Biomedical subjects

B C Chung

Publications and source records attributed to B C Chung.

At least 55 records · Page 3Linked to original sources

PubMed1999

Determination of testosterone:epitestosterone ratio after pentafluorophenyldimethylsilyl-trimethylsilyl derivatisation using gas chromatography-mass spectrometry in equine urine.

A highly specific method is described for measuring the testosterone:epitestosterone ratio in equine urine by gas chromatography-mass spectrometry (GC-MS) with stable isotope internal standards. The procedure was based on Serdolit Pad-1 resin extraction, enzymatic hydrolysis, and chemical derivatisation prior to instrumental analysis. The mixed derivatives, 3-trimethylsilyl-17-pentafluorophenyldimethylsilyl ether (3-TMS-17-flophemesyl) testosterone and epitestosterone, were found to have excellent analytical properties. The specificity of the derivatisation method exploits a unique feature of steroids: the selective exchange of the alcoholic flophemesyl ether for the trimethylsilyl ether. The sensitivity and specificity of the mixed 3-TMS-17-flophemesyl derivatives allow adequate determinations of testosterone and epitestosterone, even in urine from mares, in 5 ml samples. The repeatability of testosterone and epitestosterone was 6.2 and 5.7%, respectively, and their reproducibility was in the range of 6.4-8.7%.

Animals↗

PubMed1999

GC-MS determination of steroids related to androgen biosynthesis in human hair with pentafluorophenyldimethylsilyl-trimethylsilyl derivatisation.

An efficient method for the simultaneous determination of eight steroids, androstenedione, dihydrotestosterone (DHT), dehydroepiandrosterone (DHEA), testosterone, androsterone, etiocholanolone, progesterone and pregnenolone, in human hair by gas chromatography-mass spectrometry (GC-MS) using d3-testosterone as internal standard is described. The method involves alkaline digestion, liquid-liquid extraction and subsequent conversion to mixed pentafluorophenyldimethylsilyl-trimethylsilyl (flophemesyl-TMS) derivatives for sensitive analysis in the selected ion monitoring (SIM) mode. This method showed good overall repeatability and reproducibility of 4.88-11.24 and 3.19-9.58%, respectively. For the first time, the quantification of DHT, DHEA and pregnenolone in human hair has been achieved by GC-MS, testosterone was also quantified. The detection of four steroids in hair samples was possible in the concentration range 0.12-8.45 ng g-1. The other four steroids, androstenedione, androsterone, etiocholanolone and progesterone, were not detected. The detection limits for SIM of the steroids varied in the range 0.02-0.5 ng g-1, and the SIM responses were linear with correlation coefficients varying from 0.991 to 0.996 for most of the steroids studied. The concentrations of the four steroids detected were different in male and female hair samples.

Androgens↗

PubMed1999

Respiratory abnormalities among male foundry workers in central Taiwan.

The objectives of this study were to determine the relationship between exposure levels and respiratory abnormalities, to measure FVC and FEV1(1) changes per year based on work duties and to investigate the prevalence of and factors related to pneumoconiosis. A total of 583 male workers from 50 iron foundries in central Taiwan were investigated. First, workers' respiratory symptoms were categorized using a modified American Thoracic Society (ATS) questionnaire and then were verified by physician's examination. Next, pulmonary function tests were performed including: forced vital capacity (FVC), forced expiratory volume in one second (FEV1) and forced expiratory flow rate. A chest radiograph was used to diagnose pneumoconiosis according to ILO criteria. Furnace workers were found to have the highest prevalence of chronic phlegm, thoracic disorders and chronic bronchitis. In general, smokers had a higher prevalence of respiratory symptoms as compared with non-smokers. Pulmonary function abnormalities and pneumoconiosis were closely linked to smoking and work duration. After adjusting for age, height and smoking there was a significant decrease based on work duration in FVC and FEV1 for furnace and moulding workers compared with after-processing and administrative workers. The overall prevalence of pneumoconiosis was 8.8%, highest among furnace (16.3%) and after-processing workers (11.4%) and lowest among administrative workers (2.5%). Using multiple logistic regression, the risk of developing pneumoconiosis (as compared with the administrative workers) for furnace workers was highest (8.98 times greater risk), followed by after-processing workers (6.77 times greater risk) and moulding workers (5.41 times greater risk). Prolonged exposure to free silica, and smoking habits, can result in respiratory abnormalities among foundry workers.

