Brain Enzyme Research Articles

Regulation of Antioxidant Enzyme Levels in Rat Brain.

The whole plant of Nasturtium microphyllum is used as traditional Indian medicine to treat epilepsy. Previous studies have demonstrated that extracts of these plants were subjected to acute toxicity and then screened for antiepileptic activity on maximal electroshock (MES)- and pentylenetetrazole (PTZ)-induced seizure models in albino Wistar rats. The purpose of the present study is to investigate the effect of ethanolic (95%) extract of N. microphyllum (EENM) on antioxidant enzymes in rat brain after induction of seizures by MES and PTZ. Our aim of study was relationship between seizure activities and altered levels of antioxidant enzymes such as superoxide dismutase (SOD), glutathione peroxidase (GP), glutathione reductase (GR), catalase, and lipid peroxidation on rat brain. Superoxide dismutase, glutathione peroxidase, glutathione reductase, and catalase were decreased in rat brain due to seizure, and it was restored significantly by administration of ethanol extract of N. microphyllum on treated rats. Similar dose-dependent results were obtained in PTZ model also, whereas EERS significantly decreased lipid peroxidation in both models. The anticonvulsant activity of EERS might have antioxidant properties and it delays the generation of free radical in MES- and PTZ-induced epilepsy.

Upregulation of hemeoxygenase enzymes HO-1 and HO-2 following ischemia-reperfusion injury in connection with experimental cardiac arrest and cardiopulmonary resuscitation: Neuroprotective effects of methylene blue.

Oxidative stress plays an important role in neuronal injuries after cardiac arrest. Increased production of carbon monoxide (CO) by the enzyme hemeoxygenase (HO) in the brain is induced by the oxidative stress. HO is present in the CNS in two isoforms, namely the inducible HO-1 and the constitutive HO-2. Elevated levels of serum HO-1 occurs in cardiac arrest patients and upregulation of HO-1 in cardiac arrest is seen in the neurons. However, the role of HO-2 in cardiac arrest is not well known. In this review involvement of HO-1 and HO-2 enzymes in the porcine brain following cardiac arrest and resuscitation is discussed based on our own observations. In addition, neuroprotective role of methylene blue- an antioxidant dye on alterations in HO under in cardiac arrest is also presented. The biochemical findings of HO-1 and HO-2 enzymes using ELISA were further confirmed by immunocytochemical approach to localize selective regional alterations in cardiac arrest. Our observations are the first to show that cardiac arrest followed by successful cardiopulmonary resuscitation results in significant alteration in cerebral concentrations of HO-1 and HO-2 levels indicating a prominent role of CO in brain pathology and methylene blue during CPR followed by induced hypothermia leading to superior neuroprotection after return of spontaneous circulation (ROSC), not reported earlier.

Dietary n-9, n-6 and n-3 fatty acids modulate the oxidative stress in brain and liver of mice. Effect of trans fatty acids supplementation

BACKGROUND: Arachidonic (20:4n-6) and docosahexaenoic (22:6n-3) acids interaction affects brain structure and function. Unsaturated fatty acids (UFAs) generate oxygenated lipid-derived eicosanoids which modulate the inflammatory response. The presence of trans fatty acids (TFA) in neuronal membranes can favor to generation of pro-oxidant metabolites. OBJECTIVE: This study evaluated the effect of supplementation with TFA to diets containing different proportions of FA, on the oxidative stress (OS) generation and the inflammatory response in mice brain and liver. METHODS: CF1 mice were fed diets (16 weeks) with olive (O), corn (C) or rapeseed (R) oils. OS parameters and gene expression of some key liver and brain enzymes involved in OS production were evaluated. RESULTS: In brain and liver, lipoperoxidation was increased and catalase activity was decreased in C. In brain, glutathione was diminished by supplementation with TFA in all diets and histological sections showed lymphocytes in O and C. In liver, decreased amount of lipid vacuoles and increased of cyclooxygenase-1 (COX-1) and PPARγ mRNA levels were observed in R and Rt. IL-1b and IL-6 in serum were augmented in O and Ot. CONCLUSIONS: Rapeseed oil could have protective effects on the development of OS and inflammation, while TFA supplementation did not showed marked effects on these parameters.

