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Biomedical subjects

C C Chang

Publications and source records attributed to C C Chang.

At least 325 records · Page 18Linked to original sources
PubMed1994

Two-step cell disruption for the extraction of membrane-associated recombinant protein from Saccharomyces cerevisiae.

The use of rDNA technology to express heterologous proteins has been very successful during the last several years. Choice of an expression host is very important in order to retain the biological activity of recombinant proteins. Baker's yeast, Saccharomyces cerevisiae, is a eucaryotic GRAS organism suitable for the expression of biologically active proteins. Specifically, hepatitis B surface antigen (HBsAg) is expressed in baker's yeast. Because the yeast cells need to be disrupted for the recovery of bioactive intracellular proteins and because the protein HBsAg is hydrophobic and has a tendency to become associated with cell membranes, the use of detergent increases the recovery yield. In order to remove most of the contaminants from yeast, a two-step disruption/extraction scheme has been developed that facilitates downstream processing. Furthermore, it also has the advantage of minimizing proteolytic actions on the recombinant protein by removing most of the contaminants and proteases into the supernatant during the first disruption step, while keeping the desired protein in the pellet fraction. Final recovery is then achieved by the extraction process. Parameters affecting the disruption/extraction processes have been discussed.

Biotechnology↗
PubMed1994

Effects of estrogen and estrogen receptor in normal human melanocytes.

Normal human melanocytes were cultured selectively with F12 culture medium supplemented with growth hormones, phorbol ester and 1% of fetal calf serum. The estrogen receptors were analyzed using hydroxylapatite-column assay with tritiated 17-beta-estradiol as the binding ligand. Phenol red- free medium was used when the changes in cell numbers, melanin content and tyrosinase were assessed after incubating with physiological concentration of 17-beta estradiol (10(-12) and 10(-9) M). It was found that the melanocytes contained both cytosol (5.42 +/- 1.11 fmol/mg protein) and nuclear (59.13 +/- 17.12 fmol/mg protein) estrogen receptor. In response to estradiol, the cell number increased but both the melanin content and the tyrosinase activity decreased in a dose related pattern. These data suggested the presence of estrogen receptor with biological function in normal human melanocytes.

Animals↗
PubMed1994

A nested case-control study on association between hepatitis C virus antibodies and primary liver cancer in a cohort of 9,775 men in Taiwan.

Most studies on the association between antibodies against hepatitis C virus (anti-HCV) and primary liver cancer (PLC) were limited to case-series, or cross-sectional case-control studies leaving a controversy on causal temporality. A nested case-control study on 38 newly-developed PLC patients and 152 matched controls selected from a cohort of 9,775 men in Taiwan recruited from September, 1984, to February, 1986, was carried out to examine the relation between HCV infection and PLC. Case-control pairs were matched on age (+/- 1 year), residence, and the date at recruitment. Serum samples collected from study subjects at the initial recruitment were examined for anti-HCV by enzyme immunoassay and hepatitis B surface antigen (HBsAg) by reverse passive hemagglutination assay combined with radioimmunoassay. History of cigarette smoking, alcohol consumption, vegetable consumption, vegetarian habit, and chronic liver diseases were also obtained through standardized interviews according to a structured questionnaire at the recruitment. After adjusting for HBsAg status and other risk factors, the anti-HCV was significantly associated with the development of PLC showing a multivariate-adjusted relative risk of 88.24. The results suggest that HCV infection may play an important role in the etiology of human PLC in Taiwan.

Adult↗
PubMed1994

Infection at the site of implanted materials with and without preadhered bacteria.

A study was undertaken to examine the role of bacterial adherence in the development of infection at the site of an implant. The amount of in vitro adherence of Staphylococcus epidermidis was greatest for stainless steel, followed by polymethylmethacrylate and commercially pure titanium, and was least for polymethylmethacrylate with gentamicin. These materials then were preincubated with S. epidermidis and implanted. The number of organisms that were isolated and the rate of infection followed the same pattern as that in the in vitro studies. Materials that were not preincubated with bacteria also were implanted and bacteria were injected into the site. The number of organisms isolated from the site and the rate of infection were lower than those for the preincubated materials, but the trend was the same as in both the in vitro and the in vivo studies. The rates of infection and colonization correlated with the propensity for the organisms to adhere to a given material. Materials colonized with S. epidermidis at the time of implantation caused a high rate of infection. The ability of organisms to adhere to a material in vitro is correlated with their propensity to cause biomaterial-based infection.

