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Frequent outbreaks of avian influenza viruses (AIV), which are influenza A viruses, result in heavy economic and national damages. However, current diagnostic methods require about 5–7 days for AIV to be confirmed, which allows enough time for the widespread dissemination of the virus. Therefore, there is a need for rapid and accurate diagnosis of AIV. Here, we developed a diagnostic method using chip-based quantitative reverse transcription-polymerase chain reaction (RT-qPCR) for the rapid identification of the major subtypes H5, H7, and H9 of AIV that influence the pathogenicity of the virus. Specific primer pairs and probes were designed to target the highly conserved Matrix (M) gene regions in influenza A viruses. Specific primer pairs and probes for subtyping were developed by targeting the conserved sequences in the haemagglutinin (HA) gene specific to each subtype. For pathotyping, specific primer pairs and probes for the highly pathogenic avian influenza viruses (HPAIV) were generated by targeting the HA cleavage site, which is highly related to the pathogenicity of the virus. Using the primer pairs and probe sets, we synthesized the major sequence of each type and constructed a standard curve to confirm their detection limit. In addition, we used a chip-based RT-qPCR method to test the samples collected from the faeces of wild birds, cloacal and oropharyngeal swab samples of infected chicken, and allantoic fluids samples incubated with inoculated eggs. As a result, AIV was accurately detected in all samples. Our study shows that by using a single DNA chip in a portable chip-based RT-qPCR device and specific primer pairs and probes, rapid and accurate AIV detection and subtyping and pathotyping is possible, which will reduce the spread of the virus, thereby reducing the economic and national damages.

A novel porcine circovirus 4 has been recently identified in China and Korea. A sensitive and specific diagnostic method is urgently required to detect the virus in field samples. We developed a loop-mediated isothermal amplification (LAMP) the assay for the visual detection of PCV4 and evaluated its sensitivity, specificity, and applicability in clinical samples. This assay's results can be directly visualized by the naked eye using hydroxynaphthol blue after incubation for 40 min at 64 °C. The assay specifically amplified PCV4 DNA and no other viral nucleic acids. The sensitivity of the assay was

Diabodies are bispecific antibody fragments that have two antigen binding Fv domains. They are unique among hundreds of different formats of bispecific antibodies because they are small and rigid enough to be crystallized. Diabodies are generated by connecting variable regions of heavy and light chains by a peptide linker. Because of the short length of the linker, intramolecular association of the variable regions is not allowed. Instead, the variable regions from the different peptide chains associate together, forming a dimeric complex with two antigen binding sites. Previous crystallographic studies of diabodies demonstrate the extraordinary structural diversity of diabodies. They have also shown that the relative orientation and interaction of the two Fv domains in diabodies have substantial flexibility due to instability of the Fv interface. Introduction of site specific mutations and disulfide bridges can reduce flexibility and therefore increase rigidity and predictability of the diabody structures. These stabilized diabodies will be useful for future application to structural biology and protein nanotechnology.

Methods generating fusion proteins with rigid and predictable structures have been developed in recent years. Among them, helix fusion methods that link two proteins by connecting their terminal alpha helices into a single and extended alpha helix can be particularly useful because designing fusion helices is conceptually and technically simple. These methods have been shown crucial in obtaining crystals that diffract x-rays to high resolution or attaching large and symmetrical backbone proteins to small target proteins for cryo-EM analysis. The structural rigidity of the fusion helix is crucial for these applications, and the reduction of structural ambiguity and flexibility at the fusion sites will further enhance the usefulness of this method.

A simple reverse transcription loop–mediated isothermal amplification combined with visual detection method (vRT–LAMP) assay was developed for rapid and specific detection of porcine epidemic diarrhea virus (PEDV) in this study, which overcomes the shortcomings of previously described RT–LAMP assays that require additional detection steps or pose a risk of cross-contamination. The assay results can be directly detected by the naked eye using hydroxynaphthol blue after incubating for 40 min at 62 °C. The assay specifically amplified PEDV RNA and no other viral nucleic acids. The limit of detection of the assay was less than 50 RNA copies per reaction, which was 100 times more sensitive than conventional reverse transcription polymerase chain reaction (RT-PCR) and comparable to real-time RT-PCR (RRT-PCR). In the clinical evaluation, the PEDV detection rate of vRT–LAMP was higher than that of RRT-PCR, showing 99 % concordance, with a kappa value (95 % confidence interval) of 0.97 (0.93–1.01). Considering the advantages of high sensitivity and specificity, simple and direct visual monitoring of the results, no possibility for cross–contamination, and being able to be used as low-cost equipment, the developed vRT–LAMP assay will be a valuable tool for detecting PEDV from clinical samples, even in resource-limited laboratories.