Rapid and long-lasting antidepressant-like effects of ketamine and their relationship with the expression of brain enzymes, BDNF, and astrocytes.

Ketamine (KET) is an N-methyl-D-aspartate (NMDA) antagonist with rapid and long-lasting antidepressant effects, but how the drug shows its sustained effects is still a matter of controversy. The objectives were to evaluate the mechanisms for KET rapid (30 min) and long-lasting (15 and 30 days after) antidepressant effects in mice. A single dose of KET (2, 5, or 10 mg/kg, po) was administered to male Swiss mice and the forced swim test (FST) was performed 30 min, 15, or 30 days later. Imipramine (IMI, 30 mg/kg, ip), a tricyclic antidepressant drug, was used as reference. The mice were euthanized, separated into two time-point groups (D1, first day after KET injection; D30, 30 days later), and brain sections were processed for glycogen synthase kinase-3 (GSK-3), histone deacetylase (HDAC), brain-derived neurotrophic factor (BDNF), and glial fibrillary acidic protein (GFAP) immunohistochemical assays. KET (5 and 10 mg/kg) presented rapid and long-lasting antidepressant-like effects. As expected, the immunoreactivities for brain GSK-3 and HDAC decreased compared to control groups in all areas (striatum, DG, CA1, CA3, and mainly pre-frontal cortex, PFC) after KET injection. Increases in BDNF immunostaining were demonstrated in the PFC, DG, CA1, and CA3 areas at D1 and D30 time-points. GFAP immunoreactivity was also increased in the PFC and striatum at both time-points. In conclusion, KET changed brain BDNF and GFAP expressions 30 days after a single administration. Although neuroplasticity could be involved in the observed effects of KET, more studies are needed to explain the mechanisms for the drug’s sustained antidepressant-like effects.

The effects of NO on the urea cycle pathway in short-term intermittent hypobaric hypoxia in rats

Short-term hypoxic states can influence the health and life activities of lowlanders who travel shortly to high altitudes, in transitory situations, such as surgical ischemia-reperfusion (to one or several organs), and in some sporting activities, such as parachuting and extreme skiing, mountain rescue teams, regular commercial flight crews, in which the subject may not even notice the hypoxia. NO is an integral part of the human physiological response to hypoxia. Until recently, the urea cycle (UC) was only considered as an important mechanism for neutralizing ammonia. We are the first to reveal an interrelation in hypoxic states between the activities of NO-synthase and UC enzymes in male rats' liver, kidney and brain. In the presented work, we have shown that during short-term intermittent hypobaric hypoxia (IHH) all enzymes of UC play an important role in the maintenance of NO quantity. The results allow thinking that kidney and brain argininosuccinate synthase (ASS) and argininosuccinate lyase (ASL) and liver ASS and ASL can be different isoenzymes. It is worth mentioning that the results have revealed new sides of l-arginine metabolism in a hypoxic state in male rats.

Paradoxical Effects of a Cytokine and an Anticonvulsant Strengthen the Epigenetic/Enzymatic Avenue for Autism Research.

