Search PubMed⌕ Search

Biomedical subjects

S F Chang

Publications and source records attributed to S F Chang.

At least 37 records · Page 2Linked to original sources
PubMed1998

Assaying for structural variation in the parvovirus capsid and its role in infection.

The capsid of canine parvovirus (CPV) was assayed for susceptibility to proteases and for structural variation. The natural cleavage of VP2 to VP3 in CPV full (DNA containing) particles recovered from tissue culture occurred within the sequence Arg-Asn-Glu-Arg Ala-Thr. Trypsin, chymotrypsin, bromelain, and cathepsin B all cleaved >90% of the VP2 to VP3 in full but not in empty capsids and did not digest the capsid further. Digestion with proteinase K, Pronase, papain, or subtilisin cleaved the VP2 to VP3 and also cleaved at additional internal sites, causing particle disintegration and protein degradation. Several partial digestion products produced by proteinase K or subtilisin were approximately 31-32.5 kDa, indicating cleavage within loop 3 of the capsid protein as well as other sites. Protease treatment of capsids at pH 5.5 or 7.5 did not significantly alter their susceptibility to digestion. The isoelectric point of CPV empty capsids was pH 5.3, and full capsids were 0.3 pH more acidic, but after proteolysis of VP2 to VP3, the pI of the full capsids became the same as that of the empty capsids. Antibodies against various capsid protein sequences showed the amino termini of most VP2 molecules were on the outside of full but not empty particles, that the VP1-unique sequence was internal, and that the capsid could be disintegrated by heat or urea treatment to expose the internal sequences. Capsids added to cells were localized within the cell cytoplasm in vesicles that appeared to be lysosomes. Microinjected capsids remained primarily in the cytoplasm, although a small proportion was observed to be in the nucleus after 2 h. After CPV capsids labeled with [35S]methionine were bound to cells at 0 degrees C and the cells warmed, little cleavage of VP1 or VP2 was observed even after prolonged incubation. Inoculation of cells with virus in the presence of proteinase inhibitors did not significantly reduce the infection.

Animals↗
PubMed1998

The negative influence of endogenous opioid receptor activity on the differentiation of the rat pheochromocytoma PC12 cells induced by nerve growth factor.

We determined the effect of naloxone and morphine on the differentiation of pheochromocytoma cell, the PC12 cells, induced by nerve growth factor (NGF). PC12 cells were grown in medium containing NGF with or without the addition of naloxone or morphine for up to 10-day treatments. NGF-induced morphological differentiation of PC12 cells was manifested by an increase in the percentage of differentiated cells and the average length of neurite per cell. Co-addition of morphine with NGF did not affect both parameters as compared to that of NGF alone. On the contrary, co-addition of naloxone with NGF significantly increased the percentage of differentiated cells, but did not affect the outgrowth of neurites. This effect of naloxone was reversed by the addition of morphine, suggesting that naloxone produced its effect by inhibiting the endogenous activity of opioid receptor. This study indicates a significant functional role of opioid receptor in NGF-induced differentiation of PC12 cells.

Analgesics, Opioid↗
PubMed1998

The -104G nucleotide of the human CYP21 gene is important for CYP21 transcription activity and protein interaction.

CYP21 gene encodes the steroid 21-hydroxylase (P450c21) that is involved in steroidogenesis in the adrenal cortex. Mutations occurring on CYP21 which convert it to the neighboring pseudogene, CYP21P, are found in patients with congenital adrenal hyperplasia (CAH), an autosomal recessive disease. We previously reported that the CYP21P pseudogene had lower transcription activity when compared with the active CYP21 gene. The sequences determining the basal transcription activity of the human CYP21 gene are located within the 166 bp region upstream from the transcriptional start site. Within this region, only 4 nucleotides are different between the active CYP21 and the CYP21P pseudogene; they are located at the -117, -104, -101 and -94 positions from the start site of the gene. Here, we report that the CYP21 gene-specific G nucleotide sequence at the -104 position is required for its basal transcription activity driven by the native TATA box of the human CYP21 gene. When this single nucleotide is changed to the CYP21P sequence, the basal transcription activity decreases by approximately 80% in transient transfection assay. In addition, our data from gel retardation assay show that this sequence is also critical for interaction with nuclear proteins from adrenal cells. These results therefore suggest that the single G sequence of the human CYP21 gene is crucial for the expression of its basal transcription activity, and this may be influenced by the interaction with specific nuclear proteins from the adrenal gland.