Adult↗

PubMed1999

Tissue-specific, hormonal, and developmental regulation of SCC-LacZ expression in transgenic mice leads to adrenocortical zone characterization.

We report here the study of the human CYP11A1 promoter in driving tissue-specific, developmentally and hormonally regulated reporter gene expression. A 4.4-kb fragment containing all known regulatory elements is more efficient than a short basal promoter fused to an upstream adrenal enhancer in driving reporter LacZ gene expression both in cell culture and in transgenic mice. The LacZ gene controlled by the 4.4- and 2.3-kb promoters was expressed in the adrenal cortex, testicular Leydig cells, ovarian corpora lutea, and granulosa cells. Transgene expression in the adrenals was stimulated by ACTH, indicating the presence of ACTH-responsive sequence. Beta-galactosidase activity was first detected in the adrenal primordia at 11.5 days postcoitum. Its expression continued throughout all stages of adrenal development in a pattern similar to that of the endogenous CYP11A1, which was expressed in all zones of the adrenal cortex, but was strongest in the X zone. The X zone grew before puberty but regressed afterward, as did the levels of CYP11A1 and LacZ gene expression in the X zone. Our study of the CYP11A1 promoter in transgenic mice led to characterization of the adrenocortical zones.

Adrenal Cortex↗

PubMed1999

Function of steroidogenic factor 1 domains in nuclear localization, transactivation, and interaction with transcription factor TFIIB and c-Jun.

Normal endocrine development and function require nuclear hormone receptor SF-1 (steroidogenic factor 1). To understand the molecular mechanism of SF-1 action, we have investigated its domain function by mutagenesis and functional analyses. Our mutant studies show that the putative AF2 (activation function 2) helix located at the C-terminal end is indispensable for gene activation. SF-1 does not have an N-terminal AF1 domain. Instead, it contains a unique FP region, composed of the Ftz-F1 box and the proline cluster, after the zinc finger motif. The FP region interacts with transcription factor IIB (TFIIB) in vitro. This interaction requires residues 178-201 of TFIIB, a domain capable of binding several transcription factors. The FP region also mediates physical interaction with c-Jun, and this interaction greatly enhances SF-1 activity. The putative SF-1 ligand, 25-hydroxycholesterol, has no effects on these bindings. In addition, the Ftz-F1 box contains a bipartite nuclear localization signal (NLS). Removing the basic residues at either end of the key nuclear localization sequence NLS2.2 abolishes the nuclear transport. Expression of mutants containing only the FP region or lacking the AF2 domain blocks wild-type SF-1 activity in cells. By contrast, the mutant having a truncated nuclear localization signal lacks this dominant negative effect. These results delineate the importance of the FP and AF2 regions in nuclear localization, protein-protein interaction, and transcriptional activation.

Amino Acid Sequence↗

PubMed1999

Identification of urinary metabolites of clemastine after oral administration to man.

The metabolism of clemastine, 2-[2-[1-(4-chlorophenyl)-1-phenylethoxylethyl])-1-methylpyrrolidin e, has been studied in three adult male volunteers after a single oral dose of 20 mg as the fumarate. After enzymatic hydrolysis solvent extracts of urine were derivatized with N-methyl-N-trimethylsilyltrifluoroacetamide-ammonium iodide and analysed by gas chromatography-mass spectrometry. The structures of metabolites were determined on the basis of electron impact and chemical ionization mass spectra and the identities of some (e.g. carbinol, 4-chlorobenzophenone and 4-chlorophenylstyrene) were confirmed by use of authentic standards. The principal route of metabolism of clemastine in man involves direct oxidation, O-dealkylation (fission of the ether bond), aromatic hydroxylation, aliphatic oxidation, alcoholic dehydration, and then enzymatic hydrolysis. Of the total amount of metabolites excreted in the urine 35% was carbinol (metabolite M3, major metabolite), 15% was M1, 17% was M2, 11% was M4, 9% was M5, 8% was M6 and 5% was M7.

Acetamides↗

PubMed1998

Gas chromatographic-mass spectrometric determination of urinary oxoacids using O-(2,3,4,5,6-pentafluorobenzyl)oxime-trimethylsilyl ester derivatization and cation-exchange chromatography.

