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Biomedical subjects

B W Moore

Publications and source records attributed to B W Moore.

At least 19 recordsLinked to original sources
PubMed1992

Nonoxidative ethanol metabolism: expression of fatty acid ethyl ester synthase-III in cultured neural cells.

Alcohol metabolism in the human brain has been characterized as essentially nonoxidative in nature, with the esterification of ethanol with fatty acids via fatty acid ethyl ester synthase. This pathway of ethanol metabolism is related to end organ damage in the brain but the neural cell type expressing FAEES has not been identified. In this study human and rodent neuroblastoma and glioma cell lines are assayed for fatty acid ethyl ester synthase activity. Cells with neuronal properties demonstrated higher activity than glioma cell lines. We confirmed the presence of the mRNA for one type of synthase, fatty acid ethyl ester synthase-III in three neuronal cell lines--N1E115 cells, PC12 cells, and SK-N-MC cells. These results support the hypothesis that FAEES activity is expressed chiefly in cells with neuronal properties and suggest that non-oxidative ethanol metabolism is potentially related to the toxic effect of ethanol on the human brain.

Acyltransferases↗
PubMed1992

S100 is present in developing chicken neurons and Schwann cells and promotes motor neuron survival in vivo.

We used polyclonal antisera recognizing S100, a small acidic protein highly enriched in nervous tissue, to stain sections of embryonic chicken lumbosacral spinal cord and hindlimb. S100 immunoreactivity was detected in developing sensory neurons of the dorsal root ganglia (DRG) and motor neurons of the ventral spinal cord as early as embryonic day (E) 5, and staining persisted through hatching. In contrast, expression of S100 first became apparent in Schwann cells at E13, just before myelination, and was not detected in developing skin or muscle. Since S100 beta was present in motor and sensory neurons and is known to promote neuronal survival and neurite extension in vitro (Winningham-Major, Staecker, Barger, Coats, and Van Eldik, 1989), we tested the ability of S100 to promote neuron survival in an in ovo survival assay. Addition of S100 to chick embryos in ovo during the period of naturally occurring motor neuron cell death resulted in a significant increase in motor neuron survival, but had no effect on the in vivo survival of sensory neurons in the DRG. The findings that S100 is present in spinal motor neurons and that the addition of S100 enhances the survival of these cells in vivo are consistent with the possibility that S100 may act as a naturally occurring neuron survival factor during development.

Animals↗
PubMed1992

A simple, highly sensitive assay for measurement of digitonin during receptor solubilization.

A simple and highly sensitive assay for measuring total digitonin in biological samples is described. The assay is based on the ability of digitonin to hemolyze red blood cells. The precision and reproducibility of the assay was excellent with intra- and inter-assay variabilities of less than 1% and 6%, respectively. The assay was used to evaluate several potential methods for removing digitonin from biological samples (digitonin extracts from guinea pig brain membranes). Dialysis and G-25 Sephadex chromatography were ineffective. However, protein and digitonin can be effectively separated by ammonium sulfate precipitation followed by dialysis. The kappa 1 opioid receptor survived these procedures with no change in affinity for [3H]U-69,593. In conclusion, the hemolytic assay for digitonin appears to provide a practical means for determining detergent concentrations during receptor purification and characterization and for evaluating potential methods for detergent removal. Although an in depth analysis of the assay was carried out only for digitonin, CHAPS and deoxycholate also caused 50% hemolysis at concentrations well below those commonly used for receptor solubilization and, therefore, the general assay procedures might have applicability for measurement of these and perhaps other detergents used in receptor solubilization as well.

Animals↗
PubMed1992

Ethanol inhibits C6 cell growth: fetal alcohol syndrome model.

Maternal consumption of ethanol produces a pattern of malformations, including nervous system abnormalities, in the developing fetus, a state called Fetal Alcohol Syndrome. We report the dose-dependent inhibition by ethanol of the growth of a glioma derived cell line, C6 cells; the effects occur at ethanol concentrations commonly encountered in the blood during human intoxication. The effects occur with different morphological subtypes of the cell line and do not occur when the cells are exposed to iso-osmolar concentrations of other chemicals. The results demonstrate that C6 cells are a model for the study of the effects of ethanol on nervous system cell growth.

Astrocytes↗
PubMed1988

Isolation of a component from commercial coomassie brilliant blue R-250 that stains rubrophilin and other proteins red on polyacrylamide gels.