Background and aims To evaluate the efficacy and safety of evogliptin, a newly developed dipeptidyl peptidase-4 inhibitor, in patients with type 2 diabetes (T2D) inadequately controlled by diet and exercise. Materials and methods In this randomized, double-blind, placebo-controlled, parallel-group, multicenter, phase III study, 160 patients with T2D were assigned to either an evogliptin 5 mg or placebo group for 24 weeks. The primary endpoint was the mean change in glycated hemoglobin (HbA1c) from baseline to week 24. Results The mean baseline HbA1c levels were similar between the evogliptin and the placebo groups (7.20 ± 0.56 vs. 7.20 ± 0.63%, respectively). At week 24, evogliptin significantly reduced HbA1c levels from baseline compared to placebo (−0.23 vs. 0.05%, respectively, P < .0001). Additionally, the proportion of patients achieving HbA1c < 6.5% was significantly higher in the evogliptin group than that in the placebo group (33.3 vs. 15.2%, respectively, P = .008). The overall incidence of adverse events, including hypoglycemia, was similar between the two groups. Conclusions In this 24-week study, once daily evogliptin monotherapy significantly improved glycemic control and was well tolerated in patients with T2D.

In this study, we examined the prompted reflections of four middle school mathematics teachers after their lessons. We used Cohen and Ball``s instructional triangle (1999) to investigate teachers`` reflections. With this framework, we addressed questions of what characteristics in reflections the participant teachers have and how the reflections differ over time. Findings indicated that the teachers showed differences in the instances of assessing and changes over time in the ways they gained more insights about students`` understanding.

본 연구는 학생들의 수학에 대한 정의적 특성을 조사하고 수학 학습 주도권과 정의적 특성과는 어떤 관련이 있는지를 알아보기 위한 목적으로 진행되었다. 이를 위하여 인천 지역의 초등학교와 중학교 각 1개교의 학생들을 대상으로 설문조사 연구를 진행하였다. 수학에 대한 정의적 특성은 수학에 대한 흥미, 자신감, 가치...

Real time PCR is a standard method for identification of species. One of limitations of the qPCR is that there would be false-positive result due to mismatched hybridization between target sequence and probe depending on the annealing temperature in the PCR condition. As an alternative, fluorescence melting curve analysis (FMCA) could be applied for species identification . FMCA is based on a dual-labeled probe . Even with subtle difference of target sequence, there are visible melting temperature (Tm) shift. One of FMCA applications is distinguishing organisms distributed and consumed globally as popular food ingredients. Their prices are set by species or country of origin. However, counterfeiting or distributing them without any verification procedure are becoming social problems and threatening food safety. Besides distinguishing them in naked eye is very difficult and almost impossible in any processed form. Therefore, it is necessary to identify species in molecular level. In this research three species of squids which have 1-2 base pair differences each are selected as samples since they have the same issue. We designed a probe which perfectly matches with one species and the others mismatches 2 and 1 base pair respectively and labeled with fluorophore and quencher. In an experiment with a single probe, we successfully distinguished them by Tm shift depending on the difference of base pair. By combining FMCA and qPCR chip, smaller-scale assay with higher sensitivity and resolution could be possible, andc furthermore, enabling results analysis with smart phone would realize point-of-care testing (POCT).

The purpose of this study is to develop a framework for understanding teachers’ knowledge for teaching mathematics (MKT) by examining a hierarchy of 317 South Korean elementary teachers’ knowledge for teaching mathematics. Through a web-based survey, the teachers were asked to rank order their beliefs about and use of knowledge subdomains for teaching mathematics. Findings show that teachers’ beliefs about knowledge for teaching mathematics are divergent from current research trends. Also, their beliefs about the subdomains of knowledge for teaching mathematics were ranked differently in comparison to their actual use of the subdomains. Their hierarchy for MKT presents suggestions for the type of curriculum that might be offered to teachers to develop their knowledge at the pre-and in-service level.

