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Background: Haptoglobin (Hp), an acute-phase protein, is known as a potential diagnostic biomarker in human diseases. Two alleles of Hp (Hp1 and Hp2 ) exist in humans allowing three phenotypes (Hp1-1, Hp2-1, and Hp2-2), which influence the biophysical and biological properties of Hp. Objective: This work aimed to investigate the variation of serum level and fucosylation change among Hp phenotypes in patients with lung cancer compared to healthy donors. Methods: 44 patients with lung cancer and 26 healthy blood donors who lived in the Northern-Thailand region were investigated by the glycoproteomic procedure. Results: The phenotypic distribution of the Hp (Hp1-1:Hp2-1:Hp2-2) in healthy donors was 0.04:0.38:0.58, while the patient group was 0.09:0.52:0.39. The Hp1 allele frequency of patients with lung cancer (0.34) was higher than the healthy donor (0.23). Glycoprotein blotting technique represented that the level of serum Hp and its fucosylation were significantly higher among lung cancer patients compared to those of the healthy donors. However, a downward trend in the fucosylation level from Hp1-1 to Hp2-1, Hp2-2, was seen in the patient group, but varying in the serum Hp level. An N-linked glycan was enzymatically released from discrete Hp multimers of Hp2-1 and Hp2-2 samples. Analysis of glycan profiling by MALDI-TOF-MS showed that reduction of the fucosylated glycan was associated with the size of Hp multimers, resulting in the lower level of fucosylation in Hp2-1 and Hp2-2, respectively. Conclusion: Our finding demonstrates that the Hp phenotype is a dependent risk factor for lung cancer and should be incorporated into further clinical and biochemical investigations of diseases, including lung cancer.

Lactobacillus plantarum PS128 has been reported as a psychobiotic to improve mental health through the gut–brain axis in experimental animal models. To explore its mechanism of action in the gut, this study aimed to analyze the effects of L. plantarum PS128 ingestion on naïve and loperamide (Lop)-induced constipation mice. We found that, in the two mouse models, the weight, number, and water content of feces in the L. plantarum PS128 group were higher than those in the vehicle control group. Histological observation revealed that L. plantarum PS128 increased the level of colonic mucins including the major mucin MUC2. In addition, the charcoal meal test showed that L. plantarum PS128 significantly increased the small intestine transit in naïve mice, but not in the Lop-treated mice. Since intestinal serotonin has been found to modulate motility, we further analyzed the expression of genes related to serotonin signal transduction in the small intestine of naïve mice. The results showed that L. plantarum PS128 significantly altered the expression levels of Tph1, Chga, Slc6a4, and Htr4, but did not affect the expression levels of Tph2, Htr3a, and Maoa. Furthermore, immunohistochemistry revealed that L. plantarum PS128 significantly increased the number of serotonin-containing intestinal cells in mice. Taken together, our results suggest that L. plantarum PS128 could promote intestinal motility, mucin production, and serotonin signal transduction, leading to a laxative effect in mice.

Introduction. Antimicrobial resistance associated with animal hosts is easily transmitted to humans either by direct contact with resistant organisms or by transferring resistance genes into human pathogens. Gap statement. There are limited studies on antimicrobial resistance genes and genetic elements of multidrug-resistant (MDR) Escherichia coli in veterinary hospitals in Taiwan. Aim. The aim of this study was to investigate antimicrobial resistance genes in multidrug-resistant Escherichia coli from animals. Methodology. Between January 2014 and August 2015, 95 multidrug-resistant Escherichia coli isolates were obtained from pigs (n=66), avians (n=18), and other animals (n=11) in a veterinary hospital in Taiwan. Susceptibility testing to 24 antimicrobial agents of 14 antimicrobial classes was performed. Antimicrobial resistance genes, integrons, and insertion sequences were analysed by polymerase chain reaction and nucleotide sequencing. Pulsed-field gel electrophoresis (PFGE), and multi-locus sequence typing were used to explore the clonal relatedness of the study isolates. Results. Different antimicrobial resistance genes found in these isolates were associated with resistance to β-lactams, tetracycline, phenicols, sulfonamides, and aminoglycosides. Fifty-five of 95 E. coli isolates (55/95, 57.9 %) were not susceptible to extended-spectrum cephalosporins, and bla CTX-M-55 (11/55, 20.0 %) and bla CMY-2 (40/55, 72.7 %) were the most common extended-spectrum β-lactamase (ESBL) and AmpC genes, respectively. Both bla CTX-M and bla CMY-2 were present on conjugative plasmids that contained the insertion sequence ISEcp1 upstream of the bla genes. Plasmid-mediated FOX-3 β-lactamase-producing E. coli was first identified in Taiwan. Forty isolates (40/95, 42 %) with class 1 integrons showed seven resistance phenotypes. Genotyping of 95 E. coli isolates revealed 91 different XbaI pulsotypes and 52 different sequence types. PFGE analysis revealed no clonal outbreaks in our study isolates. Conclusion. This study showed a high diversity of antimicrobial resistance genes and genotypes among MDR E. coli isolated from diseased livestock in Taiwan. To our knowledge, this is the first report of plasmid-mediated ESBL in FOX-3 β-lactamase-producing E. coli isolates in Taiwan. MDR E. coli isolates from animal origins may contaminate the environment, resulting in public health concerns, indicating that MDR isolates from animals need to be continuously investigated.

Background: Most drug-resistant Escherichia coli isolates in dogs come from diseased dogs. Prior to this study, the prevalence and risk factors of fecal carriage drug-resistant E. coli and epidemic clone sequence type (ST) 131 (including subtypes) isolates in dogs were unknown. Methods: Rectal swabs were used for E. coli isolation from 299 non-infectious dogs in a veterinary teaching hospital in Taiwan. Antibiotic resistance and multiplex PCR analyses of E. coli for major STs were performed. Result: There were 43.1% cefazolin-resistant, 22.1% fluoroquinolone-resistant, and 9.4% extended-spectrum beta-lactamase-producing E. coli in our cohort. In the phylogenetic study, B2 was the predominant group (30.1%). The cefazolin-resistant group and ciprofloxacin-resistant group had greater antibiotic exposure in the last 14 days (p &lt, 0.05). The age, sex, and dietary habits of the antibiotic-resistant and -susceptible groups were similar. In the seven isolates of ST131 in fecal colonization, the most predominant subtypes were FimH41 and FimH22. Conclusion: Recent antibiotic exposure was related to the fecal carriage of antibiotic-resistant E. coli isolates. Three major subtypes (FimH41, H22, and H30) of ST131 can thus be found in fecal carriage in dogs in Taiwan.