Animals↗
PubMed1994

Characterization of a major cell wall antigen and potential adhesin in three strains of Candida albicans.

Selected strains of Candida albicans were examined to reveal the surface antigenicity and biochemical nature of major cell wall proteins that also were shown to serve as cellular adhesins on human buccal epithelial cells. Confirmation of the adhesive properties of these cells was made by scanning electron microscopy and immunofluorescence microscopy. Particular attention was directed at the clinical isolate KM-302. By means of indirect immunofluorescence staining, the KM-302 blastoconidia absorbed rabbit anti-C. albicans ATCC-32354 serum, revealing specific localization of surface antigens on germ tubes and pseudohyphae. Extracellular polymeric material and the cell wall extract of C. albicans KM-302 blastoconidia were found to contain a major surface antigen of 49 kDa that exhibited 42% adhesion inhibition in vitro. Of considerable significance is that immunogold localization by electron microscopy showed the antigen to be almost exclusively cell wall bound. This major antigen, identified in affinity and gel filtration chromatography fractions, was composed of 4% carbohydrate and 95.7% protein and had an isoelectric point of 6.1. The major antigen also showed a high level of similarity with that of C. albicans strain SC-5314 inasmuch as the major antigen of that strain had carbohydrate and protein compositions of 4 and 95.5%, respectively. Both of these strains also possessed the same percent of adhesion inhibition on human buccal epithelial cells.

Antigens, Fungal↗
PubMed1994

Energy transfer from tryptophan residues of proteins to 8-anilinonaphthalene-1-sulfonate.

The binding of the apolar fluorescent dye 8-anilinonaphthalene-1-sulfonate (ANS) to bovine serum albumin (BSA), phospholipase A2 (PLA2), ovalbumin, lysozyme, cobrotoxin and N-acetyltryptophanamide was used to assess the factors affecting the efficiency of energy transfer from Trp residues to the ANS molecule. We found that the efficiency of energy transfer from Trp residues to ANS was associated with the ability of proteins to enhance the ANS fluorescence. At the same molar concentration of protein, BSA enhanced ANS fluorescence most among these proteins; its Trp fluorescence was drastically quenched by the addition of ANS. Fluorescence enhancement of ANS in PLA2-ANS complex increased upon addition of Ca2+ or change of the buffer to acidic pH, resulting in a higher efficiency of energy transfer from Trp residues to ANS. There was limited ANS fluorescence enhancement with ovalbumin, lysozyme, cobrotoxin, and N-acetyltryptophanamide and a less efficient quenching in Trp fluorescence. The capabilities of proteins for binding with ANS correlated with the decrease in their Trp fluorescence being quenching by ANS. However, the microenvironment surrounding Trp residues of proteins did not affect the energy transfer. Based on these results, the factors that affected the energy transfer from Trp residues to ANS are discussed.

Anilino Naphthalenesulfonates↗
PubMed1994

Functional involvement of Lys-6 in the enzymatic activity of phospholipase A2 from Bungarus multicinctus (Taiwan banded krait) snake venom.

Phospholipase A2 (PLA2) from Bungarus multicinctus snake venom was subjected to Lys modification with 4-chloro-3,5-dinitrobenzoate and trinitrobenzene sulfonic acid, and one major carboxydinitrophenylated (CDNP) PLA2 and two trinitrophenylated (TNP) derivatives (TNP-1 and TNP-2) were separated by high-performance liquid chromatography. The results of amino acid analysis and sequence determination revealed that CDNP-PLA2 and TNP-1 contained one modified Lys residue at position 6, and both Lys-6 and Lys-62 were modified in TNP-2. It seemed that the Lys-6 was more accessible to modified reagents than other Lys residues in PLA2. Modification of Lys-6 caused a 94% drop in enzymatic activity as observed with CDNP-PLA2 and TNP-1. Alternatively, the enzyme modified on both Lys-6 and Lys-62 retained little PLA2 activity. Either carboxydinitrophenylation or trinitrophenylation did not significantly affect the secondary structure of the enzyme molecule as revealed by the CD spectra, and Ca2+ binding and antigenicity of Lys-6-modified PLA2 were unaffected. Conversion of nitro groups to amino groups resulted in a partial restoration of enzymatic activity of CDNP-PLA2 to 32% of that of PLA2. It reflected that the positively charged side chain of Lys-6 might play an exclusive role in PLA2 activity. The TNP derivatives could be regenerated with hydrazine hydrochloride. The biological activity of the regenerated PLA2 is almost the same as that of native PLA2. These results suggest that the intact Lys-6 is essential for the enzymatic activity of PLA2, and that incorporation of a bulky CDNP or TNP group on Lys-6 might give rise to the distortion of the interaction between substrate and the enzyme molecule, and the active conformation of PLA2.

