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Biomedical subjects

C M Chang

Publications and source records attributed to C M Chang.

At least 91 records · Page 5Linked to original sources
PubMed1991

Differential formation of disulfide linkages in the core antigen of extracellular and intracellular hepatitis B virus core particles.

Our understanding of the assembly of hepatitis B virus is still very limited. We present evidence to demonstrate that the HBc antigen formed oligomers through disulfide linkages in the extracellular hepatitis B virus core (HBc) particles. However, the intracellular HBV core particles did not contain disulfide-linked HBc antigens. Furthermore, the extracellular particles which had disulfide bonds were more stable than intracellular particles at pH 7.5 and 10 and in 3 M NaCl and 4 M urea. These data suggest that the formation of disulfide bonds in the HBc antigen is important for the integrity of the viral core particles.

Capsid↗
PubMed1991

Polymorphonuclear leukocytes occlude capillaries following middle cerebral artery occlusion and reperfusion in baboons.

BACKGROUND AND PURPOSE: Microvascular perfusion defects may accompany sustained occlusion and subsequent reperfusion of the middle cerebral artery; however, the nature of such "no-reflow" defects remains unclear. METHODS: In the absence of antithrombotic pretreatment, we documented lenticulostriatal microvascular flow integrity following 3-hour middle cerebral artery occlusion and 1-hour reperfusion in a baboon occlusion/reperfusion model by two methods identifying 1) microvascular occlusion and 2) microvascular patency. RESULTS: Microvascular "no-reflow" involved capillaries (vessels of 4.0-7.5 microns diameter) of the lenticulostriatal territory. Capillary reflow included 27-39% of all capaillaries in two subjects, indicating a significant reduction of perfusion from normal (2p = 0.045). In identical experimental preparations, single polymorphonuclear leukocytes completely occluded 4.7% of microvessels of capillary diameter in randomly selected fields, partially occluded 3.5% of postcapillary venules, and occluded 40% (four of 10) of capillaries in linear reconstruction along a 110 microns length. Circumferential contact between polymorphonuclear leukocytes and the luminal endothelial cell membranes was documented, with an intrecellular gap of, at most, 160 nm. Fibrin was found with degranulated platelets when the latter were associated with granulocytes, but not with polymorphonuclear leukocytes alone. CONCLUSIONS: The finding of capillary-obstructing polymorphonuclear leukocytes in the microvascular bed following middle cerebral artery reperfusion in focal ischemia in this model satisfies an essential requirement for postulating their role in early microvascular injury and the "no-reflow" phenomenon.

Animals↗
PubMed1991

In vivo stimulation of myelopoiesis in cyclophosphamide-treated mice by purified human GM-CSF.

Human granulocyte-macrophage colony-stimulating factor (hGM-CSF) secreted by a hepatoma cell line, HA22T/GVH, was purified and assessed for its effects in vivo on blood leukocytes and bone marrow granulocyte-macrophage progenitor cells (CFU-GM) in ICR mice pretreated with a sublethal dose of cyclophosphamide (cytoxan). The hGM-CSF preparations were natural and had no detectable endotoxin. Five days after the administration of 300 mg/kg cytoxan, severe leukopenia with marked myelopoietic suppression was induced. The cytoxan-treated mice were then injected intraperitoneally with 10,000 units of purified hGM-CSF/mouse daily for three days. Leukopenia was totally abrogated and the leukocyte number greatly increased to a level 2- to 3-fold higher than in GM-CSF-uninjected mice. Differential white cell count showed that the subpopulations of leukocytes responsive to hGM-CSF stimulation were mainly of neutrophils and monocytes, while the lymphocytes remained unaffected. Meanwhile, in the bone marrow, hGM-CSF administration induced an apparent (3-fold) increase in the number of myeloid progenitor cells, CFU-GM. However, the effect in vivo of a single hGM-CSF injection could only maintain for 48 hrs. In addition, the loss in body weight caused by cytoxan was less in the mice with subsequent hGM-CSF than those without CSF. These results suggest that injection of GM-CSF can effectively reconstitute the cytotoxic drug-damaged myelopoiesis without apparent in vivo toxic reaction.

Animals↗
PubMed1991

Pattern of memory deficits in a controlled psychometric study of thalamic haemorrhage.

Twelve patients with unilateral thalamic haemorrhages were assessed psychometrically. The results were compared with control subjects matched for sex, age and years of education. The pattern of deficits and preserved abilities cannot be explained in terms of semantic/episodic distinction, but could be interpreted as manifestations of disconnection between the frontal and temporal systems.