Objectives: Tophi may occur within the knee joint causing limited knee-joint range of motion (knee motion). We investigated the relationships between knee motion, total intra-articular tophi size (tIA-tophi), and total subcutaneous tophi size (tSC-tophi) and determined knee motion improvement after continual urate-lowering therapy (ULT). Methods: A total of 26 patients with tophaceous gout and limited knee motion were enrolled. Inclusion criteria were age ≤ 60 years and preserved knee joint space on a plain radiograph. tSC-tophi were measured using a Vernier caliper and tIA-tophi were measured using magnetic resonance imaging software. All patients were re-evaluated after 12–24 months of continual ULT. Upon initial visit, knee motion was related to tIA-tophi and tSC-tophi. Results: After an average of 18.2 months of ULT, the reduction in tSC-tophi was correlated with the reduction in tIA-tophi (P = 0.014, r = 0.395) and improvement in knee motion (P = 0.038, r = 0.408). Knee motion can be eventually improved even in cases of poor initial knee motion (P = 0.000, r = –0.911) or large initial tIA-tophi (P = 0.014, r = 0.476). Tophi can occur in any location within the knee joint. In multiple lineal regression, knee motion was predicted to improve 8.39° for every 10cm of tIA-tophi reduction. Conclusions: Limited knee motion in patients with intra-articular tophi improved with medical treatment, regardless of initial severity. Furthermore, tSC-tophi can be used as an indicator of limited knee motion and their improvement as a predictor of knee motion improvement.

The quality characteristics of velvet antlers obtained from Formosan sambar deer (Cervus unicolor Swinhoi) (SDVA), harvested from 63 to 81 d during the velvet antler growth period, were evaluated by investigating the relationships between antioxidant levels; including content, activity, and content/activity ratios, and physical properties; including shear force values, color, and Ca content. The hardness of samples from base velvet antler sections increased, and that the color of these samples tended to become reddish-yellow (redder and more yellow), suggesting that the Ca content in the base section of the sample was not ossified yet. Samples from the upper sections of velvet antler showed higher superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPX) content (3.91 to 1.50 mg/mL, 2.53 to 0.90 mg/mL, and 3.95 to 1.58 mg/mL, respectively) than did samples from the middle and base sections (p

Objective : Antrodia camphorata is a type of true fungus that grows only on Cinnamomum camphora trees, also known as Cinnamomum kanehirae (“kashi”) in Taiwan. Antroquinonol is a characteristic component of A. camphorata mycelia extract and was previously shown to exhibit antitumor action and lower blood cholesterol (total cholesterol and low-density lipoprotein [LDL] cholesterol) in cellular and animal models. So, This study examined the ability of A. camphorata mycelia extract to reduce LDL cholesterol in humans. Methods : We conducted a randomized double-blind trial in 26 subjects with either borderline LDL cholesterol (120–139 mg/dL; n = 11) or mildly elevated LDL cholesterol (140–159 mg/dL; n = 15). Participants ingested tablets containing either 25 mg of A. camphorata mycelia extract (antroquinonol: 0.68 mg; n = 14) or a placebo (n = 12) for 12 weeks. Results : The test group showed a significant reduction in LDL cholesterol when compared with the placebo group after 12 weeks of tablet ingestion (p < 0.05), demonstrating the effects of A. camphorata mycelia extract on LDL cholesterol. A. camphorata mycelia extract also tended to reduce total cholesterol when compared with the placebo (p < 0.10). The borderline LDL cholesterol and mildly elevated LDL cholesterol subgroups showed a significant reduction in LDL cholesterol in subjects who ingested A. camphorata mycelia extract compared with those who ingested the placebo, again demonstrating the LDL cholesterol-lowering effect of the extract. Conclusion : A. camphorata mycelia extract lowers LDL cholesterol in individuals with somewhat high LDL cholesterol levels. This clinical trial was registered with the University Hospital Medical Information Network (UMIN no. # 000019670).

Clostridium difficile is a human and animal pathogen. Recently, the incidence of community-acquired C. difficile infection has increased, and many studies have indicated that C. difficile might be food-borne. The correlation between C. difficile infection in humans and in animals has been a topic of debate. The objective of this study was to determine the genetic relatedness of C. difficile from human and pigs in Taiwan. We investigated the molecular epidemiology of C. difficile in healthy humans and pigs from 2011 to 2015. The isolation rate of C. difficile from pigs in 13 commercial farms was 49% (100/204), and a high proportion of hypervirulent (C. difficile carrying tcdA, tcdB, and cdtA/B genes and a 39-bp deletion in the tcdC gene) ribotype 078 lineage isolates (90%, 90/100; including 078, 126, 127, and 066-like isolates) were identified. In addition, the C. difficile ribotype 127 isolates from pigs typically exhibited moxifloxacin resistance (37/43; 86%). In healthy humans, the isolation rate was 4.3% (3/69), and all healthy human isolates were non-toxigenic. In particular, we compared the porcine isolates with two patient strains (ribotype 127) obtained from two hospitals in central Taiwan. The multilocus variable number tandem repeat analysis revealed a high genetic relatedness between ribotype 127 from patients and pigs. This study indicated that isolates of the ribotype 078 lineage, and especially ribotype 127, were widely distributed in pig farms and showed a high frequency of moxifloxacin resistance. The closely related ribotype 127 from patients and pigs may have had a common origin or low diversity. In conclusion, C. difficile ribotype 127 is a noteworthy pathogen in pigs and poses a potential public health threat.

In this study, we investigated whether tetracosanol, an aliphatic alcohol isolated from Saccharum sinense, enhances insulin receptor kinase activity to exhibit an insulin synergistic effect in vitro and in vivo. The insulin plus tetracosanol enhanced insulin receptor kinase showed that AKT activity was down-regulated by S961 in differentiated L6 myotubes. Meanwhile, insulin plus tetracosanol restored the ability of glucose transporter translocation and glucose uptake in differentiated myotubes with S961-induced insulin resistance in vitro. The modification of carbon chain lengths and the hydroxyl group of tetracosanol showed that it served as a critical chemical structure for the glucostasis effect in vivo. This study provides new evidence to show that tetracosanol can improve glycaemic control via insulin receptor kinase activity induced and leads to the enhancement of glucose transporter translocation to improve glucose uptake. The hydroxyl group of tetracosanol plays a critical role for insulin receptor kinase activity.

A clinical isolate of Citrobacter freundii (JA99) obtained from a bile culture of a Taiwanese patient was found to produce a plasmid-encoded β-lactamase conferring resistance to oxyimino-cephalosporins and cephamycins. Resistance arising from production of the β-lactamase could be transferred by conjugation with an IncW plasmid (pJA99) into Escherichia coli J53. The substrate and inhibition profiles of this enzyme resembled that of an AmpC β-lactamase. The resistance gene of pJA99, cloned and expressed in E. coli DH5α, was shown to contain an open reading frame showing 92% amino acid identity with the plasmid-encoded enzyme CFE-1 of E. coli KU6400. DNA sequence analysis also identified a gene upstream of ampC in pJA99 whose sequence was 95.0% identical to the ampR gene from E. coli KU6400. In addition, orf1, the fumarate operon (frdABCD), blc, lolB and repB surrounding the ampR-ampC genes in C. freundii were identified. This DNA fragment was absent in other Citrobacter spp. Therefore, we describe a new plasmid-encoded AmpC β-lactamase, named CFE-2. This study highlights the emergence of broad-spectrum cephalosporin resistance in C. freundii owing to a new type of AmpC β-lactamase.