Amino Acid Sequence↗
PubMed1994

The essentiality of B chain in stabilizing the structure of the A chain in beta 1-bungarotoxin from Bungarus multicinctus venom.

The dynamic of Trp residue in beta 1-bungarotoxin (beta 1-Bgt), the A chain of beta 1-Bgt and phospholipase A2 (PLA2) was assessed by fluorescence measurement. Acrylamide quenching studies showed that the exposure degree of the Trp in PLA2 is higher than the Trp in beta 1-Bgt. The Trp of beta 1-Bgt had a higher accessibility for iodide, reflecting that the basic nature of the B chain might exert an attractive electrostatic force for iodide and increase the susceptibility of Trp in the A chain to iodide. Removal of the B chain of beta 1-Bgt did not significantly affect the exposure degree of Trp in the A chain. Alternatively, the polarity of the environment around the Trp and the hydrophobic character of ANS and substrate binding sites in the separated A chain changed. Measurement of Trp fluorescence with increasing temperature showed that the stability of structure of beta 1-Bgt was higher than those of the separated A chain and PLA2. These results suggest that the B chain might interact with the A chain and stabilize the conformation of the A chain in beta 1-Bgt.

Animals↗
PubMed1994

Somatic cell genetic and biochemical characterization of cell lines resulting from human genomic DNA transfections of Chinese hamster ovary cell mutants defective in sterol-dependent activation of sterol synthesis and LDL receptor expression.

We have isolated several non-leaky mutant Chinese hamster ovary (CHO) cell clones (M4, M19, and M21) requiring cholesterol and unsaturated fatty acid for growth. These mutants belong to the same complementation group as the mutant M1 cells previously reported from this laboratory. M19 cells reverted to lipid prototrophy at very low frequency and were chosen as recipients to perform DNA-mediated gene-transfer experiments using total human genomic DNAs. Biochemical characterization of these transfectant clones indicated that, unlike their parental M19 cells, they were able to exhibit activation of cholesterol biosynthesis and LDL receptor expression in response to sterol removal from the growth medium. RNA blotting analysis indicated that these transfectants were able to increase HMG-CoA synthase gene transcripts in response to sterol removal. From the genomic DNAs of a representative secondary transfectant cells, we cloned a unique human DNA fragment (designated as h lambda 2) and showed that h lambda 2 closely linked with the presumptive human M1 gene.

Amphotericin B↗
PubMed1994

Localization of acyl coenzyme A:cholesterol acyltransferase gene to human chromosome 1q25.

Acyl coenzyme A:cholesterol acyltransferase (ACAT) is an intracellular enzyme that catalyzes the formation of cholesterol esters from cholesterol and long-chain fatty acyl-coenzyme A. It is believed that ACAT plays a key role in lipoprotein metabolism and atherogenesis. Recently our laboratory succeeded in molecular cloning and functional expression of human macrophage ACAT cDNA. We have now mapped the ACAT gene to chromosome 1, band q25 by using fluorescence in situ hybridization to metaphase chromosomes, and by Southern blotting analysis of human--hamster somatic cell hybrid panels.

Animals↗
PubMed1994

Urinary epidermal growth factor receptor-binding growth factors in patients with cancers of the digestive tract.

