G protein-coupled receptors (GPCRs) belong to cell membrane protein family, which regulate various physiological process such as reproduction, behavior and immune etc. In other to identify the GPCRs in pheromone gland of Maruca vitrata, we carried out transcriptome analysis from both females. Transcriptome analysis in the pheromone gland yielded approximately 22Gb and 47,528 transcripts showed positive FPKM value. 48 Genes involved in GPCRs were identified such as pheromone biosynthesis activating neuropeptide receptor (PBANr), prostaglandin receptors, neuropeptide receptor, 5-hydroxytryptamine receptor, galanin receptor, calcitonin gene-related peptide receptor, diuretic hormone receptor, gonadotropin-releasing hormone receptor, frizzled and orphan receptors, etc. Various expression of GPCRs in the pheromone gland indicates the role of pheromone gland may not be limited to the production of pheromone.

The aim of the study was to investigate the effects of incentive spirometry and Ujjayi breathing technique on the pulmonary function of smokers. Subjects were individuals who had a smoking habit of at least a year. Subjects were randomly divided into 3 groups: the incentive spirometry group (n=8), Ujjayi breathing technique (n=9), and a group applying both incentive spirometry as well as Ujjayi breathing technique (n=8). Each intervention was performed twice a day, 5 times a week, for a total of 8 weeks whereupon the change in pulmonary function was evaluated. A spirometer was used to measure FVC, FEV1, and FEV1/FVC. The survey used for this study included the Fagerström Test of Nicotine Dependence (FTND) and the Shortness of Breath Questionnaire (SOBQ). Study results for the comparison within groups showed that in the group that performed both the incentive spirometry and Ujjayi breathing technique, FEV1 improved with statistical significance (p<.05). Furthermore, within this comparison the FEV1/FVC improved with statistical significance. Comparison amongst the groups showed no statistically significant differences in all areas. Following, to effectively increase pulmonary function in young adult smokers, both incentive spirometry and Ujjayi breathing technique should be employed together.

The purpose of this study is to improve practical skill evaluation method of mushroom - trained certified technician's practical skill evaluation which is one of national qualification tests based on national incompetency standards. One of the current National Competency Standards (NCS), mushroom - trained certified technician uses NCS based practical assessment method. In order to improve the current practical evaluation method, we try to improve practical evaluation method based on field customized problem solving ability and improve the practical evaluation method, various evaluation methods should be constructed. On the purpose of identify the diversity and problems of the evaluation method, the experts of the group consultation, the mushroom-related research institute and the related industry collaborated to identify the problems of the actual mushroom - trained certified technician practicum test, This study on the evaluation improvement method was carried out. In this study, the contents of practical test of the current mushroom traits were analyzed and the trends of the latest mushroom industry were widely reflected.

An experiment was conducted to study the potential of thermal treated oak sawdust(steaming and steam explosion) as horticultural medium component in plug seedlings production of Chinese cabbage(Brassica campestris L.). This study involves the chemical, physical characterization and growth test of thermal treated oak sawdust(steaming and steam explosion) in order to evaluate their use as components of horticultural media. A commercial peat moss and oak sawdust were used as control. The total carbohydrate, C/N ratio, pH, phenolic compound, total porosity and water holding capacity were 45.1g/100g dry wt, 425.1, 4.4, 141.8mg/g wt, 82.5%, 47.1% in oak sawdust and 39.2g/100g dry wt, 300.3, 4.7, 131.7mg/g wt, 84.9% and 49.2% in steamed oak sawdust and 30.3g/100g dry wt, 247.8, 5.7, 40.8mg/g wt, 92.3% and 51.7% in steam exploded oak sawdust, respectively. The mixtures of the horticultural media were prepared using different substrate as peat moss, oak sawdust, steamed oak sawdust, steam exploded oak sawdust and perlite to grow Chinese cabbage in a greenhouse. The seed germination, stem height and leaf area were 68%, 2.2cm, 1.1cm2 in OSP(containing 90% oak sawdust and 10% perlite) and 69%, 2.5cm, 1.5cm2 in SMP(containing 90% steamed oak sawdust and 10% perlite) and 87%, 3.0cm, 2.2cm2 in SEP(containing 90% steam exploded oak sawdust and 10% perlite), respectively. The leaf area SEP(containing 90% steam exploded oak sawdust and 10% perlite) was higher than that of PP(containing 90% peat moss and 10% perlite). This research indicates that steam exploded oak sawdust may be utilized as a suitable replacement for peat moss in horticultural media component for Chinese cabbage.

