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Biomedical subjects

W C Chang

Publications and source records attributed to W C Chang.

At least 181 records · Page 10Linked to original sources

Stress and adenosine: II. Adenosine analogs mimic the effect of inescapable shock on shuttle-escape performance in rats.

In 3 experiments, the authors examined the role of adenosine regulation in escape deficits produced by earlier exposure to inescapable shock in rats (learned helplessness). Adenosine analogs injected before escape testing mimicked the effect of earlier inescapable shock, with the magnitude of the deficit varying with dose and drug specificity for A2 adenosine receptors. Agonist-induced and stress-induced escape deficits were eliminated by pretest treatment with the centrally acting adenosine receptor antagonist theophylline but not the peripheral antagonist 8-[p-sulfophenyl]-theophylline. Finally, preexposure to an ineffective number of inescapable shocks interacted in synergy with an ineffective pretest injection of adenosine agonist to maximize deficits in escape performance. These data implicate energy regulation and a central compensatory action by adenosine in the aspects of helplessness related to conservation-withdrawal.

Adenosine↗

A very low intake of fat is required to decrease fecal bile acid concentrations in rats.

The purpose of this study was to determine the effect of different amounts of dietary fat on colonic cell proliferation and fecal bile acid concentrations. Thirty-nine male Sprague-Dawley rats were randomly assigned to three diets (13 rats per diet) containing 5, 10 and 20 g butter/100 g diet. Diets were fed for 3 wk. As fat intake increased, total fecal fat excretion remained constant. When dietary fat was decreased from 20 to 10 g/100 g diet, total fecal bile acid concentrations tended to increase 14.5%. However, a further reduction to 5 g butter/100 g diet significantly decreased fecal total bile acid concentration by 48% from the concentration in feces of rats fed 10 g butter/100 g diet. The concentration of deoxycholate (considered a highly promotive bile acid) was not reduced unless the amount of fat in the diet was reduced to 5 g/100 g. Labeling index was used as an intermediate marker for colon carcinogenesis. For deoxycholate, a decrease of fat intake to 5 g/100 g diet decreased the cecal labeling index relative to those of rats fed 10 or 20 g butter/100 g diet. These data indicate that decreasing the dietary fat from 20 to 10 g/100 g does not decrease fecal bile acid concentration or colonic cell proliferation, but some effects are seen in rats fed 5 g butter/100 g diet.

Animals↗

Protective effects of arginine-vasopressin on aspirin-induced gastric mucosal damage in anaesthetized dogs.

The protective effects of graded doses of arginine-vasopressin (AVP) on acidified acetylsalicylic acid (ASA) solution-induced changes in gastric prostaglandin E2 (PGE2) secretion, mucus production, acid back-diffusion and mucosal damage were studied in the bilateral truncal vagotomized stomach of anaesthetized dogs. After 1-4 h intragastric irrigation of the stomach with AVP (1-100 ng kg-1) plus 20 mM acidified ASA solution, a significant (P < 0.05) inhibition in gastric mucosal lesions and acid back-diffusion produced by acidified ASA solution was observed. The reduction in the gastric PGE2 secretion and in mucus production provoked by the same dose of acidified ASA solution was also diminished. Furthermore, a correlation (r = 0.883; P < 0.01) between AVP-induced inhibition in ASA-provoked reduction in gastric PGE2 secretion and in mucus production was found. During the experiment, the heart rate, the peripheral arterial blood pressure and the gastric arterial blood pressure were not altered by AVP (1-100 ng kg-1). Thus, intragastric AVP protects gastric mucosa against ASA-induced damage without producing cardiovascular side effects. The inhibitory effects of AVP (100 ng kg-1) on acidified ASA-induced reduction in PGE2 and mucus secretion, as well as on ASA-induced enhancement in acid back-diffusion and erosion production were dose-dependently reversed by a specific V1 antagonist, 1-(beta-mercapto-beta,beta-cyclopenta-methylene-propionic acid), 2-(o-methyl)tyrosine-Arg8-vasopressin.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Pulsatile flow through a bifurcation with a cerebrovascular aneurysm.

