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T M Chang

Publications and source records attributed to T M Chang.

At least 73 records · Page 4Linked to original sources
PubMed1997

Cloning and sequence analysis of the gene (eprA1) encoding an extracellular protease from Aeromonas hydrophila.

A gene (eprA1) encoding the extracellular protease of Aeromonas hydrophila AH1 has been cloned and sequenced. Nucleotide sequence analysis of eprA1 predicted a single open reading frame (ORF) of 1038 bp encoding a 346 amino acid (aa) polypeptide, with a potential 21-aa signal peptide. When the eprA1 gene was expressed in minicells, one major band of approx. 37 kDa was identified, while protease activity staining experiments identified a caseinolytic band of approx. 29 kDa determined by SDS-PAGE analysis of the minicells. The deduced C-terminal aa region (Arg-290 to Gly-313) showed sequence homology to partial C-terminal sequences of other zinc metalloproteases including Penicillium citrinum metalloprotease (PlnC), Aspergillus oryzae metalloprotease (NpII), Aspergillus flavus metalloprotease (MepA), and Aspergillus fumigatus metalloprotease (Mep20), particularly with respect to zinc-binding residues.

Aeromonas hydrophila↗
PubMed1997

Cloning and characterization of the gene (empV) encoding extracellular metalloprotease from Vibrio vulnificus.

A gene (empV) encoding the extracellular metalloprotease of Vibrio vulnificus CKM-1 has been cloned and sequenced. When the empV gene was expressed in minicells, a unique peptide of approx. 46 kDa was identified. Protease activity staining experiments also indicated a similar M(r) for the protease. The empV gene product (EmpV) is secreted into the periplasm of Escherichia coli, but not out of it. The crude enzyme prepared from the periplasmic fraction of recombinant E. coli was inhibited by a metalloprotease inhibitor and Zn2+ is essential for its protease activity. Nucleotide sequence analysis predicted a single open reading frame (ORF) of 1818 bp encoding a 606 amino acid (aa) polypeptide, with a potential 24 aa signal peptide followed by a long 'pro' sequence consisting of 172 aa. The N-terminal 20 aa sequence for the elastolytic protease (EepV), purified from the culture supernatant of V. vulnificus ATCC 29307, completely identified the beginning of the predicted mature protein within the deduced aa sequence except for 1 aa residue difference. The estimated pI and molecular weight of the predicted mature protein were 5.86 and 44.3 kDa, respectively, which are nearly identical to those of V. vulnificus L-180 extracellular neutral metalloprotease (EnmV) and of strain ATCC 29307 EepV. The estimated molecular weight also closely matches that determined by SDS-PAGE analysis of the minicells and by protease activity staining. The deduced aa sequence of EmpV showed high homology to V. anguillarum metalloprotease (EmpA), V. cholerae HA/protease (HprC), and V proteolyticus neutral protease (NprP), particularly with respect to active-site residues, zinc-binding residues, and cysteine residues.

Amino Acid Sequence↗
PubMed1997

Mechanism of gastric acid hypersecretion in patients with islet cell tumor without hypergastrinemia: studies in rats.

BACKGROUND & AIMS: A nongastrin acid-stimulating peptide (NGASP) has been found in ulcerogenic pancreatic tumor syndrome without hypergastrinemia. The mechanism of gastric acid hypersecretion by NGASP was investigated in rats. METHODS: In vivo, gastric acid secretion and in vitro histamine release from enterochromaffin-like (ECL) cells in responses to tumor extract (TE) and synthetic human gastrin-17 I or pentagastrin (PG) were studied. Whether the 2 secretagogues potentiate each other was determined. RESULTS: TE dose-dependently stimulated histamine release, which was not blocked by a cholecystokinin (CCK)-B receptor antagonist. When TE was incubated with trypsin, the activity was abolished but was not affected by antibody. However, when rats were pretreated with antigastrin serum or CCK-B receptor antagonist, the acid secretion by TE was virtually abolished. The dose response of acid secretion to TE in the rats receiving PG in a threshold dose was significantly greater than that achieved by TE alone. Similarly, the dose response to PG combined with a threshold dose of TE was significantly greater than that produced by PG alone. CONCLUSIONS: NGASP stimulates histamine release from ECL cells, but the release is not mediated via CCK-B/gastrin receptor. NGASP and gastrin may potentiate each other to produce acid hypersecretion in ulcerogenic pancreatic tumor syndrome.