Adrenal Cortex↗
PubMed1998

Diagnostic pitfalls of Merkel cell carcinoma and dramatic response to chemotherapy.

Merkel cell carcinoma (MCC) is an unusual malignant tumor that arises from neuroendocrine cells with features of epithelial differentiation. We describe a MCC patient with unusual clinical, histopathological, and immunohistochemical features. Although the microscopic, immunohistochemical, and ultrastructural characteristics of MCC have been well defined, diagnostic difficulties remain, particularly in distinguishing it from lymphoma involving the skin, as suggested by our case. This is an unusual case in which dense lymphoid infiltration masked the true tumor. All the immunohistochemical markers of MCC except neuron-specific enolase (NSE) were negative. The dramatic response to primary chemotherapy was also very noteworthy.

Adult↗
PubMed1998

Detailed characterization of the binding site of the lipoprotein lipase-specific monoclonal antibody 5D2.

Monoclonal antibody (MAb) 5D2 recognizes lipoprotein lipases (LPL) from different species but not related lipases. This MAb is a unique reagent, used world-wide, because it differentiates between monomeric inactive and dimeric active LPL, inhibits human LPL enzyme activity, and binds to C-terminal LPL sequences involved in interactions with lipoproteins, lipoprotein receptors, and heparin. In this study we have analyzed the fine specificity of the MAb epitope recognition in order to better understand its functional properties and species-specific LPL immune reactivity. In peptide scan assays, MAb 5D2 reacted with all, except two, 13 amino acid-long peptides located between positions 380 and 410. Peptides from the amino terminal end of this region reacted more strongly than those from the carboxyl terminal end. Furthermore, only a peptide from the amino terminal end competed effectively with the binding of MAb 5D2 to native LPL bound to microtiter plates or nitrocellulose. A systematic peptide mutagenesis study indicated that 8 amino acids of the reactive region, mainly located in the amino terminal end, are critical for binding and probably directly interact with MAb 5D2. The experimentally determined antigenicities of species-specific LPL peptides and of the corresponding denatured full-length LPL proteins on immunoblots were consistent with these findings. According to a proposed 3D-model for LPL, only the amino terminal end of the antigenic region is easily surface-accessible. These data combined with 3D-modelling of monoclonal antibody (MAb)-lipoprotein lipase (LPL) protein interaction provide new insight into the known biological effects of MAb 5D2 on LPL and the antigenic determinants that are recognized.

Amino Acid Sequence↗
PubMed1997

Hepatitis B virus X protein and p53 tumor suppressor interactions in the modulation of apoptosis.

We have reported previously that the hepatitis B virus oncoprotein, HBx, can bind to the C terminus of p53 and inhibit several critical p53-mediated cellular processes, including DNA sequence-specific binding, transcriptional transactivation, and apoptosis. Recognizing the importance of p53-mediated apoptosis for maintaining homeostasis and preventing neoplastic transformation, here we further examine the physical interaction between HBx and p53 as well as the functional consequences of this association. In vitro binding studies indicate that the ayw and adr viral subtypes of HBx bind similar amounts of glutathione S-transferase-p53 with the distal C terminus of HBx (from residues 111 to 154) being critical for this interaction. Using a microinjection technique, we show that this same C-terminal region of HBx is necessary for sequestering p53 in the cytoplasm and abrogating p53-mediated apoptosis. The transcriptional transactivation domain of HBx also maps to its C terminus; however, a comparison of the ability of full-length and truncated HBx protein to abrogate p53-induced apoptosis versus transactivate simian virus 40- or human nitric oxide synthase-2 promoter-driven reporter constructs indicates that these two functional properties are distinct and thus may contribute to hepatocarcinogenesis differently. Collectively, our data indicate that the distal C-terminal domain of HBx, independent of its transactivation activity, complexes with p53 in the cytoplasm, partially preventing its nuclear entry and ability to induce apoptosis. These pathobiological effects of HBx may contribute to the early stages of hepatocellular carcinogenesis.

Adult↗
PubMed1997

Genomic deletion and p53 inactivation in cervical carcinoma.