Multidrug-resistant Escherichia coli can contaminate food meat during processing and cause human infection. Phenotypic and genotypic characterization of the antimicrobial resistance were conducted for 45 multidrug-resistant E. coli isolates from 208 samples of beef carcasses. The mechanisms of resistance were evaluated using polymerase chain reaction and sequencing methods, and the clonal relationship among isolates was evaluated using multilocus sequence typing (MLST) and pulsed-field gel electrophoresis (PFGE). Different variants of bla, tet, flo, dfrA, and aadA genes were detected in most of the strains resistant to β-lactam, tetracycline, chloramphenicol, sulfonamides, and aminoglycosides, respectively. Extended-spectrum β-lactamase (ESBL)-producing E. coli was found in 42.2% of the 45 E. coli isolates and the most commonly detected ESBL genotypes were CTX-M group 1 and 9. Class 1 integrons with nine different arrangements of gene cassettes were present in 28 of 45 E. coli isolates. Twenty-nine PFGE groups and 24 MLST types were identified in their clonal structure. This study revealed that E. coli isolates from beef contained high diversity of antimicrobial resistance genes, integrons, and genotypes. These results highlighted the role of beef meat as a potential source for multidrug-resistant E. coli strains and the need for controlling beef safety.

Stability of neuronal connectivity is critical for brain functions, and morphological perturbations are associated with neurodegenerative disorders. However, how neuronal morphology is maintained in the adult brain remains poorly understood. Here, we identify Wnt5a, a member of the Wnt family of secreted morphogens, as an essential factor in maintaining dendritic architecture in the adult hippocampus and for related cognitive functions in mice. Wnt5a expression in hippocampal neurons begins postnatally, and its deletion attenuated CaMKII and Rac1 activity, reduced GluN1 glutamate receptor expression, and impaired synaptic plasticity and spatial learning and memory in 3-mo-old mice. With increased age, Wnt5a loss caused progressive attrition of dendrite arbors and spines in Cornu Ammonis (CA)1 pyramidal neurons and exacerbated behavioral defects. Wnt5a functions cell-autonomously to maintain CA1 dendrites, and exogenous Wnt5a expression corrected structural anomalies even at late-adult stages. These findings reveal a maintenance factor in the adult brain, and highlight a trophic pathway that can be targeted to ameliorate dendrite loss in pathological conditions.

The role of Wnt5a has been extensively explored in various aspects of development but its role in cerebellar development remains elusive. Here, for the first time we unravel the expression pattern and functional significance of Wnt5a in cerebellar development using Wnt5a−/− and Nestin-Cre mediated conditional knockout mouse models. We demonstrate that loss of Wnt5a results in cerebellar hypoplasia and depletion of GABAergic and glutamatergic neurons. Besides, Purkinje cells of the mutants displayed stunted, poorly branched dendritic arbors. Furthermore, we show that the overall reduction is due to decreased radial glial and granule neuron progenitor cell proliferation. At molecular level we provide evidence for non-canonical mode of action of Wnt5a and its regulation over genes associated with progenitor proliferation. Altogether our findings imply that Wnt5a signaling is a crucial regulator of cerebellar development and would aid in better understanding of cerebellar disease pathogenesis caused due to deregulation of Wnt signaling.

From 2007 to 2009, we collected a total of 83 bacteraemic isolates of Escherichia coli with reduced susceptibility or resistance to third-generation cephalosporins (TGCs). Isolates were genotyped by PFGE and multilocus sequence typing (MLST). The PFGE patterns revealed two highly correlated clusters (cluster E: nine isolates; cluster G: 22 isolates) associated with this prolonged clonal spreading. Compared with cluster E isolates, cluster G isolates were significantly more likely to harbour aac(6')-Ib-cr (PPE. coli cases caused by cluster E and G isolates were significantly associated with having stayed in our hospital’s respiratory care ward (Paac(6')-Ib-cr and bla CTX-M-14 positivity, and were more likely to belong to ST68/S68-like (all PE. coli ST68 associated with a stay in a long-term care facility. Using epidemiological investigations and PFGE and MLST analyses, we have identified long-term clonal spreading caused by E. coli ST68, with extra antimicrobial-resistance genes possibly acquired during the prolonged spreading period.

This study was intended to investigate the trend in vancomycin susceptibility and correlation with molecular characteristics of methicillin-resistant Staphylococcus aureus (MRSA) causing invasive infections. A total of 670 MRSA isolates were collected from patients with invasive infections as part of bacterial collection in the Tigecycline in vitro Surveillance in Taiwan (TIST) from 2006 to 2010. MICs of the isolates to vancomycin were determined using the agar dilution method. Characteristics of staphylococcal cassette chromosome mec (SCCmec), mec-associated hypervariable region (dru), and accessory gene regulator (agr) of the isolates were identified by polymerase chain reaction methods. MRSA isolates with SCCmec types I, II, and III were molecularly defined as hospital-associated MRSA (HA-MRSA), and those with SCCmec types IV, V, and VT were assigned as community-associated MRSA (CA-MRSA). All but 1 MRSA isolates exhibited vancomycin MICs ≤1 mg/L. A declining trend in vancomycin MICs among MRSA isolates was noted, which was associated with the decline in proportion of HA-MRSA. The percentage of CA-MRSA increased from 25.6% in 2006 to 46.0% in 2010. An increase in the geometric mean of vancomycin MICs was found in MRSA with particular molecular types such as SCCmec types II and III, agr groups I and II, and dru10-14. A significant correlation among particular molecular types was found, including SCCmecII-agr group II-dru4, SCCmecIII-agr group I-dru11-14, SCCmecIV-agr group II-dru9, and SCCmecVT-agr group I-dru9 and dru11. There was no vancomycin creep among MRSA isolates, and the declining trend of vancomycin MIC against MRSA was attributed to the increasing prevalence of CA-MRSA over time.

To reduce effectively the reading anxiety of learners while reading English articles, a C4.5 decision tree, a widely used data mining technique, was used to develop a personalized reading anxiety prediction model (PRAPM) based on individual learners' reading annotation behavior in a collaborative digital reading annotation system (CDRAS). In addition to forecasting immediately the reading anxiety levels of learners, the proposed PRAPM can be used to identify the key factors that cause reading anxiety based on the fired prediction rules determined by the developed decision tree. By understanding these key factors that cause reading anxiety, instructors can apply reading strategies to reduce reading anxiety, thus promoting English-language reading performance. To assess whether the proposed PRAPM can assist instructors in reducing the reading anxiety of learners, this study applies the quasi-experimental method to compare the learning performance of three learning groups, which are, respectively, supported ...

