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Biomedical subjects

H C Tseng

Publications and source records attributed to H C Tseng.

At least 19 recordsLinked to original sources
PubMed1998

Natural methylamine osmolytes, trimethylamine N-oxide and betaine, increase tau-induced polymerization of microtubules.

The natural osmolyte trimethylamine N-oxide (TMAO) at 200 mM increases the extent and the rate of formation of polymerized microtubule (MT) complex caused by tau. TMAO at this concentration has no effect on tubulin alone. Urea at 200 mM blocks tubulin assembly caused by tau, but this inhibition can be reversed by an equal amount of TMAO. Besides TMAO, betaine, another natural osmolyte, was found to have the same effects on MT as TMAO. On the contrary, glycerol (a carbohydrate osmolyte) and glycine (an amino acid osmolyte) do not increase tau-induced MT assembly. The mechanism by which TMAO and betaine enhance tau's effectiveness is not known, but physical studies suggest that the secondary structure of tau is not appreciably changed by 200 mM TMAO. This is the first report showing that natural osmolytes, TMAO and betaine, at a near physiological concentration are able to stimulate tau-induced tubulin assembly.

Animals↗
PubMed1997

The endocrine secretion of human insulin and growth hormone by exocrine glands of the gastrointestinal tract.

The exocrine pancreas, liver, and submandibular glands of the rat were used to express and secrete two exogenous, human protein hormones (growth hormone and insulin) into blood at physiological concentrations. Transfection, expression, and secretion were achieved by the in vivo retrograde injection of plasmid DNA into the secretory ducts of these glands. Pancreatic acinar cells secreted physiological concentrations of growth hormone into the circulation, and its secretion was enhanced by cholinergic stimulation. A human insulin gene was engineered to allow normal processing of insulin in non-beta cells. With this gene, the secretion of human insulin by the exocrine pancreas normalized elevated blood glucose levels in diabetic rats. These in vivo observations demonstrate the utility of retrograde ductal administration of naked DNA into exocrine organs as a novel method for the regulated systemic delivery of protein-based pharmaceuticals.

Animals↗
PubMed1996

Comparison of stored and secreted rat pancreatic digestive enzymes by mass spectrometry: alpha-amylase.

As part of a continuing effort to better understand the mechanisms of protein secretion, we compared the mass of pancreatic digestive enzymes, in resting and stimulated states, both in secretion and in the zymogen granule to determine whether their secretion is accompanied by chemical modification. Mass spectra were obtained applying the electrospray method on samples separated by reverse-phase HPLC. We report here our results for alpha-amylase (1,4-alpha-D-glucan glucanohydrolase EC 3.2.1.1). The data illustrate structural differences between states and compartments for this enzyme. Multiple isozymes were identified from the mass spectra, varying roughly from 52 to 60 kDa. On the basis of mass comparisons, not all of the products seen in the zymogen granule were found in secretion, nor were all secreted isoforms in the granule. Stimulation of protein secretion with a cholinergic agonist, led to time-dependent changes in the number and masses of isoforms in secretion, leaving only one of five resolvable forms in the granule. Only one form, 55.5 kDa, was found in all samples, granule and secretion. In addition to these differences, microheterogeneities of 400 Da or less were observed. The data suggest the differential or non-parallel release of different amylase forms and their chemical modification during the secretion process. As such, release appears to involve a third, intermediate compartment, between zymogen granule to ductal space, such as the cytoplasm, in which chemical modification takes place.

Animals↗
PubMed1995

Characterization of the manganese-resistant mutants derived from Vibrio parahaemolyticus.

The virulence and some related factors of Vibrio parahaemolyticus are regulated by the level of iron. In this study, five Mn-resistant mutants were selected after N-methyl-N'-nitrosoguanidine treatment and two transfers in medium containing high levels of manganese chloride. Production of siderophores and the 77-kDa iron-regulated outer-membrane protein and the bacterial growth in these Mn-resistants were deregulated, as compared with the wild-type strain. In addition, the regulation of these phenomena was partially or completely restored by the introduction of Escherichia coli fur gene. Also, the total cellular protein profiles of the wild-type and mutants showed that production of some proteins were positively or negatively regulated by iron, and expression of some of these proteins remained unaffected in these mutants. These results suggested the presence of a complicated iron regulation system, similar to the Fur system of E. coli, in this pathogen.

