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Y S Wu

Publications and source records attributed to Y S Wu.

At least 19 recordsLinked to original sources
PubMed2006

A combined phage display ScFv library against Myxobolus rotundus infecting crucian carp, Carassius auratus auratus (L.), in China.

Immunological methods have been developed for the diagnosis of Myxobolus rotundus but their use has been limited for the prevention and therapy of this serious parasitic pathogen. Phage display antibody libraries are a powerful technique for the development of antibodies to molecules of interest and have advantages over traditional hybridroma approaches. In the present study, four antigen fractions related to M. rotundus were prepared and a combined phage display single-chain antibody fragments (ScFv) library was constructed against this parasite. Preliminary analysis indicated that a combined antibody library of about 2.08 x 10(5) individual clones and high diversity was generated. After four rounds of screening (bio-panning) against soluble spore protein prepared from lysed, intact, mature M. rotundus spores, a strain monoclonal phage display ScFv, termed pCAN-6H9, with better affinity, was isolated. The pCAN-6H9 gene fragment was sequenced and analysed. The specificity of pCAN-6H9 was further demonstrated by dot-blot. In competition enzyme-linked immunosorbent assay, both the original and enriched phage-displayed ScFv repertoire showed significant inhibition of mouse anti-M. rotundus serum binding to coated antigen, while the inhibition rate of monoclonal pCAN-6H9 phage particles was only 11.83%.

Animals↗
PubMed2006

Relationship between common functional polymorphisms of the p22phox gene (-930A > G and +242C > T) and nephropathy as a result of Type 2 diabetes in a Chinese population.

OBJECTIVE: Genetic determinants are important in diabetic nephropathy (DN). Oxidative stress has also emerged as an important pathogenic factor in DN and vascular NADH oxidase is a major source of reactive oxygen species (ROS). Previous small studies reported a strong but contradictory association between functional genetic variation of p22(phox), an important subcomponent of NADH oxidase, and DN. We investigated the association between two common functional single nucleotide polymorphisms (SNPs) (-930 A > G and +242 C > T) and DN in a much larger group of Chinese patients with Type 2 diabetes mellitus (T2DM). RESEARCH DESIGN AND METHODS: Case-control study of Chinese subjects with long-standing T2DM (> 10 years). Cases (n = 306) were subjects with a spot urinary albumin : creatinine ratio (ACR) of > 113 mg/mmol or elevated serum creatinine. Control subjects (n = 306) had ACR < 3.3 mg/mmol and normal serum creatinine. Genotyping was carried out by standard PCR and restriction fragment length polymorphism analysis. RESULTS: Gender distribution, age, duration of diabetes and HbA(1c) were similar in cases and control subjects. Distribution of genotypes in the control subjects for both SNPs was consistent with the Hardy-Weinberg equilibrium. Distribution of genotypes did not differ significantly between cases and control subjects for both polymorphisms-+2424C > T: cases CC 84.6%, CT 15.0%, TT 0.4% and control subjects CC 87.6%, CT 11.8%, TT 0.6% (P = 0.45); -930 A > G: cases AA 40.5%, AG 41.8%, GG 17.7% and control subjects AA 38.2%, AG 49.0%, GG 12.8% (P = 0.12). Distribution of alleles was also similar-+2424 C > T: cases C 92.2%, T 7.8% and control subjects C 93.5%, T 6.5% (P = 0.66); -930 A > G cases A 61.4%, G 38.6% and control subjects A 62.7%, G 37.3% (P = 0.38). We estimated that our study has approximately 80% power to detect a relative risk of 1.65 (for +242 C > T) and 1.35 (for -930 A > G) conferred by the minor allele, respectively. CONCLUSIONS: In contrast with previous small studies, our data suggest that these SNPs do not confer significantly increased susceptibility to DN secondary to T2DM in Chinese subjects.

Aged↗
PubMed2005

Humoral immune responses of the grouper Epinephelus akaara against the microsporidium Glugea epinephelusis.

