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T M Chang

Publications and source records attributed to T M Chang.

At least 163 records · Page 9Linked to original sources
PubMed1990

Conversion of ammonia or urea into essential amino acids, L-leucine, L-valine, and L-isoleucine using artificial cells containing an immobilized multienzyme system and dextran-NAD. L-lactic dehydrogenase for coenzyme recycling.

A multienzyme system consisting of leucine dehydrogenase (EC 1.4.1.9), L-lactic dehydrogenase (EC 1.1.1.27), urease (EC 3.5.1.5), and dextran-NAD+ was microencapsulated within artificial cells. This system could convert ammonia and urea into essential amino acids, L-leucine, L-valine, and L-isoleucine. L-lactate acted as a cosubstrate for the regeneration of dextran-NADH. Greater concentrations of L-lactate favored the higher conversion ratios. The effects of ammonium salts and urea on reaction rate were also studied. The relative reaction rates in ammonium salts solutions were 44.6-78.8% of those in urea solutions. More than 90% of the original activity was retained when artificial cells were kept at 4 degrees C for 6 wk.

Amino Acid Oxidoreductases↗
PubMed1990

Effect of pancreatic proteases on plasma cholecystokinin, secretin, and pancreatic exocrine secretion in response to sodium oleate.

The effect of pancreatic proteases or juice on the sodium oleate-stimulated pancreatic secretion and plasma concentrations of secretin and cholecystokinin in anesthetized rats was investigated. Each rat received sodium oleate in a dose of 0.12 mmol.h-1 via a duodenal tube. Sodium oleate infusion significantly increased pancreatic secretion (volume and protein output) compared with the saline given the control group. The increase in pancreatic secretion paralleled significant elevations of plasma concentrations of secretin and cholecystokinin. To determine a possible role of pancreatic proteases on the responses induced by sodium oleate, saline, chymotrypsin, and trypsin, a combination of chymotrypsin and trypsin or pancreatic juice was infused into the duodenum. The pancreatic secretion was significantly reduced by pancreatic proteases or pancreatic juice, and the reduction paralleled the decreases in plasma concentrations of the two hormones. These agents suppressed both pancreatic secretion and plasma hormone levels in the following order of magnitude: (pancreatic juice or chymotrypsin + trypsin) greater than (trypsin) greater than (chymotrypsin). The reduction of pancreatic secretion by pancreatic proteases was reversed by intravenous administration of secretin and cholecystokinin in physiological doses. It is concluded that negative-feedback regulation of pancreatic secretion is operative in the intestinal phase in rats and that both secretin and cholecystokinin are involved in the regulation.

Animals↗
PubMed1990

An important role of endogenous insulin on exocrine pancreatic secretion in rats.

We have investigated a physiological role of endogenous insulin on exocrine pancreatic secretion stimulated by a liquid meal as well as exogenous secretin and cholecystokinin octapeptide (CCK-8) in conscious rats. Each rat was prepared with a chronic pancreatic fistula and an indwelling catheter in a jugular vein. Oral ingestion of a liquid meal (5 ml) resulted in significant increases in pancreatic secretion, including volume, bicarbonate, and amylase output, in these rats. A rabbit anti-insulin serum (1.0 ml) given intravenously completely blocked the postprandial exocrine pancreatic secretion, whereas a normal rabbit serum did not influence the pancreatic secretion in the same rats. When pancreatic secretion was stimulated by intravenous administration of both secretin and CCK-8 in three different doses, including 0.015, 0.03, and 0.06 clinical unit and microgram.kg-1.h-1, respectively, volume, bicarbonate, and amylase output increased significantly in a dose-dependent manner. This increase in pancreatic secretion was also completely blocked by a rabbit anti-insulin serum, whereas it was not influenced by a normal rabbit serum. The amount of the antiserum employed abolished the postprandial increases in plasma insulin concentration. We conclude that endogenous insulin plays an important role on the regulation of postprandial pancreatic secretion in rats. Furthermore, for the stimulatory action of the two intestinal hormones secretin and CCK-8 on the pancreatic exocrine secretion, endogenous insulin is need.

