Porcine Circovirus Type2 (PCV2), a single-stranded DNA virus associated with Postweaning multisystemic wasting syndrome(PMWS) of swine, has two major open reading frames, ORF1 and ORF2. The genomic size and molecular weight of ORF2 is respectively 699bp, 28kDa. ORF2 encodes the capsid protein (structural protein) that has type-specific epitopes and is very immunogenic and associated with the induction of neutralizing antibodies, suggesting its potential use in diagnostic assays as well as vaccine development. For efficient production of the capsid proteins, we expressed the PCV2 ORF2 gene with baculovirus in the insect cells. In this study, PCV2 ORF2 was appropriately ligated into the baculovirus transfer vector, pBacPAK9 and pB9-Acpol19-110-EK. Sf21 cells were transfected with a mixture of the purified recombinant transfer vector and bAcGOZA. We generated and purified recombinant viruses containing PCV2 ORF2, and named rAc-B9-PCV2ORF2 and rAc-B9-19-110-EK-PCV2ORF2, respectively. Expression levels of capsid fusion proteins with a partial polyhedrin region of AcNPV more increased than recombinant proteins from non-fusion expressed. Also, expression efficiency increased over time and differed at MOI. As a results, fusion expression of porcine circovirus type2 ORF2 using baculovirus could be utilized as an alternative expression method to produce recombinant antigen against PCV2 infection and is worthy of further investigation.
Aujeszky's disease (AD), also called pseudorabies, is an infectious viral disease caused by an alpha herpes virus and has domestic and wild pigs, as well as a wide range of domestic and wild animals, as the natural host. Aujeszky's disease virus (ADV) virions contain several envelope glycoproteins. Among them, glycoproteins gB, gC and gD are regarded as the major immunogenicity proteins and the antibodies induced by them can neutralize virus in vitro or in vivo. In this study, we investigated expression of these glycoproteins using the bacterial and baculovirus expressionn system. Successful expression of ADV glycoproteins in E. coli was confirmed by SDS-PAGE and Western blot analysis and their optimal expression condition was determined. However, the recombinant proteins generated in the bacterial expression system which lacks glycosylation process frequently lose their biological activity. We tried to express the ADV glycoproteins using the baculovirus expression vector system. The recombinant gB, gC and gD were detected at approximately 100, 60 and 50 kDa on SDS-PAGE and Western blotting, respectively. The optimal expression conditions were determined for MOI(multiplicity of infection) and post-infection days. One MOI and 4 or 5 days post-infection were the best conditions for the expression of the ADV glycoproteins in Sf21 cells. We are currently investigating the antigenicity of recombinant proteins using experimental animals.
Bacillus thuringiensis (Bt) is characterized by its ability to synthesize crystal toxins and also able to produce bacteriocins such as thuricin, tochicin, entomocin and bacthuricin. The present work, for the first time, describes the biological activity of bacteriocins from B. thuringiensis subsp. cameroun (Btc). Supernatant which was produced from a liquid culture of Btc had antimicrobial activity against Bacillus cereus, ending up to making a inhibition zone on an agar medium. A significant reduction in antimicrobial activity was observed when the supernatant was exposed to heat at 75~100°C for 15 min. Proteins were separated from the supernatant by a fast protein liquid chromatography (FPLC) given the thermal instability. A group of FPLC fractions had antimicrobial activity against Bt subsp. palmanyolensis, israelensis, 1-3, morrisoni, toguchini and kurstaki, and B. cereus ACTC21768, ATCC14579 and NRRLB-569. Interestingly, when the supernatant was individually incorporated into the liquid cultures of Bt subsp. israelensis (Bti) and mogi (Btm) with mosquitocidal activity, a vegetative cell growth was observed only in the Btm culture 10 h post-incubation. A possible recovery of vegetative Btm cell growth was observed, compared to a control without the supernatant. These results suggest that Btc produced proteinous antimicrobial substances, one of which may be used as a selection marker to separate Btm after possibly conjugating the two mosquitocidal strains.