Maternal exposure to the valproate short-chain fatty acid (SCFA) during pregnancy is known to possibly induce autism spectrum disorders (ASDs) in the offspring. By contrast, case studies have evidenced positive outcomes of this anticonvulsant drug in children with severe autism. Interestingly, the same paradoxical pattern applies to the IL-17a inflammatory cytokine involved in the immune system regulation. Such joint apparent contradictions can be overcome by pointing out that, among their respective signaling pathways, valproate and IL-17a share an enhancement of the “type A monoamine oxidase” (MAOA) enzyme carried by the X chromosome. In the Guided Propagation (GP) model of autism, such enzymatic rise triggers a prenatal epigenetic downregulation, which, without possible X-inactivation, and when coinciding with genetic expression variants of other brain enzymes, results in the delayed onset of autistic symptoms. The underlying imbalance of synaptic monoamines, serotonin in the first place, would reflect a mismatch between the environment to which the brain metabolism was prepared during gestation and the postnatal actual surroundings. Following a prenatal exposure to molecules that significantly elicit the MAOA gene expression, a daily treatment with the same metabolic impact would tend to recreate the fetal environment and contribute to rebalance monoamines, thus allowing proper neural circuits to gradually develop, provided behavioral re-education. Given the multifaceted other players than MAOA that are involved in the regulation of serotonin levels, potential compensatory effects are surveyed, which may underlie the autism heterogeneity. This explanatory framework opens up prospects regarding autism prevention and treatment, strikingly in line with current advances along the gut microbiome–brain axis.

Metabolomic and Imaging Mass Spectrometric Assays of Labile Brain Metabolites: Critical Importance of Brain Harvest Procedures

Metabolomic technologies including imaging mass spectrometry (IMS; also called mass spectrometry imaging, MSI, or matrix-assisted laser desorption/ionization-mass spectrometry imaging, MALDI MSI) are important methods to evaluate levels of many compounds in brain with high spatial resolution, characterize metabolic phenotypes of brain disorders, and identify disease biomarkers. ATP is central to brain energetics, and reports of its heterogeneous distribution in brain and regional differences in ATP/ADP ratios reported in IMS studies conflict with earlier studies. These discordant data were, therefore, analyzed and compared with biochemical literature that used rigorous methods to preserve labile metabolites. Unequal, very low regional ATP levels and low ATP/ADP ratios are explained by rapid metabolism during postmortem ischemia. A critical aspect of any analysis of brain components is their stability during and after tissue harvest so measured concentrations closely approximate their physiological levels in vivo. Unfortunately, the requirement for inactivation of brain enzymes by freezing or heating is not widely recognized outside the neurochemistry discipline, and procedures that do not prevent postmortem autolysis, including decapitation, brain removal/dissection, and 'snap freezing' are commonly used. Strong emphasis is placed on use of supplementary approaches to calibrate metabolite abundance in units of concentration in IMS studies and comparison of IMS results with biochemical data obtained by different methods to help identify potential artifacts.

Evaluation of Anticonvulsant and Anti-oxidant Potentials of Basella Alba by Influencing the Brain Enzyme Levels in Laboratory Animals

Epilepsy and convulsions constitute a significant class of symptoms that are commonly seen in many neurological diseases. It is understood that there is an apparent alteration in the levels of the enzymes which present brain during epilepsy. Anti-oxidant drugs are known to elevate the protective enzyme levels in the body and restore them to ensure the proper functioning is done. But when the enzyme levels are not managed properly, they can cause further damage and deterioration of the tissue. Basella alba is one of such drugs, which is rich in anti-oxidant chemical constituents. Many researchers concluded that many components like vitamin A, C and flavonoids, polyphenols are being reported. In the study, Basella alba was investigated for effect on an anti-oxidant enzyme in the brain. The extracts showed a better activity and prevented the brain tissue damage from any oxidative free radical generation. The enzyme levels were healthy at the dose of 400mg/kg of extract.

Protective effect of Cyperus esculentus (tiger nut) extract against scopolamine-induced memory loss and oxidative stress in mouse brain.