Amylases↗
PubMed1990

Hormonal mechanism of sodium oleate-stimulated pancreatic secretion in rats.

We have investigated the role of two intestinal hormones, secretin and cholecystokinin (CCK), on the pancreatic exocrine secretion stimulated by sodium oleate in anesthetized rats. Each rat was prepared with a polyethylene tube in the proximal duodenum and ligation of the pylorus. To collect pancreatic juice, the common bile-pancreatic duct was cannulated near the duodenal wall while bile was diverted to the exterior. Intraduodenal infusion of sodium oleate at doses of 0.03, 0.06, 0.12, and 0.24 mmol/h resulted in significant increases in pancreatic secretion including fluid, bicarbonate, and protein output. The increases of the three parameters were dose dependent and were correlated well with the increases in plasma secretin and CCK concentrations. To further clarify their hormonal roles, we have repeated identical experiments under intravenous administration of a rabbit anti-secretin serum (0.1 ml) or CR 1409 (4 mg.kg-1.h-1), a CCK-receptor antagonist, or a combination of both the antiserum and CR 1409. The antiserum significantly suppressed volume flow and bicarbonate secretion with a minor inhibitory effect on protein secretion, whereas a normal rabbit serum did not. CR 1409 significantly suppressed all three parameters. The combined treatment with both the antiserum and CR 1409 almost completely abolished the pancreatic secretion. Atropine given intravenously significantly inhibited the protein output but did not influence volume flow or bicarbonate output in response to sodium oleate. We thus conclude that, in rats, fat-stimulated pancreatic secretion of volume flow and bicarbonate depends entirely on the circulating endogenous secretin and CCK but that the protein output appears to be under control of both hormonal and cholinergic controls.

Animals↗
PubMed1990

Mechanism of acid-induced release of secretin in rats. Presence of a secretin-releasing peptide.

In fasting rats, intraduodenal infusion of dilute hydrochloric acid results in significant increases in both pancreatic exocrine secretion and plasma concentration of secretin. To test the hypothesis that acid-induced release of secretin is mediated by a secretin-releasing factor (S-RF), anesthetized rats were prepared with pyloric ligation, duodenal and jejunal cannulas, and pancreatic duct cannulas. Donor rats were infused intraduodenally with 0.01 N HCl, 0.15 M NaCl, or a combination of 0.01 N HCl and 0.05 N NaHCO3 at 0.3 ml/min for 1.5 h, and the perfusates were collected via jejunal cannulas. The perfusates with pH adjusted to 6.0 were concentrated threefold and infused into the duodena of recipient rats. The concentrate of acid perfusate (CAP) significantly increased both pancreatic volume flow and bicarbonate output and plasma concentration of secretin, whereas concentrates of the saline perfusate (CSP) or the perfusate of a combination of 0.01 N HCl and 0.05 N NaHCO3 (CABP) did not influence pancreatic secretion or plasma concentration of secretin. The increased pancreatic secretion by CAP was attributed to increased circulating secretin because when secretin was immunoneutralized by a rabbit antisecretin serum, CAP-stimulated pancreatic secretion was abolished. The bioactivity of CAP was trypsin-sensitive and heat stable. The active substance in CAP had a molecular weight of less than 5,000 and greater than 1,000, as determined by ultrafiltration and bioassay. In conclusion, dilute HCl releases an S-RF into the upper small intestinal lumen to stimulate release of secretin. This substance, with molecular weight of less than 5,000, is heat stable and trypsin sensitive. Thus, the acid-stimulated release of secretin is mediated by a secretin-releasing peptide in the upper small intestinal lumen.

Animals↗
PubMed1990

Whole complement hemolytic activity (CH50) following infusion of stroma-free and polyhemoglobin solutions in rats.