Adenoma, Islet Cell↗
PubMed1997

Cloning and sequence analysis of a novel hemolysin gene (vllY) from Vibrio vulnificus.

A gene (vllY) encoding a novel hemolysin of Vibrio vulnificus CKM-1 has been cloned and sequenced. When the vllY gene was expressed in minicells, a unique peptide of approximately 40 kDa was identified. Subcellular fractionation of Escherichia coli cells carrying the vllY gene indicated that the VllY protein was distributed in both the cytoplasmic and the periplasmic fractions, with the notable ability to appear in the latter compartment. Nucleotide sequence analysis predicted a single open reading frame of 1,071 bp encoding a 357-amino acid polypeptide with an estimated pI of 5.02. The deduced amino acid sequence of VllY showed high similarity to the sequence of legiolysin, responsible for hemolysis, pigment production, and fluorescence in Legionella pneumophila. The enzyme also exhibited sequence homology to the MelA protein sequence of Shewanella colwelliana and the sequences of 4-hydroxyphenylpyruvate dioxygenase family proteins from various organisms. PCR screening and Southern blotting of V. vulnificus strains revealed that all of the 41 V. vulnificus clinical isolates contained vllY-like genes.

Amino Acid Sequence↗
PubMed1997

Kinetic aspects of polyelectrolyte adsorption: adsorption of chitin derivatives onto liposomes as a model system.

Carboxymethylchitin (CMC) and Carboxymethyl/Glycolchitin (CO) have been adsorbed onto liposomes at physiological ionic strength (I) and pH using phosphate buffer saline (PBS, I = 154 mM, pH = 7.4). Adsorption isotherms at different polymer weight average molecular weights (Mw), for Positive ([+]) or high surface affinity liposomes (DSPC:CHOL:DMTAP, 5:4:1), and Neutral ([N]) or low surface affinity liposomes (DSPC:CHOL, 1:1), have been obtained at T = 25 degrees C. For all CMCs, the adsorbed amount increases with polymer concentration ([P]zero) and no true plateau is observed. The CMC Mw = 4.19 x 10(5) adsorbed on a positive surface fits Langmuir kinetics with maximal coverage gamma max = 325.4 micrograms/mg (polymer/lipid), and inverse of equilibrium constant K* = 4.743 x 10(-4) mg/ml. For all isotherms the predicted amount of polymer needed to coat the entire surface based on the Radius of Gyration (Rg) is inferior to the amount adsorbed. This fact in conjunction with the dependence of gamma max on the number of adsorption shots, suggests that the adsorption is not thermodynamically but kinetically controlled.

Adsorption↗
PubMed1997

Effect of cisapride and renzapride on gastrointestinal motility and plasma motilin concentration in dogs.

The effects of cisapride and renzapride (BRL 24924), on plasma concentration of motilin and gastroduodenal motility were studied in seven dogs with implanted force transducers in the antrum and duodenum. In the interdigestive state, the i.v. administration of cisapride (5 mg) or renzapride (5 mg) administered in phase I resulted in a prompt and marked increase in plasma motilin concentration and in gastroduodenal motility. Mean plasma motilin levels during the first 30 min after cisapride and after renzapride injection were 85.0 +/- 6.5 (+/- S.E.) and 96.1 +/- 6.3 pM., respectively. These values were significantly greater (P < .001) than those for the corresponding time period of the control cycle, 52.2 +/- 5.6 and 57.4 +/- 5.3 pM (mean phase III level, 120 +/- 8.1 pM), respectively. The increases in the motilin level after cisapride or renzapride coincided with significant increases in contractile activities of the antrum to 43.2 +/- 5.3% and 44.9 +/- 4.6% and of the duodenum to 28.4 +/- 3.1% and 34.2 +/- 2.2% of phase III activity (100%) from that in the corresponding control period, 0.7 +/- 0.4% and 0.2 +/- 0.1%, respectively. The changes in both plasma motilin and motility in response to the two drugs were abolished completely by the i.v. administration of atropine. The drugs also enhanced the meal-induced contractile activities of the antrum as well as the duodenum but failed to influence the postprandial plasma motilin concentration. We conclude that cisapride and renzapride have similar effects on plasma motilin and gastroduodenal motility: 1) the two drugs increase plasma motilin levels and stimulate gastroduodenal motility in the interdigestive state, and 2) in the digestive state, both drugs enhance motility without influencing the plasma motilin levels.