Laser-Doppler velocimetry measurements and flow visualization were complementarily made in pulsatile and steady flow in a cerebrovascular aneurysm model with bifurcation angles of 60, 90, and 140 deg, and volume-flow rate ratios between the branches of 1 to 1 and 3 to 1. The mean, peak, and minimal Reynolds numbers based on the bulk average velocity and diameter of the parent vessel were 600, 800, and 280, respectively. For uneven branch flow, it is found that the flow activity inside the aneurysm and the stresses acting on the aneurysmal wall increase with increasing bifurcation angle. More importantly, the present angle suggests the presence of a critical bifurcation angle below which the aneurysm is prone to thrombosis, whereas above which the aneurysm is susceptible to progression or rupture. For evenly distributed branch flow, the intra-aneurysmal flow is sluggish and therefore prone to thrombosis for all studied bifurcation angles.

Aneurysm, Ruptured↗

cDNA and protein sequences coding for the precursor of phospholipase A2 from Taiwan cobra, Naja naja atra.

The cDNA sequence encoding phospholipase A2 (PLA2) was determined by analysis of polymerase-chain-reaction (PCR) product amplified from total cDNA mixture which had been constructed from the poly(A)+RNA of venom glands obtained from Taiwan cobras. Two oligonucleotide segments corresponding to the 5'- and 3'-noncoding regions of sea-snake PLA2 gene were used as primers for PCR-amplified reaction. Plasmids of transformed E. coli strain JM109 containing amplified PLA2 cDNA were purified and prepared for nucleotide sequencing by dideoxynucleotide chain-termination method. Sequencing more than five clones containing about 0.5 kb DNA inserts revealed two isoforms with complete reading frames of 468 base pairs each covering a precursor for phospholipase A2 with a deduced mature protein sequence of 119 amino acids and a 27 amino-acid signal peptide. These two enzymes of Group I PLA2 differ in six nucleotide residues at the gene level and three amino acids along the whole polypeptide chain, each consisting of 14 cysteine residues similar to all reported PLA2 of different snake venoms. The signal peptides and hydropathy profiles of Group I PLA2 reported here are distinctly different from those of Group II PLA2 in viperid snakes.

Amino Acid Sequence↗

Induction of 12-lipoxygenase expression by epidermal growth factor is mediated by protein kinase C in A431 cells.

Epidermal growth factor (EGF) increases 12-lipoxygenase mRNA by about 2-fold with a lag period of 4 to 8 hr, which precedes the increase in 12-lipoxygenase activity by 2 to 4 hr in human epidermoid carcinoma A431 cells. Induction of 12-lipoxygenase expression in human erythroleukemia cells by phorbol 12-myristate 13-acetate (PMA) has been reported previously. The present report describes a study of the involvement of protein kinase C (PKC) in EGF-induced 12-lipoxygenase expression in A431 cells. EGF-induced 12-lipoxygenase expression was inhibited by methyl 2,5-dihydroxycinnamate, a tyrosine kinase inhibitor. Staurosporine and calphostin C, which are two PKC inhibitors, inhibited EGF-induced enzyme activity and mRNA expression of 12-lipoxygenase. 1,2-Dioctanoyl-sn-glycerol (a membrane-permeant diacylglycerol) and PMA significantly induced enzyme activity and mRNA expression. Simultaneous treatment of cells with EGF and PMA did not exhibit an additive effect, suggesting that EGF and PMA share a common biochemical pathway in 12-lipoxygenase induction. Expression of mRNA for PKC alpha, delta and zeta was detected in A431 cells, whereas no mRNA expression for PKC beta 1, gamma and epsilon was observed. Taken together, these results suggest that EGF-induced 12-lipoxygenase expression is at least in part mediated by the PKC signal transduction pathway.

