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Biomedical subjects

W C Lee

Publications and source records attributed to W C Lee.

At least 19 recordsLinked to original sources
PubMed2003

Bilateral parotid and submandibular gland enlargement: rare features of Wegener's granulomatosis.

Wegener's granulomatosis is a potentially fatal disease of unknown origin affecting mainly the upper and lower respiratory tracts and kidneys. Prompt recognition of the more unusual presentations of the disease is necessary to ensure early treatment. We present a case of a 46-year-old female with bilateral submandibular and parotid gland enlargement. Parotid or submandibular salivary gland enlargement is a rare presenting feature of Wegener's granulomatosis. Common to 80 per cent of these cases is nasal involvement, while ear pathology or lung lesions may occur in the remaining cases. The diagnosis is both clinical and pathological, biopsy of suspicious tissue, serum c-ANCA levels and a chest X-ray are valuable investigations. Treatment with immunosuppressive therapy is essential, and usually ensures a long-term remission.

Female↗
PubMed2002

Effects of single nucleotide polymorphisms in the 5'-flanking region of heat shock protein 70.2 gene on semen quality in boars.

This study aims to elucidate the effects of single nucleotide polymorphisms (SNPs) in the 5'-flanking region of porcine heat shock protein 70.2 gene (HSP70.2) on semen quality in boars. Genomic DNA isolated from 55 boars (41 Duroc, nine Landrace, and five Yorkshire) was subjected to PCR amplification of the 5'-flanking region of HSP70.2. The nucleotide sequences were determined by automated sequencing. Five SNPs (sites 44, 232, 250, 345, and 393) were detected in this region. Semen quality was evaluated in terms of sperm motility, percentage of normal sperm, percentage of sperm with proximal plasma droplet, percentage of abnormal sperm, sperm concentration, semen volume per ejaculate and total sperm number per ejaculate. The effect of the SNPs on semen quality was evaluated based on breed-corrected data within a season. During the cool season, the sperm motility of boars with AA genotype at the 232 site was significantly higher than that of boars with CC genotype (P<0.05). Meanwhile, boars with AC genotype at the 232 site had higher total sperm number per ejaculate than did those with CC genotype. In the hot season, heterozygotes at both the 232 and 250 sites had significantly higher total sperm number of per ejaculate than AA homozygotes (P<0.05). Semen volume of boars with TT and TC genotypes at the 345 site was significantly larger than that of those with CC genotype (P<0.05). Meanwhile, semen quality for boars with TT genotype at the 345 site was significantly higher than that of boars with TC or CC genotype (P<0.05), that is the semen contained higher percentages of normal sperm and lower percentages of abnormal sperm or sperm with proximal plasma droplets. Results herein suggest that the SNPs in the 5'-flanking region of porcine HSP70.2 are associated with semen quality traits in the hot season.

Animals↗
PubMed2002

Hemodynamic effects of a combination of octreotide and terlipressin in patients with viral hepatitis related cirrhosis.

BACKGROUND: Terlipressin or octreotide given alone has been used as the first-line pharmacological treatment for acute variceal bleeding. In portal hypertensive animals, pre-infusion of octreotide followed by the addition of terlipressin has an additive or complementary effect on splanchnic hemodynamics. The current study was aimed at evaluating such a combination treatment in patients with cirrhosis and portal hypertension. METHODS: Patients were randomly assigned to receive either a placebo (n = 11) or an intravenous infusion of octreotide 100 microg/h after an initial bolus of 100 microg (n = 13). Thereafter, each patient received an intravenous injection of terlipressin 2 mg. Hemodynamic values were measured basally, 30 min after octreotide or placebo, and 60 min after terlipressin. RESULTS: Placebo administration did not affect any of the hemodynamic values. Terlipressin administration resulted in expected changes in hepatic venous pressure gradient, hepatic blood flow and systemic hemodynamics. In contrast, octreotide administration significantly decreased hepatic blood flow but did not affect other hemodynamic values. After terlipressin administration, significant hemodynamic changes were observed that were similar to the hemodynamic changes with terlipressin alone. The magnitude of changes in hepatic venous pressure gradient, cardiac index and systemic vascular resistance were no different between the two groups of patients. The heart rate was significantly lower in patients receiving octreotide plus terlipressin than those receiving terlipressin alone. CONCLUSION: The current study showed that a combination of octreotide and terlipressin did not exert an additive effect in reducing hepatic venous pressure gradient in patients with cirrhosis. In addition, the systemic hemodynamic changes were comparable between the two groups.

