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Biomedical subjects

W S Wu

Publications and source records attributed to W S Wu.

At least 19 recordsLinked to original sources
PubMed2003

Enhanced PCE dechlorination by biobarrier systems under different redox conditions.

The industrial solvent tetrachloroethylene (PCE) is among the most ubiquitous chlorinated compounds found in groundwater contamination. The objective of this study was to evaluate the (1) feasibility of enhancing PCE biodegradation using cane molasses and sludge cakes as the primary substrates under methanogenic and iron reducing conditions, and (2) potential of installation a sludge cake/cane molasses biobarrier to clean up PCE-contaminated aquifers. The biodegradability of sludge cake (from secondary wastewater treatment system) and cane molasses was tested using bioavailability experiments. Results show that biodegradable materials were released from sludge cake/cane molasses and utilized by microbial consortia. Based on the chemical oxygen demand (COD) tests, approximately 28 and 248 mg of biodegradable COD can be released from 1g of sludge cake and 1g of cane molasses under anaerobic conditions, which have the potential to convert 70 and 620 mg of PCE to ethylene (ETH), respectively. Reductive dechlorination was evaluated using microcosms containing primary substrates (sludge cake/cane molasses) and inocula (aquifer sediments). Results indicate that sludge cake and cane molasses can serve as the diffusion sources of primary substrates, and enhance the reductive dechlorination of PCE under methanogenic processes. However, results from this study were not sufficient enough to show that reductive dechlorination of PCE would occur under iron-reducing conditions. This indicates that more studies need to be performed to further evaluate the role of iron reduction on the PCE dechlorination. Results reveal that it is feasible and applicable to install a sludge cake or cane molasses biobarrier to clean up PCE contaminated aquifers. From an engineering point of view, the sludge cake/cane molasses biobarrier has the potential to become an environmentally and economically acceptable technology for PCE bioremediation.

Biodegradation, Environmental↗
PubMed2001

Involvement of p-15(INK4b) and p-16(INK4a) gene expression in saikosaponin a and TPA-induced growth inhibition of HepG2 cells.

Saikosaponin a, a purified ingredient of Chinese herb with known antitumor activity, can inhibit cell growth and DNA synthesis of hepatoma cell line HepG2. Both mRNA and protein of the CDK inhibitor p-16(INK4a) and p-15(INK4b) in HepG2 were greatly induced by saikosaponin a while that of p-21(CIP), p-27(KIP) and other cell cycle related genes were not. In addition, reduced phosphorylation of RB protein is observed in saikosaponin a-treated HepG2. Staurosporin, one of the PKC inhibitors, significantly prevented the saikosaponin a induced growth inhibition suggesting PKC pathway be involved. On the other hand, the phorbol ester tumor promoter TPA (12-O-Tetredecanolyphorbol 13-acetate) also inhibited HepG2 growth and specifically induced p-16(INK4a) and p-15(INK4b) mRNA expression. The results suggest that both saikosaponin a and TPA-induced HepG2 growth inhibition are associated with p-15(INK4a) and p-16(INK4b) gene expression and might be mediated by PKC signaling pathway.

Antineoplastic Agents, Phytogenic↗
PubMed2001

The growth suppressor PML represses transcription by functionally and physically interacting with histone deacetylases.

The growth suppressor promyelocytic leukemia protein (PML) is disrupted by the chromosomal translocation t(15;17) in acute promyelocytic leukemia (APL). PML plays a key role in multiple pathways of apoptosis and regulates cell cycle progression. The present study demonstrates that PML represses transcription by functionally and physically interacting with histone deacetylase (HDAC). Transcriptional repression mediated by PML can be inhibited by trichostatin A, a specific inhibitor of HDAC. PML coimmunoprecipitates a significant level of HDAC activity in several cell lines. PML is associated with HDAC in vivo and directly interacts with HDAC in vitro. The fusion protein PML-RARalpha encoded by the t(15;17) breakpoint interacts with HDAC poorly. PML interacts with all three isoforms of HDAC through specific domains, and its expression deacetylates histone H3 in vivo. Together, the results of our study show that PML modulates histone deacetylation and that loss of this function in APL alters chromatin remodeling and gene expression. This event may contribute to the development of leukemia.