Objectives The juice extract of Cyperus esculentus commonly known as tiger nuts (TINUT) is widely used for its numerous health promoting effects including alleviation of symptoms associated with neurological disorders. Herein, we investigated the influence of the aqueous extract of C.esculentus on cognitive disorder and the underlying changes in acetylcholinesterase (AChE) activity and oxidative stress biomarkers in mice exposed to scopolamine. Methods C.esculentus (50-200mg/kg) or saline (10mL/kg) was given alone or with scopolamine 30min after, to male Swiss mice (6/group) daily for seven days. We evaluated the cognitive performance using Y-maze and object recognition on day seven post-treatment. Brains of the animals were afterwards processed for spectrophotometric determination of AChE activity and contents of oxidative stress biomarkers (malondialdehyde [MDA], glutathione [GSH], catalase, superoxide dismutase and nitrite). Results The extract improves cognitive function and also upturned scopolamine amnesia in mice. The extract markedly reduced brain AChE, MDA, and nitrite contents in mice injected with scopolamine (p<0.05). It also attenuated scopolamine-induced deregulated GSH contents and antioxidant enzymes in mouse brain. Conclusions The results of this study suggest that regular consumption of TINUT might offer beneficial effects in memory-related disorders.

Discovery of Novel Inhibitors of a Critical Brain Enzyme Using a Homology Model and a Deep Convolutional Neural Network.

Rare neglected diseases may be neglected but are hardly rare, affecting hundreds of millions of people around the world. Here, we present a hit identification approach using AtomNet, the world's first deep convolutional neural network for structure-based drug discovery, to identify inhibitors targeting aspartate N-acetyltransferase (ANAT), a promising target for the treatment of patients suffering from Canavan disease. Despite the lack of a protein structure or high sequence identity homologous templates, the approach successfully identified five low-micromolar inhibitors with drug-like properties.

Effectiveness of melatonin to restore fish brain activity in face of permethrin induced toxicity

Present study demonstrates permethrin induced oxidative damage in fish brain and explores effectiveness of melatonin to ameliorate brain function. Adult female Notopterus notopterus were exposed to nominal permethrin concentrations at 1/20th (0.34 μg/l) and 1/10th (0.68 μg/l) of LC50 for 15 days. The measured permethrin concentrations using gas chromatography (GC-ECD) were 0.28 μg/l and 0.57 μg/l, respectively. Some fish were sacrificed to collect brain tissue after 15 days of exposure. Remaining fish from both groups were administered exogenous melatonin (50 μg/kg, 100 μg/kg body weight) for 7 days and brain tissues were collected. Brain enzymes, ntioxidant factors, HSP70, HSP90, nuclear factor-kappa binding (NFkB), melatonin receptor (MT1R) proteins were measured. Permethrin treatment significantly (P < 0.05) decreased the levels of glutathione and brain enzymes. Malondialdehyde (MDA), xanthine oxidase (XO), HSPs increased at each concentration of permethrin. However, superoxide dismutase, glutathione s-transferase levels increased at low permethrin concentration followed by sharp decrease at higher concentration. Expression of NFkB and MT1R increased significantly (P < 0.05). Melatonin administration reinstated activity of brain enzymes, reduced MDA, XO levels and modulated HSPs. Melatonin also increased expression of NFkB and MT1R. Exogenous melatonin improves oxidative status in permethrin stressed fish brain. Melatonin modulates expression of HSPs that enables brain to become stress tolerant and survive by initiating NFkB translocation. Melatonin could act through melatonin receptor protein to induce synthesis of antioxidant proteins. Therefore the study successfully evaluates the potential of melatonin application for better culture and management of fish against pesticide toxicity.

Effects of curcumin on lipid peroxidation and antioxidant enzymes in kidney, liver, brain and testis of mice bearing Ehrlich Solid Tumor