Alkaloids↗

Individual differences in vulnerability to inescapable shock in rats.

The present study determined whether individual differences in neophobia during an open-field pretest predict vulnerability to inescapable electric shock, as measured by 2 tests of learned helplessness in rats. Shuttle-escape latencies and saccharin finickiness increased across groups that had received increasing numbers of inescapable shocks 24 hr earlier. Dispersion in the test measure as well as the percentage of variance explained by pretest neophobia were greater when no or few shocks were delivered in the interpolated stress phase. Pretest neophobia was positively related to stress vulnerability in both tests under these conditions. Further increments in stressor severity overwhelmed even the most stress-resistant rats, thereby decreasing dispersion in the test measure and eliminating the predictive value of pretest neophobia. This pattern of outcomes was more robust for the shuttle-escape measure of helplessness.

Animals↗

The change of penile blood thromboxane B2 and prostacyclin after intracavernous injection of vasoactive drugs for the treatment of arteriogenic impotence.

Penile hypercoagulability during erection may predispose to aging vascular changes and, eventually, arteriogenic impotence. The relationship of penile blood thromboxane B2 and 6-keto-prostaglandin F1 alpha in both psychogenically and arteriogenically impotent patients after intracavernosal treatment with papaverine plus phentolamine or prostaglandin E1 was evaluated. No significant change in penile blood thromboxane B2 was observed with treatment of these vasoactive drugs. On the other hand, penile blood 6-keto-prostaglandin F1 alpha was significantly increased with the injection of 30 mg of papaverine plus 0.5 mg of phentolamine, and of 20 micrograms prostaglandin E1. Furthermore, the prostacyclin-to-thromboxane A2 ratio for the patient who received papaverine plus phentolamine was significantly lower than that for the same individual receiving prostaglandin E1. Our preliminary findings suggest that penile blood prostacyclin may participate in the pathogenesis of arteriogenic impotence and priapism.

6-Ketoprostaglandin F1 alpha↗

Effects of sodium fluoride and cobalt chloride on the enantioselectivity of microsomal and cytosolic esterases in rat intestinal mucosa.

The effects of sodium fluoride (NaF) and cobalt chloride (CoCl2) on the enantioselective hydrolysis of racemic oxazepam 3-acetate (rac-OXA) by microsomal and cytosolic esterases in rat intestinal mucosa were studied. Microsomal and cytosolic esterases hydrolyzed S-OXA and R-OXA in approximately 1:19 and 4:1 ratios, respectively. The hydrolysis of R-OXA by microsomal esterases was inhibited by NaF with an IC50 of 13.4 +/- 1.5 mM. Hydrolyses of both S-OXA and R-OXA by cytosolic esterases were inhibited by NaF with a similar IC50 value (approximately 3 mM). The hydrolysis of S-OXA by cytosolic esterases was inhibited by CoCl2 (IC50 = approximately 5 mM), whereas the hydrolysis of R-OXA by cytosolic esterases was stimulated by approximately 10% in the presence of 1 mM CoCl2. In comparison, the hydrolysis of R-OXA by microsomal esterases was stimulated by approximately 55% in the presence of 1 mM CoCl2. These results not only revealed the effects of NaF and CoCl2 on the catalytic activities of enantioselective cytosolic and microsomal esterases, but also indicated that microsomal and cytosolic esterases that selectively hydrolyzed R-OXA were distinctly different protein entities.

Animals↗

Induction of arachidonate 12-lipoxygenase mRNA by epidermal growth factor in A431 cells.