Animals↗
PubMed2000

G-33A mutation in the promoter region of thrombomodulin gene and its association with coronary artery disease and plasma soluble thrombomodulin levels.

Thrombomodulin is an endothelial glycoprotein that decreases thrombin activity and activates protein C. A recent study has shown that G-33A promoter mutation of the thrombomodulin gene occurs particularly in Asians. In this study, we analyzed the distribution of G-33A mutation in the promoter region of the thrombomodulin gene in the Chinese population and determined whether the mutation might be a risk for coronary artery disease (CAD). In addition, the influence of this mutation on plasma soluble thrombomodulin levels in patients with CAD was also examined. We studied 320 consecutive patients (mean age 63 years; 73% men) with CAD and 200 age- and sex-matched control subjects. Screening for thrombomodulin G-33A promoter mutation was conducted using polymerase chain reaction, single-strand conformation polymorphism, and direct deoxyribonucleic acid sequencing. The frequency of the G-33A mutation (GA+AA genotypes) was significantly higher in the CAD group (23.8% vs 15.5%, odds ratio [OR] 1.70, p = 0.031). Multiple logistic regression analysis showed that the mutation was an independent risk factor (OR 1.81, p = 0.016) for CAD, as was hypertension (OR 1.44, p = 0.040), diabetes mellitus (OR 2.50, p <0.001), and smoking (OR 2.15, p <0.001). In CAD patients with GG genotype, the soluble thrombomodulin level increased with the extent of CAD (36 +/- 15 vs 47 +/- 18 vs 55 +/- 36 ng/ml in 1-, 2-, or 3-vessel CAD, p <0.001). However, in CAD patients with G-33A mutation, there was no difference between the levels of soluble thrombomodulin (39 +/- 17 vs 37 +/- 15 vs 42 +/- 18 ng/ml, p = NS) in 1-, 2-, or 3-vessel CAD. Our observations suggest that there is a significant association of the G-33A mutation in thrombomodulin gene with CAD, and this mutation may influence the soluble thrombomodulin levels in patients with CAD.

Asian People↗
PubMed2000

Qualitative determination of urinary morphine by capillary zone electrophoresis and ion trap mass spectrometry.

A rapid, sensitive method for the determination of morphine and amphetamine was developed using capillary zone electrophoresis coupled with electrospray interface (ESI), ion-trap tandem mass spectrometry (CE-ESI-MS2). Morphine and amphetamine were separated in 20 mM ammonium acetate buffer (pH 6.6) and detected by ion-trap mass detector in different analytical time segments (0-6.25 min for amphetamine and 6.25-12.0 min for morphine) in which the tune file for each compound was used separately. Molecular ions of morphine (m/z 286) and amphetamine (m/z 136) were detected at 5.77 and 6.83 min, respectively, while product ions of MS2 for each compound (m/z 229, 201 for morphine and m/z 119 for amphetamine) were detected almost exactly at the same time with their parent compounds. The limits of detection (LOD) for MS2 determination were 30 and 50 ng/mL for amphetamine and morphine, respectively, with an S/N ratio of 3. For more sensitive detection of morphine, the sample was injected for a longer time (i.e., 80 s) and hydrodynamically transported into a CE capillary for MS detection. Morphine and its product ion appear at 0.36 and 0.39 min on the ion chromatogram, respectively, with a five-fold increase of detection sensitivity (LOD, 10 ng/mL). The CE-MS system thus established was further applied for forensic urine samples screened as morphine-positive by fluorescence polarization immunoassay (FPIA). These results indicated the feasibility of CE-ESI-MS2 for confirmative testing of morphine in urine sample.

Amphetamine↗
PubMed2000

Concentration of vascular endothelial growth factor in the subretinal fluid of retinal detachment.