Animals↗
PubMed1997

Mouse skin damage caused by a recombinant extracellular metalloprotease from Vibrio vulnificus and by V. vulnificus infection.

Vibrio vulnificus wound infection is characterized by an intense acute cellulitis which spreads along the subcutaneous tissue with severe tissue destruction. Toxins produced by the bacteria appear to be involved in the pathogenesis of skin necrosis and to facilitate microbial dissemination in vivo. We report microscopic studies of mouse skin damage caused by a single intradermal injection of V. vulnificus or of an extracellular metalloprotease isolated from this organism. The gene encoding this metalloprotease has been cloned and expressed in Escherichia coli. The crude metalloprotease extract obtained from the periplasmic fraction by osmotic shock was used in this study. Intradermal injection of this preparation resulted in marked edematous changes of the skin, which extended throughout the dermis and subcutaneous tissues and into adjacent muscles. The collagen and elastic fibers degenerated and some muscle cells even appeared necrotic. Blood vessels became congested, with moderate perivascular neutrophil and mononuclear cell infiltration. There was no evidence of necrotic changes in the blood vessel itself. These results show that the damage to the connective tissues caused by V. vulnificus is at least partly attributable to its protease. Therefore, the protease seems to be an important virulence factor of this microbe.

Animals↗
PubMed1997

Allelic loss at BRCA1, BRCA2, and adjacent loci in relation to TP53 abnormality in breast cancer.

Cells with abnormal TP53 lose cell cycle checkpoints, resulting in genomic instability and neoplastic transformation. However, the evidence linking the tumor-specific targets of genomic alteration to an abnormal TP53 is limited. The present study tested the hypothesis that TP53 abnormalities are correlated with an increased frequency of deletion of breast cancer susceptibility loci (17q and 13q) in breast carcinomas. Tumors from 90 patients were examined for TP53 abnormality and loss of heterozygosity (LOH) at 11 loci on 17q (17q11.2-21) and 13q (13q12-14), including the loci for BRCA1 and BRCA2. A higher frequency of LOH was consistently found at 17q or 13q loci in tumors with an abnormal TP53. The increased LOH in relation to TP53 abnormality was statistically significant at the BRCA1, D17S588, and D13S267 loci (P < 0.05) but not at the locus for BRCA2 (P = 0.64). These observations imply a possible link between an abnormal TP53 and specific genomic deletions of breast cancer susceptibility loci, which may provide clues to the role of TP53 during breast tumorigenesis.

Adult↗
PubMed1996

Microencapsulated genetically engineered live E. coli DH5 cells administered orally to maintain normal plasma urea level in uremic rats.

Safety concerns about introducing genetically engineered cells into the body have prevented their use in medical treatments. To solve this problem, we prepared polymeric membrane artificial cells (semipermeable microcapsules) containing genetically engineered live cells from the bacteria Escherichia coli DH5. When given orally, the cells remain at all times in the microcapsules and are finally excreted in the stool. During their passage through the intestine, small molecules like urea diffuse rapidly into the microcapsules and are acted on by the genetically engineered cells. This lowers the high plasma urea level to normal in uremic rats with induced kidney failure, and has exciting implications for the use of this and many other types of genetically engineered cells in a number of medical applications.

Administration, Oral↗
PubMed1996

Pituitary adenylate cyclase-activating polypeptide stimulates cholecystokinin secretion in STC-1 cells.