Commercially available Coomassie Brilliant Blue R-250 (C.I. 42660) is a popular and useful dye that stains most proteins blue on polyacrylamide gels. Some proteins from brain (rubrophilin), collagens, histones and parotid gland proteins are distinctly red when stained with Coomassie Blue. Commonly used Coomassie Brilliant Blue R-250 preparations may contain more than 30 distinct colored and fluorescent components that can be separated on silica gel chromatographic columns. A specific component has been isolated on silica gel columns that stains rubrophilin and other proline-rich proteins a reddish color. Fast atom bombardment mass spectrometry of the isolated rubrophilin staining principle indicates a molecular weight of 634 as compared to 826 for the major dye in the original Coomassie Brilliant Blue R-250. Infrared spectrometry is consistent with a difference between the rubrophilin staining principle and Coomassie Brilliant Blue R-250 of a toluene sulfonic acid residue.

Acrylic Resins↗
PubMed1988

Identification of calcium binding proteins from brain.

A procedure was worked out for purification and identification of calcium-binding proteins from bovine brain using Ca2+-dependent, reversible binding to a hydrophobic support, phenyl-Sepharose, as the method of isolation. These proteins could be visualized during and after their separation by running them on non-denaturing polyacrylamide gels, blotting to Zeta-probe paper, and autoradiographing with 45Ca2+. About 24 polypeptides could be seen in this fraction on SDS (Laemmli) gels and about 8-10 native, Ca2+-binding proteins could be seen on non-denaturing gels and on blots of their 45Ca2+ autoradiographs. Some of these proteins could be purified further by chromatography on DEAE-Sephacel and still retain their 45Ca2+-binding activity.

Animals↗
PubMed1988

Conformational and hydrophobic properties of rat and bovine S-100 proteins.

The binding of Ca2+ to rat or bovine S-100 proteins, in the absence of ligands, showed a dissociation constant (in 60 mM K+) of 0.5 to 1.0 mM as measured by the effects of Ca2+ on binding of S-100 to phenyl-Sepharose, reactivity of sulfhydryl groups, and difference spectra for PHE, TYR, and TRP residues. Binding of the ligands, "Stainsall" and chlorpromazine lowered the dissociation constant of S-100 for Ca2+ by 2- to 10-fold as measured by the same parameters. The conformational change, in response to Ca2+ binding, probably occurs by exposure to solvent of the hydrophobic region of alpha and beta subunits of S-100 at residue positions 74-93.

Animals↗
PubMed1988

A new preparation of S-100 protein from rat and bovine brains.

The S-100 nervous system protein was purified from bovine and rat brains by a modification of the original procedure. The main modification consisted in substituting a step of calcium-dependent binding of S-100 to a phenyl-Sepharose column for the original step of chromatography on G-200 Sephadex. The proteins were pure as determined by SDS gel electrophoresis. HPLC on a reversed phase and on a size-separation column, and by immunological criteria. The bovine S-100 behaved as previously described, during calcium binding, by displaying a conformational change as evidenced by increase in native fluorescence.

Animals↗
PubMed1987

Purification and properties of rubrophilin: a novel brain specific membrane polypeptide.

Rubrophilin, a unique brain specific polypeptide, was purified to apparent homogeneity from microsomal fractions of bovine brains. The peptide stains pink with Coomassie Brilliant Blue R-250 (C.I. No. 42660) under specific conditions, has an apparent Mr of 53,000, and is acidic with an apparent pI of 4.9. The purification involves initial solubilization of delipidated microsomes in sodium dodecyl sulfate, followed by ammonium sulfate fractionation, reversed ammonium sulfate gradient elution from diatomaceous earth, gel filtration on polyacrylamide (Biogel P-200), gradient elution chromatography from hydroxylapatite, and reverse-phase chromatography from phenyl-Sepharose. A yield of about 5 mg of rubrophilin was obtained from 9 g of microsomal proteins. Amino acid analysis shows that rubrophilin contains only nine amino acids with residues/mol as follows: alanine (102), glutamic acid (97), lysine (65), proline (55), aspartic acid (48), glycine (44), serine (37), threonine (35), and valine (10). Cysteine, methionine, tryptophan, tyrosine, isoleucine, phenylalanine, histidine, and arginine could not be detected. Relative rubrophilin content of vertebrate brains was as follows: mammals greater than birds greater than reptiles greater than fishes. It is present in mouse retina and human neuroblastoma cell cultures but could not be detected in octopus optic lobe or in cultured C-6 rat glioma cells.

Amino Acids↗
PubMed1987

Failure of effervescent zinc acetate lozenges to alter the course of upper respiratory tract infections in Australian adults.