The purpose of this study was to investigate the release of vascular endothelial growth factor (VEGF) within the subretinal fluid in eyes with retinal detachment. Subretinal fluid was collected from patients with retinal detachment undergoing surgery for scleral buckling. Serum samples were also collected. The concentration of VEGF in the subretinal fluid and serum was investigated by enzyme-linked immunospecific assay. The average concentration of VEGF in serum samples was 168 +/- 153 pg/ml (mean +/- standard deviation). It was lower than the VEGF concentration in the subretinal fluid (485 +/- 570 pg/ml) in the same 18 patients with retinal detachment (p < 0.05). The average concentration of VEGF was 355 +/- 373 pg/ml in 31 samples of the subretinal fluid collected from simple rhegmatogenous retinal detachment. It was lower than the average concentration of 901 +/- 385 pg/ml in 8 samples of the subretinal fluid from retinal detachment with proliferative vitreoretinopathy (p < 0.05). The results suggest that the relative retinal ischemia in detached retina increases the release of VEGF into the subretinal space. Also, the concentration of VEGF within the subretinal fluid in proliferative vitreoretinopathy was higher than in simple rhegmatogenous retinal detachment.

Adolescent↗
PubMed1999

Analysis of variable region of T-cell receptor beta chain usage in the human anti-porcine xenoresponse.

BACKGROUND: The human xenoreactive T-cell receptor (TCR) repertoire is not well documented. The aim of this study was to analyze the TCR repertoire in human anti-porcine xenoresponses. METHODS: Peripheral blood lymphocytes (PBLs) from healthy donors were used to generate human T-cell lines against two different haplotypes of inbred Yucatan miniature swine (y/y and z/z). The variable region of TCR beta-chain (Vbeta) gene usage was determined by fluorescence CDR3 spectrotyping. RESULTS: TCR Vbeta usage of an established human antiporcine T-cell line analyzed at weeks 5, 7, and 9 showed a sequential increase in Vbeta 1, 2, 6.2, 11, and 19 as compared to unprimed peripheral blood lymphocytes, whereas the usage of other Vbetas decreased. The selection of limited Vbeta genes correlated with the sequential increase in the specific lysis of the T-cell line, suggesting a non-random clonal selection and expansion of T-cell clones that recognized porcine targets. Different Vbeta restriction was found using the same peripheral blood lymphocytes against a different haplotype of swine, indicating this selection of Vbeta gene was swine leukocyte antigen-dependent. CONCLUSIONS: There is restricted TCR Vbeta usage in the human anti-porcine response, suggesting that a limited number of xenogeneic epitopes are recognized by human T cells. The selection of particular TCR Vbeta clonotypes depends on the swine leukocyte antigen background.

Animals↗
PubMed1999

Analysis of morphine and morphine-3beta-D glucuronide in human urine by capillary zone electrophoresis with minimal sample pretreatment.

The presence of two of the metabolites of heroin, free morphine and morphine 3-beta-D-glucuronide (MO3G) in acidified urine samples was simultaneously determined by capillary zone electrophoresis (CZE). In a run buffer containing 50 mM sodium borate and 250 mM boric acid (pH 8.6), free morphine migrates before a group of neutral compounds (peak N) in urine, which move with the velocity of electro-osmotic flow. In contrast, the glucuronidated form is negatively charged and migrates behind peak N. Both analytes can be precisely identified within their respective analytical window by their migration time with respect to peak N. The on-line multi-wavelengths scanning of the peak permits further confirmation. The detection sensitivity of both analytes was increased three-four fold if the samples were introduced with electro-injection as compared with hydrodynamic injection. Limits of detection (LOD) of free and conjugated morphine using electro-injection were 200 ng/mL and 500 ng/mL, respectively, determined at a 3:1 signal to noise ratio. A dramatic increase of free morphine was observed after acid hydrolysis of the urine concomitantly with the decrease of the glucuronidated form. We conclude that CZE is a rapid, simple, sensitive and useful screening technique for detection of heroin metabolites in the urine.

Calibration↗
PubMed1999

A full genome scan for late onset Alzheimer's disease.