We introduced a new combined method to isolate, purify and quantify oxoacids in human urine. Preparation of O-(2,3,4,5,6-pentafluorobenzyl) oximes of oxoacids at pH 2 to 3 was followed by cation-exchange column chromatography for removing the biological interferences. The effluent with water was extracted with ethyl acetate and the oxoacids were quantitatively converted into their trimethylsilyl derivatives for detection by gas chromatography-mass spectrometry. Good quality control data were obtained through precision and accuracy tests. Analytical recoveries (53.5-99.8%) were quantitative for a wide variety of oxoacids. This method was used for the measurement of 18 oxoacids in the urine of healthy volunteers.

Acids↗

PubMed1998

Estrogens and polyamines in breast cancer: their profiles and values in disease staging.

The urinary concentrations of 16 estrogens and 11 polyamines were quantitatively determined by gas chromatography-mass spectrometry and gas chromatography with nitrogen-phosphorus detection. Samples from patients with stages I-IV of breast cancer (35 cases, aged 27-65 years) as well as from age-matched normal female subjects (25 cases, aged 22-61 years) were tested. Also, the ratios of precursor to product metabolite including 16alpha-OH E1 to 2-OH E1, which are linked to estrogen and polyamine biosynthetic pathways, were determined to explore enzyme involvement in breast cancer and to evaluate the potential usefulness of these ratios and concentrations as disease staging markers. It was confirmed that major estrogens and 16a-OH E1 were positively associated with breast cancer and catechol estrogens including 2-OH E1 were inversely associated with breast cancer. The ratios of N1-acSp/Spd and 16alpha-OH E1/2-OH E1 might be a useful dual marker for staging of breast cancer. From the variation of the relative ratios of polyamines, it is suggested that alteration in polyamine oxidase (PAO) activity may play an important role in the development of breast cancer.

Adult↗

PubMed1998

Prevalence of and factors related to pneumoconiosis among foundry workers in central Taiwan.

The objective of this study was to investigate the prevalence of and factors related to pneumoconiosis in foundry workers. Seven hundred and eighteen workers from 50 foundries in central Taiwan were interviewed using a constructed questionnaire. A full-sized PA chest radiograph was used to diagnose pneumoconiosis, according to ILO criteria. Overall, pneumoconiosis was found in 7.5% of the workers. The highest prevalence was found among furnace workers (15.9%) and molding workers (8.40%). All foundry workers except those in administrative positions had a significantly increased risk of developing pneumoconiosis. Using a multiple logistic regression, compared to administrative workers, furnace workers had the highest risk (10.63 times), followed by post-treatment workers (6.63 times), and molding workers (5.41 times). In conclusion, the prevalence of pneumoconiosis was significantly related to high concentrations of dust, especially with a high proportion of free silica, however, smoking and length of exposure were also contributing factors.

Adult↗

PubMed1998

Function of steroidogenic factor 1 (SF1) ligand-binding domain in gene activation and interaction with AP1.

The nuclear receptor SF1 is an essential mediator in ventromedial hypothalamus-pituitary-gonadal development. As with other nuclear receptors, SF1 possesses a DNA-binding domain composed of two zinc fingers and a ligand-binding domain containing a ligand-dependent activation sequence termed AF2. To dissect the domain function of SF1, we examined various SF1 mutants in mouse adrenocortical Y1 cells and human placental JEG3 cells. Destruction of the AF2 structure removed 73-90% transactivation activity, suggesting that AF2 is indispensable for transactivation. Mutants carrying the DNA-binding domain but lacking the AF2 or the ligand-binding domain blocked the activity of normal SF1. Disrupting the zinc finger diminished the dominant negative effect of mutant. Cotransfection of SF1 with AP1 showed that the two transcription factors cooperated to activate gene expression. Some mutants lost the synergistic action with AP1, while some retained partial activity. These experiments delineate the functional domains of SF1.

Animals↗

PubMed1998

Polyamine profiles in the urine of patients with leukemia.