Aged↗
PubMed2002

Transmission/disequilibrium test when neither parent is available in some families: a non-iterative approach.

BACKGROUND: The transmission/disequilibrium test (TDT) is a simple method of detecting linkage between a marker locus and a disease-susceptibility locus. The test requires genotype data from the affected individuals (the probands) and their parents, the 'case-parents triads'. However, missing data from parents often complicate the situations. Several non-iterative methods, such as the sibling-based TDT, the reconstruction-combined TDT, and the 1-TDT have been proposed to deal with the problem. But when the father, the mother, and other non-proband siblings in a family are all unavailable for study, that family has to be excluded from analysis by any of the above methods. METHODS: The author proposes a new non-iterative method to deal with the situation when the data contain some 'orphaned' probands. The method utilises the complete data from case-parents triads to infer probabilistically the missing parental genotypes, under the assumption that the missingness is independent of genotype and ethnic origin. The methods can be tailored to specific genetic models, such as the gene-dose, dominance, and recessive alternatives. RESULTS: Simulation analysis shows that, under the null hypothesis of no linkage, the proposed method is not prone to produce an excess of false positive results, even in complex situations such as population stratification or assortive mating. Under the linkage alternative, the new method recaptures much of the information that would be lost by discarding the orphaned probands. DISCUSSION: The method provides a way for dealing with the situation where neither parents nor siblings are available in a TDT study.

Algorithms↗
PubMed2001

Chromatographic resolution of the enantiomers of phenylpropanolamine by using molecularly imprinted polymer as the stationary phase.

In this study molecular imprinting technology was employed to prepare a specific affinity sorbent for the resolution of phenylpropanolamine, a chiral drug. The molecularly imprinted polymer (MIP) was prepared by non-covalent molecular imprinting with either (-)- or (+)-phenylpropanolamine as the template. Methacrylic acid and ethylene glycol dimethacrylate were copolymerized in the presence of the template molecule. The bulk polymerization was carried out in chloroform with 2,2'-azobisisobutyronitrile as the initiator, at 4 degrees C and under UV radiation. The resulting MIP was ground into powders, which were slurry packed into analytical columns. After removal of template molecules, the MIP-packed columns were found to be effective for the resolution of (+/-)-phenylpropanolamine racemates. The separation factor for the enantiomers ranged between 1.8 and 3.8 when the column was packed with MIP prepared with (+)-phenylpropanolamine as the template. A separation factor ranging from 2.1 to 3.6 could be achieved from the column packed with MIP, prepared with (-)-phenylpropanolamine as the template. Although the separation factor was higher with that previously obtained from reversed-phase column chromatography following derivatization with a chiral agent, elution peaks were broader due to the heterogeneity of binding sites on MIP particles and the possible non-specific interaction.

Chromatography, Liquid↗
PubMed2001

Predicting the elution behavior of proteins in affinity chromatography on non-porous particles.

Affinity chromatography on non-porous particles of microsize is particularly useful for the rapid analysis and micropreparative separation of proteins. The elution behavior of proteins in an affinity column packed with non-porous copolymerized particles of styrene, methyl methacrylate and glycidyl methacrylate was investigated both theoretically and experimentally, using the lysozyme-Cibacron Blue 3G-A affinity system. Equations used to predict the elution profiles, resulting from the elution by increasing the ionic strength (NaCl concentration) in the mobile phase, were obtained. The maximum adsorbate concentration, desorption rate constant and equilibrium constant under elution conditions were determined by matching experimental data with predicted elution profiles. Based on the parameters determined at a flow-rate of 0.5 ml/min and with 1 M NaCl in the elution buffer, the model equations could predict the elution profiles for other experimental runs, where different flow-rates and sodium chloride concentrations were used. Both the experimental and predicted results revealed that the affinity interaction kinetics are not significantly influenced by the flow-rate and, hence, the film mass transfer. To elute bound lysozyme from immobilized dye ligand, a higher value of the ionic strength leads to a faster elution and a sharper elution peak. The influence of elution conditions on the kinetic and thermodynamic parameters and, consequently, on the elution peak profiles was evaluated. The model equations can also predict the behavior of protein elution from an affinity column by changing the pH of the mobile phase, according to a previous study.

Adsorption↗
PubMed2001

Heat shock proteins in canine transmissible venereal tumor.