We have genotyped 292 affected sibling pairs (ASPs) with Alzheimer's disease (AD) according to NINCDS-ADRDA diagnostic criteria and with onset ages of >/=65 years using 237 microsatellite markers separated by an average distance of 16.3 cM. Data were analysed by SPLINK and MAPMAKER/SIBS on the whole sample of 292 ASPs and subsets of 162 ASPs where both members possessed an apolipoprotein E (APOE)straightepsilon4 allele and 63 pairs where neither possessed anstraightepsilon4 allele. Sixteen peaks with a multipoint lod score (MLS) >1 either in the whole sample, the straightepsilon4-positive or -negative subgroups were observed on chromosomes 1 (two peaks), 2, 5, 6, 9 (two peaks), 10 (two peaks), 12, 13, 14, 19, 21 and X (two peaks). Simulation studies revealed that these findings exceeded those expected by chance, although many are likely to be false positives. The highest lod scores on chromosomes 1 (MLS 2.67), 9 (MLS 2.38), 10 (MLS 2.27) and 19 (MLS 1.79) fulfilLander and Kruglyak's definition of 'suggestive' in that they would be expected to occur by chance once or less per genome scan. Several other peaks were only marginally less significant than this, in particular those on chromosomes 14 (MLS 2.16), 5 (MLS 2.00), 12, close to alpha2-macroglobulin (MLS 1.91), and 21, close to amyloid precursor protein (MLS 1.77). This is the largest genome scan to date in AD and shows for the first time that this is a genetically complex disorder involving several, perhaps many, genes in addition to APOE. Moreover, our data will be of interest to those hoping to identify positional candidate genes using information emerging from neurobiological studies of AD.

Age of Onset↗
PubMed1998

Genetic studies on chromosome 12 in late-onset Alzheimer disease.

CONTEXT: The only genetic locus universally accepted to be important as a risk factor for late-onset Alzheimer disease (AD) is the apolipoprotein E (APOE) locus on chromosome 19. However, this locus does not account for all the risk in late-onset disease, and a recent report has suggested a second locus on chromosome 12p11-12. OBJECTIVE: To look for evidence of linkage on chromosome 12 and to test for the presence of the new locus in an independent sample of familial late-onset AD cases. DESIGN: Retrospective cohort study. As part of a 20-centimorgan genome screen (approximately equal to 200 markers), we tested a series of 18 genetic markers on chromosome 12 and carried out multipoint, nonparametric lod score and exclusion analyses. SETTING: Clinic populations in the continental United States selected from the National Institute of Mental Health AD Genetics Consortium. PATIENTS: We selected samples for DNA analysis from affected sibling pairs, 497 subjects from 230 families with 2 or more affected individuals with probable or definite AD with onset ages older than 60 years (mean+/-SD, 75+/-6 years). Within the families, we used the 2 probable or definitely affected individuals. In families with more than 2 such cases available, we used all of them; in families with only 2 such cases in which unaffected individuals were available, we also sampled the oldest unaffected individual and used genotype data from this unaffected individual to check for nonpaternity and genotyping errors. MAIN OUTCOME MEASURE: Presence of linkage or locus on chromosome 12. RESULTS: Although linkage analyses confirmed the presence of a genetic susceptibility factor at the APOE locus in these families with late-onset AD, we were unable to confirm the presence of a locus close to the marker D12S1042. A moderate lod score (1.91) was found near D12S98 close to the alpha2-macroglobulin locus in the affected pairs in which both members did not possess an APOE epsilon4 allele. CONCLUSIONS: APOE remains the only locus established to be a risk factor for late-onset AD. We were unable to confirm that a locus on chromosome 12p11-12 has a major effect on risk for late-onset AD, although an effect smaller than that for APOE cannot be excluded.

Age of Onset↗
PubMed1998

Retinoic acid-induced apoptosis is prevented by serum albumin and enhanced by Lipiodol in human hepatoma Hep3B cells.

