Phosphorus-31 NMR studies of anchorage-dependent neural-type cells in tissue culture.
Item
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Title
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Phosphorus-31 NMR studies of anchorage-dependent neural-type cells in tissue culture.
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Identifier
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AAI8821083
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identifier
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8821083
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Creator
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Glynn, Paul.
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Contributor
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Adviser: Richard L. Chappell
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Date
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1988
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Language
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English
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Publisher
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City University of New York.
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Subject
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Biology, General | Chemistry, Biochemistry
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Abstract
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A {dollar}\sp{lcub}31{rcub}{dollar}P NMR probe and microcarrier-based life-support system were built to measure NMR spectra of anchorage-dependent cells in a tissue-culture environment and to study development and metabolic processes in N1E-115 neuroblastoma, C6 glioma, dissociated rat brain cells (DRBC) and C3HT10{dollar}1\over2{dollar} fibroblast cells. Development changes were found in the phosphoethanolamine (PEt) and phosphocholine (PCh). Creatine kinase activity was found in DRBC, C6 glioma cells, and in differentiated and undifferentiated cells. The uptake of creatine into these cells was extremely slow under physiological conditions.;A major finding of this work centered around the creatine kinase reaction and the observed changes in the cell Pi, PCr, and ATP when the cells were subjected to an external pH jump. Measurements of these values, and determination of the PCr/Cr ratio from {dollar}\sp1{dollar}H and {dollar}\sp{lcub}31{rcub}{dollar}P spectra of the cell extracts allowed the phosphate potential to be calculated. This important parameter had not been unambiguously measured in live, intact cells in tissue culture.;DRBC showed a higher creatine kinase activity in the range of 0.07 to 0.09 S{dollar}\sp{lcub}-1{rcub}{dollar} comparable to in vivo measurements made on rat brains. C6 glioma cells were found to have creatine kinase activity and to take up extracellular creatine. Undifferentiated neuroblastoma cells with high PCr when stressed with an ischemic episode showed that depleted ATP and PCr concentrations recovered to near preischemic value in a fixed pattern dependent on the time of the ischemic episode. Stress of depolarization resulting from addition of potassium chloride (final concentration 90 mM) to the extracellular medium were also studied.
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Type
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dissertation
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Source
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PQT Legacy CUNY.xlsx
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degree
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Ph.D.