Introduction: Cancer is the second most common cause of death in the world. Several natural products have been studied for anticancer activity and for prevention or repair of oxidative injury. Curcumin is one of the natural products of high medicinal interest. This study was performed to investigate effects of curcumin on lipid peroxidation and antioxidant enzymes in tissues of mice bearing Ehrlich solid tumor. Materials and Methods: Forty mice were distributed to four groups as healthy control and treatments that received 1x106 Ehrlich ascites tumor (EAT) cells and EAT cells plus 25 mg/kg/day or 50 mg/kg/day curcumin with a single subcutaneous injection. The liver, kidney, brain and testis tissues were collected for the MDA, SOD and CAT analyses. Results: Tumor development increased MDA levels in liver (p=0.001), kidney (p&lt;0.001) and testis (p&lt;0.01) and curcumin reduced liver MDA. Liver and kidney SOD activities were increased by both levels of curcumin (p=0.001) but 50 mg/kg/day curcumin increased brain SOD activity (p&lt;0.001). The kidney CAT activity was increased by 50 mg/kg/day curcumin (p&lt;0.001). Discussion: This study showed that curcumin suppresses tumor progression, and alleviates the lipid peroxidation and improves antioxidant status in the tissues of solid tumor-bearing mice.

Effects of Long-Term Supplementation with Aluminum or Selenium on the Activities of Antioxidant Enzymes in Mouse Brain and Liver

The aim of this study was to investigate the effects of aluminum (Al) or selenium (Se) on the “primary” antioxidant defense system enzymes (superoxide dismutase, catalase, and glutathione reductase) in cells of mouse brain and liver after long-term (8-week) exposure to drinking water supplemented with AlCl3 (50 mg or 100 mg Al/L in drinking water) or Na2SeO3 (0.2 mg or 0.4 mg Se/L in drinking water). Results have shown that a high dose of Se increased the activities of superoxide dismutase and catalase in mouse brain and liver. Exposure to a low dose of Se resulted in an increase in catalase activity in mouse brain, but did not show any statistically significant changes in superoxide dismutase activity in both organs. Meanwhile, the administration of both doses of Al caused no changes in activities of these enzymes in mouse brain and liver. The greatest sensitivity to the effect of Al or Se was exhibited by glutathione reductase. Exposure to both doses of Al or Se resulted in statistically significant increase in glutathione reductase activity in both brain and liver. It was concluded that 8-week exposure to Se caused a statistically significant increase in superoxide dismutase, catalase and glutathione reductase activities in mouse brain and/or liver, however, these changes were dependent on the used dose. The exposure to both Al doses caused a statistically significant increase only in glutathione reductase activity of both organs.

Anxiety and Depression Are Related to Higher Activity of Sphingolipid Metabolizing Enzymes in the Rat Brain.

Changes in sphingolipid metabolism have been suggested to contribute to the pathophysiology of major depression. In this study, we investigated the activity of acid and neutral sphingomyelinases (ASM, NSM) and ceramidases (AC, NC), respectively, in twelve brain regions of female rats selectively bred for high (HAB) versus low (LAB) anxiety-like behavior. Concomitant with their highly anxious and depressive-like phenotype, HAB rats showed increased activity of ASM and NSM as well as of AC and NC in multiple brain regions associated with anxiety- and depressive-like behavior, including the lateral septum, hypothalamus, ventral hippocampus, ventral and dorsal mesencephalon. Strong correlations between anxiety-like behavior and ASM activity were found in female HAB rats in the amygdala, ventral hippocampus and dorsal mesencephalon, whereas NSM activity correlated with anxiety levels in the dorsal mesencephalon. These results provide novel information about the sphingolipid metabolism, especially about the sphingomyelinases and ceramidases, in major depression and comorbid anxiety.

Environmentally Available Very Low Doses of a Dioxin (2,3, 7,8 TCDD) Affects to Membrane Bound Ion Dependent ATPases in Mice Brain

