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Biomedical subjects

C H Chang

Publications and source records attributed to C H Chang.

At least 271 records · Page 15Linked to original sources
PubMed1996

Improved cyclic urea inhibitors of the HIV-1 protease: synthesis, potency, resistance profile, human pharmacokinetics and X-ray crystal structure of DMP 450.

BACKGROUND: Effective HIV protease inhibitors must combine potency towards wild-type and mutant variants of HIV with oral bioavailability such that drug levels in relevant tissues continuously exceed that required for inhibition of virus replication. Computer-aided design led to the discovery of cyclic urea inhibitors of the HIV protease. We set out to improve the physical properties and oral bioavailability of these compounds. RESULTS: We have synthesized DMP 450 (bis-methanesulfonic acid salt), a water-soluble cyclic urea compound and a potent inhibitor of HIV replication in cell culture that also inhibits variants of HIV with single amino acid substitutions in the protease. DMP 450 is highly selective for HIV protease, consistent with displacement of the retrovirus-specific structural water molecule. Single doses of 10 mg kg-1 DMP 450 result in plasma levels in man in excess of that required to inhibit wild-type and several mutant HIVs. A plasmid-based, in vivo assay model suggests that maintenance of plasma levels of DMP 450 near the antiviral IC90 suppresses HIV protease activity in the animal. We did identify mutants that are resistant to DMP 450, however; multiple mutations within the protease gene caused a significant reduction in the antiviral response. CONCLUSIONS: DMP 450 is a significant advance within the cyclic urea class of HIV protease inhibitors due to its exceptional oral bioavailability. The data presented here suggest that an optimal cyclic urea will provide clinical benefit in treating AIDS if it combines favorable pharmacokinetics with potent activity against not only single mutants of HIV, but also multiply-mutant variants.

Administration, Oral↗
PubMed1996

Mice lacking the MHC class II transactivator (CIITA) show tissue-specific impairment of MHC class II expression.

CIITA activates the expression of multiple genes involved in antigen presentation and it is believed to be required for both constitutive and IFN gamma-inducible expression of these genes. To understand the role of CIITA in vivo, we have used gene targeting to generate mice that lack CIITA. CIITA-deficient (-/-) mice do not express conventional MHC class II molecules on the surface of splenic B cells and dendritic cells. In addition, macrophages resident in the peritoneal cavity do not express MHC class II molecules upon IFN gamma stimulation nor do somatic tissues of mice injected with IFN gamma, in contrast with wild-type mice. The levels of Ii and H-2M gene transcripts are substantially decreased but absent in CIITA (-/-) mice. The transcription of nonconventional MHC class II genes is, however not affected by CIITA deficiency. A subset of thymic epithelial cells express MHC class II molecules. Nonetheless, very few mature CD4 T cells are present in the periphery of CIITA (-/-) mice despite MHC class II expression in the thymus. Consequently, CIITA(-/-) mice are impaired in T-dependent antigen responses and MHC class II-mediated allogeneic responses.

Animals↗
PubMed1996

Using MTT viability assay to test the cytotoxicity of antibiotics and steroid to cultured porcine corneal endothelial cells.

Intracameral injection (ICI) of antibiotics and steroid is an effective method to deliver drugs into eyeballs, and rapidly achieve therapeutic concentrations. The high intraocular concentrations, however, that occur by such injections can harm the corneal endothelium. The MTT assay, MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide], has been used to test cytotoxicity of five antibiotics (including amphotericin-B, colistin-M, sulbenicillin, amikacin, cephradine) and a steroid, betamethasone, to cultured porcine corneal endothelial cells. The third passage of porcine corneal endothelial cells were plated into 96-well microtitration plates for drugs exposure. Cytotoxicities of the six drugs in different concentrations were compared, using exposure time as an independent variable. In its original and 10-fold ICI dose, only amphotericin-B among the six tested drugs showed significant cytotoxicity; the other drugs were considered safe. In a 100-fold ICI dose, amphotericin-B, colistin-M, and sulbenicillin were all toxic. Although a cell culture system utilizing the MTT assay is an in vitro method of testing for drug toxicity to the corneal endothelium, it nevertheless offers the advantages over in vivo methods. It is rapid, convenient, and economical. Large numbers of toxic compounds can be compared simultaneously, so that relative toxicities among different drugs and concentrations can be obtained, and be a guide for determining ICI dose, or as a reference for further in vivo testing.

Animals↗
PubMed1996

Identification and characterization of novel dermal Thy-1 antigen-bearing dendritic cells in murine skin.