Secretion of cholecystokinin (CCK) from the endocrine cells of small intestinal mucosa and the murine intestinal tumor cell line STC-1 is known to involve both adenosine 3',5'-cyclic monophosphate (cAMP)-and Ca(2+)-dependent signal transduction pathways. However, the endogenous stimulant(s) that acts through the cAMP-dependent cascade has not been identified. We determined the effect of pituitary adenylate cyclase-activating polypeptide (PACAP) on CCK secretion and cAMP production and its interaction with other CCK secretagogues in STC-1 cells. At concentrations > 10 nM, PACAP-27 stimulated the release of large intestinal CCK from STC-1 cells in a time- and dose-dependent manner. The stimulatory effect of PACAP-27 was enhanced by the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (IBMX). PACAP-27, PACAP-38, and vasoactive intestinal polypeptide (VIP), with or without IBMX, were equally effective and potent to elicit CCK release with similar half-maximal doses and maximal levels of stimulation. Both forms of PACAP and VIP stimulated a transient but not significant increase in the cellular cAMP level. In the presence of IBMX, all three peptides increased significantly the cellular cAMP level between 2 and 5 min, but PACAP produced a two times higher level than VIP. The stimulatory effect of PACAP-27 on CCK release was also potentiated by bombesin and KCl but without a synergistic production of cAMP. With or without IBMX, PACAP-27-stimulated CCK secretion was not affected by the Ca2+ channel blocker diltiazem (1 microM), the cell-permeable Ca2+ chelator 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid acetoxymethyl ester (BAPTA-AM; 25 microM), or by downregulation of protein kinase C. The stimulatory effects of KCl and bombesin were either reduced or abolished by these treatments. The synergistic effect of bombesin with PACAP was abolished by diltiazem and BAPTA-AM but not by downregulation of protein kinase C, whereas KCl remained synergistic with PACAP after these treatments. Taken together, these results indicate that PACAP may be a neuromodulator of CCK secretion that acts through activation of adenylate cyclase and may function as a coregulator with other CCK secretagogues that are known to increase intracellular Ca2+ concentration.

Animals↗
PubMed1996

Inhibition of acid secretion by electrical acupuncture is mediated via beta-endorphin and somatostatin.

Electroacupuncture (EAP) was shown to inhibit basal gastric acid secretion in dogs and sham feeding-stimulated acid secretion in humans. However, its effect on a meal-stimulated acid secretion in dogs and the mechanisms involved remain unclear. In five dogs prepared with gastric cannulas, gastric acid secretion was determined by a dye-dilution technique for 60 min after intragastric administration of 200 ml of 4% mixed amino acid meal in six different experiments: study 1, no acupuncture; study 2, sham acupuncture (SAP); study 3, EAP; study 4, EAP plus naloxone; study 5, naloxone alone; and study 6, intravenous infusion of somatostatin (SS) and vasoactive intestinal peptide (VIP) at doses of 0.5 and 1.0 micrograms.kg-1.h-1, respectively. EAP was performed on three different points including Pishu, ZusanLi, and Neiguan. Biphasic electrical pulse (25-100 Hz, 12-16 mA) was applied continuously via needles for 75 min starting 15 min before meal. SAP on nonacupoints in hind- and forelegs was performed with the same electrical pulse. Plasma SS, VIP, beta-endorphin, and gastrin were determined by specific radioimmunoassays. EAP significantly inhibited acid secretion (75%; P < 0.01), which coincided with significant increases in plasma SS, VIP, and beta-endorphin and a significant decrease in plasma gastrin. Naloxone completely reversed EAP-induced inhibition of acid secretion and changes in plasma concentration of peptides. SAP also significantly suppressed acid output (30%; P < 0.05), with a modest but significant increase in plasma beta-endorphin. However, the inhibition by EAP on the acid output was significantly greater than that by SAP (P < 0.01). Furthermore, exogenous SS (0.5 microgram.kg-1.h-1) significantly inhibited acid output (78%), whereas VIP failed to inhibit gastric acid secretion. We conclude that, in dogs, EAP significantly inhibits meal-stimulated acid secretion. This acid inhibition is mediated by the release of beta-endorphin and somatostatin, and an endogenous opiate or opiates appear to play an important role in the release of SS, VIP, and beta-endorphin.

Acupuncture Therapy↗
PubMed1996

Submicron polymer membrane hemoglobin nanocapsules as potential blood substitutes: preparation and characterization.