The Riptortus (stinkbug) has a specialized symbiotic organ, M4 midgut, to harboring symbiont Burkholderia. M4 midgut is located in abdomen and surrounded with insect hemolymph. Recently our group demonstrated that symbiotic Burkholderia showed different physiology after adapting in M4 gut compare with in vitro cultured Burkholderia. And population of symbiotic Burkholderia in the M4 midgut is regulated by special organ. However, the molecular mechanism to prevent spreading and migrating symbiont bacteria to other host tissues from symbiotic organ is not clear. Therefore, we assumed that symbiont Burkholderia are susceptible to host humoral immunity after established infection in M4 midgut to prevent spreading and migrating into the other host tissues through Riptortus hemolymph. To prove this assuming, we tested the susceptibility and survival rate of symbiont Burkholderia in hemolymph of Riptortus in vitro and in vivo. We also examined the susceptibility of symbiont Burkholderia using purified antimicrobial peptides (AMP), pyrrhocoricin-like, thanatin-like and defensin-like AMPs. Finally, we tested inducing ability for AMPs by systemic infection of symbiotic Burkholderia. Gene expression of purified AMPs was not different after systemic infection of both symbiont and in vitro cultured Burkholderia. Surprisingly, in vitro cultured Burkholderia resisted on bacteria injected hemolymph and purified AMPs but symbiont Burkholderia were highly susceptible in bacteria injected hemolymph and purified AMP. These results suggest that symbiont Burkholderia can't survive in the hemolymph after escaping symbiotic organ. Moreover, humoral immunity of host Riptortus is important to prevent spreading and migrating symbiont Burkholderia into the other host tissue or organ from symbiotic organ.

The fact that flip-flops, one of many different types of unstable shoes, are light and relatively easy to put on, accounts for their popularity among people. But because flip-flops rely heavily on the support of a single thong between your first and second toes, they impose a huge amount of pressure onto lower leg. Thus in the following experiment we tried to examine the different effects of flip-flops and running shoes in terms of their effect on muscle activity and fatigue of tibialis anterior and gastrocnemius during walking. In order to measure an electromyogram we used Free EMG system. 10 men and 10 women in running shoes ran on treadmills for 15 minutes at 4.8km/h, 2 days later the same experiment was carried out, but this time, in flip-flops. p value turned out to be greater than .05 and thus there was no considerable difference between the effects of flip-flops and running shoes on muscle activity and fatigue during walking. Therefore we conclude that despite the fact that flip-flops are considered unstable, their effects on muscle activity and fatigue of tibialis anterior and gastrocnemius are negligible.

Most traditional genome sequencing projects involving infectious viruses include culturing and purification of the virus. This can present difficulties as an analysis of multiple populations from multiple locations may be required to acquire sufficient amount of high-quality DNA for sequence analysis. The electrophoretic method provides a strategy whereby the genomic DNA sequences of the Korean isolate of Pieris rapae granulovirus (PiraGV-K) were analyzed by purifying it from host DNA by pulsed-field gel electrophoresis, thus simplifying sampling and labor time. The genomic DNA of infected P. rapae was embedded in agarose plugs, digested with a restriction nuclease and methylase, and pulsed-field gel electrophoresis (PFGE) was used to separate PiraGV-K DNA from the DNA of P. rapae, followed by mapping of fosmid clones of the separated viral DNA. The double-stranded circular genome of PiraGV-K encodes 120 open reading frames (ORFs), covering 92% of the sequenced genome. BLAST and ORF arrangement showed the presence of 78 homologs to other genes in the database. The mean overall amino acid identity of PiraGV-K ORFs was highest with the Chinese isolate of PiraGV (~99%), followed up with Choristoneura occidentalis ORFs at 58%. PiraGV-K ORFs were grouped, according to function, into 10 genes involved in transcription, 11 involved in replication, 25 structural protein genes, and 15 auxiliary genes. Genes for Chitinase (ORF 10) and cathepsin (ORF11), involved in the liquefaction of the host, were found in the genome. The recovery of PiraGV-K DNA genome by pulse-field electrophoretic separation from host genomic DNA had several advantages, compared with its isolation from particles harvested as virions or inclusions from the P. rapae host. We have sequenced and analyzed the 108,658 bp PiraGV-K genome purified by the pulsed field electrophoretic method. The method appears to be applicable to the analysis of genomes of large viruses. The chitinase, identified by PiraGV-K genome sequence, was functionally characterized by quantitative PCR, Western blot analysis, immunohistochemistry and transmission electron microscopy.

The purpose of this study is to analysis of muscle fatigue in the upper trapezius and splenius capitis muscles according to therapy table height variation. The subjects were consisted of 15 healthy adults(10 males, 5 females) who had no medical history of neurological and musculoskeletal problems. In experiment, wireless electrode EMG system was measured for each the upper trapezius and splenius capitis muscles during the treatment performed on table. the differences in the muscle fatigue was compared for 4 types of table height(-6cm, -3cm, 0, +3cm from elbow in 90° flexion position). Muscle fatigue according to therapy table height were significant difference except for left upper trapezius. And muscle fatigue of right upper trapezius and splenius capitis showed significant decrease in +3cm table height compared to -6cm table height(p<.05). Muscle fatigue of right upper trapezius and splenius capitis were the highest in -6cm table height, but those were the lowest in +3cm table height. This study propose to change therapy table height higher than +3cm from elbow in 90° flexion position, if you hope to reduce muscle fatigue.