Aged↗
PubMed1990

Interactions at the nucleic acid binding site of the avian retroviral nucleocapsid protein: studies utilizing the fluorescent probe 4,4'-bis(phenylamino)(1,1'-binaphthalene)-5,5'-disulfonic acid.

The structural and functional properties of the nucleocapsid (NC) protein of the avian myeloblastosis virus were examined by steady-state fluorescence and fluorescence anisotropy measurements of the complex between the NC and the extrinsic fluorophore 4,4'-bis(phenylamino)(1,1'-binaphthalene)-5,5'-disulfonic acid (bis-ANS). The intrinsic fluorescence of bis-ANS is enhanced many fold upon forming a complex with the NC. Between 2 and 10 molecules of bis-ANS bind strongly to the NC, with an overall Kd of less than 10(-6) M. The emission of bis-ANS in the complex can also be induced by excitation at 298 nm, indicating that energy is transferred from Trp 80, the sole tryptophan in the NC protein, to bis-ANS. The energy transferred between the Trp 80 and bis-ANS was analyzed to yield a calculated distance of separation between these fluorophores of 28 +/- 3 A; thus, Trp 80 is well removed from the nearest bound bis-ANS. The fluorescence emission of bis-ANS in the NC.bis-ANS complex is efficiently quenched by added salts and by poly(A), suggesting that salt (presumably anions), nucleic acid, and bis-ANS bind to the same, positively charged region on the NC protein. A site size of six nucleotides was determined for nucleic acid binding to the NC protein, with an estimated Kd of less than 10(-6) M. Salt (anion) binding is strong, but nonspecific, with a Kapp of 4 mM, raising the possibility that anion binding to the NC protein might regulate the interaction of the NC with viral RNA inside the host cell.

Amino Acid Sequence↗
PubMed1990

The formation of bile canaliculi in human hepatoma cell lines.

Hepatocytes, known as polarized epithelial cells, are composed of sinusoid, basolateral and bile canalicular domains. Each domain contains proteins specific for it. Our studies indicate that the well-differentiated human hepatoma cell lines HepG2 and HuH-7 formed bile canaliculi in tissue culture, whereas the poorly differentiated hepatoma cell lines HA22T/VGH and SK-HEP-1 did not. We also used the 9B2 monoclonal antibody, previously shown to be specific for the human bile canalicular domain, to study formation of bile canaliculi in these human hepatoma cell lines. All four cell lines synthesize the 140-kD 9B2 antigen. Studies using peroxidase-antiperoxidase staining and immunoelectron microscopy revealed that the 9B2 antigen was first detected in cytoplasm and packaged in microvilli-lined vesicles, then vectorially transported to the cell surface and eventually fused with microvilli-lined vesicles from neighboring cells to form bile canaliculi in well-differentiated hepatoma cell lines. However, the 9B2 antigen of poorly differentiated lines was synthesized in cytoplasm, then transported directly to and evenly distributed on the cell membrane. These results lead us to conclude that human hepatoma cell lines could serve as a good in vitro model to study the formation of bile canaliculi in human hepatocytes. The bile canaliculi of human hepatocytes may be preformed and assembled in the intracellular, microvilli-lined vesicles, then vectorially transported to the cell surface, where they form the bile canaliculi through vesicles fusion. Finally, formation of bile canaliculi and transport of 9B2 antigen may be related to the differentiation of hepatocytes or progression stages of human hepatoma cells.

Antibodies, Monoclonal↗
PubMed1990

Effect of pancreas allografts on the ultrastructure of sciatic nerves in diabetic rats.

In a long-term study using cyclosporin-A (Cy-A) as immunosuppressant in a dose of 10 mg/kg/day, pancreas-duodenum was transplanted from Brown-Norway donors to alloxan-diabetic Lewis rats (n = 190). The pancreas transplants (PT) were performed after 1, 3, 6, 9, 12, and 15 months of diabetes. Recipient rats were sacrificed between 3 and 12 months of graft retainment. The mean axonal cross-sectional area and relative percentage of small, medium, and large myelinated fibers was evaluated. Also studied were unmyelinated fiber, ovoid body, and glycogen inclusion densities. Control rats consisted of non-diabetic rats (n = 36), similar rats receiving Cy-A (n = 42), diabetic rats (n = 103), and diabetic rats receiving Cy-A (n = 45). It was found that PT had a beneficial effect on the axonal cross-sectional area of myelinated nerves, the relative percentage of the various sizes of nerve fibers, and the ovoid body density, especially so in early diabetes. The effects in late diabetes were less spectacular. PT did not prove beneficial to the glycogen inclusion and unmyelinated fiber densities.