Bacterial Outer Membrane Proteins↗
PubMed1994

Selective determination of adenine-containing compounds by capillary electrophoresis with laser-induced fluorescence detection.

Capillary electrophoresis coupled with laser-induced fluorescence detection provides a selective analysis of mixtures of adenine-containing compounds (adenine, adenosine, cAMP, AMP, ADP, and ATP) that are derivatized using chloroacetaldehyde as a fluorogenic reagent. The components can be detected with linear response over the concentration range of 10(-4) to 10(-9) M, and the detection limit corresponds to 5 to 10 x 10(-19) mol for each nucleotide. Analysis is achieved in less than 10 min with high separation efficiencies (up to 400,000 theoretical plates) and shows good reproducibility for migration times (0.5 to 1.0%) and peak heights (2.8 to 7.7%). To demonstrate the rapid analysis of small-volume samples, the adenine-containing compounds present in an oocyte from Xenopus laevis are determined.

Adenine↗
PubMed1993

Urinary N-acetyl-glucosaminidase excretion and environmental lead exposure. Green Cross Health Service Association Study Group.

To understand the relationship between chronic low-level lead exposure and renal function, residents living nearby a lead battery factory for more than 10 years were selected and entered in this cross-section study. The residents living in the 1st village, within 500 m from the factory, were grouped in group 1; those in the 2nd village, within 1,000-1,500 m, in group 2, and those in the 3rd village, far from any lead-contaminated sources, in group 3. Twenty-four-hour urinary N-acetyl-glucosaminidase (NAG) was detected as early indicator of renal damage, and an ethylenediamine-tetraacetic acid mobilization test was performed to estimate total body lead burden of lead-exposed persons. Blood lead level (BLL) showed a significant difference among the three study groups. The further the distance between the group and the factory, the higher BLL. The results showed a significant high prevalence of abnormal urine NAG excretion in the chronic lead-exposed group, although BLL and body lead burden of these persons were within the 'normal' range. A significant correlation between body lead burden less than 200 micrograms and 24-hour urine NAG excretion and a dose-response relationship between them were found. These observations suggested that lead was the possible cause of abnormal renal tubular function in persons with chronic low-level lead exposure, but this effect became blunt when body lead burden was more than 200 micrograms. The possible explanation may be that high body lead burden from long-term exposure will deplete the kidney of NAG or render it insensitive to the effects of lead exposure.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetylglucosaminidase↗
PubMed1991

A cloned ompR-like gene of Streptomyces lividans 66 suppresses defective melC1, a putative copper-transfer gene.

Expression of tyrosinase in Streptomyces requires functional MelC1 protein, which is postulated to transfer copper to apotyrosinase. We have previously isolated a mutant of Streptomyces lividans, HT32, that phenotypically suppressed mutations in cloned melC1 (H.-C. Tseng and C. W. Chen, in preparation). Plasmid pLUS132, containing an ATG to ATA transition at the initiation codon of melC1, was used for cloning the suppressor gene from HT32. A 1687 bp suppressor DNA was isolated that contained two characteristic Streptomyces coding sequences: a 217-amino-acid open reading frame (cutR) and a truncated open reading frame (cutS) downstream. Subcloning analysis attributed the phenotypic suppression activity to the putative cutR gene from HT32. The putative CutR exhibited similarity to the response regulator OmpR of the osmoregulatory signal-transduction system in Escherichia coli. The truncated CutS resembled, to a lesser degree, the N-terminus of EnvZ, the histidine protein kinase counterpart of OmpR. DNA hybridizing to the cloned cutR-cutS sequence was detected in 16 other Streptomyces species. We postulate that the putative cutR-cutS operon regulates copper metabolism in Streptomyces.

Amino Acid Sequence↗
PubMed1991

Neural regulation of lysozyme secretion from tracheal submucosal glands of ferrets in vivo.

