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Biomedical subjects

J Chou

Publications and source records attributed to J Chou.

At least 91 records · Page 5Linked to original sources
PubMed1983

The effect of ingestion of meat on hepatic extraction of insulin and glucagon and hepatic glucose output in conscious dogs.

The effect of ingestion of protein on hepatic extraction of insulin and glucagon and hepatic glucose output were investigated in conscious dogs. The ingestion of meat stimulated both insulin and glucagon secretion but the glucagon response was much more rapid and greater than that of insulin. Secretion of glucagon demonstrated a biphasic pattern while insulin release was monophasic. The fractional hepatic extraction of glucagon increased gradually from the basal value of 15 +/- 3% to a peak of 36 +/- 5% at 90 min, and that of insulin increased from the basal level of 41 +/- 2% to 54 +/- 4% at 45 and 60 min. The increased hepatic extraction of glucagon and insulin after meat ingestion may be explained by neural or hormonal signals from the gut. The blood glucose and hepatic glucose output did not increase significantly despite the significant decrease of the portal vein insulin to glucagon molar ratio as well as the significant decrease of the molar ratio of the hepatic uptake of these hormones. The absence of greater hepatic glucose production despite the augmented glucagon secretion and decreased portal vein insulin to glucagon molar ratio could reflect down regulation by glucagon.

Animals↗
PubMed1983

Substance P stimulates the release of Met5-enkephalin-Arg6-Phe7 and Met5-enkephalin from rat spinal cord.

Substance P (10(-6) M) injected in the fluid perfusing the subarachnoidal spaces of the spinal cord can release met5-enkephalin (YGGFM) and met5-enkephalin-Arg6-Phe7 (YGGFMRF) from spinal cord. This effect is Ca2+ dependent and can be blocked by the substance P antagonist, D-pro2, -D Trp7,9-substance P. The release of YGGFM and YGGFMRF was not observed when substance P was replaced by substance P1-9. These results suggest that substance P receptors are involved in triggering the release of enkephalins.

Animals↗
PubMed1983

New versatile plasmid vectors for expression of hybrid proteins coded by a cloned gene fused to lacZ gene sequences encoding an enzymatically active carboxy-terminal portion of beta-galactosidase.

A new class of plasmid cloning vectors has been constructed with cleavage sites in a variety of translational reading phases of the promotorless lacZ gene. Fused hybrid proteins can be produced by these vectors by cloning DNA fragments containing the promoter, translation initiation site, and the amino terminal portion of a gene, all with proper orientation, into the correct translational reading frame of the lacZ gene. Enzymatically active hybrid-beta-galactosidase proteins are formed, which have amino-terminal amino acids encoded by the cloned gene segment. Another class of these vectors retains an active lac promoter and lacZ translation-initiation region, which can direct hybrid protein synthesis from DNA fragments that do not have gene initiation regions. These vectors allow transcription from the lacZ initiation region to proceed across, or to stop and restart within, an inserted fragment into the essential part of the beta-galactosidase gene. Also described is a small lacZ gene fragment (cartridge), without a plasmid replicon and without any other lac genes, which can be inserted directly into other genes to form hybrid protein fusions. Polyrestriction site sequences were easily moved into some of these vectors by incorporating drug-resistance genes that serve as markers for the selection and detection of these sequences; those markers can be easily removed afterwards.

Bacterial Proteins↗
PubMed1983

Expression of a preproinsulin-beta-galactosidase gene fusion in mammalian cells.

As an approach to the study of mammalian gene expression, the promoters and translation initiation regions of the rat preproinsulin II and the simian virus 40 early genes were fused to the structural gene of Escherichia coli beta-galactosidase, a sensitive probe for gene expression. These fusions were introduced into COS-7 cells, a simian virus 40 large tumor-antigen-producing monkey kidney cell line, where they directed the synthesis of enzymatically active hybrid beta-galactosidase proteins. Conditions for transfection were varied to optimize the expression of beta-galactosidase activity in the transfected cells. The pH optimum of this activity was found to be 7.0, the same as that of native E. coli beta-galactosidase and distinct from the major lysosomal "acid" beta-galactosidase. The fused preproinsulin-beta-galactosidase was further characterized by gel electrophoresis of nondenatured cell extracts stained by a fluorogenic substrate and by immunoprecipitation and gel electrophoresis of 3H-labeled cell proteins. These results all indicate that fully active tetrameric beta-galactosidase hybrids can be produced in mammalian cells. The expression of preproinsulin-beta-galactosidase activity was measured in the presence of high glucose, insulin, dexamethasone, or epidermal growth factor but no regulatory changes were observed.

Animals↗
PubMed1983

Absence of hepatic extraction of pancreatic polypeptide in conscious dogs.