Biodegradable polymer membrane nanocapsules containing hemoglobin have been prepared with phase separation and polymer precipitation methods. We have used different polymers including polylactic acid and polyisobutyl-cyanoacrylate. The mean diameter of the hemoglobin nanocapsules can be as low as 0.07 mu and up to 0.39 mu dependent on the method of preparation. The concentration of the hemoglobin in the suspension can be up to 11 gm/dl, as compared to 15 gm/dl in whole blood. The viscosity of the preparation is slightly lower than that of human blood. In vitro tests show that P50 of these nanocapsules containing bovine hemoglobin is identical to free bovine hemoglobin solution. The surface of the nanocapsules can be modified for example with polyethylene glycol(PEG) or with PEG 2000 PE. Preliminary in vivo tests show that surface modified nanocapsules containing hemoglobin can survive longer in the circulation.

Animals↗
PubMed1996

Microencapsulated genetically engineered E. coli DH5 cells for plasma urea and ammonia removal based on: 1. Column bioreactor and 2. Oral administration in uremic rats.

We report a novel approach for plasma urea and ammonia removal using artificial cells microencapsulated genetically engineered bacteria E. coli DH5 cells. This has been evaluated for use in a column bioreactor for removing plasma urea and ammonia. It has also been evaluated in uremic rats for urea removal by oral administration. In 30 minutes, microencapsulated E. coli DH5 in a column bioreactor in-vitro lowered plasma urea to 10.47 +/- 3.45 mg/dl from 45.85 +/- 2.98 mg/dl and lowered plasma ammonia concentration to 46.00 +/- 4.00 microM from 679 +/- 32 microM/1. The efficiency of this bioreactor for plasma urea and ammonia removal is much higher than any other available methods. Initial plasma urea and ammonia concentration does not affect the plasma urea and ammonia removal efficiency of the column bioreactor. In in-vivo studies of oral administration to uremic rats, both free and encapsulated bacteria both lowered systemic urea from the initial 52.08 (S.D.2.37) mg/dl to 10.58 (S.D.0.85) mg/dl. Unlike free bacteria, microencapsulated bacteria was not retained in the intestine.

Administration, Oral↗
PubMed1996

The importance of standardization of carboxymethylchitin concentration by the dye-binding capacity of alcian blue before adsorption on liposomes.

A new method based on the measurement of the relative dye-binding capacity of Alcian Blue to carboxymethylchitin (CMC) at various molecular weights (MW) has been developed to facilitate the standardization of the initial polyelectrolyte concentration. In the absence of standardization, non-reproducible adsorption patterns are encountered during the adsorption of the MW CMC on neutral and positively charged liposomes. This method is sensitive down to a concentration of 5 mu g/ml of polymer in water. Static Light Scattering (SLS) measurements are used to obtain the weight average molecular weight (Mw) and the size of the polyelectrolyte (Rg) and overlap concentrations (c*). The Mws are then used to determine the constants K and a of the Mark-Houwink equation which are 1.65 x 10(-2) dl/g and 0.4701, respectively, evaluated at kappa = 0.154 M, pH = 7.4 and T = 25 degrees C. The critical electrolyte concentration decreases with molecular weight for Mws ranging from 5.0 x 10(4)-1.2 x 10(6). The dye-binding capacity changes with the molecular weight distribution of the polyelectrolyte demonstrating the sensitivity of this technique to polydispersity.

Adsorption↗
PubMed1996

A case-control study of parity, age at first full-term pregnancy, breast feeding and breast cancer in Taiwanese women.

A hospital-based case-control study was designed herein to investigate the relationship between parity, breast feeding and breast cancer in Taiwan. Reproductive histories of patients with breast cancer confirmed by either pathology or cytology were assessed by questionnaire and compared with age-matched healthy controls. One hundred and fourteen cases and 228 age-matched controls were recruited for this study. According to those results, women having had more than three full-term pregnancies, age at first full-term pregnancy younger than 30, and breast feeding for more than 3 years displayed significantly protective effects against breast cancer. Following adjustment for ethnicity and menopausal status, women with more than three full-term pregnancies and younger than 30 years old at first full-term pregnancy had a decreased risk of breast cancer. However, duration of breast feeding was not related to breast cancer, after adjustment for ethnicity and menopausal status. The effect of number of full-term pregnancies on the risk of breast cancer was found to be independent of the effect of age at first full-term pregnancy. Moreover, the effect of age at first full-term pregnancy and number of full-term pregnancies was also independent of the effect of breast feeding.

Abortion, Spontaneous↗