Animals↗
PubMed1990

Effect of pancreatic allografts on vascular basement membrane thickness in the diabetic rat.

Diabetic Lewis rats received pancreaticoduodenum allotransplants from Brown-Norway donors. Cyclosporine A (Cy-A) was used in a dose of 10 mg/kg/day for immunosuppression. These transplanted rats (n = 190) were compared with nondiabetic Lewis rats (n = 36), nondiabetic Lewis rats receiving 10 mg/kg/day Cy-A (n = 42), diabetic rats (n = 103), and diabetic rats receiving 10 mg/kg/day Cy-A (n = 45). The percentage area of periodic acid-Schiff (PAS) positive basement membrane (BM) of rectus muscle microvasculature was compared in each of the groups. It was found that the percentage area of PAS positive BM increased markedly over 15 months of uncontrolled diabetes. Cy-A did not have a significant effect on either normal or diabetic skeletal muscle vascular BM. Rats with established diabetes showed some reversal in the percentage area of PAS positive BM, when pancreas transplantation was performed at 9, 12, and 15 months of diabetes. Pancreas transplantation may appear beneficial even after the development of BM thickening of skeletal muscle vascular BM.

Abdominal Muscles↗
PubMed1990

Encapsidation of truncated human hepatitis B virus genomes through trans-complementation of the core protein and polymerase.

Mutational analyses and complementation tests were used to analyze the strategy of packaging and of replication of human hepatitis B virus (HBV). By creating new restriction enzyme sites and by varying the genome length of HBV mutants, we identified that the mutated genomes could be encapsidated through trans-complementation of the polymerase and/or core protein. This study demonstrates that the polymerase of HBV, similar to that of duck hepatitis B virus (DHBV), is synthesized de novo instead of through a core-polymerase fusion protein. The results also indicate that both the polymerase and the core protein can be supplied in trans during viral packaging, and that the complementation is not due to recombination between the cotransfected plasmids. Furthermore, HBV genome deleted down to 2.4 kb is still able to be encapsidated, as measured by the endogenous polymerase reaction. Taken together, these results provide a basis for using HBV as a vector to deliver foreign genes into hepatocytes and for defining the location of the packaging signal on the HBV genome.

Capsid↗
PubMed1990

Acrylonitrile-induced sister-chromatid exchanges and DNA single-strand breaks in adult human bronchial epithelial cells.

The ability of acrylonitrile to induce cytotoxicity, sister-chromatid exchanges and DNA single-strand breaks was studied in cultured human bronchial epithelial cells. The toxic effect as determined by cloning efficiency was observed at a dose of 600 micrograms/ml but not at doses of both 150 and 300 micrograms/ml. The frequency of sister-chromatid exchange in untreated cells was 3.7 +/- 1.3 per cell. In contrast, cells treated with acrylonitrile at 150 and 300 micrograms/ml exhibited 6.6 +/- 1.3 and 10.7 +/- 1.7 sister-chromatid exchanges per metaphase, respectively. DNA single-strand breaks were induced by acrylonitrile at dose levels of 200 and 500 micrograms/ml. The genotoxic effects on human bronchial epithelial cells that were directly exposed to acrylonitrile are of interest in relation to evidence for the higher lung cancer incidence of acrylonitrile workers in epidemiological studies.

Acrylonitrile↗
PubMed1990

A human hepatocellular carcinoma cell line with multiple copies of structurally normal chromosomes.

At Veterans General Hospital (VGH), a cell line, HA59T(HA59T/VGH), was established from a primary hepatocellular carcinoma of a 52-year-old Chinese male patient. G-banded metaphases were analyzed at passages 12 and 62. Of the 200 cells counted, 68% had 100-110 chromosomes/cell at passage 12, and 74% had 90-100 chromosomes/cell at passage 62. The presence of multiple copies of a structurally normal chromosome in a single cell was common for most of the chromosomes. Both X and Y sex chromosomes were normally present in the majority of the cells. This cell line is karyotypically different from those reported in other literature.

Carcinoma, Hepatocellular↗
PubMed1990

[A feasibility study on teaching evaluation system in medical education].