Bacillus thuringiensis (Bt) strain K4 was isolated from fallen leaves which had been collected at a forest stand in Mungyeong city, Republic of Korea. The flagellated vegetative cells of Bt K4 were agglutinated with the H3 reference antiserum among 55 reference H-antisera. In a further test to identify subfactors, 3b and 3d monospecific antisera were reactive to the cells, followed up with introducing a novel serogroup of 3a3b3d, designated as serovar mogi. The strain K4 had mosquitocidal activity against Dipteran larvae, Anopheles sinensis and Culex pipiens pallens, with no Lepidopteran toxicity observed. The SDS-PAGE profile of K4 crystal protein, ovoidal-shaped, included several bands ranging from 30-75 kDa. Four putative peptides, Cry19Ba, Cry40ORF2, Cry27Aa and Cry20Aa were detected from the bands by a nano-LC-ESI-IT MS analysis. Through a thermal asymmetric interlaced PCR, cry19Ba, cry40ORF2 and cry27Aa genes were partially cloned from K4 strain. Three cry genes were further found in the strain by a 454 pyrosequencing, ending up to showing 58%, 39% and 84% homology in amino acids with Cry56Aa, Cry8Ba and Cry39ORF2 toxins, respectively. This novel 3a3b3d type strain, B. thuringiensis subsp. mogi, can be used as a good resource for studying unknown mosquitocidal cry genes.
From our previous studies in a farm in Songcheon, Andong where a series of crops was cultivated in 2008 and 2009, occurrence patterns of Riptortus pedestris (Fabricius) (Hemiptera: Alydidae) and its egg parasitism by Ooencyrtus nezarae Ishii (Hymenoptera: Encyrtidae) and Gryon japonicum (Ashmead) (Hymenoptera: Scelionidae) were reported. With an objective to verify the trend of the occurrence pattern, a year long monitoring of R. pedestris and its egg parasitism was continued in 2010 in the same location where barley, sesame, and soybean were cultivated in series. We placed four aggregation pheromone traps added with 50 refrigerated eggs of R. pedestris each along the perimeter of the field from 11 April to 31 October. Weekly occurrence patterns of R. pedestris and its parasitism in 2010 were generally similar to the patterns of previous years. Riptortus pedestris population first appeared in the last week of April, peaked after the third week of August, and declined in October. Parasitism by G. japonicum was recorded up to 64% in July, 2010 which was much higher than previous years. This study verifies that G. japonicum is the first colonizer that appears as early as May and remains active until September. Ooencyrtus nezarae, however, starts to occur late from the last week of August and exist in the field until October.
The four genetically distinct isolates have been identified previously from Bombyx mori nucleopolyhedroviruses (BmNPVs) isolated in Korea. To further understand the complex of viruses infecting Bombyx mori, the genome of BmNPV-K1 and K4 strains was completely sequenced and analyzed in comparison with the genome of other sequenced baculoviruses including previously reported BmNPV. BmNPV-K1 consisted of 127,542 bp and 133 open reading frames (ORFs) of 150 nucleotides or longer with minimal overlap have been identified. In contrast, BmNPV-K4 consisted of 128,615 bp and 134 open reading frames (ORFs). Although gene arrangement is virtually identical, the genome of BmNPV-K4 is 1,073 bp longer than BmNPV-K1. This was related to the more existence of bro genes in BmNPV-K4. To investigate the relationship between BmNPV-K1 and K4, phylogenetic analysis with each member of the paired ORFs was performed. The sequence data suggest that BmNPVK1 and BmNPV-K4 are closely related but have diverged and evolved into two separate strains. This was study to identify highly related but separately evolving viruses in the same insect host and geographic location. We are currently comparing the differences of these BmNPV genomes to elucidate characteristics of each virus.
Polyhedrin is the major component of the nuclear viral occlusions produced during replication of the baculovirus Autographa californica multicapsid nuclear polyhedrosis virus (AcMNPV). To enhance the expression level of baculovirus vector system, we constructed several fusion vectors using various fragments of the polyhedrin. The polyhedrin fragments were genetically fused to the enhanced green fluorescent protein (eGFP) under the control of polyhedrin promoter, and their expressions were analyzed in Sf21 insect cells. Expression of the fusion protein was identified by SDS-PAGE and Western blot analysis using anti-GFP and anti-Polyhedrin. The expression level of eGFP was markedly increased by the fusion of partial polyhedrin. Also, the fluorescence intensity of fusion proteins was higher than that of non-fusion protein. Confocal laser scanning microscopy demonstrated that fusion proteins were localized to the cytosol or nucleus of insect cells. In additional, the glycoprotein E2 (gE2) of classical swine fever virus (CSFV) expressed by the these vectors was dramatically increased and its immunogenicity was proofed using experimental animal guinea pigs that were immunized with the partial polyhedrin containing gE2. This study provides a new option for the higher expression of useful foreign recombinant protein by using the partial polyhedrin in BEVS.
