Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:1.4.1.2 (glutamate dehydrogenase)
 4,380 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Cytochemical methods for the demonstration of enzyme activities in blood and bone marrow cells were systematically improved by the addition of an inert polymer, polyvinyl alcohol (PVA), to the incubation medium and by using optimized reaction media. The methods investigated were tetrazolium salt methods for lactate, glucose-6-phosphate, succinate and glutamate dehydrogenase, the indoxyl-tetrazolium salt method for alkaline phosphatase, the diaminobenzidine method for peroxidase, and diazonium salt methods for chloroacetate esterase, beta-glucosaminidase, beta-glucuronidase, acid phosphatase, and dipeptidylpeptidase II and IV. PVA in the media preserved the morphology of cells very well and prevented leakage of large molecules such as enzymes from the cells. Therefore, fixation or long periods of air-drying prior to incubation leading to substantial loss of enzyme activity could be avoided. A brief period of drying (2 min at 37 degrees C) of the cell preparations just before the incubation was sufficient for making the cells permeable. Localization of enzyme activities was very precise and precipitation of the final reaction product was confined to sites which are known to contain the enzyme under study (granules, mitochondria, lysosomes). These advantages advocate the use of PVA in haematological enzyme cytochemistry and especially for diagnosis of leukemia.
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PMID:Enzyme cytochemistry of unfixed leukocytes and bone marrow cells using polyvinyl alcohol for the diagnosis of leukemia. 280 89

Replication-defective Moloney murine leukemia virus expressing the GAD67 gene under the control of the GFAP promoter was produced using selected clones of a fibroblast-packaging cell line. A spontaneously immortalized astrocyte cell line was infected with this virus and cellular clones expressing GAD67 selected. Astrocyte and fibroblast clones expressed functional GAD (detected by glutamic acid decarboxylation), but only fibroblasts were able to also produce GABA in the extracellular medium. When exposed to 200 microM glutamate, despite an observed difference in the rates of glutamate accumulation in control and GAD67-expressing astrocytes, similar proportions of glutamate taken up were detected. In GAD67-expressing astrocytes, the glutamate was mainly converted into GABA, suggesting GAD transgene activity to be dominant over other glutamate metabolic pathways, such as glutamine synthetase and glutamate dehydrogenase. Moreover, rapid GABA release into the cell medium was also observed, suggesting the involvement of reverse GABA transporters. The use of the GFAP promoter might be able to take advantage of its activation in response to factors inducing reactive gliosis observed in pathological insults. GAD67-expressing astrocytes might therefore be used for future grafting in pathological situations in which an excess of glutamate results in neuronal dysfunction or cell death.
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PMID:Glutamate-modulated production of GABA in immortalized astrocytes transduced by a glutamic acid decarboxylase-expressing retrovirus. 943 90