Eleven urinary polyamine levels were determined in controls (32 cases) and 43 patients with varying stages of leukemia including initial, relapse and complete remission, using gas chromatography nitrogen-phosphorus detection. Also, to indirectly evaluate the possible involvement of enzymes, precursor to product concentration ratios were compared between controls and patients with each stage of leukemia. As a result, it is confirmed that the ratio of N1-acSpd/N8-acSpd could be used as a diagnostic marker and the level of N1,N12-diacetylspermine could be used for determining disease stage and as a malignancy marker for leukemia. An altered metabolic pathway related to leukemia is also proposed in which N1,N12-diacetylspermine can be produced directly from spermine and N1,N12-diacetylspermine is a major source of N1-acetylspermidine.

Adolescent↗

PubMed1998

Determination of four anabolic steroid metabolites by gas chromatography/mass spectrometry with negative ion chemical ionization and tandem mass spectrometry.

A gas chromatography/mass spectrometry method is described which uses negative ion chemical ionization and tandem mass spectrometry for the determination of anabolic steroid metabolites. Four anabolic steroid metabolites to be derivatized by Pentafluoropropionic anhydride (PFPA) were determined using gas chromatography/mass spectrometry (GC/MS) with negative chemical ionization (NCI) and NCI/MS/MS. The repeatability and reproducibility of this procedure were in the range of 5.3-9.7% and 6.1-10.2%, respectively. This method of derivatization with PFPA for NCI and NCI/MS/MS was useful to determine four metabolites of nandrolone, dromostanolone, methenolone and boldenone. The derivatized metabolites of boldenone could be detected to 2 ppb and the other three steroids could be detected to 25 ppb in urine at a signal-to-noise ratio of S/N = 3.

Anabolic Agents↗

PubMed1998

Identification of four novel mutations in the CYP21 gene in congenital adrenal hyperplasia in the Chinese.

Congenital adrenal hyperplasia (CAH) is a common autosomal recessive disorder mainly caused by defects in the steroid 21-hydroxylase (CYP21) gene. We have analyzed CYP21 gene sequences in 65 CAH families in Taiwan. All ten exons of the CYP21 gene were analyzed by differential polymerase chain reaction followed by single-strand conformation polymorphism electrophoresis and the amplification-created restriction site method. About 95% (123 chromosomes) contain mutations due to conversion of DNA sequences into its neighboring homologous pseudogene, CYP21P. Four novel mutations representing 5% of the total chromosomes have also been identified. The mutations were confirmed by sequencing an aberrant DNA fragment. These four mutations included a base change of the splicing donor site at intron 2 from GT to AT, a base substitution of C to T at codon 316, deletion of ten bases (TCCAGCTCCC) at codons 330-333 of exon 8, and duplication of 16 bases (CCTGGATGACACGGTC) at codons 393-397 of exon 9. The loss of the splicing donor site at intron 2 and the premature stop at codon 316 may result in aberrant splicing to reduce enzyme activity and a truncated protein with no enzyme activity, respectively. Likewise, both the duplication and the deletion forms create a frameshift and premature stop during translation. The resulting proteins lack the heme-binding domain and hence are expected to lose enzymatic activity. Since these mutations are not found in the neighboring CYP21P pseudogene, gene conversion should not be the cause of these novel mutations.

Adrenal Hyperplasia, Congenital↗

PubMed1998

Altered profile of endogenous steroids in the urine of patients with prolactinoma.

OBJECTIVE: The aim of this study was to investigate the possible effect of prolactin on the metabolism of androgens and estrogens in patients with prolactinoma. To accomplish this, prolactin, urinary androgen, and estrogen metabolite levels were determined. In order to indirectly evaluate the possible involvement of enzymes, the concentration ratios of precursor metabolite to product metabolite were also compared with controls. METHODS: Urine samples were obtained from 27 female patients with prolactinoma (macro, micro, and idiopathic) and from 31 age-matched normal female subjects. Urinary metabolites of 21 androgens and corticoids and 20 estrogens were analyzed by a gas chromatography-mass spectrometry system. RESULTS: In patients with prolactinoma, urinary 17-ketosteroids, and all estrogen metabolite concentrations were elevated. The ratios of delta 5/delta 4-steroids and 5 beta/5 alpha-hydrogensteroids were higher in the patients with prolactinoma than in normal female controls. In addition, no significant differences between patients and controls were observed in the precursor metabolite to product metabolite ratios relating to estrogen metabolism. CONCLUSION: Our data suggests that enhanced PRL levels have a direct effect on urinary steroid secretion and metabolism, probably due to lowered activities of 5 alpha-reductase and 3 beta-hydroxysteroid dehydrogenase in the patients with prolactinoma.