In 15 conscious dogs basal portal vein pancreatic polypeptide (PP) (306 +/- 8 pg/ml) exceeded both hepatic vein PP (255 +/- 8 pg/ml) and arterial PP (244 +/- 9 pg/ml) and increased rapidly 10 min after oral glucose administration. In contrast to oral glucose, intraportal glucose infusion decreased PP levels from 45 min until the end of the infusion. Meat ingestion rapidly and promptly increased PP. During the basal state, hepatic extraction of total immunoreactive PP was 10 +/- 4%, not significantly different from zero, of the 150 +/- 14 ng/min presented to the liver. Fractional hepatic extraction of PP did not change after oral glucose, meat ingestion, or intraportal glucose infusion. Chromatographic analysis showed at least four different components of immunoreactive PP. The first peak eluted with the void volume, the second peak between the void volume and authentic PP, and the third peak coincided with authentic PP. When present, the fourth peak eluted after authentic PP. More than half of the immunoreactive PP in the basal state eluted with authentic PP, whereas about one-third was found in the second peak. The marked alterations in PP after meat, oral glucose, or intraportal glucose and atropine reflected primarily changes in the third peak. The fractional hepatic extraction of all the components was very similar.

Animals↗
PubMed1983

Differential effects of oral, peripheral intravenous, and intraportal glucose on hepatic glucose uptake and insulin and glucagon extraction in conscious dogs.

The effect of equal (1.1 +/- 0.1 g/kg body wt) amounts of glucose administered orally, or by peripheral intravenous or intraportal infusion on hepatic glucose uptake and fractional hepatic extraction of insulin and glucagon was studied in conscious dogs with chronically implanted Doppler flow probes on the portal vein and hepatic artery and catheters in the portal vein, hepatic vein, carotid artery, and superior mesenteric vein. Portal vein and hepatic vein plasma flow increased only after oral glucose administration. Arterial plasma glucose increased equally to 150-160 mg/100 ml after all three routes of glucose administration. Portal vein glucose was similar after oral (195 +/- 15 mg/100 ml) and intraportal glucose infusion (215 +/- 11 mg/100 ml) and significantly higher than after peripheral intravenous glucose. Hepatic glucose uptake after oral (68 +/- 4%) and intraportal glucose administration (65 +/- 7%) significantly exceeded that after peripheral intravenous glucose infusion (23 +/- 5%). The amount of insulin above basal presented to the liver during the 180 min after oral glucose was 7.6 +/- 1.3 U, 4.3 +/- 0.6 U after intraportal glucose, and 4.1 +/- 0.6 U after peripheral intravenous glucose. Hepatic extraction of insulin increased significantly after oral glucose (42 +/- 3 to 61 +/- 4%), but was unchanged after intraportal and peripheral intravenous glucose administration. When the portal vein glucose levels achieved during peripheral intravenous glucose infusion for 90 min were maintained by a subsequent 90-min intraportal glucose infusion, hepatic glucose uptake was significantly greater during the intraportal glucose infusion. Glucagon secretion was suppressed equally after oral glucose, intraportal glucose, and peripheral intravenous glucose administration; fractional hepatic extraction of that hormone, which was significantly less than that of insulin, was unchanged. These results indicate that hepatic glucose uptake is significantly greater after oral and intraportal glucose administration than after peripheral intravenous glucose infusion. This difference is not simply related to the amount of glucose or insulin presented to the liver and the increased hepatic glucose uptake did not depend solely upon the augmented fractional hepatic extraction of insulin. Hepatic extraction of insulin and hepatic glucose uptake appear to be regulated independently.

Administration, Oral↗
PubMed1982

Discovery rate of dysplasia and carcinoma of the uterine cervix in an urban medical center serving patients at high risk.

File analysis based on 98,970 Pap tests on 58,053 patients from the Martland Medical Center of CMDNJ and its clinics was performed. Discovery rates, period prevalence and incidence rates were calculated for categories of mild to moderate dysplasia through invasive carcinoma. An incidence rate of 27/100,000 for invasive carcinoma was obtained, which is lower than the national average. Period prevalence and incidence rates of dysplasias are both high and similar. This indicates that epidemiologic parameters may need to be studied further. The mean age for the mild to moderate dysplasia was 25.7 years, for moderate to severe dysplasia, 29.29 years and for CIS, 33.25 years. These data may imply that younger women, especially in the urban areas, are at much higher risk than previously expected.

Adolescent↗
PubMed1982

Overproduction of the Tn3 transposition protein and its role in DNA transposition.

Five single base pair mutations that increase expression of the tnpA (transposase) gene of the Tn3 transposon approximately 30-fold, but which still allow the gene to be regulated, have been isolated by using a generally applicable procedure that involves distally linked lac gene fusions. The mutations, which are all located in a region controlling initiation of translation of the tnpA gene, do not affect normal repression of tnpA by the tnpR gene product, and yield up to a 9000-fold increase in tnpA protein production when combined with a tnpR mutation and placed on a high copy number plasmid. The mutation yielding the highest expression level was separated from the fused lac gene segment by homologous recombination and was found to increase the rate of transposition without altering the nature of the transposition product; in cells defective in both the E. coli recA gene and the tnpR gene of tn3, cointegrate transposition-intermediate structures occur with the overproducing--as well as with the wild-type--tnpA gene. In the presence of a functional Tn3 tnpR gene or the related transposon delta gamma, such cointegrate structures are resolved into the final products of transposition.