To investigate whether the urinary excretion of epidermal growth factor (EGF) and transforming growth factor-alpha (TGF-alpha) is increased in patients with cancer of the digestive tract, EGF and TGF-alpha were determined in 109 cancer patients and 40 healthy controls. Excluding EGF in hepatocellular carcinoma (HCC) and TGF-alpha in pancreatic cancer, both growth factors in each cancer group were significantly higher than in the control group. A receiver operating characteristic curve and likelihood ratio were applied to obtain the best diagnostic efficiency. Both EGF and TGF-alpha had high specificity (100%) in all cancer group. The high sensitivity of EGF in gastric cancer (100%) and metastatic liver cancer (93.3%), moderate sensitivity of TGF-alpha in metastatic liver cancer (86.6%), colon cancer (80.0%), and HCC (61.7%) suggested that they might be helpful in identifying these cancers. In conclusion, urinary excretion of EGF and TGF-alpha increased in most cancers of the digestive tract. They may be used as tumor markers.

Adult↗
PubMed1994

Effects of antimalarials and protease inhibitors on plasmodial hemozoin production.

Malarial hemozoin may play an important role as a target for antimalarial drugs and in disease pathogenesis. A new assay for hemozoin was developed in which the hemozoin was separated from cells by filtration. Trophozoites have substantially more hemozoin than rings, but there are relatively small differences between chloroquine-sensitive and chloroquine-resistant strains. The effects of hemozoin content of chloroquine and artemisinin, two antimalarial drugs, and E64 and Pepstatin A, two protease inhibitors, were measured. At concentrations at which hypoxanthine incorporation was unaffected, the hemozoin content of rings was decreased by E64, but not by the other three compounds. Artemisinin and Pepstatin A also had little effect on the hemozoin content of trophozoites. Chloroquine and E64 inhibited trophozoite hemozoin formation, but inhibited hypoxanthine uptake to a similar or greater extent. When either rings or trophozoites were exposed to several higher concentrations of chloroquine, hemozoin content was diminished, but significantly less than hypoxanthine uptake. Various concentrations of E64, in contrast, inhibited hemozoin production by both rings and trophozoites significantly more than hypoxanthine incorporation, suggesting that hemozoin production may be directly affected by E64.

Animals↗
PubMed1994

Immunological neutralization of cobrotoxin by its homologous precipitin and non-precipitin antibodies.

Anti-cobrotoxin antibodies can be separated into precipitin and non-precipitin antibodies. The precipitin antibody possesses the same binding affinity to cobrotoxin as non-precipitin antibody, but the neutralizing capability of the latter is superior to that of the former in blocking cobrotoxin binding to nicotinic acetylcholine receptor (nAChR). After preincubation with antibodies, cobrotoxin completely lost its binding activity to nAChR. The dissociation of cobrotoxin-nAChR complex by the antibodies was low, and 60% of the complex formation appeared to be irreversible. These results indicate that the neutralization of cobrotoxin by the antibody may predominantly involve unbound, receptor-free cobrotoxin. The relationships of neutralization capacity and binding affinity as well as bond strength between cobrotoxin and its antibodies are incongruous. Different local conformational changes of a unique Trp in cobrotoxin on binding with the precipitin and non-precipitin antibodies seem to lead to different accessibility for fluorescence quenchers. Characterization of the binding domains by immunoprecipitation with the antibodies correlated with the quenching results. Thus, the binding topography of cobrotoxin may play an important role over the binding affinity and bond strength in neutralization by cobrotoxin antibody.

Animals↗
PubMed1994

BCL-6 and the molecular pathogenesis of B-cell lymphoma.

The results presented identify the first genetic lesion associated with DLCL, the most clinically relevant form of NHL. Although no proof yet exists of a role for these lesions in DLCL pathogenesis, the feature of the BCL-6 gene product, its specific pattern of expression in B cells, and the clustering of lesions disrupting its regulatory domain strongly suggest that deregulation of BCL-6 expression may contribute to DLCL development. A more precise definition of the role of BCL-6 in normal and neoplastic B-cell development is the goal of ongoing study of transgenic mice engineered either to express BCL-6 under heterologous promoters or lacking BCL-6 function due to targeted deletions. In addition to contributing to the understanding of DLCL pathogenesis, the identification of BCL-6 lesions may have relevant clinical implications. DLCL represent a heterogeneous group of neoplasms which are treated homogeneously despite the fact that only 50% of patients experience long-term disease-free survival (Schneider et al. 1990). The fact that BCL-6 rearrangements identify biologically and clinically distinct subsets of DLCL suggests that these lesions may be useful as markers in selection of differential therapeutic strategies based on different risk groups. Furthermore, the BCL-6 rearrangements can be used to identify and monitor the malignant clone with sensitive PCR-based techniques. Since clinical remission has been observed in a significant fraction of DLCL cases, these markers may serve as critical tools for sensitive monitoring of minimal residual disease and early diagnosis of relapse (Gribben et al. 1993).