Background Clonorchis sinensis causes a major food-borne helminthic infection. This species locates in mammalian hepatobiliary ducts, where oxidative stressors and hydrophobic substances are profuse. To adapt to the hostile micromilieu and to ensure its long-term survival, the parasite continuously produces a diverse repertoire of antioxidant enzymes including several species of glutathione transferases (GSTs). Helminth GSTs play pertinent roles during sequestration of harmful xenobiotics since most helminths lack the cytochrome P-450 detoxifying enzyme. Methods We isolated and analyzed the biochemical properties of two omega-class GSTs of C. sinensis (CsGSTo1 and CsGSTo2). We observed spatiotemporal expression patterns in accordance with the maturation of the worm’s reproductive system. Possible biological protective roles of CsGSTos in these organs under oxidative stress were investigated. Results The full-length cDNAs of CsGSTo1 and 2 constituted 965 bp and 1,061 bp with open reading frames of 737 bp (246 amino acids) and 669 bp (223 amino acids). They harbored characteristic N-terminal thioredoxin-like and C-terminal α-helical domains. A cysteine residue, which constituted omega-class specific active site, and the glutathione-binding amino acids, were recognized in appropriate positions. They shared 44 % sequence identity with each other and 14.8–44.8 % with orthologues/homologues from other organisms. Bacterially expressed recombinant proteins (rCsGSTo1 and 2) exhibited dehydroascorbate reductase (DHAR) and thioltransferase activities. DHAR activity was higher than thioltransferase activity. They showed weak canonical GST activity toward 1-chloro-2,4-dinitrobenzene. S-hexylglutathione potently and competitively inhibited the active-site at nanomolar concentrations (0.63 and 0.58 nM for rCsGSTo1 and 2). Interestingly, rCsGSTos exhibited high enzyme activity toward mu- and theta-class GST specific substrate, 4-nitrobenzyl chloride. Expression of CsGSTo transcripts and proteins increased beginning in 2-week-old juveniles and reached their highest levels in 4-week-old adults. The proteins were mainly expressed in the elements of the reproductive system, such as vitelline follicles, testes, seminal receptacle, sperm and eggs. Oxidative stressors induced upregulated expression of CsGSTos in these organs. Regardless of oxidative stresses, CsGSTos continued to be highly expressed in eggs. CsGSTo1 or 2 overexpressing bacteria demonstrated high resistance under oxidative killing. Conclusions CsGSTos might be critically involved in protection of the reproductive system during maturation of C. sinensis worms and in response to oxidative conditions, thereby contributing to maintenance of parasite fecundity. Electronic supplementary material The online version of this article (doi:10.1186/s13071-016-1622-2) contains supplementary material, which is available to authorized users.

A new concept for rapid, label-free cysteine sensing method is proposed via possible naked eye-detection of red-to-blue emission color change. Intermolecular exciton migration in conjugated polyelectrolyte-based assay complex is adopted to enhance selectivity and sensitivity for cysteine sensing by formation and dissociation of polymer-Hg(2+)-thymine assay complex in the absence and presence of cysteine, respectively. The assay complex shows red emission due to cooperative aggregation of conjugated polyelectrolyte, thymine, and Hg(2+). Upon exposure to cysteine, the assay complex dissociates into individual molecules showing transparent, blue-emitting solution, because cysteine extracts Hg(2+) from the assay complex via more favorable binding between cysteine and Hg(2+).

Teaching and learning in schools can be influenced by the ways in which curriculum is understood in terms of its characteristics and range. Viewing curriculum as encompassing all activities pertaining to teaching and learning allows us to examine various aspects of education through curriculum studies. In this respect, this study explores the trends in research on K-12 mathematics curriculum by examining prior studies` research topics categorized into five themes in curriculum studies. In addition, implications for future research are discussed based on the findings of this study.

The purpose of this study was to better understand teacher tensions in professional development. The population under study was practicing mathematics teachers engaged in a week-long professional development institute. Data sources included observations, interviews, and teacher products (such as registration forms, surveys, journals, and notebooks). The data were analyzed to determine what tensions were evident in the institute and how those tensions arose. In this paper, we examine participants’ tensions in a professional development situation when their expectations did not match reality. We have done so by presenting a theoretical model of professional development based on an instructional rhombus that illuminates the actual and applicatory aspects of teachers’ professional development experiences. The main tensions expressed by the teachers were related to the content and pedagogy of the institute. This study points toward the importance of teachers’ expectations in professional development.

