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J Burnside

Publications and source records attributed to J Burnside.

30 records · Page 2Linked to original sources
PubMed1988

Isolation and characterization of the gene encoding the alpha-subunit of the rat pituitary glycoprotein hormones.

The gene encoding the common alpha subunit of the rat pituitary glycoprotein hormones was isolated from a rat genomic DNA library. The gene spans approximately 8 kb, and contains four exons and three intervening sequences of 5.4 kb, 1.1 kb and 0.6 kb. Blot hybridization of restriction enzyme digests of rat genomic DNA suggests that the alpha gene is present in a single copy. The coding region and 424 bp of the 5'-flanking region of the gene were sequenced. Primer extension and S1 nuclease analyses revealed a single transcriptional start point downstream from consensus promoter elements. The organization of the rat alpha-subunit gene is similar to that of the human and bovine genes including the sizes and locations of the four exons and three introns. In addition, a region of strong sequence similarity has been identified in the 5'-flanking region of the rat, human and bovine genes. This region includes sequences which are similar to a putative triiodothyronine regulatory element and the previously identified cAMP regulatory region; such sequences may mediate the known effects of these factors on alpha-subunit gene expression.

Animals↗
PubMed1988

Differential carbohydrate processing and secretion of thyrotropin and free alpha subunit. Effects of 1-deoxynojirimycin.

In pulse-chase experiments we compared the kinetics of early carbohydrate processing and subsequent secretion of thyroid-stimulating hormone (TSH) and free alpha subunit under control conditions and after treatment with 1-deoxynojirimycin, an inhibitor of glucosidases I and II. Under control conditions TSH achieved resistance to endo-beta-N-acetylglucosaminidase H (endo H) more rapidly than free alpha (t1/2 0.3 h versus 0.9 h); however, free alpha was secreted more rapidly than TSH (t1/2 2.2 h versus 3.4 h). With 1-deoxynojirimycin, oligosaccharides co-migrating with G3Man9GlcNAc and G2Man9GlcNAc were demonstrated on TSH for the first time, suggesting that previous pulse-chase studies did not disclose these intermediates due to rapid removal of glucose residues from the common G3Man9GlcNAc2 precursor. 1-Deoxynojirimycin delayed the rate of attainment of endo H resistance for both TSH and free alpha, but there was no effect on subunit combination. With 5 mM 1-deoxynojirimycin the amount of secreted free alpha was reduced to 65% of control; secreted TSH was reduced markedly to 17% of control without intracellular accumulation, suggesting increased intracellular degradation. There was no significant toxicity from these doses of 1-deoxynojirimycin on the production or secretion of the two major nonglycosylated pituitary proteins, growth hormone and prolactin, or on at least 10 other secretory proteins. Basal differences in the relative rates of TSH and free alpha processing and secretion as well as differential sensitivity to 1-deoxynojirimycin suggest separate secretory pathways for these two closely related proteins.

1-Deoxynojirimycin↗
PubMed1985

Pretranslational regulation of ectopic hCG alpha production in ChaGo lung cancer cells by sodium butyrate.

Ectopic production of hCG and its free alpha subunit by ChaGo lung cancer cells is stimulated by sodium butyrate. To investigate pretranslational regulation in this system, we examined the response of the hCG alpha and beta subunit mRNAs in ChaGo-K1 cells, a clone that produces free hCG alpha but no hCG or hCG beta in the basal state. When a Northern blot of total RNA from ChaGo cells was hybridized to a [32P]- labeled hCG alpha cDNA probe, a single band was detected that was identical in size (approximately 850 bases) to placental hCG alpha mRNA. RNA from butyrate-stimulated (5 mM, 24 h) ChaGo cells contained 7.7 times as much hCG alpha mRNA as RNA from control ChaGo cells. This increase appeared to be relatively selective since no difference in total polyA-containing mRNA levels was detected between butyrate-treated and control cells by [32P]oligo(dT) hybridization. In addition, no hCG beta mRNA was detected when Northern and dot blots were hybridized to an hCG beta cDNA probe. In a time course experiment, hCG alpha mRNA accumulation in butyrate-treated cells increased significantly by 8 h with a maximum increase of 6.1-fold at 24 h compared to control values. Major differences in immunoactive hCG alpha accumulation were not apparent, however, until after 24 h. These studies show that stimulation of ChaGo hCG alpha production by butyrate can be completely accounted for by pretranslational events and that failure to detect hCG or free hCG beta production by these cells is not due to poorly translatable RNA or post-translational protein degradation. Thus, exclusive ectopic production of only one of the hCG subunits is likely to be due to selective genomic expression.

Butyrates↗
PubMed1985

Differential regulation of hCG alpha and beta subunit mRNAs in JEG-3 choriocarcinoma cells by 8-bromo-cAMP.