12(S)-Hydroxyeicosatetraenoic acid is biosynthesized from arachidonic acid by the microsomal fraction of human epidermoid carcinoma A431 cells, and the microsomal 12-lipoxygenase activity is enhanced by about 2-fold by epidermal growth factor (EGF) with a 10-h lag period (Chang, W.C., Ning, C.C., Lin, M.T., and Huang, J.D. (1992) J. Biol. Chem. 267, 3657-3666). The microsomal 12-lipoxygenase in A431 cells was only 3% active with linoleic acid as compared with arachidonic acid. The enzyme was immunoprecipitated by a monoclonal antibody against human platelet 12-lipoxygenase but not by that against porcine leukocyte enzyme. A 3.1-kilobase mRNA was detected in A431 cells by Northern blot analyses using cDNA probe of human platelet 12-lipoxygenase. EGF could increase the 12-lipoxygenase mRNA level by about 2-fold with a lag period of 10 h, which was well parallel with the increase in the enzyme activity. The induction of the 12-lipoxygenase mRNA by EGF was completely blocked by 35 microM cycloheximide, if present in culture medium during EGF treatment, indicating that a de novo protein biosynthesis was essential for EGF-induced 12-lipoxygenase mRNA expression. Our data provide the first evidence for the inducibility of human 12-lipoxygenase gene expression by a growth factor.

Antibodies, Monoclonal↗

Public attitudes in Edmonton toward assisted reproductive technology.

OBJECTIVE: To determine public attitudes toward the use and possible limitations of assisted reproductive technology (ART). DESIGN: Mail survey based on telephone numbers selected at random by computer. SETTING: Edmonton. PARTICIPANTS: A total of 602 Edmonton residents aged 16 years or more (57% of eligible subjects) reached by telephone agreed to participate. Completed questionnaires were received from 455 subjects (76%). MAIN OUTCOME MEASURES: Attitudes toward egg donation, sperm donation, selective fetal reduction, embryo freezing and experimentation, and surrogacy, as determined through responses to five cases. Comments and demographic data were also solicited. MAIN RESULTS: Overall, 66% and 63% respectively of the respondents would donate an egg or sperm to a sibling; the corresponding rates for donation to a stranger were 41% and 44%. Selective fetal reduction was supported by 47% of the respondents, although only 24% would support fetal reduction to eliminate fetuses of an undesired sex. Most (64%) thought that live embryo freezing should be permitted by law. A total of 74% agreed with surrogacy if done for medical reasons, but 85% opposed its use for reasons of convenience. Overall, 72% of the respondents thought that ART should be regulated. A total of 58% felt that physicians should be primarily responsible for determining the allowable limits of this technology, and 38% felt that the public should be primarily responsible. Only 21% agreed with public funding of ART. Religious affiliation strongly influenced attitudes toward ART. CONCLUSIONS: Public support for ART varies depending on the circumstances of its use. Education is needed to make the general community aware of the various aspects of ART. The results of this survey should help physicians and governing bodies make informed decisions about the future directions of ART in Canada.

Adolescent↗

Partial reversion of transformed phenotype of B104 cancer cells by antisense nucleic acids.

We used antisense RNA to inhibit the expression of oncogene neu and investigated the effects of diminished neu expression on the phenotypes of B104 cells containing activated oncogene neu. Antisense MT-neu and pSV-neo plasmids were cotransfected into neuroblastoma B104 cells. Southern analysis showed the integration of anti-neu DNA into B104 cells. The expression of neu was inhibited up to 90% as quantitated by immunoprecipitation. The growth rate and the potential to differentiate in these transfectants were not affected as compared to the parental cell lines. The ability to grow in soft agar was inhibited more than 90% in these transfectants. Our results indicated that antisense-RNA against a specific oncogene can decrease the tumorigenicity of tumor cells but may not be able to revert it to normal cells completely.

Animals↗

Purification of growth hormones by reversed-phase high-performance liquid chromatography.