SDS-PAGE, Western blot analysis and immunohistochemical staining were used to detect heat shock proteins (HSPs) 60, 70 and 90 in canine transmissible venereal tumor (CTVT). Tissues tested for HSPs included: (1) tissues from different growth phases of CTVT tumors artificially induced in dogs; (2) tissues from other canine tumors; (3) normal dog tissues. Our results indicate that HSP 60 was consistently higher in CTVT cells in regressing phase than those in progressing phase. However, no detectable antibody response specific to the tested HSPs was found in the sera from CTVT-laden dogs in different growth phases. Although levels of the HSPs were all detectable in CTVT cells, only 60 and 70 were higher in CTVT cells than in normal tissues. In addition, none of the HSPs were detected in cells from five other canine tumors. These data suggest that canine HSP 60 and 70 are potential markers for CTVT and HSP 60 is appear to be involved in CTVT regression.PCR was used to confirm the existence of CTVT cells using primers designed to cover the sequence between the 5' end of c-myc near the first exon and the 3' end outside the LINE gene. Only CTVT samples were positive for this sequence; samples from other tumors and normal tissues were negative. The sequenced PCR products indicated that CTVT from Taiwan and other countries exhibited over 98% sequence homology. This reconfirms that, worldwide, all CTVT cells are very similar.

Animals↗
PubMed2001

Evidence that ligand and metal ion binding to integrin alpha 4beta 1 are regulated through a coupled equilibrium.

We have used the highly selective alpha(4)beta(1) inhibitor 2S-[(1-benzenesulfonyl-pyrrolidine-2S-carbonyl)-amino]-4-[4-methyl-2S-(methyl-[2-[4-(3-o-tolyl-ureido)-phenyl]-acetyl]-amino)-pentanoylamino]-butyric acid (BIO7662) as a model ligand to study alpha(4)beta(1) integrin-ligand interactions on Jurkat cells. Binding of [(35)S]BIO7662 to Jurkat cells was dependent on the presence of divalent cations and could be blocked by treatment with an excess of unlabeled inhibitor or with EDTA. K(D) values for the binding of BIO7662 to Mn(2+)-activated alpha(4)beta(1) and to the nonactivated state of the integrin that exists in 1 mm Mg(2+), 1 mm Ca(2+) were <10 pm, indicating that it has a high affinity for both activated and nonactivated integrin. No binding was observed on alpha(4)beta(1) negative cells. Through an analysis of the metal ion dependences of ligand binding, several unexpected findings about alpha(4)beta(1) function were made. First, we observed that Ca(2+) binding to alpha(4)beta(1) was stimulated by the addition of BIO7662. From solution binding studies on purified alpha(4)beta(1), two types of Ca(2+)-binding sites were identified, one dependent upon and the other independent of BIO7662 binding. Second, we observed that the metal ion dependence of ligand binding was affected by the affinity of the ligand for alpha(4)beta(1). ED(50) values for the metal ion dependence of the binding of BIO7762 and the binding of a lower affinity ligand, BIO1211, differed by 2-fold for Mn(2+), 30-fold for Mg(2+), and >1000-fold for Ca(2+). Low Ca(2+) (ED(50) = 5-10 microm) stimulated the binding of BIO7662 to alpha(4)beta(1). The effects of microm Ca(2+) closely resembled the effects of Mn(2+) on alpha(4)beta(1) function. Third, we observed that the rate of BIO7662 binding was dependent on the metal ion concentration and that the ED(50) for the metal ion dependence of BIO7662 binding was affected by the concentration of the BIO7662. These studies point to an even more complex interplay between metal ion and ligand binding than previously appreciated and provide evidence for a three-component coupled equilibrium model for metal ion-dependent binding of ligands to alpha(4)beta(1).

Benzoates↗
PubMed2001

Affinity chromatography of proteins on non-porous copolymerized particles of styrene, methyl methacrylate and glycidyl methacrylate.

Non-porous particles having an average diameter of 2.1 microm were prepared by co-polymerization of styrene, methyl methacrylate and glycidyl methacrylate, which was abbreviated as P(S-MMA-GMA). The particles were mechanically stable due to the presence of benzene rings in the backbone of polymer chains, and could withstand high pressures when a column packed with these particles was operated in the HPLC mode. The polymer particles were advantaged by immobilization of ligands via the epoxy groups on the particle surface that were introduced by one of the monomers, glycidyl methacrylate. As a model system, Cibacron Blue 3G-A was covalently immobilized onto the non-porous copolymer beads. The dye-immobilized P(S-MMA-GMA) particles were slurry packed into a 1.0 cm x 0.46 cm I.D. column. This affinity column was effective for the separation of turkey egg white lysozyme from a protein mixture. The bound lysozyme could be eluted to yield a sharp peak by using a phosphate buffer containing 1 M NaCl. For a sample containing up to 8 microg of lysozyme, the retained portion of proteins could be completely eluted without any slit peak. Due to the use of a shorter column, the analysis time was shorter in comparison with other affinity systems reported in the literature. The retention time could be reduced significantly by increasing the flow-rate, while the capacity factor remained at the same level.