The effects of retinoic acid (RA) on the cell growth and viability of human hepatoma Hep3B cells were examined. We showed that removal of serum in the presence of RA results in cell death in a dose-dependent manner in human hepatoma Hep3B cells. Time-course cell death analysis showed that RA at a dose of 10 microM induces a rapid (48-72 h) fall in cell viability (>95%). The drug-induced cell death was RA-specific, since three RA analogs (retinol, retinal and retinol acetate) did not show any cytocidal activity at an equimolar dose. Fluorescence microscopy and DNA fragmentation analysis showed that Hep3B cells treated with RA underwent a death process highly reminiscent of apoptosis, with chromatin condensation, nuclear fragmentation and the presence of a 180-200 bp DNA fragment ladder. Additionally, we found that RA-induced apoptosis was reduced by 70-80% when the medium was supplemented with serum albumin (human and bovine) at a concentration of 0.05%. However, a variety of known growth factors were ineffective in preventing RA-induced apoptosis. Preincubating serum and serum albumin with Lipiodol restored the apoptotic effects of RA demonstrated in serum-free systems. These data suggest that the binding of RA by serum albumin may have reduced the bioavailability of RA, restricting its apoptotic effects on Hep3B cells. Blocking RA-albumin interactions with a lipid lymphographic contrast medium (Lipiodol) may improve the bioavailability of RA and significantly enhance its apoptotic effect on human hepatoma Hep3B cells.

Apoptosis↗
PubMed1998

Simultaneous determination of paraquat and diquat in urine by capillary electrophoresis.

Qualitative and quantitative detection of paraquat and diquat in urine was performed by capillary zone electrophoresis (CZE). In non-extracted urine, these two closely related herbicides were hydrodynamically or electrokinetically injected into the capillary and well separated in a acidic (pH 3.0) buffer (10 mM glycin, 50 mM NaCl and 20% methanol) within 8 min. The analytes can be identified by their respective migration time with no interfering endogeous compounds around. The peaks were further confirmed by their respective multi-wavelength scanning spectra. Limits of detection (LOD) were 0.2 and 0.1 mg/ml for PQ and DQ, respectively, in nonextracted urine using hydrodynamic injection. Detection sensitivity was greatly improved by solid phase extraction of the compounds followed by CZE using electro-injection under which LODs were 15 and 8 ng/ml for paraquat and diquat respectively. We conclude CZE is a useful method for laboratory diagnosis of herbicide intoxication.

Diquat↗
PubMed1997

Application of tryptamine as a derivatizing agent for the determination of airborne isocyanates. Part 7. Selection of impinger solvents and the evaluation against dimethyl sulfoxide used in US NIOSH Regulatory Method 5522.

The application of tryptamine to derivatize airborne isocyanates has been evaluated and adopted by the National Institute for Occupational Safety and Health (NIOSH) as the latest isocyanates regulatory method (Method 5522) in the USA. Method 5522 uses dimethyl sulfoxide (DMSO) as the impinger solvent in which tryptamine is dissolved to sample isocyanates for analysis. Since DMSO is both extremely hygroscopic and corrosive, it is not a satisfactory solvent for impinger air sampling of isocyanates. The high boiling and freezing points also present some distinct drawbacks. In a search for a suitable impinger solvent, the efficiencies of various solvents, which all were potentially more suitable for sampling isocyanates viz., N,N'-dimethylformamide, butyl acetate, isobutyl acetate, sec-butyl acetate, tert-butyl acetate and octane, were investigated. Simulated air sampling was conducted on two commonly used isocyanates in industry, hexamethylene diisocyanate (HDI) and toluene diisocyanate (TDI), which were vaporized and sampled into impingers containing dissolved tryptamine. Butyl acetate and octane were found to be most suitable for using as impinger solvents. Recoveries of isocyanates at two concentration levels of HDI and TDI were 93.4-108% in comparison with those of using DMSO.

Air Pollutants, Occupational↗
PubMed1997

DNA polymorphisms and mutations of the tumor necrosis factor-alpha (TNF-alpha) promoter in Langerhans cell histiocytosis (LCH).