Among hemipteran insects which is the most important insect vector of plant viruses, small brown planthopper, Laodelphax striatellus, transmits the rice stripe virus (RSV) causing rice stripe disease. For effective control of RSV, it is important to understand interaction between RSV and L. striatellus. Therefore, in this study, expressed sequence tag (EST) databases were generated based on 454 GS-FLX pyrosequencing for comparative transcriptome analysis between nonviruliferous and RSV-viruliferous L. striatellus. By comparing the two EST libraries, we showed that 108 host genes were significantly up-regulated and 28 host genes were significantly down-regulated in viruliferous insects. Interestingly, genes encoding ribosomal proteins were mainly up-regulated in viruliferous L. striatellus, whereas genes related to translation were concentrated in the downregulated cohort. These RSV-dependently regulated genes may have important function in the behavior of planthopper or the transmission of RSV.

Bacillus thuringiensis serovar mogi of a novel serogroup (H3a3b3d), which showed mosquitocidal activity against Anopheles sinensis and Culex pipiens pallens, was isolated from fallen leaves in Mungyeong city, Republic of Korea. In contrast to the complicated plasmid profiles of B. thuringiensis H3 serotype strains, the B. thuringiensis serovar mogi contained only megaplasmid (> 30 MDa) on which the toxin genes were occasionally located. Sequence analysis using 454-pyrosequencing revealed that the megaplasmid harbored at least seven putative cry genes, showing about 84%, 75%, 73%, 58%, 84%, 39% and 75% similarities in amino acid sequences with Cry27Aa, Cry19Ba, Cry20-like, Cry56Aa, Cry39ORF2, Cry8Ba and Cry40ORF2, respectively. These cry genes were cloned to the Escherichia coli-B. thuringiensis shuttle vector, pHT1K, and then introduced into an acrystalliferous B. thuringiensis Cry-B strain for further molecular characterization. To investigate the role of these genes in crystal production, the expression profiles of these toxin genes were analyzed by quantitative real-time PCR (qrtPCR) from the wild type strain as well as transformant strains. The results clearly indicate that the cry39orf2 was the dominant ingredient in the crystal. This novel 3a3b3d type strain, B. thuringiensis serovar mogi, could be used as a good resource for studying unknown mosquitocidal cry genes.

The baculovirus expression system is one of the most popular methods used for the production of recombinant proteins but has several complex steps which have proved inherently difficult to meet a multi-parellel process. We have developed a novel recombinant bacmid, bEasyBm that enabling easy and fast generation of pure recombinant virus without any purification step. In the bEasyBm, attR recombination sites were introduced to facilitate the generation of recombinant viral genome by in vitro transposition. Moreover, extracellular RNase gene from bacillus amyloliquefaciens, barnase, was expressed under the control of Cotesia plutellae bracovirus early promoter. Therefore, only when the barnase gene was replaced to gene of interest, the bEasyBm could replicate in host insect cells. When the bEasyBm was transposed with pDualBac-EGFP and pDualBac-LUC respectively, there were no non-recombinant backgrounds were detected from unpurified BmEasy-EGFP or BmEasy-LUC stocks. In addition, the resulting recombinant virus, BmEasy-EGFP, showed comparable level of EGFP expression efficiency with the plaque-purified recombinant virus, BmEGFP, which was constructed using bBmGOZA system. Based on these results, high-throughput condition for generation of multiple recombinant viruses in a time was established.

Varieties of Bacillus thuringiensis (Bt) crystal proteins, Cry proteins, have so far been found as one of the most successful biological control agents which are safe to natural environments for a long time. Recently, cry genes encoding these Cry proteins have been widely applied for construction of transgenic crops resistant to pest insects. In this study, through the 3D structure prediction and accompanying mutagenesis study for the Mod-Cry1Ac, 7 and 16 amino acid residues from domain I and II, respectively, responsible for its insecticidal activity against larvae of Spodoptera exigua and Ostrinia furnacalis were identified. To construct novel cry genes with improved insecticidal activity, we randomly mutated these 23 amino acid sequences by in vitro muti site-directed mutagenesis, resulting in totally 24 mutant cry genes. For further characterization, these mutant cry genes were expressed as a fusion protein with polyhedrin using baculovirus expression system. SDS-PAGE analysis of the recombinant polyhedra revealed that expressed Cry proteins was occluded into polyhedra and activated stably to 65 kDa by trypsin. In the further study, we plan to investigate their insecticidal activity against Plutella xylostella, S. exigua and O. furnacalis larvae.

ORF11 (ac11) of Autographa californica multiple nucleopolyhedrovirus (AcMNPV) is a highly conserved baculovirus gene whose homologs are found in all lepidoteran Group I NPV, but its function is unknown so far. To determine the role of ac11 in baculovirus life cycle, ac11 knock-out mutant, Ac11KO, was constructed using the plasmid capture system (PCS). Real-Time PCR analysis showed that ac11 transcript was first detected at 6 h post-infection (p.i.) and accumulated to maximum at 48 h p.i., indicating that ac11 is belong to late gene. When the genomic DNA of Ac11KO was transfected into Sf9 cells, viral replication was restricted to a cell transfected originally. While viral transmission of the Ac11KO was not observed in Sf9 cells, production of budded virus (BV) in Sf9 cells transfected with Ac11KO was observed by transmission electron microscopy (TEM). These results suggest that the ac11 is essential for AcMNPV to produce infective BV.