Effervescent lozenges containing 10 mg of zinc acetate were evaluated as a treatment of upper respiratory tract infections in a double-blind randomized trial by using a placebo which was indistinguishable to most observers in taste and appearance from the active material. Of the 70 treatment courses used by 55 individuals in 34 families, 63 (33 zinc and 30 placebo) were considered evaluable, in that the volunteer used the medication at least four times daily for at least 3 days, the average utilization being 5.4 days at an average dose of six lozenges daily. Six users of zinc reported nausea (versus no placebo users), and eight reported an unpleasant taste or aftertaste (versus one placebo user). No benefit was observed among the users of zinc acetate. The mean duration of symptoms in users of the zinc was 12.1 days, compared with 7.7 days in those who used the placebo. Nor was any beneficial effect of zinc evident among the four zinc-treated versus the two placebo-treated individuals from whom rhinovirus was grown.

Acetates↗
PubMed1986

Prophylactic efficacy of intranasal alpha 2-interferon against rhinovirus infections in the family setting.

In a double-blind evaluation of alpha 2-interferon as prophylaxis against naturally acquired respiratory infections, 120 adult members of 46 Australian families used 325 courses of intranasal spray during a six-month period, applying 5 million IU to the anterior nasal mucosa daily for seven days when respiratory symptoms developed in another member of the family. Used in this way, the alpha 2-interferon was well tolerated, and the rate of minor nasal bleeding (12 percent) did not increase with repeated courses. By comparison with the control group of 109 members of 49 families who used 319 seven-day courses of placebo spray, the users of alpha 2-interferon experienced 33 percent fewer days with nasal symptoms and 41 percent fewer episodes of "definite" respiratory illness. The users of alpha 2-interferon who were exposed to rhinovirus infections experienced 76 percent fewer days with symptoms and 86 percent fewer "definite" illnesses than their counterparts who used placebo. All of the observed clinical benefits, which suggested prevention of 6.8 "definite" respiratory illnesses per 100 courses of medication used, could be explained by a protective effect against illness associated with rhinoviruses that was not demonstrated for influenza A or B or coronavirus 229E.

Administration, Intranasal↗
PubMed1986

Protein organization of rat synaptic plasma membranes and synaptic vesicles: a one- and two-dimensional study.

The protein organization of rat brain synaptic plasma membranes (SPM) and synaptic vesicles (SV) was investigated by surface iodination and one- and two-dimensional electrophoresis. Polypeptides of molecular weights (MWs, in Kilodaltons) 170 K, 135 K, 96-86 K, 68-64-61 K, 56 K, 52 K, 38 K, 35-33 K, and 18 K are predominantly or exclusively exposed on the extracellular side of synaptosomes. Several polypeptides of MW between 70 K and 40 K are exclusively exposed on the cytoplasmic side of SPM. The use of two-dimensional electrophoresis allowed to recognize that, for some classes of MW, there are polypeptides of nearly the same MW and different isoelectric points exposed on both sides of SPM. The synaptosomal membrane shows a predominance of acidic proteins on the extracellular side and more neutral and basic proteins on the cytoplasmic side. With respect to SPM, SV are particularly enriched with polypeptides of MW 71 K, 56 K, 39-38 K, 32 K, 16 K, and 15 K. One of them, a doublet of MW 39-38 K, is the most highly labeled species upon surface iodination and is similar, but not identical, with a doublet located on the cytoplasmic side of SPM.

Animals↗
PubMed1986

Adult onset Still's disease or coxsackie polyarthritis?

The clinical manifestations and laboratory findings of two patients with a presumptive diagnosis of coxsackie B4 virus infection are described. A striking feature was the similarity with adult onset Still's disease, with spiking fever, evanescent macular rash, and severe polyarthritis. This latter feature persisted for many weeks and required steroids to control the symptoms. Review of the literature has supported the proposition that many cases of adult onset Still's disease may be due to coxsackie B4 or other viral infection and it is suggested that these agents should be actively sought in future cases.

Adult↗
PubMed1985

Mortality from influenza and allied infections in South Australia during 1968-1981.

Numbers of deaths from pneumonia and influenza and other causes were analysed for successive four-week periods in South Australia during 1968-1981. An overall excess in deaths from pneumonia or influenza of 74% was evident during the winter months and early spring, compared with summer and early autumn. An accompanying excess of 18% occurred for deaths assigned to other causes. There was a strong association between numbers of deaths from pneumonia and influenza and other deaths, suggesting that influenza may have a broad impact on mortality. This mostly applied to individuals aged 60 years and over. There is the need for medical practitioners to provide prophylactic care to protect aged patients against the effects of influenza. This should be done in autumn, and special attention should be given to individuals with underlying conditions.

Adolescent↗
PubMed1982

Chemistry and biology of the S-100 protein.