Mammary analogue secretory carcinoma (MASC) is a recently described malignancy of the salivary glands characterized by an ETV6-NTRK3 (EN) fusion gene. Morphologically, MASC is sometimes difficult to distinguish from acinic cell carcinoma. Consequently, identifying the chromosomal translocation is essential for diagnosis. We present a case of parotid gland MASC in a 13-year-old boy. To the best of our knowledge, this is the youngest case reported in the literature. Histologic evaluation showed a tumor composed of microcysts, tubular structures, solid nests, or papillary architecture, with secretions within the lumens of the cysts or tubules. Immunohistochemically, tumor cells showed diffuse positive staining of S-100 protein, cytokeratin 19, and vimentin. ETV6 rearrangement was detected by fluorescence in situ hybridization and EN fusion transcripts were verified by reverse transcription (RT-PCR) assay.

The purposes of this study were to assess the improvement of fatty liver induced by ethanol with animal liver and bile extracts. This research aimed to increase the economic values of animal liver and bile extracts and used these to reduce damage of ethanol-induced fatty liver. Extracts came from animal liver and bile, including pig bile powder, pig liver extract, a mixture of pig bile powder and pig liver extract, chicken bile powder, chicken liver extract, and a mixture of chicken bile powder and chicken liver extract, and these were fed to Long-Evans rats. The results showed that rats fed ethanol for long terms could increase values of aspartate transaminase, cholesterol, γ-glutamy-transferase and alkaline phosphatase. Pig bile powder could decrease the values of aspartate transaminase, cholesterol and γ-glutamy-transferase. The significances also decreased on aspartate transaminase, γ-glutamy-transferase and aspartate transaminase, which were carried out with the pig liver extract treatment. These results suggest pig bile and liver extracts have high potential to improve rats' ethanol-induced fatty liver with serum biochemical parameters.

Clostridium difficile causes antibiotic-associated diarrhea in both humans and animals. The ribotype 078, predominant in food animals, is associated with community-acquired C. difficile infection, and C. difficile is suggested to be a foodborne pathogen. Recently, the C. difficile ribotype 078 lineage emerged in patients and pigs in Taiwan. This study aimed to investigate the prevalence and molecular characterization of C. difficile isolated from a pig slaughterhouse, retail meat, ready-to-eat meals, and humans in Taiwan. We collected samples from one slaughterhouse (n=422), 29 retail markets (raw pork, n=62; ready-to-eat pork, n=65), and one hospital (non-diarrheal humans, stool, n=317) in 2015. The isolated C. difficile were subjected to ribotyping and multilocus variable-number tandem-repeat analysis (MLVA). In the slaughterhouse, the isolation rate from carcasses was high (23%, 21/92) and ribotype 126 dominated. Scalding water was found to have C. difficile contamination (44%, 4/9), and two of the seven isolates were ribotype 126. The isolation rates from raw pork and ready-to-eat pork were between 20% and 29%. Ribotypes 126, 127, and 014 were found in raw pork, whereas ribotype 078 was not identified in this study. Eight isolates-seven non-toxigenic isolates and one ribotype 017-were found in non-diarrheal human samples. Notably, MLVA showed that ribotype 126 isolates from the slaughterhouse, pig stool, colons, carcasses, and scalding water were closely genetically related, indicating serious risk for cross-contamination. However, the genetic evidence of foodborne transmission from carcasses to food and humans is still lacking.

Among the 219 vancomycin-resistant Enterococcus faecium isolates collected in 20 Taiwanese hospitals from 2006 to 2010, all were susceptible to linezolid and daptomycin, and 98.6% were susceptible to tigecycline. There was a shift toward higher tigecycline MIC values (MIC 90 s) from 2006-2007 (0.06 μg/ml) to 2008–2010 (0.12 μg/ml). The MIC 90 s of daptomycin and linezolid remained stationary. Although pulsotypes among the isolates from the 20 hospitals varied, intrahospital spreading of several clones was identified in 13 hospitals.

The Tigecycline In Vitro Surveillance in Taiwan (TIST) study, initiated in 2006, is a nationwide surveillance program designed to longitudinally monitor the in vitro activity of tigecycline against commonly encountered drug-resistant bacteria. This study compared the in vitro activity of tigecycline against 3,014 isolates of clinically important drug-resistant bacteria using the standard broth microdilution and disk diffusion methods. Species studied included methicillin-resistant Staphylococcus aureus (MRSA; n = 759), vancomycin-resistant Enterococcus faecium (VRE; n = 191), extended-spectrum β-lactamase (ESBL)-producing Escherichia coli ( n = 602), ESBL-producing Klebsiella pneumoniae ( n = 736), and Acinetobacter baumannii ( n = 726) that had been collected from patients treated between 2008 and 2010 at 20 hospitals in Taiwan. MICs and inhibition zone diameters were interpreted according to the currently recommended U.S. Food and Drug Administration (FDA) criteria and the European Committee on Antimicrobial Susceptibility Testing (EUCAST) criteria. The MIC 90 values of tigecycline against MRSA, VRE, ESBL-producing E. coli , ESBL-producing K. pneumoniae , and A. baumannii were 0.5, 0.125, 0.5, 2, and 8 μg/ml, respectively. The total error rates between the two methods using the FDA criteria were high: 38.4% for ESBL-producing K. pneumoniae and 33.8% for A. baumannii . Using the EUCAST criteria, the total error rate was also high (54.6%) for A. baumannii isolates. The total error rates between these two methods were E. coli . For routine susceptibility testing of ESBL-producing K. pneumoniae and A. baumannii against tigecycline, the broth microdilution method should be used because of the poor correlation of results between these two methods.

Changs-f, Yangrs, Chencm, Huangcy & Yangrj (2011) Journal of Nursing and Healthcare of Chronic Illness 3, 445–455 Cost-effectiveness and effectiveness of applying intelligent osteoporosis health risk assessment system in community-dwelling women: a novel evidence-based study Background. Few studies have explored how to assist at risk females understand the status of osteoporosis without invasive data. Aims. To develop an intelligence osteoporosis health risk evaluation system and used for a preliminary empirical analysis of Asian women. Setting. This study is focused on community-dwelling women who had accepted dual-energy X-ray absorptiometry examination in health evaluation centre of a medical centre in north Taiwan from 2005–2006. Design. A longitudinal, evidence-based study was conducted. Method. The first stage is to establish the data base. This part included information from random samples (n = 220), including data for bone mineral density, osteoporosis risk factor, knowledge, belief and behaviour. The second stage is to detect the accuracy of this system including sensitivity, specificity, positive predictive value and negative predictive value. The third part is to evaluate the preliminary effect of nursing evidence-based study. Participants (n = 90) determined the relative risks of osteoporosis with system were randomly grouped into experimental or control group. Results. The content validity index among knowledge, beliefs and behaviours were 0·99, 0·89 and 0·95, respectively. In Cronbach’s alpha value, osteoporosis prevention knowledge belies and behaviours were 0·84, 0·79 and 0·77, respectively. The sensitivity and specificity of the analytical tools were 75%. In preliminary empirical study, this study showed the knowledge and perceived benefits of osteoporosis could be promoted by individual nursing instruction and could obtain economical and effective results. Conclusions. This study can be applied in different diseases health risk evaluation in the future and provides community residents a good reliability, good validity, practical, convenient and economic self-health management model. Relevance to clinical practice. This study developed a useful approach for providing Asia women with a reliable, valid, convenient and economical self-health management model to increase the effectiveness of osteoporosis prevention programs for at risk women.