The purpose of this study is to develop an evaluation instrument with high feasibility and acceptability, and to quantify the outcome of evaluation, in order to set up an efficient evaluation system. Teaching evaluation with questionnaire by students has been carried out in National Yang-Ming Medical College for two years. With the support of both teachers and students, the system has been established and conducted on a regular basis. The most important purpose of evaluation is to improve the quality of teaching. During the two academic years (Sep. 1986-June 1988) of the program, the overall response rate was 44.5%, the Department of Nursing had the highest response rate, followed by Dentistry, Medicine-Post Graduate, Medical Technology, and Medicine. Taking into consideration of the year and the class size, the regression analysis found that higher year or smaller size of the class had better response rate. The response rates dropped significantly after the first academic year regardless of department or year. A total of 23 classes were included in the evaluation program and 99 courses were evaluated. All questions in the questionnaire used a 0 to 4 ordinal scale, in which 0 (improvement needed) was the low end and 4 (excellent) the high end. The mean score of the seven questions of teaching evaluation was 2.47. As a whole, the students were satisfied with the teaching. As to the categories of courses, clinical courses had better mean score than basic medical courses, and basic medical courses had better mean score than common required courses. To evaluate the effectiveness of the teaching, students' achievement was used as the outcome variable. The most important predictive variable was the method of instruction, followed by the content of lecture such as degree of difficulty of the lecture and cognitiveness of the contents. The above 3 variables explained 76% of the variation of the students' achievement. However, the significant of teachers' speech, performance and attitude were not so influential. Analysis based on the characteristics of the teachers (sex, age, position, and teaching experience), the characteristics of students (department and year), teaching environments (time and place), and the 3 categories of courses (clinical, basic medical and common required courses) showed that all the above variables only explained less then 10% of the variation of the students' achievement.

Education, Medical↗
PubMed1990

Pharyngeal tuberculosis.

A case of pharyngeal tuberculosis is reported in a 54-year-old Chinese man. This is an uncommon condition and is often associated with pulmonary tuberculosis as in our patient.

Antitubercular Agents↗
PubMed1989

Insulin suppresses hepatitis B surface antigen expression in human hepatoma cells.

The human hepatoma Hep3B cells contain integrated hepatitis B viral genome and continually secret hepatitis B surface antigen (HBsAg). The production of HBsAg (but not alpha-fetoprotein) was suppressed by addition of low concentrations (0.1-1 nM) of insulin into serum-free medium. In addition, the suppression of HBsAg production by insulin was paralleled with the decrease in HBsAg mRNA abundance. Insulin also cause a rapid rate of disappearance of HBsAg mRNA (t 1/2, 2 h) in Hep3B cells. The Hep3B cells carry specific receptor with high affinity for insulin (Kd = 1.8 nM). The receptor showed an insulin-dependent protein tyrosine kinase activity. The half-maximal insulin concentration for the activation of the receptor kinase was about 5 nM. Only very high concentrations of insulin-like growth factor I and human proinsulin can compete for the insulin receptor binding and suppress HBsAg production, this suggests that insulin may act through its receptor binding to suppress HBsAg expression in human hepatoma Hep3B cells.

Blotting, Northern↗
PubMed1989

Cytomegalovirus enhances lysis of HIV-infected T lymphoblasts.

A T4+ lymphoblastoid cell line (CR-10) persistently infected with the human immunodeficiency virus (HIV) and designated CR-10/NIT was superinfected with cytomegalovirus (CMV) isolated from peripheral blood mononuclear cells of a patient with AIDS. A productive CMV cycle in the CR-10/NIT lymphoblasts was demonstrated by fluorescent antibody staining (IF) using a monoclonal antibody (MAb) to the 150-kDa major capsid protein, by infectivity assays and by electron microscopy (EM). Two-color IF analysis showed that a small percentage of the CR-10/NIT cells were producing both CMV and HIV at any one time. EM studies revealed that all doubly infected cells were lysed whereas most cells infected only with HIV appeared intact. Cell lysis appeared 24 hr after superinfection of the CR-10/NIT cells with CMV and progressed to complete destruction of the cell culture between days 9 and 10. Our results suggest that CMV may convert a mildly cytopathic HIV infection of T lymphoblasts into a highly lytic process.

Antigens, Viral↗
PubMed1989

Visualization of membrane-associated folate transport proteins.

Transport of Methotrexate (MTX) into cells, via the "reduced folate" transport system, is a critical factor in the effectiveness of the drug in cancer chemotherapy, and defective transport is one of the principal types of resistance to MTX. Probes capable of detecting membrane-associated folate transport proteins (ftp's) in individual cells are potentially useful for identifying structural and functional domains and for investigating mechanisms of substrate translocation. Polyclonal antibody to highly purified ftp from Lactobacillus casei, in conjunction with a second, gold-labeled antibody, has been used to visualize, via electron microscopy, the protein in Triton-treated membrane fragments and in the membrane and cytoplasm of spheroplasts. To visualize ftp in L1210 cells, the substrate-binding site was first labeled covalently with activated fluorescein-Methotrexate, and the cells were then treated with anti-fluorescein antibody and the gold-labeled antibody.

Animals↗