We developed a method to determine whole complement hemolytic activity (CH50) in rat plasma using a modification of the method of Mayer(1). Plasma CH50 level in 30 male Sprague Dawley rats was 1000 +/- 200 units. Following infusions with human and rat stroma-free hemoglobin or polyhemoglobin solutions, CH50 did not change significantly in rats when compared to albumin infused rats. On the other hand, CH50 significantly decreased in the group receiving membrane stroma and bacterial endotoxins.

Animals↗
PubMed1990

Effects of homologous and heterologous stroma-free hemoglobin and polyhemoglobin on complement activation, leucocytes and platelets.

This is a study of the effects of stroma-free hemoglobin(Hb) and polyhemoglobin on C3 and C3a levels and on blood cell counts. The effects of membrane stroma from red cells and endotoxins on complement activation were also investigated. Plasma samples from rats were incubated with hemoglobin solutions or control solutions. C3 was measured using anti-rat C3 by nephelometric method. C3a was measured using radioimmunoassay. C3 and C3a were also determined in rats receiving hemoglobin solutions, red cell stroma solutions, or a solution containing complement activators (membrane stroma and bacterial endotoxins). There were no significant differences in C3 or C3a levels between the plasma incubated with hemoglobin solutions and the control plasma incubated with saline (P greater than 0.05). Compared with the saline incubated plasma, C3 was significantly lower and C3a significantly higher in the zymosan treated plasma (P less than 0.01). In hemoglobin infused rats, there was no significant difference between pre-versus post-infusion values in C3 and C3a. There was a significant increase in C3a in the rats receiving stroma suspension (P less than 0.05) or stroma and endotoxins (P less than 0.02). Polyhemoglobin and stroma-free hemoglobin did not cause significant changes in total leucocyte, differential, or platelet counts. Our study suggests that purified stroma-free Hb and polyHb do not activate complement, and that C3a is a more sensitive parameter for monitoring complement activation than C3. On the other hand, contaminates in hemoglobin preparations, such as membrane stroma or endotoxins, activate the complement system.

Animals↗
PubMed1990

Chronotropic effects of in vitro perfusion with albumin, stroma-free hemoglobin, and polyhemoglobin solutions.

A negative chronotropic effect of bovine stroma-free hemoglobin was identified in spontaneously contracting myocardiocytes derived from neonatal rats. This model allowed for the evaluation of direct effects of hemoglobin solutions without the influence of hemodynamic reflexes. Significant slowing of myocardiocyte beating rates were observed from 0.8 to 11.6 g.dl-1 stroma-free hemoglobin solution. Polymerization of hemoglobin into soluble crosslinked polyhemoglobin markedly reduced its negative chronotrophic effects by more than 50% at most concentrations assayed. Chronotropic potency (% change in beating rate from baseline per g.dl-1 was significantly higher for stroma-free hemoglobin when compared to polyhemoglobin and albumin solutions. Colloid osmotic potency (% change in beating rate from baseline per mmHg) however, was similar for stroma-free hemoglobin and polyhemoglobin but significantly lower for albumin solutions. This negative chronotropic effect may in part contribute to the transient bradycardia observed following hemoglobin infusion.

Albumins↗
PubMed1990

Towards an artificial cornea: surface modifications of optically clear, oxygen permeable soft contact lens materials by ammonia plasma modification technique for the enhanced attachment and growth of corneal epithelial cells.

The advent of high water content, oxygen permeable contact lens materials has made the intracorneal implants more feasible. A major obstacle encountered is the regrowth of a stable epithelium over the implant. Therefore, ammonia gaseous plasma modification technique was used to modify the surface chemical properties of soft contact lens material such as poly(2-hydroxyethyl methacrylate and methacrylic acid), PHEMA-MAA copolymer, in an attempt to enhance the cell attachment and growth of rabbit corneal epithelial cells.

Ammonia↗
PubMed1990

A screening test for modified hemoglobin blood substitutes before clinical use: based on C3a complement activation in human plasma.