The objectives of this paper are to evaluate the heterogeneity of magnetic resonance imaging (MRI)-derived tumor thickness within tumors of the same T4a-staged tongue carcinoma and to elucidate the effects of tumor thickness on treatment outcomes. A sequential and prospectively maintained head and neck cancer database was retrospectively searched for newly diagnosed tongue carcinoma treated with surgery between 2003 and 2006. Fifty-eight patients with newly diagnosed T4a-staged tongue carcinoma were included in this study. Tumor thickness was obtained from preoperative magnetic resonance imaging. The mean MRI-derived tumor thickness was 22.29 mm. The patients with long tumor thickness (>26 mm) were associated with a significantly poor disease-specific survival (P = 0.015). The 2-year disease-specific survival rates were 72% in patients with short tumor thickness and only 27% in patients with long tumor thickness within the same T4a-staged disease. A substantial variation in MRI-derived tumor thickness was present within the same T4a-staged tongue carcinoma, and tumor thickness represented an important prognostic factor.

e-Learning is becoming an increasingly popular educational paradigm because of the rapid growth of the Internet. Recent studies have argued that affective modelling (ie, considering a learner's emotional or motivational state) should also be considered while designing learning activities. Many studies indicated that various learning emotions markedly impact learning outcomes. In the language education field, many studies have investigated anxiety associated with learning a second language, noting that anxiety has an adverse effect on the performance of those speaking English as a second language. Therefore, how to reduce anxiety associated with learning a second language to increase learning performance is an important research issue in the language education field. Accordingly, this study employed a sensor, signal processing, wireless communication, system-on-chip and machine-learning techniques in developing an embedded human emotion recognition system based on human pulse signals for detecting three human emotions-nervousness, peace and joy-to help teachers reduce language-learning anxiety of individual learners in a web-based one-to-one synchronous learning environment. The accuracy rate of the proposed emotion recognition model evaluated by cross-validation is as high as 79.7136% when filtering out human pulse signals that have bias. Moreover, this study applied the embedded emotion recognition system to assist instructor's teaching in a synchronous English conversation environment by immediately reporting variations in individual learner emotions to the teacher during learning. In this instructional experiment, the teacher can give appropriate learning assistance or guidance based on the emotion states of individual learners. Experimental results indicate that the proposed embedded human emotion recognition system is helpful in reducing language-based anxiety, thus promoting instruction effectiveness in English conversation classes. [ABSTRACT FROM AUTHOR]

Many countries have already become aging societies, as evidenced by annually decreasing fertility rates. Elderly individuals often live independently because their families cannot look after them. Therefore, computer-assisted nursing has received increasing attention in modern society, explaining why intelligent systems with physiology signal monitoring for e-health care is an emerging area of development, owing to the urgent needs of homecare for elderly people suffering chronic or sudden diseases at home. Importantly, a physiology signal monitoring system can help medical staff to monitor and analyze physiology signal effectively, such that they can not only monitor the patients' physiology states immediately, but also reduce medical cost and avoid having to visit doctors in hospital. Therefore, this study adopts system on chip (SOC) techniques to develop an embedded human pulse monitoring system with intelligent data analysis mechanism for disease detection and long-term health care. The proposed system can be applied to monitor and analyze pulse signal in daily life. The proposed system also has a friendly web-based interface for medical staff to observe immediate pulse signals for remote treatment. Hence, the proposed system provides aids long-distance medical treatment, exploring trends of potential chronic diseases, and urgent situations informing for sudden diseases. Moreover, this study also presents an intelligent data analysis scheme based on the modified cosine similarity measure to diagnose abnormal pulses for exploring potential chronic diseases.

Obesity is a major risk factor for the development of type 2 diabetes mellitus (T2DM). Surgery is the most effective treatment for morbid obesity, and it has been shown to lead to dramatic improvement in type T2DM. However, data concerning obese Asian patients have not been reported, and the mechanism of improving T2DM after bariatric surgery in this population remains speculative. From April 1997 to March 2006, 1,375 patients undergoing obesity surgery were recruited for our study of the effects of weight loss surgery on type 2 diabetes mellitus (T2DM). Laparoscopic vertical banded gastric partition (LVBG) was performed in 552 patients (40.1%), laparoscopic gastric bypass in 660 patients (48.0%), and laparoscopic adjustable gastric banding (LAGB) in 140 patients (10.2%). Another 23 patients (1.7%) received open or revision surgery. We evaluated the clinical data and effect of obesity surgery on T2DM, impaired fasting glucose (IFG) and compared the changes with those with normal fasting glucose. Among all the patients, 166 (12.1%) had impaired fasting glucose (IFG) and 247 (18.0%) had T2DM. Patients with IFG or DM were significantly older, more central obese, and they had higher insulin resistance and a higher HbA1C level than those with normal fasting glucose. The mean total weight loss for the population was 28.6%, 29.1%, 28.5%, 23.8%, and 24.4% (at 1, 2, 3, 5, and 7 years after surgery). There was no increase of surgical risk in T2DM or IFG patients. One year after operation, fasting plasma glucose returned to normal in 78.5% of T2DM patients and 94.7% of IFG patients. The HbA1C level returned to under 7.0 in 81.5% of T2DM and in 100% of IFG patients. Laparoscopic gastric bypass patients had a greater weight loss and a higher rate of glucose normalization (93.1%) than the LVBG patients (85.3%) and LAGB patients (73.9%), but no difference in HbA1C normalization. Obesity surgery resulted in significant and sustained weight loss with a cure rate of T2DM up to 80% in morbidly obese Asian patients. Gastric bypass surgery had a better result in T2DM cure rate than LVBG and LAGB because of better weight reduction.

A total of 34 Acinetobacter baumannii isolates from a district hospital in Taiwan were identified with carbapenem-hydrolyzing oxacillinase OXA-66/OXA-51-like. In addition, 26 of 28 carbapenem-resistant isolates harbored plasmid-encoded bla(OXA-23)-like genes. Twenty of 28 carbapenem-resistant isolates mapped to the major genotype cluster A of carbapenemase producer by pulsed-field gel electrophoresis.

A high-throughput and sensitive liquid chromatography/tandem mass spectrometry assay was established to detect total unlabeled hepatitis C virus inhibitor concentrations in replicon cells. The intracellular concentrations determined by this assay correlated well with concentrations obtained using radiolabeled compound. Some compounds accumulated inside the cells, with concentrations up to 300-fold higher than the input concentration. Confocal microscopic evaluation of two fluorescent-tagged inhibitors confirmed high accumulation inside the cells, sequestered inside vesicles within the cytoplasm. Incubation of cells with compound at 4 degrees C revealed that nonspecific binding to the outside of the cell membrane and to the cell culture plate occurred for some compounds. Therefore, the total concentration of compound extracted at 37 degrees C was reduced by the amount that was nonspecifically bound at 4 degrees C to yield the amount of compound inside the cells. A modification of the protocol was used for compounds with low intracellular concentrations in which cells were harvested with trypsin-EDTA prior to extraction. This eliminated the nonspecific binding to the cell culture plate and decreased the overall background of the assay. This assay was used to understand differences in cellular potency between compounds and the effects of serum proteins on the metabolic stability of compounds during incubation with cells.