Perovskite manganites such as RE1-xAxMnO3 (RE = rare earth, A = Ca, Sr, Ba) have been the subject of intense research in the last few years, ever since the discovery that these systems demonstrate colossal magnetoresistance (CMR). The CMR is usually explained with the double-exchange (DE) mechanism, and CMR materials have potential applications for magnetic switching, recording devices, and more. However, the intrinsic CMR effect is usually found under the conditions of a magnetic field of several Teslas and a narrow temperature range near the Curie temperature (Tc). This magnetic field and temperature range make practical applications impossible. Recently, another type of MR, called the low-field magnetoresistance(LFMR), has also been a research focus. This MR is typically found in polycrystalline half-metallic ferromagnets, and is associated with the spin-dependent charge transport across grain boundaries. Composites with compositions La0.7(Ca1-xSrx)0.3MnO3)]0.99/(BaTiO3)0.01 [(LCSMO)0.99/(BTO)0.01]were prepared with different Sr doping levels x by a standard ceramic technique, and their electrical transport and magnetoresistance (MR) properties were investigated. The structure and morphology of the composites were studied by X-ray diffraction (XRD) and scanning electronic microscopy (SEM). BTO peaks could not be found in the XRD pattern because the amount of BTO in the composites was too small. As the content of x decreased, the crystal structure changed from orthorhombic to rhombohedral. This change can be explained by the fact that the crystal structure of pure LCMO is orthorhombic and the crystal structure of pure LSMO is rhombohedral. The SEM results indicate that LCSMO and BTO coexist in the composites and BTO mostly segregates at the grain boundaries of LCSMO, which are in accordance with the results of the magnetic measurements. The resistivity of all the composites was measured in the range of 90-400K at 0T, 0.5T magnetic field. The result indicates that the MR of the composites increases systematically as the Ca concentration increases, although the transition temperature Tc shifts to a lower range.
The purpose of this study was to investigate the effects of meditation music with lumbar stabilization exercise on balance, concentration and muscle activation. Participants are divided into Group A and Group B. Group A listened to meditation music while doing 4 sets of lumbar stabilization exercise. Group B did not listen to meditation music while doing 4 sets of lumbar stabilization exercise. Surface EMG data was obtained rectus abdominis, external oblique, erector spinae, multifidus muscle during lumbar stabilization exercise. To examine the effect of before-after experiment for concentration and balance, paired t-test was adopted. Changes in EMG data of each muscles were analyzed by independence t-test. There were statistically significant increasing at concentration level and balance level in Group A. But significant difference was not shown in muscle activation. The clinical effect of the meditation music with lumbar stabilization exercise was investigated in this study with more excellent results in concentration and balance. In conclusion, these results suggest that meditation music and lumbar stabilization exercise may be useful in elderly people.
The purpose of this study is to investigate and reveal the effects that the complex exercise training consisting of aerobic exercise and strength training(sit up, push up) that everyone can easily practice regardless of a time and a place in order to manage practically the physical strength of the aged affects the difference on their body composition and the change of physical fitness level. Looking into the change of body composition of an experimental group, the weight of 2.5kg was reduced after applying complex training for 12 weeks and the body fat mass of 2.65kg was reduced. Also, the abdominal fat of 0.13% was decreased and the muscle mass of 1.56kg was increased. For the change factors of physical fitness, cardiovascular endurance, muscular strength, muscular endurance, balance and flexibility excluding agility showed significant improvement after applying complex exercise training. The improvement of health fitness of the aged under this study was significantly effective to improve specified body functions which had been lowered by aging and insufficient physical activities. So, it is regarded that their health fitness is the important factor to improve the activity competence required for daily life and to lead healthy living by the improved activity competence. Henceforth, it needs to study more the complex composition of several sports, exercise intensity and the frequency based on the previous researches and studies. In addition, it needs to develop the complex exercise training in accordance with various characteristics such as a sex of the aged, an age, a physical fitness level, environment, a disease and the program in consideration of the efficacy and safety during training.