The present communication reports very low doses of environmentally available TCDD exposed for sub acute exposure duration affects to membrane bound ATPases enzymes in mice brain. Healthy female Swiss albino mice with the same body weight of about 25-30g were considered for the experimental purposes after acclimatization for a period of 2-3 days prior to the experiment. The animals were grouped before the experiments and kept under standard conditions. The experiments were conducted according to the ethical norms approved by the CPCSEA (Letter: CP6EA/CH/RF/ACK-2003, 29-07-2003). Animals were provided with rodent diet (Keval Sales Corporation, India) and water ad libitum.Different groups of mice were received different concentrations of TCDD (0.004 mg/kg bw and 0.04 mg/kg bw) for 2, 4 and 6 days of exposure durations. Control group of mice were received corn oil as a vehicle. Two hypotheses viz., (a) invivo exposure of environmentally available low concentrations of TCDD provokes both dose and subacute exposure duration dependent effects to membrane bound ATPases system, and (b) TCDD possibly has an indirect effect on membrane bound ATPases system, were tested in this study. The results suggested predominant exposure duration dependent effects on the membrane bound ion dependent ATPases studied. The observed results were indicative of a possible indirect effect of TCDD occurred by complex chain of reactions exhibited by cell when exposed to TCDD. In conclusion of present study being a lipophilic toxic, TCDD affects to ion channels by affecting different physiological reaction. TCDD may not be affecting directly to membrane bound ion channels.

Discovery, Radiolabeling, and Evaluation of Subtype-Selective Inhibitors for Positron Emission Tomography Imaging of Brain Phosphodiesterase-4D.

We aimed to develop radioligands for PET imaging of brain phosphodiesterase subtype 4D (PDE4D), a potential target for developing cognition enhancing or antidepressive drugs. Exploration of several chemical series gave four leads with high PDE4D inhibitory potency and selectivity, optimal lipophilicity, and good brain uptake. These leads featured alkoxypyridinyl cores. They were successfully labeled with carbon-11 (t1/2 = 20.4 min) for evaluation with PET in monkey. Whereas two of these radioligands did not provide PDE4D-specific signal in monkey brain, two others, [11C]T1660 and [11C]T1650, provided sizable specific signal, as judged by pharmacological challenge using rolipram or a selective PDE4D inhibitor (BPN14770) and subsequent biomathematical analysis. Specific binding was highest in prefrontal cortex, temporal cortex, and hippocampus, regions that are important for cognitive function. [11C]T1650 was progressed to evaluation in humans with PET, but the output measure of brain enzyme density (VT) increased with scan duration. This instability over time suggests that radiometabolite(s) were accumulating in the brain. BPN14770 blocked PDE4D uptake in human brain after a single dose, but the percentage occupancy was difficult to estimate because of the unreliability of measuring VT. Overall, these results show that imaging of PDE4D in primate brain is feasible but that further radioligand refinement is needed, most likely to avoid problematic radiometabolites.

H2S-producing enzymes in anoxia-tolerant vertebrates: Effects of cold acclimation, anoxia exposure and reoxygenation on gene and protein expression.

To lend insight into the potential role of the gasotransmitter hydrogen sulfide (H2S) in facilitating anoxia survival of anoxia-tolerant vertebrates, we quantified the gene expression of the primary H2S-synthesizing enzymes, 3-mercaptopyruvate sulfurtransferase (3MST), cystathionine γ-lyase (CSE) and cystathionine β-synthase (CBS), in ventricle and brain of normoxic, anoxic and reoxygenated 21°C- and 5°C-acclimated freshwater turtles (Trachemys scripta) and 10°C-acclimated crucian carp (Carassius carassius). Semi-quantitative Western blotting analysis was also conducted to assess 3MST and CBS protein abundance in ventricle and brain of 5°C turtles and 10°C crucian carp subjected to normoxia, anoxia and reoxygenation. We hypothesized that if H2S was advantageous for anoxia survival, expression levels would remain unchanged or be upregulated with anoxia and/or reoxygenation. Indeed, for both species, gene and protein expression were largely maintained with anoxia exposure (24h, 21°C; 5 d, 10°C; 14 d, 5°C). With reoxygenation, 3MST expression was increased in turtle and crucian carp brain at the protein and gene level, respectively. Additionally, the effect of cold acclimation on gene expression was assessed in several tissues of the turtle. Expression levels were maintained in most tissues, but decreased in others. The maintenance of gene and protein expression of the H2S-producing enzymes with anoxia exposure and the up-regulation of 3MST with reoxygenation suggests that H2S may facilitate anoxic survival of the two champions of vertebrate anoxia survival. The differential effects of cold acclimation on H2S enzyme expression may influence blood flow to different tissues during winter anoxia.

