We sequenced 17,329 bp of mitochondrial genome (mitogenome) of the black dwarf honey bee, Apis andreniformis (Hymenoptera: Apidae), that lacks ~200 bp of the A+T-rich region for the completion of the genomic sequence. The gene arrangement of A. andreniformis mitogenome is identical to that of A. cerana. However, the genome contains 5 additional tRNALeu(CUN) located 4 copies between tRNAMet and tRNAGln, and 1 copy between tRNAGln and tRNAAla, along with the typical sets of genes (13 protein-coding genes, 22 tRNAs, and 2 rRNAs) including regular tRNALeu(CUN) and the A+T-rich region (at least 923 bp). Only 1 copy of tRNALeu(CUN) differed by 1 bp from other 4 copies of tRNALeu(CUN). Each additional tRNALeu(CUN) is followed by nearly identical 68-bp long repeat sequence (95.6% identity). All 13 protein coding genes have typical start codons found in insect mitochondrial PCGs (2 ATA, 9 ATT, and 2 ATG).

Cuticular proteins (CPs) and the polysaccharide chitin are the major components of the exo- and endocuticular layers or procuticle. CPs contain a conserved sequence known as the Rebers & Riddiford (R&R) motif, which may function as a chitin-binding domain that helps to coordinate the interaction between chitin fibers and the protein network. We identified two highly abundant RR-2 CPs, TcCPR18 and TcCPR27, in protein samples extracted from elytra (rigid cuticle) of Tribolium castaneum adults and determined that these two CPs are required for rigid cuticle morphology. In this study, we identified the third most abundant protein (TcCP30) extracted from the elytra, and cloned a full-length cDNA. It encodes a very unusual 171 amino acid residue protein of which 36% of the residues of the mature protein are Glu, 21% are His, 19% are Arg, and 16% are Gly, organized in a regular pattern but not R&R consensus motif. TcCPR18 and TcCPR27 genes are expressed at 4 d-old pupae, while TcCP30 is highly expressed at 5 d-old pupae (last pupal stage) and 0 d-old adults. Immunohistochemical studies revealed the presence of TcCP30 in rigid adult cuticle (e.g. elytron, pronotum and ventral abdomen) but not soft cuticle (e.g. hindwing and dorsal abdomen). Injection of dsRNA for TcCP30 into late instar larvae had no affect on larval and pupal growth and development. The subsequent pupal-adult molt, however, more than 50% adults were unable to shed their exuvium and died. In addition, the resulting adults exhibited wrinkled, warped and split elytra. TcCP30-deficient adults could not fold their hindwings properly. These results indicate that TcCP30 may play critical roles in rigid adult cuticle formation, development and insect growth and survival. This work was supported by NRF (NRF-2012R1A2A1A01006467).

Insect chitinases (CHTs), which belong to family 18 glycosylhydrolases (GH-18), have been detected in molting fluid and gut tissues and are predicted to mediate the digestion of chitin present in the exoskeleton and peritrophic matrix (PM) in the gut. Based on amino acid sequence similarity and phylogenetic analysis, insect CHT family proteins have been classified into eight groups (group I to VIII). The CHTs belonging to different groups have distinctly different developmental patterns of expression and tissue specificity, suggestive of distinct biological functions. CHT7s belong to Group III chitinase contain two catalytic domains and one chitin binding domain (CBD). The catalytic domain 1 of this group of chitinases exhibits greater sequence similarity to one another than to the catalytic domain 2 in the same protein(s), suggesting distinct functions and/or evolutionary origins for each of these two catalytic domains. This group of chitinases, unlike most insect CHTs, possesses a predicted transmembrane segment at the N-terminal region. The recombinant T. castaneum CHT7 that was expressed in Hi-5 insect cells was bound to the cell membrane. Apparently, the catalytic domains of this CHT face the extracellular space as revealed by its ability to hydrolyze an artificial chitin substrate added to the medium. DsRNA-based functional studies (RNAi) for several CHT genes in Tribolium castaneum indicated that CHTs belong to groups I (TcCHT5) and II (TcCHT10) are critical for molting and turnover of chitin in the old cuticle. In other hand, RNAi for TcCHT7 did not affect any types of molting such as larval-larval, larval-pupal and pupal-adult. The resulting pupae or adults, however, failed to wing-expansion and abdominal contraction. Immunohistochemical analysis revealed that TcCHT7 protein is localized in newly synthesized procuticle, suggesting that TcCHT7 could be released form the plasma membrane of epidermal cells by proteolysis. Chitin seems to accumulate within the assembly zone of the elytral and body wall cuticle in dsTcCHT7-treated animals. Transmission electron microscopy revealed that down-regulation of TcCHT7 transcripts resulted in disorganization of chitin laminar and vertical canals in the procuticle. These results suggest that TcCHT7 may have critical roles in the laminar assembly and synthesis and/or deposition of cuticular chitin. This work was supported by NRF (NRF-2012R1A2A1A01006467).