This article aims to provide guiding principles for English language teacher evaluation through the review of recent accounts of research and practice. Teacher effectiveness needs to be contextually defined and measured. It is suggested that education providers need to work with all stakeholders in their systems both to define criteria for effective teaching and the means by which it may be assessed. As teacher evaluation is multifaceted, a range of data needs to be collected encompassing the views of all the stakeholders as well as the reflections of teachers themselves. Teacher evaluation should not be regarded as a summative product, the result of single evaluation measures conducted upon teachers, but should be thought of as a formative process in which teachers are actively engaged. Regarding evaluator training, it should not be assumed that those charged with evaluating teachers are, because of their positions, adequately trained to perform the evaluations. Evaluator training should be carried out before any evaluation is conducted under a new system. In order for the evaluation system to be meaningful and contribute to improvements in the education system as a whole, it should be seen primarily as a means of professional development for teachers as well as for anyone else involved in the process.
Activated carbon fibers (ACFs) were prepared from cost effective commercial textiles through stabilization, carbonization, and subsequently activation by carbon dioxide. ACFs were characterized for surface area and pore size distribution by physical adsorption of nitrogen at 77 K. ACFs were also examined for various surface characteristics by scanning electron microscopy, Fourier transform infrared spectroscopy, and CHNO elemental analyzer. The prepared ACFs exhibited good surface textural properties with well developed micro porous structure. With improvement in physical strength, the commercial textile grade acrylic precursor based ACFs developed in this study may have great utility as cost effective adsorbents in environmental remediation applications.
To understand the marine algal flora on Goheung coast, Korea, marine algae at 8 points were collected from November 2008 to February 2009 and from April to June 2009. Thirty-seven species (2 species of angiosperms, 5 chlorophytes, 12 phaeophytes, and 18 r
Gudul is a traditional heating system which was created by our ancestors. This study is focused to the developments of new flooring material through the physical and chemical analyses of the Gudul rocks. The study proceeded with selecting the theme and study areas, sampling, geological survey, several analyses of the rocks, development of new floor material (a kind of porcelain). Gudul rocks are sampled from a total of twenty one cities and counties are assigned to nine tuffs, five quartz porphyries, four gneisses and three schists in rock type. From the geological study, these rocks are very similar to ones in surrounding areas. Comparing Gudul rocks to others not used in Ondol, Gudul rocks show low densities, higher porosities and brittle features. Analyzed in thermal properties, the rocks are lower in conductivity but higher in conservation and resistance in the aspects of the properties. Considering mutual relations between thermal properties and chemistries of the rocks, conservative characteristics are proportional to MgO contents but not to SiO2. In particular, minute quartz grains in the rocks were gradually cracked and rejected under torch flames. Those features may indicate that quartz grains are closely related with the thermal resistances of the rocks. On the basis of above rock characteristics, new flooring material was made. The material includes the mixed states of amphibolite sludges with high MgO contents and white china soils (bearing kaolinte). Then, the material was tested in the above thermal properties under the variable mixed ratios. At the results, 4:6 (sludge : soil) ratio was the most excellent among above tested materials. So, the textiles used in architecture and the electric heating cushion and the mugcups were produced as test goods.
The present study was to investigate the source of contamination during semen processing for in vitro uses. In the present study, frozen semen was prepared from liquid semen in our laboratory for in vitro fertilization (IVF) experiments due to lack of fresh semen. Antibiotics were added in the frozen semen extender (kanamycin and gentamicin) and in vitro culture (IVC) medium (gentamicin) for further inhibiting growth of microorganisms. Nevertheless, proliferations of microorganisms were observed in IVC culture drop during culturing of IVF embryos using frozen semen. Randomly 3 samples were taken from the liquid semen, frozen semen and egg yolk. Contaminated IVC medium, frozen-thawed semen, liquid semen and egg yolk were cultured in de Man, Rogosa and Sharpe (MRS) agar medium. Whitish colonies were detected in contaminated IVC drop, frozen-thawed semen samples and egg yolk but no colonies were formed in liquid semen samples. Gram-negative and rod-shaped identical bacteria were found in both frozen-thawed semen sample and contaminated IVC drop and egg yolk samples. Enterobacter cloacae were confirmed by API 20E kit according to manufacturer's instruction with identification value (% ID) 94.3% and T index 0.88. Antibiotic susceptibility tests were done according to Clinical and Laboratory Standards Institute (CLSI) by using ampicillin, amikacin, cephalothin, gentamicin, kanamycin, tetracycline, oxytetracycline, sulfamethoxazole trimethoprim, norfloxacin and ciprofloxacin test. Among them Enterobacter cloacae were resistant to ampicillin, amikacin, cephalothin, gentamicin, kanamycin but susceptible to tetracycline, oxytetracycline, sulfamethoxazole trimethoprim, norfloxacin and ciprofloxacin. From these findings it could be suggested that this contamination sources might be from egg yolk.