The tumor-suppressor gene p53 acts as "the guardian of the genome", sensing DNA damage and initiating protective responses. To examine the hypothesis that p53 abnormality leads to increased genomic alterations in primary tumor cells, our study utilized 51 primary tumors of cervical carcinoma and 10 microsatellite markers. These markers were mapped to the short arms of chromosomes 3 and 5, covering the regions 3p13-25 and 5p15.1-15.3. Genomic deletion on 3p and 5p was correlated with genetic or epigenetic p53 inactivation pathways, including p53 mutation, genetic deletion of p53 and cervical infection with human papillomavirus. The proportion of abnormal p53 was found to be significantly higher in the cases exhibiting loss of heterozygosity (LOH) on 5p (p < 0.001), supporting the hypothesis of the presence of a p53-dependent pathway to cervical tumorigenesis. In contrast, however, LOH on 3p was found to be independent of p53 inactivation. A common deletion region, 3p22-24, was identified in 44% of informative cases, and genomic loss at this specific region was correlated with early tumorigenic onset and poor grade of tumor differentiation. Diversity within the patterns of genomic alteration in the same form of cancer suggests different sets of risk/tumorigenic profiles, molecular pathogenesis, as well as prognosis and outcome.

Adult↗
PubMed1997

Structural and functional analysis of the promoter of the hepatic lipase gene.

Hepatic lipase (HL) gene transcription is almost exclusively limited to hepatocytes. Here we have studied sequences and transcription factors regulating basal and hepatocyte-restricted HL promoter activity. Sequencing of a cloned 3.4-kb HL promoter fragment revealed three Alu repeat sequences and a consensus hepatocyte-enriched nuclear transcription factor 1 (HNF1) binding site located upstream of one major and one minor transcription initiation site. By transfection of cell lines of hepatic and non-hepatic origin and of primary hepatocyte cultures, sequences controlling basic HL promoter activity and negative elements located downstream and upstream thereof which extinguish or enhance this activity were defined. Some HL-promoter fragments with internal deletions were active only in primary hepatocyte cultures. Human HNF1 protein was shown to bind to the HL-specific HNF1 response element and the activity of a heterologous promoter was enhanced by HL-HNF1 in rat primary hepatocyte cultures but not in the context of the authentic 3.4-kb HL promoter sequences. In cell lines the presence of HNF4 but not of HNF1 and vHNF1 mRNA was found to correlate with HL gene expression although no perfect consensus HNF4 binding motif was detected in the promoter region tested. Taken together, these data indicate that hepatocyte-specific HL gene transcription is controlled by positive and negative transcription regulatory proteins which bind to sequence motifs within and outside of the proximal 3.4-kb promoter fragment studied. For the elucidation of the control of HL promoter activity in vivo the use of primary hepatocyte cultures is essential.

Animals↗
PubMed1997

Pharmacokinetics and dose proportionality of beclomethasone from three strengths of a CFC-free beclomethasone dipropionate metered-dose inhaler.

As part of a development program to offer alternatives to chlorofluorocarbon (CFC) containing metered-dose inhalers, beclomethasone dipropionate has been formulated in a CFC-free system at three strengths: 50, 100, and 200 micrograms/actuation ex valve. To measure serum levels and dose proportionality of the beclomethasone derived from beclomethasone dipropionate, 13 mild to moderate asthmatic patients received a single dose of eight inhalations from each strength according to a double-blind crossover design. Seven patients were studied over 4 h and six patients over 12 h. For the total doses of 400, 800, and 1600 micrograms studied over 12 h, Cmax and AUC increased in a ratio of 1:1.8:3.1. A good correlation was seen between the fine-particle mass delivered and the in vivo performance of the three strengths. From a clinical point of view, the predictable increases in serum levels with an increase in dose will permit the clinician to effectively titrate a patient with this product.

Administration, Inhalation↗
PubMed1997

Novel patterns of p53 abnormality in breast cancer from Taiwan: experience from a low-incidence area.

Among 114 breast cancers in Taiwan, the prevalence of p53 mutation (22.8%) and p53 accumulation (38.3%) was similar to that in high-incidence areas. However, novel patterns of p53 abnormalities, including unique sites or types of mutation (i.e. an excessive proportion of G:C to A:T transition at CpG site), and accumulation of wild-type p53 either within nuclear or cytoplasmic compartments were noted. These may have relevance to breast cancer in Taiwan, a low-incidence area.

Breast Neoplasms↗
PubMed1997

Comparative serum estradiol profiles from a new once-a-week transdermal estradiol patch and a twice-a-week transdermal estradiol patch.