Reversed-phase high-performance liquid chromatography (HPLC) on a column of Radial-Pak C18 cartridge was utilized for the purification of a variety of growth hormone (GH) proteins from mammalian, avian, amphibian and fish pituitary glands. Recovery of GH from pituitary glands of up to 0.43% of total protein was obtained with a high degree of homogeneity as revealed by sodium dodecyl sulphate polyacrylamide gel electrophoresis. The HPLC-purified GHs show reactions of identity or near identity by immuno-diffusion studies on agar gel. This method offers a convenient and rapid purification of vertebrate GH on an analytical or preparative scale.

Amino Acids↗

Purification and characterization of reversible sperm motility inhibitors from porcine seminal plasma.

Two sperm motility inhibitors (SMI1 and SMI2) were purified from porcine seminal plasma with high performance liquid chromatography (HPLC). Their molecular weights are about 15,000 as estimated by SDS-polyacrylamide gel electrophoresis (SDS-PAGE). Both of them decreased the percentage of motile spermatozoa in a dose-dependent manner. The inhibitory effect can be abolished by addition of the porcine follicular fluid. Both SMI1 and SMI2 have similar amino acid composition, suggesting that they may be structurally related. They also have inhibitory effect on chymotrypsin.

Amino Acid Sequence↗

Release of the p53-induced repression on thymidine kinase promoter by single p53-binding sequence.

Herpes simplex thymidine kinase (TK) promoter was shown to be repressed by the wild-type p53. Using a model system that the p53-binding site was linked to the thymidine kinase promoter, we demonstrated that single p53-specific binding site was sufficient to abolish the repression. On the contrary, the mutant p53 had the opposite effects on the HSV-TK promoter in BHK cells. The results suggest that the p53-binding site may act as an enhancer to regulate the gene expression in a novel way in vivo.

Animals↗

Snake venom cardiotoxin can rapidly induce actin polymerization in intact platelets.

The action of Taiwan cobra (Naja naja atra) venom cardiotoxin on rabbit platelets at 37 degrees C was characterized by observing cytoskeletal alterations and cell lysis. At a concentration of 21.4 microM the toxin produced no cell lysis within 30 s, and less than 5% of the total lactate dehydrogenase activity of intact cells was detected in the suspending medium after the interaction had proceeded for 3 min. The extent of cell lysis was proportional to toxin concentration and interaction time. Before cell lysis, the toxin caused rapid incorporation of actin monomers into cross-linked actin filaments. The actin incorporation could be inhibited by either the presence of cytochalasin B or increased CaCl2 concentration in the suspending medium. However, addition of indomethacin did not influence the toxin-induced cytoskeletal change.

Actins↗

The effect of copper ion on arachidonic acid metabolism in the porcine corneal epithelium.

The effect of copper ion on arachidonic acid metabolism in the corneal epithelium was studied. Our results showed that when 3H-arachidonic acid (1 microCi/microgram) was incubated with homogenate of corneal epithelium at 37 degrees C for 45 min., three metabolites were formed. Metabolite A, the major metabolite, was identified as 12-HETE by HPLC and GC. An analysis with chiral phase HPLC revealed that metabolite A was a (R)-enantiomer. The production of 12(R)-HETE was stimulated by low concentrations of CuCl2 (< 0.1 mM) and inhibited by higher concentrations of CuCl2. Study on the biological activity of 12(R)-HETE showed that it could stimulate migration of endothelial cells in a concentration as low as 10 ng/ml. These results suggest that one of the effects of copper ion on the corneal neovascularization may be to regulate the level of 12(R)-HETE.

12-Hydroxy-5,8,10,14-eicosatetraenoic Acid↗

Effect of design of prefabricated post heads on core materials.

This study investigated post-core adaptation as it relates to core materials and post-head configuration. A total of 360 samples that consisted of three prefabricated post systems and three core materials of amalgam and composite resin of 1 and 3 mm post-head-covering thicknesses were made. Composite resin with a post-head covering of 3 mm was most resistant to compression, whereas amalgam was most resistant to retentive forces. The addition of titanium to composite resin as a reinforcement filler did not improve resistance to either compressive or tensile forces.

Analysis of Variance↗