vessel occlusion) no changes in glutathione or ascorbate were detected after 30 min of ischemia. During 24 h of reperfusion following such an insult no detectable change in total ascorbate, reduced ascorbate, or oxidized glutathione was noted: however, total brain glutathione declined by 25%. The findings are discussed in relation to the hypothesis that the deleterious effects of ischemia are due to an increase in free radical production which in turn leads to increased lipid peroxidation.
Animals↗
Blood and brain ammonia concentrations after portacaval anastomosis. Effects of acute ammonia loading.
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Ammonia↗