The coordinate regulation of human chorionic gonadotropin (hCG) subunit synthesis by JEG-3 choriocarcinoma cells was studied at the pretranslational level. The responses of the hCG alpha and beta mRNAs were measured during stimulation with the potent cAMP analog 8-bromo-cAMP (8-Br-cAMP) using 32P-labeled hCG alpha and beta cDNA probes. The hCG alpha mRNA (850 bases) and beta mRNA (1050 bases) from JEG-3 cells were identical in size to that of their respective mRNAs from placenta, by Northern blot analysis. After 48 h of stimulation with 2 mM 8-Br-cAMP, production of immunoreactive alpha and beta subunits increased 25- and 52-fold, respectively; corresponding levels of the alpha and beta mRNAs increased 36- and 43-fold, respectively, in a dot blot hybridization assay. Total cellular protein, DNA content, and messenger RNA pools were not altered by treatment with 8-Br-cAMP. The temporal coordination of the expression of the hCG alpha- and beta-subunit genes was examined by comparing the time course of stimulation of the respective mRNAs and the production of immunoreactive subunits. The kinetic responses of the alpha and beta mRNAs differed: the increase in hCG alpha mRNA preceded the increase in hCG beta mRNA, while levels of free alpha subunit and intact hCG increased in parallel with the increase in beta mRNA. hCG alpha mRNA levels increased rapidly between 8 and 24 h after the addition of 8-Br-cAMP, and approached a plateau by 48 h. The levels of hCG beta mRNA increased steadily throughout the 8-48 h period. These results demonstrate that the cAMP analog 8-Br-cAMP differentially regulates hCG subunit biosynthesis in JEG-3 cells at a pretranslational level, and that the stimulation by 8-Br-cAMP in this system appears to be relatively selective for hCG subunits.

8-Bromo Cyclic Adenosine Monophosphate↗
PubMed1984

NLM-CHEMSORT: an algorithm and computer program for sorting chemical names.

An algorithm is described that has been designed to sort medium-size lists of chemical names, including common, generic, trivial, and systematic names and code numbers, into a logical sequence. It successfully sorted more than 99.5% of 3767 names in its first application. Minor revisions then resulted in more than 99.9% success with the same set of names. The algorithm generates an 80-character primary sort key (alphabetic characters only) and a 16-character secondary level sort key (alphanumeric characters). These sort keys are generated de novo from the name as needed and, thus, do not require increased permanent-storage costs. Sorting on the primary sort key (and secondary sort keys when identical primary keys exist) results in logical sequences of chemical names.

Chemical Phenomena↗
PubMed1982

Characterization of the membrane proteins of rat liver lysosomes. Composition, enzyme activities and turnover.

Lysosomes prepared from the livers of untreated rats and from the livers of rats injected with either Triton WR-1339 or dextran yielded membranes that were similar in both polypeptide composition and activities of ATPase and acid 5'-nucleotidase. The administration of Triton WR-1339 (and dextran) resulted in an increase in ATPase activity of liver homogenates that was associated with a parallel increase in the ATPase activity of the lysosomal membrane. On the other hand, plasma membranes appear to be different from lysosomal membranes with respect to polypeptide composition and enzyme activities. The ATPase activity of lysosomal membranes is not affected by ouabain and suramin, inhibitors of the plasma-membrane ATPase. The plasma-membrane alkaline 5'-nucleotidase has little activity at acid pH. Pulse-labelling of lysosomal membranes with [3H]fucose and with [3H]- and [14C]-leucine occurred rapidly, faster than labelling of plasma membranes. The labelling kinetics indicate that lysosomal membranes may be assembled independently of plasma membranes. These data suggest that, in liver, little bulk transport of plasma membrane to lysosomes takes place, and lysosomal-membrane proteins may not be derived from those of plasma membranes.

Animals↗
PubMed1978

Properties of the membrane proteins of rat liver lysosomes. The majority of lysosomal membrane proteins are exposed to the cytoplasm.

Rat liver lysosomes were lysed and subfractionated by differential centrifugation through 0.2M-NaCl to yield a membranous pellet. This membrane fraction contains less than 20% of the lysosomal protein, adenosine triphosphatase activity of about 1.2mumol/min per mg of protein, 120nmol of thiol groups/mg of protein and at least 16 protein and glycoprotein bands on sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. The gel patterns of membranes isolated from lysosomes after treatment with (1) [125I]iodidehydrogen peroxide-lactoperoxidase, (2) toluene 2,4-di-isocyanate-activated bovine serum albumin, (3) trypsin and (4) subtilisin indicate that most of the membrane proteins are exposed to the cytoplasm. These exposed proteins are candidates for intracellular receptors which recognize either substances that are to be degraded or vesicles containing those substances.

Adenosine Triphosphatases↗
PubMed1976

Nocardial infections in the United States, 1972-1974.

A survey of members of the Infectious Diseases Society of America indicated that nocardial infections are not rare. Probably between 500 and 1,000 cases are recognized in the United States each year, of which 85% are serious pulmonary or systemic infections. Although nocardial infections are usually opportunistic infections in the compromised host, at least 15% of the infections in this series occurred in patients without a definable predisposing condition. Nocardial infections occurred in a random geographic distribution within this country, with affected males outnumbering females by 3:1. Most patients were between the ages of 21 and 50 years; however, the age range was broad. The number and variety of infections caused by Nocardia species other than Nocardia species other than Nocardia asteroides have been underestimated. Between 8.6% and 18.8% of pulmonary-systemic infections in this series were caused by species of Nocardia other than N. asteroides.

Adolescent↗
PubMed1973

Pyridine extraction of nocardial acid fastness.

Several strains of Mycobacterium and Nocardia were stained for acid fastness by using the method of Kinyoun. Duplicate, glutaraldehyde-fixed smears were extracted with fresh pyridine prior to staining. The results clearly demonstrated that both in vivo- and in vitro-grown Nocardia differed from Mycobacterium by becoming nonacid fast after extraction. This pyridine extraction of nocardial acid fastness should have diagnostic and taxonomic application.

Acids↗