A total of 393 isolates of A. baumannii were collected from patients treated at 19 teaching hospitals in Taiwan. Minimum inhibitory concentrations (MICs) and inhibitory zone diameters for tigecycline were determined by the broth microdilution method and the disk diffusion method, respectively. The MIC results were interpreted using the US FDA tigecycline susceptibility breakpoints for Enterobacteriaceae (susceptible [S]or=2 microg/mL; intermediate [I] 4 microg/mL; resistant [R]or=8 microg/mL). The disk diffusion results were interpreted by criteria recommended by Jones et al. (Sor=16 mm; I 13-15 mm; Ror=12 mm) and also by those recommended by the US FDA for Enterobacteriaceae (Sor=19 mm; I 15-18 mm; Ror=14 mm). The percentages of susceptible, intermediate and resistant isolates determined by the broth microdilution method were 80.9%, 12.2%, and 6.9%, respectively. The rates of susceptible, intermediate and resistant isolates by the disk diffusion method using the criteria of Jones et al. were 88.3%, 9.9% and 1.8% and using the US FDA criteria were 44.0%, 51.7% and 4.3%. Using the criteria recommended by Jones et al., the total error rate of the disk diffusion method in comparison with the broth microdilution method was 14.2% (56/393). For routine susceptibility testing of tigecycline against A. baumannii the broth microdilution method, not the disk diffusion method, should be used due to the poor correlation of results between these two methods interpreted either by the Jones et al. or US FDA criteria.

Tigecycline In-vitro Surveillance in Taiwan (TIST), initiated in 2006, is a nationwide surveillance programme designed to monitor longitudinally the in-vitro activity of tigecycline against commonly encountered resistant bacteria in Taiwan. This study, part of TIST-2006 study, aimed to compare the in-vitro activity of tigecycline against clinical isolates of Gram-positive bacteria. A total of 805 isolates of Gram-positive bacteria were collected from patients treated at 20 teaching hospitals. Minimum inhibitory concentrations (MICs) of tigecycline for these isolates were determined by the broth microdilution method according to the guidelines of the Clinical and Laboratory Standards Institute, and by the Etest as per the manufacturer's instructions. Susceptibility results were interpreted by the MIC criteria recommended by the US FDA. Agreement between the two methods was low: 80.7% for methicillin-resistant Staphylococcus aureus (MRSA), 27.2% for Streptococcus pneumoniae, 22.8% for other Streptococcus spp., and 30.8% for vancomycin-resistant E. faecium (VRE). There were no very major or major errors noted. Tigecycline exhibited excellent in-vitro activity against Gram-positive cocci, including MRSA, VRE, S. pneumoniae and other Streptococcus spp. isolates in Taiwan. Correlation between MIC values determined using the broth microdilution and Etest methods for these organisms was poor.

This study applies the oxygen/recycled flue gas (O2/RFG) combustion technology for waste incineration in a laboratory-scale fluidized bed incinerator to investigate the effects of different RFG percentages and O2 concentrations on the emission characteristics of organic pollutants (PAHs, phenol and benzene hydrocarbons). Experimental results show that most PAHs with high-ring structures were present in solid-phase and most low-ring PAHs were present in gas-phase. The major compounds of benzene and phenol hydrocarbons were benzene, toluene, trichlorobenzene and 2,4-dinitrophenol, phenol, dichlorophenol, respectively. As the O2 concentration in feed gas was increased from 21% to 40%, the emissions of solid- and gas-phase PAHs and phenol compounds were decreased but not for benzene compounds. Increasing RFG percentages would decrease the emissions of gas-phase PAHs, benzene and phenol compounds, but increased those of solid-phase pollutants. The best operating conditions of such O2/RFG combustion system to reduce the emissions of PAHs and phenol compounds were 40% O2, 35% RFG, and that for benzene compounds was 21% O2, 75% RFG. Comparing with conventional air combustion system, the best diminution efficiencies of PAHs, benzene and phenol compounds at such O2/RFG conditions were 59.54%, 70.97% and 52.60%, respectively. With proper feed gas compositions and RFG percentages, the combustion efficiency and destruction efficiency of organic pollutants can be improved by this O2/RFG combustion technology.

Maturation of the hepatitis C virus (HCV) polyprotein occurs by a series of proteolytic processes catalyzed by host cell proteases and the virally encoded proteases NS2 and NS3. Although several peptidomimetic inhibitors of NS3 protease have been published, only a few small molecule inhibitors have been reported. In an effort to improve screening efficiency by minimizing the spectral interference of various test compounds, a substrate that contains the longer wavelength fluorescence resonance energy transfer (FRET) pair, TAMRA/QSY-7, was devised. For the optimized substrate T-Abu-Q, with sequence Ac-Asp-Glu-Lys(TAMRA)-Glu-Glu-Abu-Psi(COO)Ala-Ser-Lys(QSY-7)-amide, the kinetic parameters with HCV NS3 protease are K(m)=30 microM, k(cat)=0.6s(-1), and k(cat)/K(m)=20,100s(-1)M(-1). We show that this substrate is suitable for inhibitor analysis and mechanistic studies so long as the substrate concentration is low enough (0.5 microM) to avoid complications from high inner filter effects. The substrate is especially useful with ultra-high-density screening formats, such as microarrayed compound screening technology, because there is less spectral interference from the compounds being tested than with more traditional (EDANS/DABCYL) FRET protease substrates. The merits of the new substrate, as well as potential applications of this FRET pair to other protease substrates, are discussed.

A transient subgenomic replicon-based shuttle vector system has been developed to investigate how genetic heterogeneity affects HCV replication efficiency. Individual NS5A or NS5B genes or cassettes containing both NS5A and NS5B genes were amplified from "quasispecies" pools derived from HCV genotype 1a or 1b patient sera using RT-PCR and cloned into their respective shuttle vectors. All shuttle vectors containing NS5A or NS5A-5B genes were constructed with the S2204I "adaptive" mutation because replicons lacking the S2204I mutation replicated poorly. Gene sequences of the quasispecies pools within either genotype 1a or 1b patient samples ranged from 94 to 95% in identity. The replication capacity of 1b shuttle vectors containing patient-derived NS5A or NS5B genes averaged 67 and 75%, respectively, relative to the laboratory-optimized 1b replicon. In contrast, the replication efficiencies of both 1a and 1b shuttle vectors containing patient-derived NS5A-5B gene cassettes averaged around 2% relative to the respective laboratory-optimized replicon. All patient-derived replicons were tested in a transient assay for their sensitivity to either interferon-alpha (IFN-alpha) or to the polymerase inhibitor A-782759. Despite the differences in replication efficiency, IC(50) values measured for most of the patient-derived replicons were equivalent to the respective values measured in the control laboratory strain replicons. These results demonstrate that patient sequence heterogeneity affects replication efficiency whenever patient-derived NS5A-5B genes are inserted into the laboratory-optimized replicon. The findings also demonstrate the utility of the shuttle vector system to test patient-derived gene sequences for sensitivity to IFN-alpha and to small molecule inhibitors.