The humoral immune responses of grouper Epinephelus akaara to a natural infection with Glugea epinephelusis was studied by ELISA utilizing intact mature spores as the coated antigen. Results showed that a specific humoral immune response was elicited, but the intensity of infection (in terms of the number of cysts) was not related to the antibody level in naturally infected hosts. The differences in the antigenicity of intact mature spores and soluble spore proteins derived from cracked mature spores were also analyzed. Results suggested that similar antigen epitopes existed between the 2 groups. Additionally, antigen component patterns and the distribution of antigen with immunogenicity were investigated by using the western blot and the immunofluorescent antibody technique (IFAT). The new parasitic microsporidium has specific polypeptide patterns comparable to the reported fish microsporidians. The main antigenic substances are concentrated on the surface of spores, and are mostly located on the anterior and posterior end of the spore bodies. Most surface components of the G. epinephelusis spores are soluble. The potential role of the surface components in initiating infection was also discussed.

Animals↗
PubMed2005

Modulation of I(Kr) inactivation by mutation N588K in KCNH2: a link to arrhythmogenesis in short QT syndrome.

OBJECTIVE: Short QT syndrome (SQTS) is characterized by ventricular arrhythmias and sudden death. One form of SQTS is caused by mutation N588K in human ether-a-go-go-related gene (HERG). In this study we sought to determine the potential role of N588K in arrhythmias. METHODS: We measured the characteristics of HERG current generated by wild-type (WT) KCNH2 and the N588K mutant channel expressed in mammalian TSA201 cells. RESULTS: Whole-cell patch-clamp recordings of WT HERG currents showed the usual rapid onset of inactivation (rectification) at potentials more positive than +10 mV. In contrast, N588K currents rectified at potentials over +80 mV. Over the physiological range of potentials, N588K currents do not inactivate. During an action potential clamp, WT currents displayed a "hump" like waveform with slow activation kinetics and a rapid increase during phase 3 repolarization. In contrast, N588K currents were proportional to the amplitude of the action potential and displayed a dome-like configuration and a much larger current during the initial phases in the ventricle. Purkinje cell action potentials display a more negative phase 2 repolarization than the ventricle and elicited much smaller WT and N588K currents of similar amplitudes. CONCLUSIONS: Physiologically the N588K mutation abolishes rectification of HERG currents and specifically increases I(Kr) in the ventricle with minimal effects on the Purkinje fiber action potential duration. Such preferential prolongation may explain the separation of the T and U waves observed in the ECG of SQTS patients and lead to re-excitation of the ventricle endocardium.

Action Potentials↗
PubMed2005

Molecular orientation of evaporated pentacene films on gold: alignment effect of self-assembled monolayer.

Pentacene films deposited on self-assembled monolayers (SAMs) bearing different terminal functional groups have been studied by reflection-absorption IR, grazing angle XRD, NEXAFS, AFM, and SEM analyses. A film with pentacene molecules nearly perpendicularly oriented was observed on Au surfaces covered with an SAM of alkanethiol derivative of X-(CH2)(n)-SH, with X = -CH(3), -COOH, -OH, -CN, -NH(2), C(60), or an aromatic thiol p-terphenylmethanethiol. On the other hand, a film with the pentacene molecular plane nearly parallel to the substrate surface was found on bare Au surface. A similar molecular orientation was found in thinner ( approximately 5 nm) and thicker (100 nm) deposited films. Films deposited on different surfaces exhibit distinct morphologies: with apparently smaller and rod-shaped grains on clean bare Au surface but larger and islandlike crystals on SAM-modified surfaces. X-ray photoemission electron microscopy (X-PEEM) was used to analyze the orientation of pentacene molecules deposited on a SAM-patterned Au surface. With the micro-NEXAFS spectra and PEEM image analysis, the microarea-selective orientation control on Au was characterized. The ability to control the packing orientation in organic molecular crystals is of great interest in fabricating organic field effect transistors because of the anisotropic nature of charge transport in organic semiconducting materials.

Journal Article↗
PubMed2005

Contribution of neuronal sodium channels to the cardiac fast sodium current INa is greater in dog heart Purkinje fibers than in ventricles.