Two identical open-label, randomized crossover studies were conducted to compare serum estradiol profiles from the new 12.5- and 25-cm2 once-a-week adhesive patches with those from the 10- and 20-cm2 commercially available twice-a-week Estraderm patches when applied as directed during a 1-week patch-wear period. Both studies were conducted in healthy postmenopausal women; serum estradiol levels were determined by gas chromatography/mass spectroscopy (GC/MS). Although both sizes of both patch treatments produced mean serum estradiol levels in the therapeutic range, the once-a-week patch provided more constant mean levels, avoiding large peak-to-trough fluctuations. As expected, the differences in mean serum estradiol concentrations between the two patch treatments occurred during the second application of the twice-a-week patch. Based on these results, the once-a-week drug in adhesive patch appears to be an acceptable means of hormone replacement therapy.

Administration, Cutaneous↗
PubMed1997

Sequence heterogeneity of heron hepatitis B virus genomes determined by full-length DNA amplification and direct sequencing reveals novel and unique features.

So far, only a single heron hepatitis B virus genome (HHBV-4) has been cloned and sequenced. Therefore, neither the significance of its sequence divergence from other avian hepadnaviruses nor the sequence variability of HHBV genomes in general are known. Here we have analysed the sequence heterogeneity of HHBV genome populations in several sera from naturally infected herons. A highly sensitive PCR method for full-length HHBV genome amplification was established which allowed direct sequencing of entire HHBV populations without prior cloning. Sequences of HHBV genomes from four sera were thus obtained which differed from those of HHBV-4 by up to 7%. Some of the divergent nucleotides and the corresponding amino acids of the predicted viral proteins were conserved in all four new HHBV isolates and varied only in HHBV-4. This indicates that the HHBV-4 genome is not in all aspects representative of this class of viruses. Interestingly, a highly conserved ORF upstream of the C-gene present in a position analogous to that of the mammalian hepadnavirus X-gene became apparent in all HHBV genomes. In contrast to the duck hepadnaviruses, the small (sAg-S) instead of the largest (sAg-L) envelope protein of all HHBVs has a myristylation site. These data confirm the significant sequence divergence of HHBV from other avian hepadnaviruses. Moreover, they show that HHBV has low sequence variability and indicate two new and unique features not evident in other avihepadnaviruses: an additional, highly conserved gene and potential myristylation of the sAg-S instead of the sAg-L envelope protein.

Amino Acid Sequence↗
PubMed1997

Dose-response relationship between prevalence of cerebrovascular disease and ingested inorganic arsenic.

BACKGROUND AND PURPOSE: Circulatory diseases such as ischemic heart disease and peripheral vascular disease induced by long-term arsenic exposure have been well documented in previous studies, but the dose-response relationship between cerebrovascular disease and ingested inorganic arsenic remains to be elucidated. The prevalence of cerebrovascular disease among residents of the Lanyang Basin on the northeast coast of Taiwan was surveyed to examine its association with exposure to arsenic in well water. METHODS: A total of 8102 men and women from 3901 households were recruited in this study. The status of cerebrovascular disease of study subjects was identified through home-visit personal interviews and ascertained by review of hospital medical records according to the World Health Organization criteria. Information on consumption of well water, sociodemographic characteristics, cigarette smoking, and alcohol consumption habits, as well as personal and family history of diseases, was also obtained. Arsenic concentration in the well water of each household was determined by hydride generation and atomic absorption spectrometry. Logistic regression analysis was used to estimate multivariate-adjusted odds ratios and 95% confidence intervals for various risk factors of cerebrovascular disease. RESULTS: A significant dose-response relationship was observed between arsenic concentration in well water and prevalence of cerebrovascular disease after adjustment for age, sex, hypertension, diabetes mellitus, cigarette smoking, and alcohol consumption. The biological gradient was even more prominent for cerebral infarction, showing multivariate-adjusted odds ratios of 1.0, 3.4, 4.5, and 6.9, respectively, for those who consumed well water with an arsenic content of 0, 0.1 to 50.0, 50.1 to 299.9, and > 300 micrograms/L. CONCLUSIONS: Long-term exposure to inorganic arsenic from well water was associated with an increased prevalence of cerebrovascular disease, especially cerebral infarction.

Adult↗
PubMed1997

Genetic localization of the type I trimethoprim resistance gene and its dissemination in urinary tract isolates in Taiwan.