N-nitrosamines (NAms) are well-documented for their carcinogenic potential. Human exposure to NAms may arise from the daily environment and endogenous formation via the reaction of secondary amines with nitrites or from bacteria infection. We describe the use of isotope dilution online solid-phase extraction (SPE) LC-MS/MS to quantify nine NAms in human urine. This method was validated and further applied to healthy subjects and patients with urinary tract infection (UTI). N-nitrosodimethylamine (NDMA), N-nitrosomethylethylamine (NMEA), N-nitrosopyrrolidine (NPYR) and N-nitrosomorpholine (NMOR) were analyzed with an APCI source, while N-nitrosodiethylamine (NDEA), N-nitrosopiperidine (NPIP), N-nitrosodi-n-propylamine (NDPA), N-nitrosodibutylamine (NDBA) and N-nitrosodiphenylamine (NDPhA) were quantified with an ESI source, due to their effect on the sensitivity and chromatography. NDMA was the most abundant N-nitrosamine, while NDPhA was firstly identified in human. UTI patients had three to twelve-fold higher concentrations for NDMA, NPIP, NDEA, NMOR and NDBA in urine than healthy subjects, and the NAms were significantly decreased after antibiotics treatment. NDMA concentrations were also significantly correlated with the pH value, leukocyte esterase activity or nitrite in urines of UTI patients. Our findings by online SPE LC-MS/MS method evidenced that UTI patients experienced various NAms exposures, especially the potent carcinogen NDMA, which was likely induced by bacteria infection.

To disclose the association between diabetes mellitus (DM) and cryptococcosis in HIV-uninfected patients.Case patients' diagnoses with cryptococcosis and cryptococcal meningitis were based on data from the Taiwan National Health Insurance Research Database from 2000 to 2010, and diagnoses were defined using the International Classification of Disease, Ninth Revision, Clinical Modification. A case-control study comprising inpatient and outpatient populations was conducted using 4 controls for each patient (4587 case patients and 18,348 controls) matched for age, sex, monthly income, and urbanization level.Human immunodeficiency virus (HIV)-uninfected patients with cryptococcosis were more likely to have diabetes than matched control patients (adjusted odds ratio [OR], 1.45; 95% confidence interval [95% CI], 1.36-1.64; p 0.001). A diagnosis of DM was also associated with 1-year and overall mortality from cryptococcosis (hazard ratio [HR], 1.39; 95% CI, 1.17-1.65; p 0.001, HR, 1.47; 95% CI, 1.29-1.67; p 0.001; respectively) and cryptococcal meningitis (p = 0.018).Diabetes was associated with the occurrence of cryptococcosis and cryptococcal meningitis in HIV-uninfected patients. DM was also associated with 1-year and overall mortality for these patients.

Frankfurters, in 1-link, 5-link, or 10-link packages, were surface inoculated with a five-strain mixture of Listeria monocytogenes (3.40 or 5.20 log CFU/g) after treatment with 3,000 arbitrary units (AU) or 6,000 AU of pediocin (in ALTA 2341) per link. The frankfurters were vacuum packaged, after which the 1-link and 5-link packages were irradiated at 1.2 or 2.3 kGy and the 10-link packages were irradiated at 1.4 or 3.5 kGy. L. monocytogenes was enumerated following the treatments. Selected treatments were subsequently evaluated during storage at 4, 10, and 25 degrees C for up to 12 weeks. Combination of pediocin with postpackaging irradiation at 1.2 kGy or more was necessary to achieve a 50% reduction of L. monocytogenes on frankfurters in 1-link or 5-link packages. The combination of 6,000 AU of pediocin and irradiation at 2.3 kGy or more was effective in all package sizes for inhibition of the pathogen for 12 weeks at 4 or 10 degrees C. There was a synergistic effect between pediocin and irradiation for inhibition of L. monocytogenes. Storage at 4 degrees C enhanced the antilisterial effects of the treatment combinations, with little or no growth of the pathogen in 1-link or 5-link packages during 12 weeks of storage. In general, these treatments did not affect the sensory quality of frankfurters.

Frankfurters were surfaced-inoculated with a 5-strain mixture of Listeria monocytogenes (3.40 or 5.20 log CFU/g) and vacuum-packaged (as one link, 5 links or 10 links per package) after treatments with 3,000 AU or 6,000 AU of pediocin (in ALTATM2341) per link. Treatments were evaluated for L. monocytogenes counts initially and during storage (4C, 10C and 25C)for up to 12 weeks. The populations of L. monocytogenes were reduced by addition of pediocin but were unaffected by the number of frankfurters per package. Treatments stored at 4C with pediocin effectively prolonged the time prior to growth of L. monocytogenes for 7 weeks and reduced the growth of L. monocytogenes for up to 12 weeks.

Bartonella henselae, the etiologic agent of cat-scratch disease, rarely causes back pain and is considered to be transmitted through animal scratches and bites. Here we report a cat-scratch disease case possibly with an unusual route of transmission. The patient was a 32-year-old man, and he was working as a veterinarian in a private veterinary clinic. He sought for clinical help because of unknown fever and persistent back pain for at least a month after an accidental needle puncture. Through serological testing and molecular identifications, this clinical case was confirmed to be caused by B. henselae.

Background World Health Organization (WHO) has recommended individuals with increased risk of contracting influenza A H5N1 infection to be immunized against the virus during the inter-pandemic period. Safety and immunogenicity of H5N1 vaccine among participants primed with homologous or heterologous H5N1 vaccines produced by diverse manufactures have not been reported. Methods Healthy individuals aged 20 to 60 years old were recruited and stratified into three groups: participants without priming (control group), participants primed with A/Indonesia/05/2005 vaccine, participants primed with A/Vietnam/1194/2004 vaccine and A/Indonesia/05/2005 vaccine. Enrolled participants received two doses of MF59-adjuvanted A/Vietnam/1194/2004 vaccine (study vaccine). Solicited reactions were recorded by vaccine recipients. Blood samples were obtained for hemagglutination inhibition test. Results A total of 131 participants were enrolled. No significant adverse events were recorded. Tenderness, fatigue and general muscle ache were the most common solicited reactions which alleviated within one week of immunization. Three weeks after two doses of the study vaccine, 63%, 68% and 88% were in seroprotective status in the control group, A/Indonesia/05/2005 primed group and A/Vietnam/1194/2004 and A/Indonesia/05/2005 primed group, respectively. Participants primed with A/Vietnam/1194/2004 and A/Indonesia/05/2005 showed high immune response after booster with one dose of the study vaccine. Conclusion The study vaccine did not cause severe adverse events. It elicited mostly mild to moderate reactions among participants. Participants primed with A/Vietnam/1194/2004 and A/Indonesia/05/2005 vaccine showed higher immune response than those without priming or primed with A/Indonesia/05/2005 vaccine. The report suggested those with an increased risk of influenza A H5N1 virus exposure may benefit from receiving influenza A H5N1 priming during the inter-pandemic period if the antigenicity of the pandemic influenza strain is similar to that of the priming strain. Electronic supplementary material The online version of this article (doi:10.1186/s12879-014-0587-z) contains supplementary material, which is available to authorized users.