Anionically charged fluorescent conjugated polyelectrolytes of poly{[4,7-(2,1,3-benzothiadiazole)-alt-1,4-phenyl- ene]-co-[2,5-bis(4-sulfonatobutoxy)-alt-1,4-phenylene]} (P1) and poly{[4,7-(bis(thiophen-2-yl)benzo-2,1,3-thiadiazole)-alt-1,4-phenylene]-co-[2,5-bis(4-sulfonatobutoxy)-alt-1,4-phenylenel]} (P2) were synthesized by Suzuki crosscoupling polymerization in the presence of a palladium catalyst. The conjugated polyelectrolytes with sulfonate groups, as efficient signal amplifying reporters, were carefully designed to be soluble in water over the entire pH range examined and interact with proteins through intermolecular forces. The polymers exhibited blue emission in aqueous solutions but green or red emission in solid form depending on the conjugation length due to intermolecular exciton migration. The anionic conjugated polymers exhibited blue-to-green or blue-to-red changes in fluorescence upon exposure to charged proteins, indicating that the polymers have potential applications in fluorescent array systems for protein.

A cholesteryl-based 2-(2′-hydroxyphenyl)benzoxazole (HPB) derivative 3 linked with an amide bond was prepared through an efficient synthetic pathway. The HPB, amide, and cholesteryl groups play important roles in constructing the supramolecular gel structure. UV–vis and fluorescence spectroscopy also showed that HPB and amide groups, which provide intra- and intermolecular hydrogen bonding, respectively, also contribute the recognition of fluoride anions.

A low molar mass organogelator 1 containing 2-(2'-hydroxyphenyl)benzoxazole (HPB) unit with long alkyl chain was synthesized by the reaction with HPB and octyl isocyanate in THF at room temperature. A new chelate-based organogelator 1-Zn(II) was prepared with the reaction of 1 and zinc(II) acetate in methanol and dichloromethane at mild condition. The gelation ability of organogelator was tested by heating-cooling method in various organic solvents, and the opaque gel was formed from DMF. Well-developed self-assembled structure of organogel was confirmed with field emission-scanning electron microscope (FE-SEM) and transmission electron microscope (TEM), and the optical properties of organogel upon aggregation were monitored by UV-Vis and fluorescence spectroscopy, 1-Zn(II) was self-assembled to the sheet-like structure with the thickness of fully extended length of molecules.

Electrical and thermal characteristics of a high-Tc superconducting (HTS) tape have close relation to a performance of a resistive type superconducting fault current limiter (SFCL). When an HTS coated conductor (CC) is applied to the SFCL, because the CC is a composite conductor, dimensions and material properties including electrical resistivity, thermal conductivity, and volumetric heat capacity of substrate, silver layer, YBCO, and stabilizer of the CC will affect to the fault current limiting performance and recovery time. This paper presents experimental results about fault current test of commercialized CCs to evaluate their compatibilities for resistive SFCLs and a numerical analysis using finite element method to design an HTS conductor for the SFCLs. The results from the analytical models of the CCs were compared to the experimental results. An HTS tape which is more suitable for the resistive type SFCLs was proposed as a conclusion.

The stabilizer of a YBCO coated conductor (CC) plays a very important role in bypassing the over-current and transferring the heat generated at the moment of a fault. Therefore, one of big issues is to determine the best material for the stabilizer and its dimensions for high performance in a HTS power application system. In the case of a superconducting fault current limiter (SFCL), which requires it to react immediately to any fault, the characteristics of the stabilizer are decisive in limiting the fault current and recovering the superconducting properties during and after quenching. This study examined the electrical and thermal behavior of YBCO CCs with various stabilizer thicknesses in fault mode to determine the effects of the stabilizer thickness on the quench/recovery characteristics of YBCO CCs. In particular, the electric field intensity (E) of YBCO CCs in fault mode were calculated and compared with the experimental results.