Modified hemoglobin preparations may potentially cause hypersensitivity and anaphylactic reactions, antibody-antigen reactions and other problems. Unfortunately response in animal safety studies may not reflect the same response in human. The next best test before clinical trial in human may the use of human plasma in-vitro. This paper present an in-vitro procedure based on complement activation (C3a) of human plasma. The procedure involves collecting heparinised blood and separating the plasma and freezing the heparinised plasma at -70 degrees C until use. Each 100 lambda of control or test samples is added to 400 lambda of this plasma. This is incubated at 37 degrees C, 60 rpm for 1 hour, then added to EDTA saline to stop the reaction and stored at -70 degrees C until analysed by standard radioimmunoassay for C3a. (C3 measurement is not sensitivity enough). Using the screening test procedure described above, C3a levels (ng/ml) in plasma were: control, 1,980 +/- 280; Zymosan, 20,000; Hemoglobin preparation A, 2,227 +/- 617; Hemoglobin preparation B, 4,967 +/- 153; A 75% + B 25%, 3,967 +/- 270; A 50% + B 50%, 4,553 +/- 517; A 25% + B 75%, 4,920 +/- 430. Hemoglobin preparation A did not cause significant increases in C3a complement activation. Hemoglobin preparation B caused significant increase in C3a complement activation. Serial dilution of Hemoglobin preparation B in Hemoglobin preparation A continued to cause the same degree of C3a complement activation. This is not due to C3 exhaustion because Zymosan resulted in C3a of greater than 20,000ng/ml. This appears to show that this screening test can detect complement activation even at low concentrations of the hemoglobin preparation.(ABSTRACT TRUNCATED AT 250 WORDS)

Blood Substitutes↗
PubMed1990

Conversion of ammonia or urea into essential amino acids, L-leucine, L-valine, and L-isoleucine, using artificial cells containing an immobilized multienzyme system and dextran-NAD+. 2. Yeast alcohol dehydrogenase for coenzyme recycling.

Semipermeable nylon-polyethylenimine artificial cells containing leucine dehydrogenase (EC 1.4.1.9), alcohol dehydrogenase (EC 1.1.1.1), urease (EC 3.5.1.5), and dextran-NAD+ were prepared. Artificial cells could convert ammonia or urea into L-leucine, L-valine, and L-isoleucine. For batch conversion in 20.0 mM of ammonium acetate substrate solutions, in 2 h 0.2 ml of artificial cells could produce 4.48 mumol of L-leucine, 9.98 mumol of L-valine, or 5.96 mumol of L-isoleucine. The corresponding conversion ratios were 22.4, 49.9, and 29.8%. In 20.0 mM of urea substrate solutions, 13.71 mumol of L-leucine, 16.12 mumol of L-valine, or 13.44 mumol of L-isoleucine was produced and the conversion ratios were 68.6, 80.6, and 67.2%. The substrate specificity of leucine dehydrogenase for the reductive amination was determined. Of the three branched-chain amino acids produced, the production rates of L-valine were the highest. The apparent Km values were as follows: 0.32 mM for alpha-ketoisocaproate, 1.63 mM for alpha-ketoisovalerate, and 0.73 mM for Dl-alpha-keto-beta-methyl-n-valerate. The leucine dehydrogenase multienzyme system had a good storage stability. It retained 72.0% of the original activity with artificial cells were stored at 4 degrees C for 6 weeks. The optimum conversion pH and temperature were 8.5-9.0 and 35-40 degrees C. The effects of urea and ammonium salts on conversion rate were also studied. The relative activities in ammonium salts solutions were 45.1-75.9% of those in urea solutions.

Alcohol Dehydrogenase↗
PubMed1990

In vivo effects of stroma-free hemoglobin and polyhemoglobin on coagulation factors in rats.