Chromatography, Affinity↗
PubMed2001

Genome-wide hypomethylation in hepatocellular carcinogenesis.

Aberrant genome-wide hypomethylation has been thought to be related to tumorigenesis. However, its mechanism and implications in hepatocellular carcinogenesis remain to be elucidated. Samples of hepatoma (hepatocellular carcinoma, HCC) and paired non-HCC liver tissues were obtained from 17 HCC patients. Normal liver tissues obtained from three individuals were used as controls. Compared with the paired non-HCC liver tissues, genome-wide 5-methylcytosine content in HCC was reduced in all of the tested HCC samples (P < 0.001). Conversely, genome-wide 5-methylcytosine content did not significantly differ among normal, noncirrhotic, and cirrhotic liver tissues. Moreover, the degree of reduced DNA methylation was related to late histopathological HCC grade (P = 0.005) and large tumor size (P = 0.079). Compared with the paired non-HCC liver tissues, expression of DNA methyltransferases DNMT-1, DNMT-3A, and DNMT-3B and the DNA methyltransferase-like gene, DNMT-2, was up-regulated in 53, 41, 59, and 47% of the HCC samples, respectively. Surprisingly, small amounts of LINE-1 retrotransposon transcripts were detected in HCC and non-HCC as well as normal liver tissues, and the expression levels were not significantly different in HCC compared with the paired non-HCC or normal liver tissues. Of interest, the 3' ends of these LINE-1 transcripts were truncated. Our findings suggest that genome-wide hypomethylation in HCC is a continuing process that persists throughout the lifetime of the tumor cells rather than a historical event occurring in precancer stages or in cell origins for HCC. Up-regulation of DNA methyltransferases might simply be a result of increased cell proliferation in cancer. In addition, our results did not support the hypothesis of activation of transposable elements in HCC via genome-wide hypomethylation.

5-Methylcytosine↗
PubMed2001

Molecular structure, dynamic expression, and promoter analysis of zebrafish (Danio rerio) myf-5 gene.

We isolated a 1,438 bp cDNA fragment that encoded Myf-5 myogenic factor of zebrafish. The deduced amino acid contained 237 residues, including the basic helix-loop-helix domain that is conserved in all known Myf-5. The zebrafish myf-5 transcripts were first detectable at 7.5 hpf, increased substantially until 16 hpf, and then declined gradually to an undetectable level by 26 hpf. During somitogenesis, zebrafish myf-5 transcripts were distributed mainly in the somites and segmental plates. Prominent signals occurred transiently in adaxial cells in two parallel rows but did not extend beyond the positive-signal somites. Various lengths of upstream region of zebrafish myf-5 fused with EGFP gene were used to carry out transgenic analysis. Results showed that a small, 82 bp (nucleotide positions from -82 to -1), regulatory cassette is sufficient to control the somite- and stage-specific expression of zebrafish myf-5 during early development.

Amino Acid Sequence↗
PubMed2001

A novel enzyme reactor using gluten membrane entrapping cell-associated enzyme.

A membrane enzyme reactor consisting of variable pieces of replaceable cell-immobilized membranes was proposed for the continuous production of bioproducts. To demonstrate the characteristics of the reactor, cell-immobilized membranes were prepared by the entrapment of permeabilized recombinant Escherichia coli cells containing penicillin G acylase within the gluten matrices. A stainless-steel net that was created with a mesh frame was used to support each gluten membrane so that the membranes could be filled into the rectangular-shaped reactor. The reactor equipped with either six or 12 pieces of cell-immobilized gluten membranes containing a biomass concentration of 5%, w/w was effective in catalyzing the production of 6-aminopenicillanic acid from penicillin G. In comparison with intact cells, the cell-immobilized preparation was more stable and the half-life time of the immobilized cell-associate enzyme in gluten membrane was estimated to be 36 days by a long-term operation. As the substrate solution was forced to flow through the reactor equipped with six membranes and in the direction perpendicular to the membranes, the pressure drop was determined to be less than 50 cm H(2)O with a flow-rate up to 50 mL/min. This low pressure due to the porous structure of gluten membrane would lead to a lower operational cost. Increasing either the number of membranes or the area of each cell-immobilized membrane can easily do scaling-up of this membrane reactor.