OBJECTIVE: To determine the presence and the potential contribution of neuronal sodium channels to dog cardiac function. METHODS: We used a combination of electrophysiological (patch clamp), RT-PCR, biochemical and immunohistochemical techniques to identify and localize neuronal Na(+) channels in dog heart and determine their potential contribution to the fast sodium current. RESULTS: In all cardiac tissues investigated, Na(v)1.1, Na(v)1.2 and Na(v)1.3 transcripts were detected. In immunoblots, we found Na(v)1.1 and Na(v)1.2 proteins in the ventricle (V) and in Purkinje fibers (PF). Na(v)1.3 immunoblots suggested strong proteolytic activity against this isoform in the heart. Na(v)1.6 was not found in any of the tissues tested. Confocal immunofluorescence on cardiac myocytes showed that Na(v)1.1 was predominantly localized at the intercalated disks in V and PF and around the nucleus (V). Na(v)1.2 was only present at the Z lines (V). Consistent with the immunoblot data, an intense but diffuse intracellular staining was observed for Na(v)1.3. Na(v)1.6 fluorescence staining was faint and diffuse. Surprisingly, immunoblots indicated the presence of two Na(v)beta 2 variants: a 42-kDa protein that co-localized with Na(v)1.2 at the Z lines in V and a 34-kDa protein that co-localized with Na(v)1.1 at the intercalated disks in PF. In agreement with the biochemical data, electrophysiological results suggest that neuronal sodium channels generate 10+/-5% and 22+/-5% of the peak sodium current in dog ventricle and Purkinje fibers, respectively. CONCLUSIONS: Our results suggest that neuronal NaChs are more abundant in Purkinje fibers than in ventricles, and this suggests a role for them in cardiac conduction.

Animals↗
PubMed2005

Compaction mechanism and tablet strength of unlubricated and lubricated (silicified) microcrystalline cellulose.

This paper describes the differences in compaction properties between microcrystalline cellulose (MCC) and microcrystalline cellulose co-processed with colloidal silicon dioxide (SMCC). The different compaction parameters are not only compared for the pure materials, but also for the lubricated powders with magnesium stearate. Neither magnesium stearate, nor colloidal silicon dioxide, facilitates extensively the densification of (silicified) microcrystalline cellulose during compaction. The difference in tablet relaxation of MCC and SMCC indicates a small negative effect of colloidal silicon dioxide on the interparticle bonding strength of unlubricated MCC. However, for lubricated MCC a larger increase in tablet relaxation at a high compression speed was found than for lubricated SMCC tablets. Accordingly, the decrease in tablet strength was larger for the MCC tablets than for the SMCC tablets when lubrication was applied. The examination of the tablet strengths of tablets compressed from physical mixtures of MCC with increasing concentrations of colloidal silicon dioxide proved the slightly negative influence of silicon dioxide on the tablet strength of unlubricated MCC tablets and the positive effect of colloidal silicon dioxide addition on the tensile strength of lubricated MCC tablets. Co-processing of MCC with colloidal silicon dioxide showed no extra contribution on the tablet strength of lubricated tablets above the physical mixtures. The interactions between the different materials were further supported by the interaction parameters based on partial solubility parameters.

Cellulose↗
PubMed2005

Emergency management and infection control in a radiology department during an outbreak of severe acute respiratory syndrome.

The World Health Organization classified Taiwan as a serious epidemic-stricken area when the extent of severe acute respiratory syndrome (SARS) in Taiwan became clear. As of 11 July 2003, 671 probable SARS cases had been identified in Taiwan and 7 healthcare workers had died from the disease. Radiographers were easily infected by SARS because they had close contact with suspected or probable cases while conducting chest X-ray examinations. Three radiographers had been infected by the end of May 2003. Because of the impact of SARS on the Radiology Department, the department established a SARS emergency infection control team and re-designed the department's infection-control and emergency-management procedures based on the concept of risk-grade protection. This effort included installing a radiographic room at the fever-screening station, re-allocating human resources in the Radiology Department, training the department staff in infection control, and drafting new operational procedures for radiographers conducting X-ray examinations on SARS patients. The goal of this program was to reduce the infection rate and distribute materials efficiently in the department. This article introduces the emergency-management procedure of the Radiology Department during the SARS outbreak and the infection-protection experience of the department staff.

Disease Outbreaks↗
PubMed2003

Coordinated down-regulation of KCNQ1 and KCNE1 expression contributes to reduction of I(Ks) in canine hypertrophied hearts.