Animals↗
PubMed1994

Pharmacological relevance of peripheral type benzodiazepine receptors on motor nerve and skeletal muscle.

1. Effects of agonists and antagonists of peripheral and central benzodiazepine receptors (pBZR and cBZR) on neuromuscular transmission were studied in mouse isolated phrenic nerve-diaphragm preparations. 2. Ro5-4864, a pBZR agonist, had no effect on the neuromuscular transmission but increased muscle contractility and antagonized the tetanic fade induced by neostigmine. 3. Ro5-4864 inhibited the regenerative tonic endplate depolarization caused by repetitive stimulation in the presence of neostigmine without affecting the amplitude and decay time of miniature and evoked single endplate potentials. 4. All the effects of Ro5-4864 were shared by PK11195, a pBZR antagonist, but not by clonazepam and flumazenil, a cBZR agonist and antagonist, respectively. 5. It is suggested that peripheral type benzodiazepine receptors modulate presynaptic function and muscle contraction.

Animals↗
PubMed1994

The role of modulated gap junctional intercellular communication in epigenetic toxicology.

The normal development and health of all multicellular organisms, including the human being, depend on the adaptive maintenance of the integrity of the genetic information (e.g., DNA protective and repair mechanisms), as well as of the homeostatic and cybernetic regulatory systems within and between tissues. The primary focus of the past and current toxicological studies and risk assessment practices has been to ascertain and predict the "genotoxicity" of various physical and chemical agents. The paradigm of "carcinogen as mutagen," while valuable for stimulating studies of the detection of mutagens and of their potential role in "causing" somatic and germ line diseases, has tended to blunt research on the role of nongenotoxic mechanisms in disease causation. This brief analysis will emphasize the need to consider the role of modulated gap junctional intercellular communication (GJIC) in any biological risk assessment model. It is based on the following assumptions and facts. Because gap junctions exist in all metazoans, they have been associated with the regulation of cell proliferation, development, differentiation, and the adaptive function of both excitable and nonexcitable coupled cells. A highly evolutionarily conserved family of genes codes for proteins (connexins), which, as hexameric units (connexons), form membrane-associated channels of gap junctions. Cells coupled by gap junctions will have their ions and small regulatory molecules equilibrated. Regulation of GJIC can be at the transcriptional, translational, or posttranslational levels. Transient down or up regulation of GJIC can be induced by endogenous or exogenous chemicals via many mechanisms at any of these three levels. Stable abnormal regulation has been associated with activated oncogenes, and normal regulation has been associated with several tumor suppressor genes. The dysfunction of these gap junctions might play a role in the actions of various toxic chemicals that have cell type/tissue/organ specificity. This could bring about distinct clinical consequences, such as embryo lethality or teratogenesis, reproductive dysfunction in the gonads, neurotoxicity of the central nervous system, hyperplasia of the skin, and tumor promotion of initiated tissue. Modulation of GJIC should be viewed as a scientific basis of "epigenetic toxicology" because the alteration of intercellular communication would alter the internal physiological state of the cell. The inhibition of GJIC is a necessary component of mitogenesis (a necessary component of the multistage carcinogenic process). The modulation of GJIC can have both toxicological, as well as therapeutic potential.

Animals↗
PubMed1994

Building components contributors of the sick building syndrome.

The sick building syndrome has been widely discussed from epidemiological perspectives. Although there is considerable difference in opinion regarding the concrete and objective evidence to support a distinct sick building syndrome and/or building-related illness, much data indicates that numerous variables within buildings can potentially influence human health. In this paper, we discuss in detail not only the potential and unique infectious diseases caused by Legionella, Pontiac fever, Q fever, and influenza, but also the data implicating noninfectious etiologies of sick building syndrome and building-related illnesses. In addition, the role of psychological factors, mass hysteria, and indoor pollution is discussed with respect to the nature of associations between exposure and symptoms. Finally, comparisons are made in different building construction types of old versus new buildings to highlight changes in modern construction that may have led to a putative increase in work-related symptomatology.

Air Pollution, Indoor↗