A new polybipyridine linked with oxadiazole and 2,5-didecyloxybenzene was prepared via simple condensation and ring closing reaction. The polymer P2 is basically an amorphous material with thermal decomposition temperature greater than 500°C. Introduction of didecyloxy group into the rigid oxadiazole backbone resulted in better solubility in common organic solvents such as THF, chloroform, and DMF. UV-vis and fluorescence spectroscopy were employed to investigate the optical properties both solution in the presence of a variety of metal cations, in which the polymer would be a good candidate for optical sensory material for metal ion.

A polyoxadiazole containing the 4-hydroxypyridine group, introduced to enhance the intermolecular interaction between polymer chains as well as for photosensitization, was synthesized from a corresponding precursor polyhydrazide. The absorption and emission spectra of the polymer film exhibited red-shifted maxima compared to those of the polyoxadiazole in chloroform solution, indicative of the presence of significant intermolecular interactions. The synthesized polymer showed a unique fluorescence enhancement upon UV irradiation in solution as well as in the film, presumably due to the photosensitized oxidation of hydroxypyridine groups to produce pyridone structures having different intermolecular interactions. Thus, this phenomenon enabled us to obtain a stable patterned image with enhanced fluorescence intensity in the UV-exposed area of the polymer film without any subsequent processes such as baking or etching.

For ordinal categorical tables, a weighted measure of association, , was proposed and its maximum likelihood estimator and asymptotic variance were drived. We redefined the last paradox of and proved its relation to marginal distributions. We also introduced the new paradox of and summaried the general relationships between and marginal distributions.

High temperature superconducting (HTS) magnets have been studied for insert coils of high field nuclear magnetic resonance (NMR) magnets but the temporal stability required for NMR is hard to achieve due to low index value and high joint resistance. In this research, the HTS power supply with magnets using coated conductor (CC) was investigated and tested in helium cryogenic system. All joints were conducted by soldering after etching stabilizer of the CC to minimize joint resistance. The pumping rate was determined by current amplitude and timing sequential control of heaters and the electromagnet. Operating characteristics were analyzed to enhance charging efficiency and the feasibility of temporally stable CC magnet during persistent mode was studied.

The YBCO coated conductor (CC) is one of the most promising superconductors for various superconducting applications. Although it has many advantages, joints of the YBCO CC are inevitable because there are technical difficulties in lengthening the YBCO CC. In the joining process, it is important to reduce the joint resistance between the YBCO CCs in order to operate in a persistent current mode. Several approaches have been proposed to eliminate the stabilizer of the YBCO CC. Among them, this paper introduces a chemical etching method that can reduce the damaging influences to the superconductor layer in the YBCO CC compared with physical or thermal methods, which may degrade the features of the superconductor. This paper discusses the characteristics of joined YBCO CCs in terms of the resistance and critical currents of the joints from the V - I curve.

In recent years, the YBCO coated conductor (CC) has been developed as a suitable material for power apparatuses. To fabricate a superconducting apparatus of a large capacity, the joint between the superconducting tapes are inevitable because of the need to have a long length conductor. Consequently, the joint characteristic is very important for the efficiency of the superconducting part in the apparatuses. Besides, maintaining the characteristic of the joint between the superconducting tapes even after repetitive over-current is also important for stability of power apparatuses. Therefore the joints between CC tapes were developed in differing joint length and the characteristics of the joints were compared. In this paper, we present the experimental results of several over-current experiments with the jointed CC tapes. The results of the over-current experiments and the trend of the joint characteristics are compared in point of stability of the joints. Resistance and critical current of the joints were measured from the V-I curve before and after several over-current experiments for observation of the trend of the joint characteristics. This study of repetitive over-current characteristics of the joints may be useful to the development of superconducting apparatuses in forms of magnets.

Continuous recording of daily symptoms constitutes an effective means of managing asthma patients. Daily management reduces the costs associated with hospitalization and improves the quality of patient care. We have implemented a Web-based mobile asthma management system. We used a pocket PC, mobile phone and desktop computer. The recorded items and individualized prescriptions were structured using Extensible Markup Language (XML) DTD (Data Type Definition). The mobile Web form was automatically adjusted to fit the different display resolutions of the terminal devices. The system provided reliable exchange of all relevant information between a doctor and the asthma patient using wireless mobile transmission. Email and Short Messaging Service (SMS) were used to send messages to patients, for example in the case of an automatically determined patient alert. Patients could obtain customized instructions according to their daily personal symptoms, peak expiratory flow (PEF), medications and activity restriction. The daily graph of PEF and the graphs of symptoms and medication were particularly useful for asthma patient control and self-awareness of the progress of the disease.