OBJECTIVE: In animal models of hypertrophy, electrical remodeling giving rise to QT prolongation occurs rapidly and is associated with the development of torsade de pointes (TdP) arrhythmias and sudden death. Chronic AV block (CAVB)-induced hypertrophy in dogs has been associated with a reduction in the slow component (I(Ks)) of the delayed rectifier potassium current (I(K)), which contributes to a prolongation of ventricular repolarization, the development of an acquired form of long QT, and the substrate for triggered activity and TdP. The present study was designed to probe the molecular basis for the decrease in I(Ks) by studying the characteristics of KCNE1 and KCNQ1, the putative genes responsible for formation of the channel. METHODS AND RESULTS: Using a combination of Northern blot, competitive multiplex PCR and immunoblot assays, we found that CAVB reduces KCNE1 and KCNQ1 RNA in the canine ventricles by 70 and 80%, respectively. Protein levels of KCNE1 and KCNQ1 were reduced by 60 and 50%, respectively. We also demonstrate at the molecular level the basis for inter-ventricular difference in I(Ks) density previously reported in hearts of normal dogs and show the basis for reduction of this difference in the CAVB dog. CONCLUSIONS: Our results indicate that the CAVB-induced reduction in I(Ks) is due to a down-regulation of KCNE1 and KCNQ1 transcription. The data suggest that electrical remodeling of the cardiac ventricle during hypertrophy involves regulation of the gene expression through modulation of transcriptional and translational regulatory pathways. The reduction in KCNE1 and KCNQ1 expression increases the dependence of ventricular repolarization on the rapid component of I(K) and may potentiate the action of Class III antiarrhythmic agents.

Animals↗
PubMed2003

Dimensional analysis for establishing the testing criteria of kinetic study with respirometry.

The kinetic study of a microbial system requires the determination of kinetic parameters under a set of operating variables. Previous researchers indicated that initial conditions, such as S0/X0 and S0/KS may influence the reliability of the parameter estimation. However, little study has been done to identify the sensitivity of system variables to the parameter estimation. This study proposes a novel dimensional analysis and identifies six dimensionless groups: mu(m)/fw, kd/fw, Yg, S0/KS, KS/S0, and 1/(fw theta(c) - 1). By incorporating the SP-moving algorithm proposed by Wu and coworkers in 2001, an algorithm was proposed in this study to perform a sensitivity analysis on the six dimensionless groups. Results of this analysis reveal that S0/X0 is more sensitive than S0/KS, as also evidenced by the fact that gross growth yield (Yg) is sensitive and affecting S0/X0. The analysis also suggests that the theta(c)-based wasting frequency (fw theta(c)) is more sensitive than the daily wasting frequency (fw). A critical minimum value of 1.3 for S0/X0 and a maximum value of 0.1 for S0/KS were suggested to establish the testing criteria for the kinetic study under the respirometric conditions.

Algorithms↗
PubMed2002

Functional expression of GFP-linked human heart sodium channel (hH1) and subcellular localization of the a subunit in HEK293 cells and dog cardiac myocytes.

Recent evidence suggests that biosynthesis of the human heart Na+ channel (hH1) protein is rapidly modulated by sympathetic interventions. However, data regarding the intracellular processing of hH1 in vivo are lacking. In this study we sought to establish a model that would allow us to study the subcellular localization of hH1 protein. Such a model could eventually help us to better understand the trafficking of hH1 in vivo and its potential role in cardiac conduction. We labeled the C-terminus of hH1 with the green fluorescent protein (GFP) and compared the expression of this construct (hH1-GFP) and hH1 in transfected HEK293 cells. Fusion of GFP to hH1 did not alter its electrophysiological properties. Confocal microscopy revealed that hH1-GFP was highly expressed in intracellular membrane structures. Immuno-electronmicrographs showed that transfection of hH1-GFP and hH1 induced proliferation of three types of endoplasmic reticulum (ER) membranes to accommodate the heterologously expressed proteins. Labeling with specific markers for the ER and the Golgi apparatus indicated that the intracellular channels are almost exclusively retained within the ER. Immunocytochemical labeling of the Na+ channel in dog cardiomyocytes showed strong fluorescence in the perinuclear region of the cells, a result consistent with our findings in HEK293 cells. We propose that the ER may serve as a reservoir for the cardiac Na+ channels and that the transport from the ER to the Golgi apparatus is among the rate-limiting steps for sarcolemmal expression of Na+ channels.