To investigate how central and peripheral nerves affect lysozyme secretion from tracheal submucosal glands in ferrets we injected substance P (20 nmol/kg in 200 microliters) intracisternally or intravenously into anesthetized artificially ventilated ferrets. We collected 3-ml samples from a perfused (3 ml/5 min) segment of trachea in situ during 15 min before and 45 min after injection of substance P. Content of lysozyme, a specific marker of tracheal submucosal gland serous cell secretion in ferrets, was measured spectrophotometrically in each sample. Intracisternal substance P increased peak lysozyme output threefold compared with baseline. This increase was abolished completely by cutting both superior laryngeal nerves (SLN) and was partially inhibited by atropine, phentolamine, or propranolol. Intravenous substance P increased peak lysozyme output 10-fold compared with baseline. This increase was partly abolished by cutting both SLN. We concluded that intracisternal substance P stimulated the central nervous system (CNS) and activated cholinergic, adrenergic, and nonadrenergic noncholinergic secretomotor nerves to tracheal glands and that intravenous substance P increased lysozyme secretion both by acting directly on tracheal glands and indirectly on the CNS to activate secretomotor nerves.

Animals↗
PubMed1990

The melanin operon of Streptomyces antibioticus: expression and use as a marker in gram-negative bacteria.

The melC operon of Streptomyces antibioticus contains two genes, melC1 and melC2, necessary for the production of melanin pigment. We transferred the coding sequence of melC1 and melC2 to Escherichia coli plasmid pMTL23 such that its transcription was under the control of the lac promoter and melC1 was translationally fused to the lacZ alpha fragment. E. coli cultures containing this plasmid, pIF413, produced melanin after overnight incubation on 2YT agar supplemented with 0.1 mM CuCl2, 0.36 mM IPTG (or 0.2% lactose), and 2 mM tyrosine. Erwina carotovora could also be transformed by pIF413 to produce melanin. Two shuttle vectors were constructed: pLUS415 for E. coli and Streptomyces, and pLAF413 for E. coli and Xanthomonas campestris. These vectors confer melanin pigmentation in all the hosts that harbor them. The melC sequence provides the vectors with a convenient cloning marker for insertional or replacement inactivation.

Catechol Oxidase↗
PubMed1988

Structure-activity relationships of chloride-sensitive fluorescent indicators for biological application.

The application of the quinoline derivative 6-methoxy-N-(3-sulfopropyl)quinolinium (SPQ) to the measurement of membrane transport of chloride in biological systems was reported recently (N.P. Illsley and A.S. Verkman (1987) Biochemistry 26, 1215-1219). To understand the structure-activity relationships of compounds with chloride-sensitive fluorescence properties, 19 structural analogs of SPQ having a single quaternized nitrogen heteroatom were synthesized and characterized. The effect of variations in ring structure, length of sulfoalkyl chain, position of ring substituent, and nature of ring substituent were examined. For each compound, the water solubility, octanol:water partition coefficient, absorbance and fluorescence spectra, fluorescence lifetime, and Stern-Volmer constants (Kq) for quenching by a series of anions were measured. All compounds were quenched by chloride, bromide, iodide, and thiocyanate, but not by cations, sulfate, phosphate, nitrate, or by pH (5-8); several compounds were quenched slightly by bicarbonate (Kq = 8-12 M-1). High chloride sensitivity (Kq greater than 50 M-1) required the presence of a quinoline backbone substituted with electron-donating groups such as methyl and methoxy, but did not depend on length of the sulfoalkyl chain or on the position of ring substituents (positions 2-7). All compounds with high chloride sensitivity had fluorescence excitation spectra in the ultraviolet (excitation maximum less than 350 nm) and fluorescence lifetimes greater than 15 ns. These results establish a set of guidelines for synthesis of chloride-sensitive fluorescent indicators tailored for specific biological applications.

Chlorides↗
PubMed1988

Evaluation of parenteral nutrition in the postoperative patient.

A variety of investigators have attempted to improve nitrogen balance during the postoperative period by modifying the composition of the infused nutrient solutions. This study compared the metabolic effects of administering standard amino acid solutions with a solution enriched with branched chain amino acids (BCAA). A prospective, randomized clinical study was performed in patients who had undergone subtotal gastrectomy or hemicolectomy, and subsequently cared for in the metabolic care unit. The patients were selected from specific entry criteria so that two groups of individuals were comparable. All patients underwent operation without complications. The plasma concentrations of valine and leucine were significantly increased (p less than 0.05 and p less than 0.01, respectively) two days after administration of solutions enriched with BCAA and throughout the entire postoperative period. The plasma glutamine concentrations tended to decrease in both groups; no concentration difference occurred between groups. Nitrogen balance tended to be more positive in the group receiving BCAA but there was no significant difference between groups after operation. Urinary excretion of 3-methylhistidine tended to increase postoperatively in both groups, but no difference occurred between groups. However, the urine excretion of isoleucine increased significantly in the patients receiving infusions enriched with BCAA. Both standard balanced amino acid and amino solutions enriched with BCAA were well tolerated in all patients.