Angiomyolipoma is a common tumor of the kidney but has rarely been found in the mediastinum. We report a case of angiomyolipoma of the posterior mediastinum in a 62-year-old woman. She experienced exertional dyspnea and intermittent cough at admission. Computed tomography indicated a tumor located at the left paravertebral and upper posterior mediastinum and MRI imaging demonstrated a mass with low signal intensity in T1-weighted image at T4-5 level. Thoracotomy was done for surgical removal of the tumor and histologic examination revealed a mesenchymal tumor composed of mature fat, capillaries and smooth muscle fibers. The tumor was immunohistochemically positive for CD34 and factor-VIII (for vascular component) smooth muscle actin (for smooth muscle component) and S-100 protein (for fat component). There have been four case reports about mediastinal angiomyolipoma, namely three Japanese cases and one French case. It is suggested that angiomyolipoma could be considered for the differential diagnosis of mediastinal tumors.

Background : Increased aortic stiffness (AS) is an independent risk factor of cardiovascular disease in patients with hypertension. Elevation of blood pressure (BP) or sympathetic tone increases AS. It was reported that acute changes in heart rate by artificial pacemaker did not affect AS.However, it is unknown whether changes in HR by sympathomimetics can affect AS in patients with hypertension. We investigated the effect of changes of HR by isoproterenol (ISO) on AS in 17 hypertensives (mean age: 599 years). Methods : All vasoactive drugs were discontinued at least 3 days before the study. Carotid-to-femoral pulse wave velocity (PWV) was measured by foot-to-foot method. Pulse wave recordings were done at baseline and at every increase of HR by 5 to 10 bpm with gradual increase of dose of ISO. BPs were measured simultaneously by the fluid-filled manometer system through the side arm of arterial sheath of femoral artery. HR was measured by electrocardiography. Compliance (C) and compliance index (Ci) were calculated from PWV, pulse pressure, systolic and diastolic blood pressure by established formulas. For analysis, HR, PWV, C and Ci were converted as percent changes () from the baseline values. Percent changes of parameters of AS, i.e., PWV, C, and Ci, were grouped by every 10% increase in HR. Results : There was no significant difference among groups in PWV, C and Ci (p>0.05 for each). With simple regression analysis, PWV and C showed no significant correlation with HR (r=0.18, 0.13 respectively, p>0.05 for each). Ci was weakly correlated with HR (r=0.22, pCi could be referable to HR (=0.046). Conclusion : AS was not affected by changes in heart rate by isoproterenol, suggesting that it is not necessary to consider changes of HR when measuring aortic PWV.

BACKGROUND Despite increasing importance of Acinetobacter baumannii in nosocomial infections and rapid development of multi-antimicrobial resistance in this strain, the resistance mechanisms of beta-lactam antimicrobials in A. baumannii were not clearly defined. In order to observe the resistance mechanisms against beta-lactams and carbapenem, we characterized the production of beta-lactamases and outermembrane protein (OMP) profiles for the 44 clinical isolates of A. baumannii. METHODS The MICs of antimicrobials were determined by agar dilution test. The secondary beta-lactamases were characterized by isoelectric focusing, polymerase chain reactions and nucleotide sequencing, and the production of chromosomal beta-lactamases was quantitated by spectrophotometric method. For two strains with an elevated MIC of carbapenem, outermembrane protein (OMP) profile was analyzed by ultracentrifugation of the sonicated bacteral cells and SDS-PAGE. RESULTS AND CONCLUSION Twenty two or 4 of 44 strains produced TEM-1-like beta-lactamase or PER-1 extended-spectrum beta-lactamase, respectively. However, when we analyzed the MICs of several beta-lactams with the beta-lactamase production, the resistance level of beta-lactam was mainly determined by the production of chromosomal beta-lactamase, not by the secondary beta-lactamases in the clinical isolates of A. baumannii. In two strains with an elevated MIC of imipenem, a decrease or loss of about 35 kDa and 22 kDa proteins in OMP was observed, which suggested that the change of OMP played a role in carbapenem resistance.