The S-100 protein is specific for the nervous system and, being present in all vertebrates, shows a high degree of stability of structure during evolution. In adult animals it is primarily localized to glial elements, although there is some evidence that a small proportion may be present in neuronal nuclei or their plasma membranes. During development it is synthesized rapidly at a relatively late period of differentiation of the nervous system. In glioma cell cultures there is a control mechanism that seems to involve some kind of signal at the external surface of the plasma membrane, possibly specific cell-cell contact, to stimulate S-100 synthesis. All of these biological properties of S-100 suggest that it is connected with some specific essential function that is common to the nervous system of all vertebrates. Several chemical properties of S-100 provide clues to this function. It is an unusually acidic and soluble protein and, in the absence of Ca2+, has no detectable hydrophobic regions accessible to solvent. It is capable of specifically binding Ca2+, a process that causes S-100 to undergo a conformational change that exposes a hydrophobic region to the solvent and stimulates binding of S-100 to membranes. The conformational change and the membrane-binding properties are reversible when Ca2+ is removed and are antagonized by monovalent cations such as K+ and Na+. These chemical properties suggest that S-100 may, as part of its function in the nervous system, be bound to some hydrophobic site, possibly a membrane, and that the extent of this binding is regulated by concentrations of Ca2+, K+ and Na+. If this is true, then it is important, as the next step in working out its function, to discover the exact site where S-100 binds in the nervous system.

Amino Acid Sequence↗
PubMed1980

Octopus calmodulin. Structural comparison with bovine brain calmodulin.

A protein previously isolated from octopus optic lobe is shown to have the biochemical characteristics of a calmodulin-like protein. The amino acid composition of the octopus calmodulin is similar to that of another sea invertebrate calmodulin, from Renilla reniformis, in that both contain a single residue of tyrosine which distinguishes them from the vertebrate calmodulins which contain two tyrosines. The 1H NMR spectra of the octopus calmodulin and bovine brain calmodulin are compared in their apo- and calcium-saturated conformations. A comparison of these spectra indicates that the single tyrosine of the octopus calmodulin is in a structurally homologous position to tyrosine-138 of bovine brain calmodulin. 1H NMR and UV difference spectroscopy also demonstrate that the solution conformations of the apo- and calcium-saturated forms of octopus calmodulin are very similar to those of bovine brain calmodulin. It is concluded that both proteins undergo similar calcium-induced changes in tertiary structure, which result in near identical solution conformations.

3',5'-Cyclic-AMP Phosphodiesterases↗
PubMed1980

An improved method of preparing rat brain synaptic membranes. Elimination of a contaminating membrane containing 2',3'-cyclic nucleotide 3'-phosphohydrolase activity.

Synaptosomes were prepared from rat cortex by subjecting a washed crude mitochondrial pellet to centrifugation first on discontinuous Ficoll-isotonic sucrose gradients and then on discontinuous sucrose gradients. The synaptosome fraction, collected from the 7.5-14% Ficoll band (II), was further separated into two additional fractions, designated IIA and IIB, which bank at the 0.32-1.05 M and at the 1.05-1.6 M sucrose interfaces, respectively. Electron microscopic analysis showed that fraction IIB contained synaptosomes and extra terminal mitochondria and was essentially free of membrane fragments. Further characterization showed that IIB contained 69% of the protein and 83% of the lactic dehydrogenase activity of fraction II and had a specific activity of a 2',3'-cyclic nucleotide 3'-phosphohydrolase approximately 1% of that obtained with myelin. Fraction IIA had approximately 50% the specific activity of the 2',3'-cyclic nucleotide 3'-phosphohydrolase found in myelin. Synaptic plasma membranes were prepared by lysing fraction IIB in 1 mM sodium phosphate, 0.1 mM EDTA at pH 8.5 and subjecting this preparation to centrifugation on a discontinuous sucrose density gradient. Enzymatic analysis indicated that membranes banding at the 0.6-0.8 M sucrose interface had high specific activities of plasma membrane enzymes (e.g. acetylcholinesterase, ATPase, 5'-nucleotidase). The specific activity of the (Na+ + K+)-ATPase in the purified membrane preparation was 8-fold higher than that in the original homogenate. Specific activities of various marker enzymes indicated that the composition of these membrane preparations for the most part was synaptic plasma membranes, approximately 7% mitochondrial outer membranes and 3% a membrane containing 2',3'-cyclic nucleotide 3'-phosphohydrolase activity. The polypeptide compositions of three possible contaminating membranes and of synaptic membranes were compared by electrophoresis in 6-20% gradient polyacrylamide gels in the presence of sodium dodecyl sulfate. Whereas mitochondrial and myelin membranes had distinct compositions, the compositions of the microsomal and synaptosomal plasma membranes were similar. Synaptic plasma membranes contained at least 27 polypeptides; the three major polypeptides had molecular weights of 103,000; 54,000; and 50,000. The major polypeptides of soluble synaptosomal proteins had molecular weights of 54,000 and 42,000.

2',3'-Cyclic-Nucleotide Phosphodiesterases↗