We studied the effects of rat stroma-free hemoglobin (rSFH), human stroma-free hemoglobin (hSFH), rat polyhemoglobin (rPoly), and human polyhemoglobin (hPoly) on coagulation factors in rats. Albumin and saline infused rats were controls. The infusion volume was 10% of the rat's blood volume. The concentrations of hemoglobin in this study were 7 g/dl. Measurements for prothrombin time (PT) and activated partial thromboplastin time (PTT) were at 5 minutes, 2, 6, 24 and 72 hours after infusion. Factor X, fibrinogen, plasminogen, antithrombin III, and antiplasmin were followed at 24 and 72 hours after infusion. Compared with saline infused rats PT and PTT did not change significantly in those rats infused with Hb preparations. There was a transient increase of PTT from 2 to 24 hours after infusion in albumin infused rats. Factor X, fibrinogen, antithrombin III and antiplasmin showed no significant differences between Hb infused groups and saline infused group. Twenty-four hours and 72 hours after infusion plasminogen decreased in all groups except the albumin infused rats at 24 hours after infusion when compared with normal rat plasma pool. However, there were no significant differences in plasminogen levels between the hemoglobin infused groups and the control saline group. Stroma-free and polyHb solutions (rSFH, hSFH, rPoly and hPoly) did not cause significant changes in prothrombin time and activated partial thromboplastin time in rats. The rats infused with hemoglobin solutions (rSFH, hSFH, rPoly, and hPoly) did not show significant differences in Factor X, fibrinogen, antithrombin III and antiplasmin levels compared with the control group.

Animals↗
PubMed1990

Characterization of L-histidine ammonia-lyase immobilized by microencapsulation in artificial cells: preparation, kinetics, stability, and in vitro depletion of histidine.

L-histidine ammonia-lyase (histidase) was encapsulated within cellulose nitrate artificial cells, and its kinetic parameters were evaluated. Microencapsulated histidase had an apparent activity of histidase in solution. Encapsulation did not alter the Km of histidase. The Km of histidase solution and the Km apparent of micro-encapsulated histidase were both 20 mM. Encapsulation of histidase resulted in increased stability of enzymatic activity at storage temperatures of 4 degrees C and 37 degrees C. At 37 degrees C histidase solution reached 50% of its original activity after 9.5 days of storage, while microencapsulated histidase reached the same level after 15 days. At 4 degrees C histidase solution had 63% of its original activity after 21 days of storage, while encapsulated histidase had 95%. In vitro experiments to evaluate the feasibility of microencapsulated histidase for possible experimental therapy in histidinemia were carried out. These experiments evaluated the effectiveness of encapsulated histidase in depleting histidine. Three different volume ratios of histidase loaded artificial cells to substrate solution were tested. A ratio of 1: 100 allowed 25% histidine depletion after 120 hours. A 1: 50 ratio allowed 35% histidine depletion after 72 hours. A 1: 25 ratio allowed 40% histidine depletion after 24 hours (Int J Artif Organs 1990; 13: 189-95).

Ammonia-Lyases↗
PubMed1989

Diacytosis of asialoglycoprotein in isolated hepatocytes is dependent on the structure of ligand and cellular distribution of the receptors.