Insect cuticle/exoskeleton is a first physical barrier to protect their body from multifarious environments such as desiccation, natural enemies and entomopathogenic microorganisms. Cuticle tanning (sclerotization and pigmentation) is a vital procedure for generating suitable cuticle depending on body region by sclerotization and pigmentation in insects. Insect cuticle tanning is a complex process involves hydroxylation of tyrosine to 3,4-dihydroxyphenylalanine (DOPA), decarboxylation of DOPA to dopamine, N-acylation of dopamine to N-acetyldopamine (NADA) or N-β-alanyldopamine (NBAD), oxidation of NADA and NBAD to their corresponding quinones, and reactions between the quinones or quinone derivatives with cuticle protein (CP) side chains resulting in protein cross-linking. One type of pigmentation (quinone tanning) is associated with the covalent linkage of CPs to the ring component of NBAD. In contrast, linkage of CPs to the side chain of NADA (b-sclerotization) is correlated with colorless cuticle. N-acetyltransferase (NAT) catalyzes the conversion of dopamine to N-acetyl dopamine (NADA) in cuticle tanning pathway. In this study, we studied function of TcNAT1 on adult cuticle tanning by double stranded-RNA (dsRNA) mediated gene silencing. Injection of dsTcNAT1 had no affect on animal development, growth and molting such as larva to larva, larva to pupa and pupa to adult. However, some of the resulting adults (~70%) showed split elytra that could not cover their abdomen, resulting in improper folding of their hindwings. Interestingly, body color of the mature adults (older than 3 days) was darker than that of control dsTcVer treated adults because probably due to the buildup of abnormally high levels of dopamine, which is used for dopamine eumelanin pigment synthesis (black pigment) and dopamine quinone-mediated protein crosslinking. On elytra and hindwings of these adults, darker pigments were observed around the sensory bristles that are located in the intervein regions, suggesting that NADA mediated b-sclerotization is occurred at these regions. Similarly, darker pigment was evident at veins of the hindwings of the dsTcNAT1-mature adults. These results suggest that TcNAT1 have important roles in sclerotization and pigmentation of adult body and wings (elytron and hindwing). This work was supported by NRF (NRF-2012R1A2A1A01006467).

Insects have a protective exoskeleton consisted with cuticle to adapt various environments and pathogens. Insect cuticle mainly composed of the polysaccharide chitin and numerous of cuticular proteins (CPs). CPs are important for insect cuticle formation, development, and growth because it produces proper combination of mechanical and physical properties of cuticle depend on the regions of an exoskeleton. The largest family of CPs contains a 28-residue motif known as the Rebers-Riddiford (R&R) consensus sequence. When sequences containing the R&R consensus are aligned, they fall into three groups based on sequence similarity, and these groups tend to correlate with the type of cuticle (soft or hard) from which the proteins are derived. Proteins with the RR-1 motif have been found primarily in soft cuticle, whereas many proteins from rigid cuticle have an extended region of similarity called RR-2. We recently reportedthat two major CPs, TcCPR18 and TcCPR27 belong to RR-2, are essential for formation of highly sclerotized modified-forewings (elytra) of a beetle. In this study, we performed functional genomics of TcCPR4, which encodes RR-1 motif. The transcript levels of TcCPR4 drastically increased in 3 d-old pupae at when adult cuticle synthesis appears to be begun. Immunohistochemical studies revealed that TcCPR4 protein was detected in the rigid cuticle of elyton and ventral abdomen but not in the flexible cuticle of hindwing and dorsal abdomen of T. castaneum adult. Furthermore, TcCPR4 protein was specifically present at basal side of the procuticle (near the epidermal cells) and vertical canals, whereas TcCPR27 protein was found entire procuticle. Injection of double-stranded RNA of TcCPR4 (dsTcCPR4) into late instar larvae had no effect on development and any types of molting such as larval-larval, larval-pupal or pupal-adult. Interestingly, depletion of both TcCPR4 and TcCPR27 transcripts could rescue the elytral cuticle defect and mortality produced by injection of dsTcCPR27 alone. Transmission electron microscopy analysis revealed that depletion of TcCPR4 had abnormal vertical canals in rigid adult cuticle while dsTcCPR27 injection showed less electron-dense-horizontal laminae and vertical canals. Surprisingly, co-injection of dsRNA for TcCPR4 and TcCPR27 exhibited more severe cuticle defect with thinner elytral cuticle and abnormal vertical canals and chtin laminae compared to those from insects treated with dsRNA for each gene. These results suggest that TcCPR4 as a RR-1 is essential structural component in the rigid cuticle of T. castaneum adult.