Using immunofluorescence on dermal sheets of mouse ear, we identified novel Thy-1+ dermal dendritic cells in murine skin. These cells are resistant to topical corticosteroid treatment. The number of these dermal Thy-1+ dendritic cells is not altered with aging of the mice or with 3 d of culture. Dermal Thy-1+ dendritic cells have melanosomes in their cytoplasm; hence, phagocytic activity is likely to be present. Double immunofluorescence revealed that most of these dermal Thy-1+ dendritic cells express CD45. Furthermore, conventional immunofluorescence and confocal laser scanning microscopic findings showed that most of these Thy-1+ dermal dendritic cells express gamma delta and V gamma 3 T-cell receptors. The relationship between these dermal Thy-1+ dendritic cells and dendritic epidermal T cells is discussed.

Animals↗
PubMed1996

Pleiotropic phenotypes caused by genetic ablation of the receiver module of the Agrobacterium tumefaciens VirA protein.

The VirA protein of Agrobacterium tumefaciens is a transmembrane sensory kinase that phosphorylates the VirG response regulator in response to chemical signals released from plant wound sites. VirA contains both a two-component kinase module and, at its carboxyl terminus, a receiver module. We previously provided evidence that this receiver module inhibited the activity of the kinase module and that inhibition might be neutralized by phosphorylation. In this report, we provide additional evidence for this model by showing that overexpressing the receiver module in trans can restore low-level basal activity to a VirA mutant protein lacking the receiver module. We also show that ablation of the receiver module restores activity to the inactive VirA (delta324-413) mutant, which has a deletion within a region designated the linker module. This indicates that deletion of the linker module does not denature the kinase module, but rather locks the kinase into a phenotypically inactive conformation, and that this inactivity requires the receiver module. These data provide genetic evidence that the kinase and receiver modules of VirA attain their native conformations autonomously. The receiver module also restricts the variety of phenolic compounds that have stimulatory activity, since removal of this module causes otherwise nonstimulatory phenolic compounds such as 4-hydroxyacetophenone to stimulate vir gene expression.

Agrobacterium tumefaciens↗
PubMed1996

Resection and mutagenesis of the acid pH-inducible P2 promoter of the Agrobacterium tumefaciens virG gene.

Transcription of the virG gene initiates from two tandem promoters, designated P1 and P2, that are located 50 nucleotides apart. Transcription of the P2 promoter is induced by extracellular acidity. cis-acting sites required for P2 activity were identified by constructing and assaying a series of 5' and 3' resections and site-directed nucleotide substitutions. Nucleotides between positions -9 and -37 were sufficient for regulated promoter activity. Within this region, nucleotide substitutions at the predicted -10 and -35 regions strongly reduced P2 expression. In addition, alterations in the region between nucleotides -24 and -32 also eliminated or strongly reduced promoter activity. These data suggest that this promoter may be regulated by a positive transcription factor that binds to nucleotide residues in this interval.

Agrobacterium tumefaciens↗
PubMed1996

Human immunodeficiency virus type 1 membrane fusion mediated by a laboratory-adapted strain and a primary isolate analyzed by resonance energy transfer.

Previous studies of human immunodeficiency virus type 1 (HIV-1) envelope glycoprotein-mediated membrane fusion have focused on laboratory-adapted T-lymphotropic strains of the virus. The goal of this study was to characterize membrane fusion mediated by a primary HIV-1 isolate in comparison with a laboratory-adapted strain. To this end, a new fusion assay was developed on the basis of the principle of resonance energy transfer, using HeLa cells stably transfected with gp120/gp41 from the T-lymphotropic isolate HIV-1LA1 or the macrophage-tropic primary isolate HIV-1JR-FL. These cells fused with CD4+ target cell lines with a tropism mirroring that of infection by the two viruses. Of particular note, HeLa cells expressing HIV-1JR-FL gp120/gp41 fused only with PM1 cells, a clonal derivative of HUT 78, and not with other T-cell or macrophage cell lines. These results demonstrate that the envelope glycoproteins of these strains play a major role in mediating viral tropism. Despite significant differences exhibited by HIV-1JR-FL and HIV-1LAI in terms of tropism and sensitivity to neutralization by CD4-based proteins, the present study found that membrane fusion mediated by the envelope glycoproteins of these viruses had remarkably similar properties. In particular, the degree and kinetics of membrane fusion were similar, fusion occurred at neutral pH and was dependent on the presence of divalent cations. Inhibition of HIV-1JR-FL envelope glycoprotein-mediated membrane fusion by soluble CD4 and CD4-IgG2 occurred at concentrations similar to those required to neutralize this virus. Interestingly, higher concentrations of these agents were required to inhibit HIV-1LAI envelope glycoprotein-mediated membrane fusion, in contrast to the greater sensitivity of HIV-1LAI virions to neutralization by soluble CD4 and CD4-IgG2. This finding suggests that the mechanisms of fusion inhibition and neutralization of HIV-1 are distinct.

Antigens, CD↗
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

Nitric oxide mediates mitogenic effect of VEGF on coronary venular endothelium.