Bacterial Proteins↗
PubMed1982

Amino-terminal sequence of the Tn3 transposase protein.

The amino-terminal sequence of the Tn3 transposase protein was determined to be Pro-Val-Asp-Phe-Leu-Thr-Thr-Glu-Gln-Val-Glu-Ser.... This was determined both from an active transposase protein purified from a transposase overproducing mutant strain and from a hybrid transposase-beta-galactosidase fusion protein. The amino acid sequence corresponded to the DNA sequence of the transposase gene beginning at an ATG initiation codon, as previously predicted from the analysis of transposase-beta-galactosidase gene fusions.

Amino Acid Sequence↗
PubMed1981

Purification of the Tn3 transposase and analysis of its binding to DNA.

The transposase encoded by the tnpA gene of Tn3 is a protein specifically required for Tn3 transposition. We have purified it to homogeneity from an Escherichia coli strain containing a mutant Tn3 that overproduces transposase. About a 10-fold additional increase in transposase resulted from growth into stationary phase. The initial purification was guided by the presence of a protein band with the electrophoretic mobility of the tnpA gene product. The identity of the purified protein was proven by the agreement of five NH2-terminal amino acids with the nucleotide sequence of the A gene; this, in turn, fixed the initiation codon. Transposase formed large aggregates in the absence of Mg2+ at salt concentrations of 0.1 M or less. In nonaggregating conditions, it had 1 or 2 copies of 113,000-dalton protomers. Subsequent purifications exploited the rapid and simple assay of transposase-mediated retention of labeled DNA to a nitrocellulose filter. Transposase bound tightly to single-stranded DNA but weakly to intact duplex DNA. DNA binding did not require Mg2+ and was highly salt-resistant. Binding did not require specific sequences, because poly(dT) was as good a substrate as phi X174 viral DNA. The high DNA binding constant of 4 X 10(9) M-1 is about the same as for some single-stranded DNA binding proteins.

Binding, Competitive↗
PubMed1980

In vitro gene fusions that join an enzymatically active beta-galactosidase segment to amino-terminal fragments of exogenous proteins: Escherichia coli plasmid vectors for the detection and cloning of translational initiation signals.

We report the construction and use of a series of plasmid vectors suitable for the detection and cloning of translational control signals and 5' coding sequences of exogenously derived genes. In these plasmids, the first eight codons of the amino-terminal end of the lactose operon beta-galactosidase gene, lacZ, were removed, and unique BamHI, EcoRI, and SmaI (XmaI) endonuclease cleavage sites were incorporated adjacent to the eighth codon of lacZ. Introduction of deoxyribonucleic acid fragments containing appropriate regulatory signals and 5' coding sequences into such lac fusion plasmids led to the production of hybrid proteins consisting of the carboxyl-terminal segment of a beta-galactosidase remnant plus a peptide fragment that contained the amino-terminal amino acids encoded by the exogenous deoxyribonucleic acid sequence. These hybrid peptides retained beta-galactosidase enzymatic activity and yielded a Lac+ phenotype. Such hybrid proteins are useful for purifying peptide sequences encoded by exogenous deoxyribonucleic acid fragments and for studies relating the structure and function of specific peptide segments.

Base Sequence↗
PubMed1979

Identification and characterization of a self-regulated repressor of translocation of the Tn3 element.

Gene fusions that bring expression of the lacZ gene under control of transcriptional and trnaslational signals within the transposable element Tn3 have been used to study regulation of Tn3-specified proteins. A gene encoding a 21,355-Mr peptide that represses translocation of Tn3 and acts at the level oquenced; amber, missense, and cis-dominant (operator-constitutive) point mutations in this gene have been isolated and characterized.

Bacterial Proteins↗
PubMed1978

Simple method for estimating glycosylated hemoglobins, and its application to evaluation of diabetic patients.

Recent reports have suggested that determination of glycosylated hemoglobin may serve as a clinical aid for long-term blood glucose control in diabetes mellitus. We describe a simple procedure for measuring it by ion-exchange chromatography. Hemolysates were subjected to Bio-Rex 70 chromatographic separation on small columns. Percentages in the normal group ranged from 4.7 to 8.8% of total hemoglobin; the mean +/- standard error was 6.61 +/- 0.31%. Values in the diabetic group ranged from 6.9 to 17.4%; the mean was 10.83 +/- 0.34. Plasma glucose concentrations after fasting, plotted vs. the percent of glycosylated hemoglobin, revealed a linear relationship at normal or moderately high glucose concentrations. However, the values for glycosylated hemolgobin approached a plateau with grossly higher plasma glucose concentrations after fasting. Our results support the view that, due to its long half-life, the estimation of glycoylated hemoglobin reflects the integrated glucose concentrations to which the erythrocytes have been previously exposed.

Adult↗