Langerhans cell histiocytosis (LCH) is a clonal proliferation of dendritic histiocytes expressing elevated levels of tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN-gamma) granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin-1 (IL-1), and leukemia inhibitory factor (LIF). The cause of the increased cytokine levels is unknown, but DNA sequence changes in promoters could alter expression. The TNF-alpha and IFN-gamma promoter DNA sequences of 12 LCH patients were studied and compared with normal individuals by dideoxy fingerprinting and DNA sequencing. Functional consequences of polymorphic or mutated sequences were assessed by cloning altered and control promoter sequences into a luciferase reporter gene vector. Electrophoretic mobility shifts (EMSA) after binding of nuclear extracts from a macrophage cell line (U-937) by mutated promoters were compared with controls. Five of 12 LCH patients had alterations in the TNF-alpha promoter DNA sequence. None were found in the IFN-gamma gene promoter. Of the 5 with TNF-alpha DNA alterations, 2 were at position -308, which has been described as a G-A polymorphism associated with upregulation of TNF-alpha in some patients with infections or immune-mediated diseases. The polymorphism at -308 but not the other TNF-alpha promoter mutations caused a 3-fold to 7-fold increased production of the luciferase reporter gene. EMSA showed that the -308 mutant promoters bound fewer nuclear proteins than normals. Polymorphisms of the TNF-alpha promoter in LCH patients could increase the production of that cytokine.

DNA↗
PubMed1997

Sodium/proton antiporter in the euryhaline crab Carcinus maenas: molecular cloning, expression and tissue distribution.

Gill epithelial cells of euryhaline crustaceans demonstrate net inward transport of sodium ions, possibly via apical Na+/H+ antiporters, Na+/K+/2Cl- cotransporters or Na+ channels working in series with the basolateral Na(+) + K(+)-ATPase. We have identified and sequenced the cDNA coding for a crustacean Na+/H+ antiporter, starting with mRNA isolated from gills of the euryhaline green shore crab Carcinus maenas. The complete 2595-base-pair cDNA includes an open reading frame coding for a 673-amino-acid protein. A search of GenBank revealed more than 20 high-scoring matches, all Na+/H+ antiporter sequences from mammalian, amphibian, teleost and nematode species. Injection of Xenopus laevis oocytes with cRNA transcribed from the cloned crab sequence substantially enhanced Na(+)-dependent H+ efflux from the oocytes. Analysis of crab tissue antiporter mRNA levels by semi-quantitative reverse transcription-polymerase chain reaction revealed that posterior and anterior gills of Carcinus maenas expressed this antiporter the most strongly, followed in decreasing order by skeletal muscle, hepatopancreas, hypodermis and heart. Hydropathy and transmembrane alpha-helix analysis suggested a 10-helix membrane-spanning topology of the antiporter protein. It is clear from this study that Carcinus maenas gills vigorously transcribe a gene coding for a Na+/H+ antiporter. Whether these gills also express a gene coding for an epithelial Na+ channel or Na+/K+/2Cl- cotransporter remains to be demonstrated.

Amino Acid Sequence↗
PubMed1996

Induction of c-jun protooncogene expression by hydrogen peroxide through hydroxyl radical generation and p60SRC tyrosine kinase activation.

The mechanisms of signal transduction of c-jun induction by hydrogen peroxide are elucidated in NIH3T3 cells by using trapping agents of hydroxyl free radical or inhibitors of various protein kinases. Pre-treatment of the cell with hydroxyl radical scavenger dimethyl sulfoxide (DMSO) abolishes the H2O2-induced c-jun expression. Hydroxyl radical generation can be detected and quantified in cells treated sequentially with DMSO and H2O2 for 30 min respectively by methane sulfinic acid (MSA) production, especially that from particulate fraction. Induction of c-jun by H2O2 is also dramatically reduced by pretreating the cells with biological antioxidant vit. E. Protein tyrosine kinase activity of membrane fraction is induced by H2O2 within 5 to 10 min, which can be prevented by DMSO pre-treatment. Inhibitor of non-receptor type tyrosine kinase, herbimycin A, has inhibitory effect on H2O2-induced c-jun expression while the inhibitor of receptor type tyrosine kinase, tyrphostin 23 or inhibitor of cyclic AMP dependent protein kinase, KT 5720, has not. TPA pre-treatment that depletes protein kinase C (PKC) has no influence on the c-jun induction by H2O2. Our results suggest that the highly reactive species HO is generated after H2O2 enter cells and mediate the signal responses of H2O2 including c-jun induction and the activation of p60src tyrosine kinase might be one of the molecular events associated with the c-jun induction pathway.

3T3 Cells↗