Vascular endothelial growth factor (VEGF) is a secreted protein that is a specific growth factor for endothelial cells. We have recently demonstrated that nitric oxide (NO) donors and vasoactive peptides promoting NO-mediated vasorelaxation induce angiogenesis in vivo as well as endothelial cell growth and motility in vitro; in contrast, inhibitors of NO synthase suppress angiogenesis. In this study we investigated the role of NO in mediating the mitogenic effect of VEGF on cultured microvascular endothelium isolated from coronary postcapillary venules. VEGF induced a dose-dependent increase in cell proliferation and DNA synthesis. The role of NO was determined by monitoring proliferation or guanosine 3',5'-cyclic monophosphate (cGMP) levels in the presence and absence of NO synthase blockers. The proliferative effect evoked by VEGF was reduced by pretreatment of the cells with NO synthase inhibitors. Exposure of the cells to VEGF induced a significant increment in cGMP levels. This effect was potentiated by superoxide dismutase addition and was abolished by NO synthase inhibitors. VEGF stimulates proliferation of postcapillary endothelial cells through the production of NO and cGMP accumulation.

Cell Line↗
PubMed1996

Silicone cream occlusive dressing--a novel noninvasive regimen in the treatment of keloid.

BACKGROUND: The use of silicone oil in the treatment of hypertrophic scars has recently been much discussed. Silicone gel and silicone cream containing 20% silicone oil have been demonstrated to effectively treat hypertrophic scars. However, the results in treating keloid are variable. OBJECTIVE: The purpose of this study was to evaluate the clinical efficacy of silicone cream with occlusive dressing in the treatment of keloid. METHODS: An open study was performed in 15 Chinese patients with single or multiple keloids. Nineteen keloids were randomly selected and treated with silicone cream occlusive dressing (SCOD). The effectiveness was assessed using a scoring system involving elevation, redness, hardness, itching and tenderness or pain of the lesions. RESULTS: Fifteen out of 19 keloids (79%) showed significant improvement after using SCOD for 6 months. None of the keloids progressed. CONCLUSIONS: We conclude that SCOD is easy to apply, noninvasive, painless, almost free of side effects. It is especially useful in children and those who cannot tolerate the pain or inconvenience of other treatment procedures. The action mechanism of SCOD is still unclear, though hydration and occlusion rather than silicone itself may play an important role.

Adult↗
PubMed1996

Estimating local-area needs for psychiatric care: a case study.

BACKGROUND: Different approaches to estimating local catchment-area needs for psychiatric services are illustrated and compared. METHOD: Data from an epidemiological morbidity survey of a random sample of 496 adults were available, as were actual service utilisation rates. Four types of utilisation were modelled (i.e. overall, out-patient, in-patient, emergency clinic) using social indicators available from Statistics Canada census-tract data. Finally, a case-control study compared out-patients from a deprived and an affluent catchment area, matched case by case for primary diagnosis, age, sex and residential status (n = 52). RESULTS: Modelling proved highly predictive of utilisation, the overall-use model accounting for 73% of the variance. The case-control study indicated a higher rate of Axis II traits, substance abuse and needs for social care in the deprived catchment area. CONCLUSIONS: Resource allocation based on the social indicators modelling method was more consistent with sensible distribution of human resources. None of the methods, however, appear to reflect adequately the severity of caseloads evidenced in the case-control study.

Adolescent↗
PubMed1996

Growth hormone (GH) and insulin-like growth factor I responses after treatments with an orally active GH secretagogue L-163,255 in swine.

L-163,255 is a potent orally active spiropiperidine GH secretagogue. When administered iv or orally, L-163,255 caused GH to be increased in a dose-related manner, with a return to baseline by 90 min. After iv administrations of saline and L-163,255 at 1, 3, and 10 micrograms/kg, GH areas under the curves (GH AUCs) over 120 min were 377 +/- 136, 1151 +/- 413 (P < 0.05), 795 +/- 413 (P = NS), and 1770 +/- 416 ng.min/ml (P < 0.01), and peak GH concentrations were 8 +/- 3, 16 +/- 7 (P = NS), 17 +/- 5 (P = NS), and 43 +/- 12 ng/ml (P < 0.01), respectively. No changes in plasma cortisol concentrations were noted. After oral administrations at 3, 10, and 30 micrograms/kg, GH AUCs over 180 min were 1133 +/- 154, 1246 +/- 129 (P = NS), and 1551 +/- 210 ng.min/ml (P = NS), and peak GH concentrations were 7 +/- 2, 11 +/- 3 (P = NS), and 23 +/- 6 ng/ml (P < 0.01), respectively. After administration in feed, L-163,255 caused a dose-related increase in GH, with an initial peak observed at 60 min for both 30 and 300 micrograms/kg dose groups, and remained elevated above baseline through 180 min for the high dose group only. GH AUCS for 180 min posttreatment were 929 +/- 134 and 1897 +/- 244 ng.min/ml, and peak GH concentrations were 9 +/- 2 and 22 +/- 4 ng/ml for the 30 and 300 micrograms/kg doses prepared in 150 g feed, respectively. When provided in feed ad libitum over the 72-h period, mean plasma insulin-like growth factor I levels increased 15%, 62% (P < 0.01), and 109% (P < 0.01) in the untreated, treated with L-163,255 at 360 ppm, or treated with porcine somatotropin groups, respectively. Repeated iv administration of L-163,255 at 1 mg/kg once daily over 14 days resulted in an initial marked GH response, followed by a much reduced, but significantly elevated, GH response over the saline control values on subsequent treatment days. Repeated iv treatments with L-163,255 also resulted in an elevated insulin-like growth factor I level (approximately 60%) over that in saline controls. Compared to those in saline controls, plasma cortisol concentrations tended to be increased after the initial dose of L-163,255, but no significant increases were noted on days 7 and 14 in the L-163,255 group. The results of these studies indicate that L-163,255 is an orally active GH secretagogue suitable for long term efficacy studies in swine.