Animals↗
PubMed2001

Determination of pyruvate and lactate in primary liver cell culture medium during hypoxia by on-line microdialysis-liquid chromatography.

A microdialysis sampling device was constructed for the measurement of pyruvate and lactate in primary liver cell culture medium during hypoxia. It was composed of a Petri dish, a dialysis membrane and two transmission tubes within a hypoxia chamber. The dialysis membrane was located in the Petri dish such that it was immersed in the culture medium. Dialysates were collected and introduced by an on-line injector to a liquid chromatographic system for analysis of pyruvate and lactate. The detection limit of this assay was 0.2-2.0 microM with acceptable intra- and inter-assay reproducibilities. In order to validate the assay, primary liver cells were incubated in the Petri dish within a hypoxia chamber in an incubator. The baseline concentrations of pyruvate and lactate in primary liver cell culture medium were 10.6+/-5.6 and 607+/-143 microM, respectively. These levels drastically changed during hypoxia and reperfusion. In conclusion, the present assay provides a sensitive, direct measurement of pyruvate and lactate in culture medium while minimizing pretreatment procedures for sample preparation.

Animals↗
PubMed2001

High-performance chiral separation of fourteen triazole fungicides by sulfated beta-cyclodextrin-mediated capillary electrophoresis.

In this paper, sulfated beta-cyclodextrin-mediated capillary electrophoresis (CE) is evaluated as a new approach for the chiral separation of triazole-type fungicides. The 14 fungicides investigated were bitertanol, cyproconazole, difenoconazole, diniconazole, flutriafol, hexaconazole, myclobutanil, paclobutrazol, penconazole, propiconazole, tebuconazole, tetraconazole, triadimefon and triadimenol. Under the optimal conditions, excellent enantioseparation was achieved for all the 14 fungicides, including those fungicides containing two chiral centers. To our knowledge, this is the only system to date that offers outstanding enantiodiscrimination towards all triazole-type fungicides. The impact of the molecular structures of the triazole compounds on their migration behavior was studied. Similar to other chemical systems involving host-guest complexation, the interaction between sulfated beta-cyclodextrin and the triazole compounds was found to be affected by a variety of factors, including electrostatic force, hydrogen bonding, steric effect and hydrophobicity. These factors, coupled with the countercurrent electroosmotic flow (EOF), were believed to be the major forces behind the exceptional chiral selectivity.

Cyclodextrins↗
PubMed2001

The study of Central Taiwan particles concentration variations during earthquake period.

Ambient particle concentration was taken on the traffic sampling site over the Chung-Chi Road over the bridge (CCROB) in front of Hungkuang Institute of Technology (HKIT). The sampling time was from August 1999 to December 1999. During the sampling period, Taiwan's biggest earthquake in more than a century registered 7.3 on the Richter scale (Taiwan Chi-Chi Earthquake). Besides, there are more than 20,000 aftershocks following the Taiwan Chi-Chi Earthquake within 3 months. Thus, the mass concentration of particles with aerodynamic diameters smaller than 2.5 microm (PM2.5) and PM2.5-10 was also collected then compared with the total mass concentration of suspended particles (TSP) in this study. The average TSP, PM2.5-10, and PM2.5 concentrations are 106, 24.6, and 58.0 microg/m3, respectively, after the Taiwan Chi-Chi Earthquake. The average TSP concentrations before and after Taiwan Chi-Chi Earthquake were 69.6 and 127 microg/ m3, respectively. In addition, statistical analysis of the PM10 data from this study and EPA in 1999 yielded a Tstatistic of 0.147, which is smaller than t(0.975,18) = 2.101. It is indicated that there was no significant difference. So, the PM10 concentrations measured after Taiwan Chi-Chi Earthquake in this study were also greater than those data previously obtained from Taiwan EPA in the same region of this area. The relationships between TSP, PM10, PM2.5-10, and PM2.5 particle concentrations and wind speed (R2) are .77, .59, .58, .58, respectively. And the ratios of PM2.5/PM25-10, PM2.5/PM10, and PM10/TSP are 221%, 67.2%, 58.0%, respectively. The average ratios of PM2.5/PM2.5-10 and PM2.5/PM10 increase by about 120% and 17%. It indicated that the fine-particles concentration increases compared to the coarse-particles concentration after 921 Taiwan Chi-Chi Earthquake. And the proposed reasons are that local motor vehicle emissions combined the fine particles transported from the Chi-Chi epicenter. More importantly, the wind direction was mainly blown from southeastern part. These two main factors enhance the fine-particles concentration in this area.