The Trichoderma disease, commonly referred to as green mold, has been previously considered as a problem in mushroom production, because it typically occurred in association with low-quality compost or poor hygiene. We studied growth of Trichoderma spp. (Trichoderma aggressivum, Trichoderma atroviride, Trichoderma harzianum, Trichoderma koningii, Trichoderma viride) is under the control of environment factors ; 15℃ - 35 ℃ of incubation temperature, pH 3 - pH 11 of medium and 55% - 75% moisture content of rice straw medium. At the temperature of incubation, Trichoderma spp. had the highest growth rate at 25 ℃ and had no growth at 35 ℃. At the pH of medium, Trichoderma spp. had the highest growth rate at pH 5 and had the lowest growth rate at pH 11. In the Moisture content of rice straw medium, Trichoderma spp. had the highest growth at 75% moisture content and had the lowest growth rate at 55% moisture content.
Essential oil from Chamaecyparis obtusa leaves and water extract from Pseudotsuga menziesii sawdust were examined for antifungal activity against Trichoderma spp. (Trichoderma aggressivum, Trichoderma atroviride, Trichoderm harzianum. Trichoderma koningii, Trichoder-ma viride). Generally, essential oil and water extract inhibited the growth of Trichoderma spp. by about 25% and 33%, respectively, at 1000 ppm. Antifungal activity of water extract was slightly higher than essential oil at 1000 ppm. Antifungal activity was increased with increasing concentration of essential oil and water extract. This study was carried out the investigation of the chemical composition of essential oil from Chamaecyparis obtusa leaves and water extract from Pseudotsuga menziesii sawdust. The essential oil was analyzed by GC-MS and water extract analyzed by GC-MS/HS. Thirty-eight components were identified, of which terpinyl acetate (13.41%), elemol (11.86%) and isobornyl acetate (7.89%) were the main compounds in essential oil. Twenty-four components were identified, of which 2-Isopropoxy-ethylamine (46.45%), Epifluorohydrin (8.62%) and Trans-2,3-Dimethyloxirane (7.78%) were the main compounds in water extract. Based on all results described above, we conclude that the essential oil and water extact may have a potent in vitro antifungal activity against Trichoderma spp.
Phenotypic traits (physiological characteristics and somatic incompatibility) and genotypic traits (target region amplification polymorphism TRAP) were used to survey the diversity in Chinese Auricularia auricula systematically, which consisted of 32 main cultivated strains. The 27 important and stable physiological indexes were evaluated; somatic incompatibility test (SIT) reaction was described from three aspects: type, pigment, and intensity while intensity clustering alone revealed the phenotypic diversity among the 32 strains;16 pairs of TRAP primer combinations produced 535 unambiguous and reproducible DNA fragments, of these 524 (97.9%) were polymorphic. The phylogenetic trees were constructed by Unweighted Pair-group Method with Arithmetic Averages (UPGMA), which distributed the test strains into four to six major groups respectively. The principal coordinate analysis (PCO) of the three methods (physiological characteristics, SIT intensity and TRAP) exhibited highly similar clustering patterns, revealing that all the test strains can be divided into three groups considered significantly correlated with geographical regions. Results revealed the occurrence of relatively low diversity of A. auricula in the study. The cultivated strains in the same region are more similar physiologically and some strains are suspected to be synonymous. This study suggests that the value for estimating diversity are represented by TRAP>SIT reaction intensity>physiological characteristics in A. auricula, and PCO analysis can provided more effective and visible information than the UPGMA clustering dendrogram. Our finding will facilitate future germplasm resources management and breeding program of A. auricula in China.