In a total of 425 urinary isolates of E. coli, Enterobacter spp., and Klebsiella spp. selected, there were 169 (45.4%) isolates harbouring type I dihydrofolate reductase (DHFR) gene among 374 trimethoprim-resistant isolates. In these 169 isolates, only 17.2% hybridized with the Tn7 probe. According to another probe specific for the integrase gene of integron, 87.6% showed a positive reaction. Further analysis by restriction mapping proved that the type I DHFR gene was inserted into a integron-like structure. These results indicate that the type I DHFR gene that was initially observed in association with transposable element Tn7 is becoming associated with an integrase function similar to integrons in most instances. Further analysis of the distribution of Tn21-like integrase gene in clinical isolates indicated that the prevalence rates were 86.4%, 84.8%, and 76.7% respectively in E. coli, Enterobacter spp., and Klebsiella spp.. Furthermore, the integrase gene found in our clinical isolates proved to be mediated by a plasmid, demonstrated by Southern hybridization. Thus, the trimethoprim-resistant gene that developed under selective pressure from the double drug trimethoprim and sulphonamide was transmitted by insertion into integron-like structure and then mediated by plasmid transfer for dissemination.

Animals↗
PubMed1997

Allelic loss at BRCA1, BRCA2, and adjacent loci in relation to TP53 abnormality in breast cancer.

Cells with abnormal TP53 lose cell cycle checkpoints, resulting in genomic instability and neoplastic transformation. However, the evidence linking the tumor-specific targets of genomic alteration to an abnormal TP53 is limited. The present study tested the hypothesis that TP53 abnormalities are correlated with an increased frequency of deletion of breast cancer susceptibility loci (17q and 13q) in breast carcinomas. Tumors from 90 patients were examined for TP53 abnormality and loss of heterozygosity (LOH) at 11 loci on 17q (17q11.2-21) and 13q (13q12-14), including the loci for BRCA1 and BRCA2. A higher frequency of LOH was consistently found at 17q or 13q loci in tumors with an abnormal TP53. The increased LOH in relation to TP53 abnormality was statistically significant at the BRCA1, D17S588, and D13S267 loci (P < 0.05) but not at the locus for BRCA2 (P = 0.64). These observations imply a possible link between an abnormal TP53 and specific genomic deletions of breast cancer susceptibility loci, which may provide clues to the role of TP53 during breast tumorigenesis.

Adult↗
PubMed1996

The common I172N mutation causes conformational change of cytochrome P450c21 revealed by systematic mutation, kinetic, and structural studies.

We have investigated the structure and function of P450c21 with regard to a conserved site around Ile-172 by site-directed mutagenesis making single amino acid substitutions of residues 169 173. Substitutions of Ile-171 and -172 resulted in production of mutant proteins with dramatic reductions in enzymatic activities, indicating the importance of these two residues in maintaining the structure and function of P450c21. The I171N protein was present at a slightly lower level, due to a decreased rate of protein synthesis. The I172N apoprotein was synthesized at the normal rate, but its heme-bound P450 form was present at a much lower level. This I172N protein was tightly integrated into the membrane of endoplasmic reticulum, similar to the wild type P450c21, as shown by immunofluorescence detection, alkaline extraction, and cellular fractionation. Kinetic studies indicated that I172N had a lower Vmax value. In addition, the I172N protein was more sensitive to proteinase K digestion, indicating a possible alteration of conformation. This conformational change may result in the lower yield of the I172N hemoprotein and the reduced catalytic activity.

Amino Acid Sequence↗
PubMed1996

Molecular epidemiology of cytomegalovirus infection in kindergarten children.

This study was conducted to determine the source of cytomegalovirus (CMV) excretion in children who had persistent viruria. A total of 287 children, ages 3-5 years, enrolled in two kindergartens were followed for 9 months, and 28.8% of 139 CMV-infected children were found to have shed virus persistently in urine for > or = 3 months. A polymerase chain reaction (PCR)-based method provided direct evidence to demonstrate differences in CMV strains shed by 32 persistent shedders selected at random. The glycoprotein B (gB) nucleotide regions of CMV were amplified and analyzed by restriction enzyme digestion. The CMV strains shed by most of these children showed different restriction profiles compared with strains from their playmates. It is considered that persistent reactivation of endogenous CMV is the most plausible explanation.

Antibodies, Viral↗