Effects of the C1473G Polymorphism in the Tryptophan Hydroxylase 2 Gene and the Length of Daylight on Behavior in Mice

Seasonal depressive disorder occurs in conditions of seasonal reductions in the duration of daylight and is a significant medical, social, and economic problem. The risk of seasonal depressive disorders is linked with impairments to the serotonin system of the brain. Tryptophan hydroxylase 2 (TPH2) is the key enzyme in serotonin synthesis in the brain. The C1473G polymorphism in the gene encoding TPH2 in mice leads to a twofold decrease in the activity of this enzyme in the mouse brain. The aim of the present work was to study the effects of the length of daylight on the behavior of mature males of the congenic strains B6-1473C and B6-1473G, which have high and low levels of TPH2 activity, respectively. Adult males of these strains were kept for one month in conditions of long (14 h) or short (4 h) daylight durations. Being kept in conditions of short daylight had no effect on mean day-round movement activity, food and water consumption, or anxiety in the open field and elevated plus maze tests in either mouse strain, but decreased mean sleep duration (p < 0.002) and active resistance in the forced swimming test (p < 0.01), changes being similar in both strains. Thus, the C1743G polymorphism had no effect on changes in behavior induced in mice by short daylight.

Purification, characterization and identification of rat brain cytosolic tyrosine transaminase as glutamine Transaminase-K

The current study was undertaken to investigate the spectrum of tyrosine transaminases enzymes in a cytosolic fraction of rat brain and to specifically purify and characterize a previously identified cytosolic brain enzyme possessing tyrosine/glyoxylate transaminase activity. Based upon extensive biochemical and immunochemical characterization of purified brain tyrosine/glyoxylate transaminase, we concluded the purified enzyme is glutamine transaminase-K (EC 2.6.1.64). This conclusion was based on: 1.) a concurrent enrichment in the tyrosine/glyoxylate and glutamine/phenylpyruvate transaminase activities during purification, 2.) demonstration of a co-substrate specificity for amino acids and α-keto acids that was highly consistent with published information for glutamine transaminase-K, 3.) results from detailed kinetic analysis, 4.) glutamine was a potent inhibitor of in vitro tyrosine/glyoxylate transamination, 5.) biochemical characterization, including pH optimum of 8.5 and spectrophotometric analysis and 6.) immunoanalytical analysis using a specific antiserum to rat renal glutamine transaminase-k. In addition, immunochemical characterization of a crude soluble extract of whole brain suggests that the in vitro tyrosine transaminase activity for several different α-keto acid co-substrates likely reflect the activity of glutamine transaminase-K. In conclusion, this investigation confirmed the presence of multiple tyrosine transaminase enzymes in a cytosolic extract of rat brain. Moreover, we concluded glutamine transaminase-K represents a predominant cytosolic enzyme in rat brain that's capable of catalyzing in vitro transamination of p-tyrosine and other aromatic amino acids, including the neurotransmitter precursors L-dopa and 5-hydroxytryptophan. The purified transaminase possesses a broad co-substrate specificity with preferential reactivity with α-keto acids derived from neutral aliphatic and aromatic amino acids. Lastly, we identified a heterogeneous regional distribution of tyrosine/glyoxylate transaminase (glutamine transaminase-K) in rat brain with a significantly higher level of in vitro activity in cerebellum.

A Study With [18F]MNI-1054 to Determine Lysine -Specific Demethylase 1A (LSD1) Brain Enzyme Occupancy of TAK-418 After Single-Dose Oral Administration in Healthy Participants

A Study With [18F]MNI-1054 to Determine Lysine -Specific Demethylase 1A (LSD1) Brain Enzyme Occupancy of TAK-418 After Single-Dose Oral Administration in Healthy Participants