Anti-Bacterial Agents↗
PubMed2001

Portopulmonary hypertension: distinctive hemodynamic and clinical manifestations.

Portopulmonary hypertension is now recognized as one of the pulmonary complications of chronic liver disease. However, previous studies reported that the incidence ranged from 0.25% to 2%, excluding fortuitous coincidence. In this study, we aimed to determine the variant hemodynamic and clinical features of portopulmonary hypertension in an area with a high prevalence of viral cirrhosis. After reviewing the hemodynamic data of 322 patients with portal hypertension admitted to the Taipei Veterans General Hospital between 1987 and 1999, we found 10 with portopulmonary hypertension. The overall incidence was, therefore, 3.1% in all patients with portal hypertension. Most of the patients with portopulmonary hypertension experienced exertional dyspnea. The survival times ranged from 2 to 86 months. In our series, most of the patients who died, died of complications related to cirrhosis and portal hypertension, but not of complications related to pulmonary hypertension. This study suggested that portopulmonary hypertension was not a frequent complication in cirrhotic patients and was not associated with an adverse outcome.

Adult↗
PubMed2001

Incidence, transmission, and clinical significance of hepatitis G virus infection in hemodialysis patients.

A high prevalence of hepatitis G virus (HGV) infection has been noted in patients receiving chronic hemodialysis (HD) therapy, yet the incidence rate and transmission route have rarely been reported. Serum samples from 160 chronically uremic patients in a HD unit were initially collected at the time chronic HD therapy was begun, and thereafter annually in July and, finally, in November 1999. Serum HGV RNA was detected using nested reverse transcription polymerase chain reaction, and HGV E2 antibody was determined using an enzyme immunoassay. Nucleotide sequences of the 5'-noncoding region were studied in the HD patients with HGV viremia. Forty healthy staff members were also enrolled as control subjects. Three of the 40 (7.5%) healthy staff members were positive for HGV RNA or HGV E2 antibodies, in contrast to 40 of the 160 (25%) HD patients, including 14 (8.8%) who were positive for HGV RNA only, 25 (15.6%) who were positive for HGV E2 antibody only, and 1 (0.6%) who had both markers. HGV exposure did not correlate with gender, age, duration of HD therapy, or history of blood transfusions. At least 20 of the 40 (50%) patients with HGV exposure had been infected before the start of chronic HD therapy. Nevertheless, at least nine (22.5%) patients acquired new HGV infections after starting chronic HD therapy, with an incidence rate of > or = 2.6% per year. Three patients with newly acquired HGV viremia after HD therapy was started and two with pre-existing HGV viremia before HD therapy was started had the same nucleotide sequences. HGV and HCV infections (with a prevalence of 14.4%) might have been transmitted independently in HD patients. In addition, HGV infection was not found to cause significant elevation of alanine aminotransferase levels in the group exposed to HGV. To conclude, the incidence of new HGV infections was at least 2.6% per year. In addition to transmission through blood transfusion, HGV may have been transmitted nosocomially patient-to-patient within the HD unit. The compliance with standard universal precautions should be carefully re-examined, but it is not necessary to routinely screen for HGV infection among patients on chronic HD.

Adult↗
PubMed2001

Effects of high-cholesterol diet on the interendothelial clefts and the associated junctional complexes in rat aorta.

The arterial endothelial intercellular cleft (AEC) and its associated junctional complex (JC) are the determinants of permeability to macromolecules. This study analyzed frequencies of AEC and JC profile types in the rat thoracic aorta at 1 and 12 months after feeding the animals with a normal or a high-cholesterol diet. Rats on either a normal diet or high-cholesterol diet for 12 months showed more of the simple 'end to end' or 'overlap' types (P < 0.01) but fewer complex 'interdigitating' type (P < 0.01) of AEC compared to the 1 month group. With regard to JC, the frequencies of gap junctions were decreased (P < 0.01) while the tight junctions and the normal junctionless complex were increased (P < 0.01) after 12 months of normal diet as compared with 1 month on the normal diet. These changes in frequencies for gap junction and tight junction were even greater for the high-cholesterol diet than for the normal diet treatment. Moreover, the incidence of open junctions was also noticeably increased after 12 months of high-cholesterol diet. These findings suggest that the proportions of the AEC and JC were highly responsive to aging whereas those of JC were more susceptible to the high-cholesterol diet treatment.

Aging↗