Diacytosis, degradation and retention of 125I-labeled asialoorosomucoid (ASOR), its reduced and carboxymethylated N-terminal cyanogen bromide-cleaved fragment (RC-ASCNBr-I) and asialofetuin preloaded into isolated rat or rabbit hepatocytes for various periods of time were compared. In rat hepatocytes preloaded with a saturating concentration (3.10(-8) M) of the ligands, the proportion of the preloaded ligands distributed to degradation and diacytosis was fairly constant during 4 h of preincubation. In addition, a small portion of the preloaded ligands was neither diacytosed nor degraded, but was retained intracellularly. Diacytosis of 125I-ASOR (29%) was greater than that of either 125I-RC-ASCNBr-I (23%) or 125I-asialofetuin (15%). Diacytosis of 125I-ASOR, when preloaded in the presence of 5 microM colchicine, was significantly enhanced by 79% (increasing from 29% to 52%), whereas those of 125I-RC-ASCNBr-I and 125I-asialofetuin were not significantly altered (with average increases of 14% and 19%, respectively). The fraction of the preloaded 125I-asialofetuin (69%) and 125I-RC-ASCNBr-I (68.6%) that was degraded was slightly higher than that of 125I-ASOR (64%) and all was decreased by colchicine. The fraction of all three ligands retained by the cells was increased 2- to 4-fold by colchicine. The extents of retention of 125I-asialofetuin and 125I-ASCNBr-I were greater than that of 125I-ASOR, particularly after preloaded for more than 2 h. Preloading of the cells with ligands at a non-saturating concentration (6.5.10(-10) M) did not change these patterns of ligand distribution. Conjugation of diphtheria toxin fragment A (DTA) to ASOR or RC-ASCNBr-I also did not significantly alter the pattern of ligand distribution. In rabbit hepatocytes containing more asialoglycoprotein receptors than rat cells, 125I-ASOR was diacytosed to a greater extent (50%, -colchicine; 61%, + cholchicine) but degraded to a lesser extent (33%, -colchicine; 13%, + colchicine) than was observed in rat cells. The extent of retention of 125I-ASOR in rabbit cells was also greater than that in rat cells. A similar pattern of differences between rabbit and rat hepatocytes was observed for 125I-DTA-ASOR. These results indicate that intracellular sorting of internalized asialglycoproteins between diacytosis and degradation is dependent on both the structure of the ligand and the distribution of the cellular receptors.

Animals↗
PubMed1989

Study on the variables affecting toxicity of hybrid toxins. The effect of different target cell receptor distribution and toxin binding chain.

Disulfide conjugates of diphtheria toxin (DT) and its fragment A (DTA) to asialoorosomucoid (ASOR) were prepared. The toxicity of the conjugates were compared with DT in isolated rat, rabbit and guinea pig hepatocytes containing different concentration of asialoglycoprotein receptors (Biochim. Biophys, Acta 942, 57, 1988). In rat hepatocytes DTA-ASOR was highly toxic with half-maximal inhibitory concentration (IC50) of protein synthesis occurring at 4 +/- 3.10(-11) M (n = 7) which was much lower than that of DT DT (7.8 +/- 9.8.10(-9) M, n = 7). In rabbit hepatocytes toxicity of the conjugate (IC50 = 5.4 +/- 4.9.10(-10) M, n = 7) was higher than that of DT (IC50 = 5 +/- 4.10(-11) M, n = 7). In guinea pig hepatocytes, DTA-ASOR was not toxic at concentration below 10(-8) M, although DT was highly toxic (IC50 = 1.8 +/- 1.4.10(-10), n = 3). In the presence of 5 microM colchicine, the toxicity of DTA-ASOR in rat and rabbit hepatocytes increased by 10-fold, while in guinea pig hepatocytes it became detectable with an IC50 of 1.2 +/- 0.8.10(-9) M (n = 3). The toxicity of DT in the rat cells was also enhanced 10-fold by colchicine, but not at all in either the rabbit or the guinea pig cells. Addition of isolated diphtheria toxin fragment B (DTB) did not affect significantly the toxicity of DTA-ASOR in all three hepatocytes and that of DT in rat hepatocytes, but reduced toxicity of DT more than 20-fold in the rabbit and guinea pig cells. Toxicity of DT-ASOR in rat hepatocytes was the same as DTA-ASOR both in the absence and presence of colchicine, and abolished completely by excess ASOR, but not by DTB. Toxicity of DT-ASOR in rabbit hepatocytes was 40-times higher than DTA-ASOR, enhanced 10-fold by cochicine and reduced more than 30-fold by excess ASOR, but only slightly by DTB. These results indicate that entry of DTA from DTA-ASOR involve a DTB-independent translocation mechanism which can be as efficient as the DTB-dependent mechanism used by DT in the rabbit and guinea pig cells. The entry of both conjugates appeared to be mediated by the asialoglycoprotein receptors. However, the DTB moiety of DT-ASOR could function only in the DT-sensitive cells indicating the lack of a DTB-mediated translocation in the DT-resistant cells.

Animals↗