Adult↗

PubMed1998

Acute gastroparesis in Duchenne's muscular dystrophy.

Duchenne's muscular dystrophy (DMD) is an X-linked recessive disease. Clinical descriptions of the disorder focus principally on skeletal muscle degeneration. Another manifestation, which involves the gastrointestinal tract, may be fatal. But its prevalence remains undefined. We report here a case of acute gastroparesis associated with Duchenne's muscular dystrophy. In our case, the patient's symptoms were improved by prokinetic agents and timely decompression in life-threatening acute gastric dilatation.

Acute Disease↗

PubMed1997

Urinary polyamine evaluation for effective diagnosis of various cancers.

With a newly modified analytical method, the concentrations of free and acetylated urinary polyamines were simultaneously determined in a control group (32 cases) and patients with various types of cancers (104 cases, 20 males and 84 females) by gas chromatography-nitrogen-phosphorus detection. Significant concentration differences between normal subjects and various cancer patients were found. The various types of cancers (advanced gastric carcinoma, ovarian cancer, acute myelocyte leukemia, non-Hodgkin's lymphoma) gave unique patterns of urinary polyamine profile as well as significant differences of concentration. To indirectly evaluate the possible involvement of enzymes, precursor-to-product concentration ratios were compared between controls and patients with various types of cancers.

Adult↗

PubMed1997

An improved process for the production of sterile modified haemoglobin solutions.

The process for manufacturing bulk quantities of sterile solutions of human haemoglobin (Hb) cross-linked between the alpha chains (alphaalphaHb) with bis(3,5-dibomosalicyl) fumarate (DBBF) was modified to: (1) improve product purity; (2) increase product yield; (3) eliminate non-United States Pharmacopoeia materials; (4) reduce reagent costs; and (5) reduce production time. These process modifications were the result of increased scientific understanding of the Hb cross-linking chemistry and were in the form of engineering and procedure controls that reflect current good manufacturing practices (cGMP). Purity, as reflected in the fractional yield of the desired alphaalphaHb product, has increased from 60% to 90+% of total Hb, and uncross-linked Hb was virtually eliminated. Impurities such as pyrogens, methaemoglobin, phospholipid, and free iron were reduced. The net yield of alphaalphaHb was increased from 33% to 58% of starting Hb content. Production time, the use of overtime, the consumption of expensive reagents and filters, and losses because of contamination have all been reduced. As a result, cost per gram of alphaalphaHb produced has decreased 60%. With this improved process, efficient production of very pure alphaalphaHb is possible.

Chemistry, Pharmaceutical↗

PubMed1997

Transcriptional regulation of the CYP11A1 and ferredoxin genes.

The first step in the synthesis of all steroids is the cleavage of cholesterol side chain, catalyzed by an electron transport system located in mitochondria consisting of ferredoxin reductase, ferredoxin, and cytochrome P450scc. These proteins are present in adrenal, gonad, placenta, and some parts of the brain. In addition, ferredoxin and ferredoxin reductase are also found in the kidney and liver. Whereas ferredoxin reductase levels remain constant in the cell, ferredoxin and P450scc levels are stimulated by trophic hormones using cAMP as an intracellular messenger. The ferredoxin promoter is relatively simple, consisting of a TATA box and two Sp1-binding sites. This simple module is enough to direct cAMP-dependent transcription in a steroidogenic cell-specific fashion. The regulatory region for the P450scc gene is more complex, containing many protein binding sites for different regulation purposes. Its TATA box directs cAMP-dependent transcription in a cell-type-specific manner. A transcription factor, steroidogenic factor 1 (SF1), activates P450scc gene expression. The tissue-specific expression of the P450scc gene is probably accomplished through the interaction of SF1 with other protein factors located further upstream of the control region. SF1 may also be involved in the cAMP response. An upstream region binding to cAMP-Responsive Element Binding Protein CREB and AP1 can respond to cAMP for gene activation. These analyses of regulatory elements provide the structural architecture for transcriptional regulation of the ferredoxin and the CYP11A11 gene.

8-Bromo Cyclic Adenosine Monophosphate↗