Administration, Oral↗
PubMed1996

Video-assisted cardiac surgery. Preliminary experience in reoperative mitral valve surgery.

OBJECTIVES: Video-assisted endoscopic techniques had been applied in the surgical correction of patent ductus arteriosus, vascular ring, or coronary artery disease. However, it has been used only recently in the correction of reoperative mitral valve lesions. DESIGN: Video-assisted cardiac operations were performed on four patients who had received surgical interventions on their mitral valves and needed emergent reoperation. PATIENTS: Four patients (3 men and 1 woman) received emergency surgery from September to December 1995 for thrombosis of mechanical mitral prosthesis (2 patients) and severe mitral regurgitation with previously failed mitral valve repair (2 patients). Six previous operations had been performed on these mitral valves. Patient ages ranged from 26.7 to 68.1 years (mean, 47.3 years). Preoperatively, acute pulmonary edema occurred in two patients, cerebral emboli occurred in one patient, and sepsis was found in one patient. Mechanical ventilatory support was used in two patients before operation. INTERVENTION: The operations were performed through right anterior minithoracotomy, guided by video-assisted endoscopic techniques with femoro-femoral extracorporeal circulation. The operative procedures were thrombectomy of mitral prosthesis in two patients, mitral valve repair in one patient, and mitral valve replacement in one patient. RESULTS: The duration of extracorporeal circulation was 166 to 320 min (222 +/- 67 min) and the operation time was 4.6 to 6.8 h (6.1 +/- 1.0 h). All patients recovered from the operations rapidly with uneventful postoperative courses except 1 patient who had sepsis preoperatively and died 2 months later. CONCLUSION: Our experience demonstrates that video-assisted cardiac surgery is technically feasible and could be performed in reoperation of the mitral valve.

Adult↗
PubMed1996

Prognostic factors of primary aldosteronism.

BACKGROUND: Primary aldosteronism (PA) is a rare but potentially curable cause of hypertension. Between October 1982 and November 1994, 30 patients of PA received unilateral adrenalectomy with a long-term follow up (mean:60.3 months). Nineteen (63.3%) cases were cured (Group 1); 11(36.7%) cases were improved (Group 2). The purpose of this study was to determine prognostic factors after surgery in patients with PA. METHODS: A retrospective analysis was performed regarding age and sex of the patients, duration of hypertension, family history of hypertension, preoperative blood pressure, plasma aldosterone concentration (PAC), plasma renin activity (PRA) and efficacy of spironolactone on blood pressure between both groups. End-organs (including kidney, heart, retina and brain) involvement was evaluated and compared. Adrenalectomy and renal biopsy specimen for pathology were similarly evaluated. RESULTS: The duration of hypertension was longer in Group 2 than in Group 1 (8.18 +/- 4.94 vs 5.21 +/- 4.24 years). The efficacy of spironolactone on blood pressure (BP) was positive in 81.8% of Group 1 and 16.7% of Group 2. Adrenal cortical adenoma in 24 cases with a cure rate of 70.8% (17/24) and adrenal cortical macronodular hyperplasia in 6 cases with a cure rate of 33.3% (2/6) were noted. Group 2 had more end-organs involvement than Group 1. The severity of histopathological change of the renal biopsy was similar. CONCLUSIONS: This study suggests that preoperative response of blood pressure to spironolactone administration predicts the postoperative prognosis of hypertension in patient with PA. Long duration of hypertension and involvement of two or more end-organs were poor prognostic factors. Excellent results can be achieved by unilateral adrenalectomy in adrenal cortical adenoma and fair results, in adrenal cortical macronodular hyperplasia.

Adolescent↗