Air Pollutants↗
PubMed2001

On the selection of primary variables in numerical formulation for modeling multiphase flow in porous media.

Selecting the proper primary variables is a critical step in efficiently modeling the highly nonlinear problem of multiphase subsurface flow in a heterogeneous porous-fractured media. Current simulation and ground modeling techniques consist of (1) spatial discretization of mass and/or heat conservation equations using finite difference or finite element methods; (2) fully implicit time discretization; (3) solving the nonlinear, discrete algebraic equations using a Newton iterative scheme. Previous modeling efforts indicate that the choice of primary variables for a Newton iteration not only impacts computational performance of a numerical code, but may also determine the feasibility of a numerical modeling study in many field applications. This paper presents an analysis and general recommendations for selecting primary variables in simulating multiphase, subsurface flow for one-active phase (Richards' equation), two-phase (gas and liquid) and three-phase (gas, water and nonaqueous phase liquid or NAPL) conditions. In many cases, a dynamic variable switching or variable substitution scheme may have to be used in order to achieve optimal numerical performance and robustness. The selection of primary variables depends in general on the sensitivity of the system of equations to the variables selected at given phase and flow conditions. We will present a series of numerical tests and large-scale field simulation examples, including modeling one (active)-phase, two-phase and three-phase flow problems in multi-dimensional, porous-fractured subsurface systems.

Air↗
PubMed2001

Full-scale evaluation of heat balance for autothermal thermophilic aerobic treatment of food processing wastewater.

A full-scale autothermal thermophilic aerobic treatment (ATAT) of food-processing wastewater was evaluated in this study. The wastewater was rich in oil and grease at concentrations of 1,500-2,000 mg/L. The system has been operated for more than one and a half years since the startup. Under steady state conditions, the ATAT process was capable of spontaneous reaction at temperatures of 45-55 degrees C without the addition of external heat. Treatment efficiency was as high as 95% in COD reduction at a volumetric COD loading of 4.1 kg/m3-d. A mathematical heat balance model was developed based on the theoretical considerations of heat sources and losses for the ATAT process. A computer algorithm was established to evaluate specific heat potential (Hs) of the wastewater under steady state conditions. Six months of steady-state data were used for the evaluation. The result shows that on average the wastewater had a specific heat potential (Hs) of 4,720 kcal/kg-COD removed and the biological heat contributed 41.4% of the total heat input. A net heat flux of 4,270 kcal/min and volumetric heat intensity (Hv) of 38.0 kcal/L was necessary to maintain reaction temperature at 48.2 degrees C for the ATAT process. The full-scale ATAT process showed the typical characteristics of high removal rate, low sludge yield, and poor solids settleability for thermophilic aerobic treatment reported in the literature.

Aerobiosis↗
PubMed2001

[Expression and immunocompetence characterization of Plasmodium falciparum lactate dehydrogenase].

OBJECTIVE: To express lactate dehydrogenase (LDH) gene of Plasmodium falciparum FCC1/HN in the E. coli TG1 and analyse its immunocompetence. METHODS: The LDH gene of the P. falciparum was specifically amplified by polymerase chain reaction, and the recovered gene fragment was cloned into pGEX-4T-1 vector for expression of fusion protein with glutathione S-transferase(GST). The recombinant plasmid was transformed into the E. coli TG1. Four mice (Kunming strain) were immunized with purified expressed protein(antigen) and the polyclonal antibodies were collected. The immunocompetence of recombinant protein was analysed by ELISA and Western-blot. RESULTS: The LDH gene of P. falciparum was successfully expressed in the E. coli TG1. The expressed protein exhibited a specific reaction with immune sera obtained from rabbits immunized with P. falciparum. The specific humoral responses were induced in mice and the titer of the specific antibody was 1:16 by two-dimensional diffusion assay. CONCLUSION: The LDH gene of P. falciparum has been successfully expressed in the E. coli TG1 and the expressed protein has high antigencity.

Animals↗