Amino Acids, Branched-Chain↗
PubMed1985

Insulinoma. An immunocytochemical and morphologic analysis of 95 cases.

One hundred twenty-seven insulinomas from 95 cases (1 malignant and 94 benign) were studied pathologically. Thirty-six tumors (35 cases) were examined by electron microscopy. Typical beta-cell secretory granules of crystalloid-form cores and/or atypical secretory granules were discerned in all tumors examined. A new type of secretory granule, with high electron-dense crystalloid-form cores and moderate electron-dense granular substance filling the space between the core and the limiting membrane, were observed in two cases. Among 68 insulinomas (67 cases) subjected to immunocytochemical investigations with ten peptide hormones (insulin, glucagon, somatostatin, pancreatic polypeptide (PP), gastrin, motilin, secretin, vasoactive intestinal polypeptide (VIP), gastric inhibitory polypeptide (GIP), and neurotensin), 42 were found to be multihormonal, varying from two to four peptides secreted. The hormones contained were insulin, glucagon, PP, somatostatin, and gastrin in different combinations. One patient had hyperinsulinemia and hypergastrinemia concurrently, and two islet tumors were excised at an interval of 10 months. Both electron microscopy and immunocytochemistry confirmed the presence of beta- and alpha-cells in the first tumor, whereas the second tumor revealed only G-cells by electron microscopy, and G- and beta-cells on immunocytochemical staining. The morphologic and immunocytochemical characteristics of the insulinomas in this series are discussed.

Adenoma, Islet Cell↗
PubMed1984

Regulation of digestion. I. Effects of glucose and lysine on pancreatic secretion.

Previous cell-free studies have shown that glucose selectively elicits the release of amylase from pancreatic zymogen granules, whereas lysine promotes the selective release of trypsinogen. To investigate the expression of these effects in situ, glucose or lysine was injected into the celiac artery of anesthetized rats, either alone or together with the pancreatic secretagogue cholecystokinin, to evaluate their effects on the secretion of amylase and trypsinogen by the pancreas. When given alone neither substance significantly changed the output of either enzyme. However, when given with cholecystokinin, each altered the effect observed with injection of cholecystokinin alone. The injection of glucose resulted in a twofold increase in both peak and total amylase output without significantly increasing trypsinogen secretion, whereas lysine increased both peak and total trypsinogen output by about 50%, leaving amylase output unchanged. These findings provide in situ confirmation for the selective enzyme release produced by glucose and lysine in cell-free studies and suggest that such end products of digestion can regulate the digestive process by modifying the secretory response of the pancreas to cholecystokinin.

Animals↗
PubMed1984

Regulation of digestion. II. Effects of insulin and glucagon on pancreatic secretion.

The endocrine islet-cell hormones insulin and glucagon are secreted at high concentrations into an intrapancreatic portal circulation and have been reported to affect the secretion of digestive enzyme by the exocrine pancreas. In the present experiments, insulin and glucagon were injected into the celiac artery of anesthetized rats to evaluate their effects on the secretion of amylase and trypsinogen by the pancreas. Neither hormone when given alone significantly changed the output of either enzyme. However, when given with the pancreatic secretagogue cholecystokinin, each altered the effect of injection of cholecystokinin. In a dose-dependent fashion insulin increased trypsinogen output without affecting amylase output, whereas glucagon inhibited amylase output and left trypsinogen output unchanged. Thus, both hormones produced a more trypsinogen-dominant pancreatic juice than that observed with cholecystokinin alone, although in different ways. These findings suggest that the endocrine hormones insulin and glucagon may regulate secretion of digestive enzymes by the pancreas by modulating the response to stimuli of overall protein secretion such as cholecystokinin.

Animals↗