This study was conducted to explore the epidemiological and molecular differences between carbapenem-susceptible Acinetobacter baumannii (CSAB) and carbapenem-resistant A. baumannii (CRAB) isolates. Thirty-two CSAB and 55 CRAB isolates were collected in 2010. By multilocus sequence typing analysis, 31 (56%) CRAB isolates and 11 (34%) CSAB isolates belonged to ST2. Twenty-one (38%) CRAB isolates, and 4 (13%) CSAB isolates belonged to a new type, ST129. The blaIMP, blaVIM, and blaOXA-58-like were not detected in our study isolates. blaOXA-23 and blaOXA-24/40-like were not detected in all CSAB isolates. On the contrary, blaOXA-23 was detected in 51 (93%) CRAB isolates. Class 1 integron was detected in 19 (35%) CRAB isolates and 8 (25%) CSAB isolates (p>0.05). In conclusion, the ST2 and ST129 were the major sequence types in both CSAB and CRAB isolates. The blaOXA-23 is the primary carbapenem-resistance gene in CRAB isolates from hospitalized patients and the specimens collected from hospital environment.

( R )-9-[4-Hydroxy-2-(hydroxymethy)butyl]guanine (H2G) is a potent and selective inhibitor of herpesvirus replication. It is a nucleoside analog, and its triphosphate derivative (H2G-TP) is a competitive inhibitor of herpesvirus DNA polymerases. In this study, the antiviral activities of H2G and acyclovir (ACV) and the development of viral resistance to these agents were compared in varicella-zoster virus (VZV)-infected cells. In plaque reduction assays, the 50% effective concentration of H2G for VZV was 60- to 400-fold lower than that of ACV, depending on the virus strain and the cell line tested. The enhanced efficacy of H2G against VZV can be accounted for in part by the fact that the intaracellular H2G-TP level (>170 pmol/10 6 cells) is higher than the intracellular ACV-TP level (6 cells). In addition, H2G-TP has extended half-lives of 3.9 and 8.6 h in VZV-infected MRC-5 and MeWo cells, respectively. To assess the emergence of H2G-resistant VZV in vitro, VZV was passaged in the presence of increasing concentrations of H2G. Earlier in the passage, when the concentration of H2G was relatively low, the predominant variant had the (A)76 deletion in the viral thymidine kinase (TK) gene. This mutant was identical to an ACV-resistant mutant generated in parallel experiments. However, higher concentrations of H2G appeared to favor a novel mutant, which had deletions of two consecutive nucleotides at positions 805 and 806 of the TK gene. All of these changes introduced frameshift mutations in the TK gene resulting in the expression of truncated polypeptides. H2G-resistant viruses were cross-resistant to ACV, and vice versa.

We have cloned Staphylococcus aureus DNA gyrase and topoisomerase IV and expressed them in Escherichia coli as polyhistidine-tagged proteins to facilitate purification and eliminate contamination by host enzymes. The enzyme preparations had specific activities similar to previously reported values. Potassium glutamate (K-Glu) stimulated the drug-induced DNA cleavage activity and was optimal between 100 and 200 mM for gyrase and peaked at 100 mM for topoisomerase IV. Higher concentrations of K-Glu inhibited the cleavage activities of both enzymes. Using a common buffer system containing 100 mM K-Glu, we tested the enzyme-mediated DNA cleavage activities of both gyrase and topoisomerase IV with oxolinic acid, norfloxacin, ciprofloxacin, trovafloxacin, clinafloxacin, and the 2-pyridone ABT-719. As expected, all drugs tested demonstrated greater potency against topoisomerase IV than against gyrase. In addition, cleavage activity was found to correlate well with antibacterial activity.

To reduce effectively the reading anxiety of learners while reading English articles, a C4.5 decision tree, a widely used data mining technique, was used to develop a personalized reading anxiety prediction model (PRAPM) based on individual learners' reading annotation behavior in a collaborative digital reading annotation system. In addition to forecasting immediately the reading anxiety levels of learners, the proposed PRAPM can be used to identify the key factors that cause reading anxiety based on the fired prediction rules determined by the developed decision tree. By understanding these key factors that cause reading anxiety, instructors can apply reading strategies to reduce reading anxiety, thus promoting English-language reading performance. To assess whether the proposed PRAPM can assist instructors in reducing the reading anxiety of learners, this study applies the quasi-experimental method to compare the learning performance of three learning groups, which are supported by a collaborative digital reading annotation system with different learning mechanisms to reduce reading anxiety. The control group, experimental group A and experimental group B conducted the same English reading activity. However, each group was given a collaborative digital reading annotation system with individual annotations, cooperative annotations, and cooperative annotation with the instructor's support to reduce reading anxiety by proposed PRAPM. Experimental results indicate that the average correct prediction rate of the proposed PRAPM in identifying the reading anxiety levels of learners was as high as 70%. The online instructor who applied reading assistive strategies based on the mining factors that affect reading anxiety from the proposed PRAPM can significantly reduce the reading anxiety of male learners in the experimental group B, showing that gender difference existed, and the online instructor's interaction with the male learners of the experimental group B indeed helped reduce the reading anxiety.

The complete nucleotide sequence of the gene encoding the protease and ICP35 capsid protein of herpes simplex virus (HSV) type 2 strain G has been determined. The gene consisits of a 1908 bp open reading frame capable of encoding 636 amino acids. Comparative nucleotide sequence analysis of the gene with published HSV-1 sequences revealed that the HSV-2 nucleic acid segment most likely encodes two distinct proteins. The first protein is the HSV-2 protease, which is required for the assembly of the herpes virus capsid. The second protein has been previously designated as ICP35 in HSV-1, and belongs to the family of herpesvirus capsid proteins.

Products Processing Division, LRI, COA, Tainan, Taiwan. Accepted 1 August, 2012 The purpose of this study was to analyze the antioxidation activities and total phenolic contents of Toona sinensis . An extractive method of the antioxidant activities of local T. sinensis leaf extracts and various antioxidation models were analyzed. The effects of various solvent concentrations to extract the T. sinensis leaf on the antioxidation activity were compared. The results showed that the T. sinensis leaf extracts of various solvent concentrations had good antioxidant activities. The chelating abilities of ferrous ion in the T. sinensis leaf extracts obtained from various solvent concentrations were above 80%. It is concluded that T. sinensis leaf extracts could be used as biopreservatives of food products. The total antioxidant activities, chelating abilities of ferrous ions and reducing capacities of the T. sinensis leaf extracts might provide a substitute for a natural antioxidant. Keywords: Antioxidative activity, bio-preservatives, extracts, Toona sinensis