온실에서 재배되는 수입 드라세나묘목 및 게발선인장에서 채집된 뿌리깍지벌레 2종 Ripersiella multiporifera Jansen 및 Rhizoecus albidus Goux를 우리나라에서 처음으로 보고한다. 이들은 수입된 식물을 통해 온실에 도입된 것으로 추정되며 우리나라에 추가적인 유입과 정착을 방지하기 위해 이들 종의 진단형질, 사진자료, 기주 및 분포정보를 제공하고자 한다.

Effective treatment for community-acquired pneumonia (CAP) requires administration of appropriate empirical therapy based on etiologic, clinical, and radiological fea- tures. However, in Korea, CAP is poorly characterized, and data on viral CAP are particularly sparse. Therefore, im- proper use of antibiotics is common, and is detrimental the potential for development of bacterial. Thus, we investigated clinical and radiological findings for discrimination of viral CAP from bacterial CAP. Etiologic, clinical, and radiologi- cal data from 467 patients with CAP at Chungbuk National University Hospital from October 2010 to September 2011 were analyzed retrospectively. Viruses were identified in 23 cases (11.4%); the influenza virus A was the most common virus detected (N=18, 25.4%), followed by the respiratory syncytial virus A (N=14, 17.9%). Bacteria were identified in 48 cases (23.8%); Streptococcus-pneumonia was the most common (N=24, 25.5%), followed by Staphylococcus aureus (N=20, 21.3%). Depending on hospitalization time, the fol- lowing significant differences were observed between viral and bacterial CAP: on admission, (1) high fever (≥ 38.5°C), (2) purulent sputum, (3) white blood cell count, (4) C- reactive protein levels, (5) and bilateral lung involvement on chest X-ray were higher in bacterial CAP; and at discharge, (1) duration of high fever and (2) radiologic improvement within three days were higher in viral CAP. Regarding sea- sonal patterns, both viruses and bacteria have been identi- fied with relative frequency in the winter season. This study described the etiological, clinical, and radiological findings of viral and bacterial CAP. Conduct of additional large- scale, prospective investigations will be required in order to improve the appropriate treatment of CAP.

There is increasing interest in zirconia as a dental material due to its aesthetics, as well as the exceptionally high fracture toughness and high strength that are on offer when it is alloyed with certain oxides like yttria. In recent years, many solution based chemical synthesis methods have been reported for synthesis of zirconia, of which the sol-gel method is considered to be best. Here, we synthesize zirconia by a sol gel assisted precipitation method using either PEG or PVA as a stabilizing agent. Zirconia sol is first synthesized using the hydrothermal method. We used NaOH as the precipitating agent in this method because it is easy to remove from the final solution. Zirconium and yttrium salts are used as precursors and PEG or PVA are used as stabilizers to separate the metal ions. The resulting amorphous zirconia powder is calcined at 900˚C for 2 h to get crystallized zirconia. XRD analysis confirmed the partially stabilized zirconia synthesis in all the synthesized powders. SEM was taken to check the morphology of the powder synthesized using either PEG or PVA as a stabilizing agent and finally the transparency was calculated. The results confirmed that the powder synthesized with 10 % PVA as the stabilizing agent had highest percentage of transparency among all the synthesized powder.

The distribution patterns of estuarine copepods were investigated in the Seomjin River estuary of southern Korea after heavy rains in August 2006. Tidal influence extended 16 km from the estuary mouth. Each estuary zone (Oligohaline salinity＜ 5, mesohaline salinity 5~18, polyhaline salinity ＞18) changed within a range of about 5~6 km between low and high tides. A total of ten species were recorded, of which Pseudodiaptomus koreanus, Sinocalanus tenellus, and Tortanus dextrilobatus were predominant in the oligohaline zone; Acartia ohtsukai and Acartia forticrusa in the mesohaline zone; and A. erythraea, Calanus sinicus, Centropages dorsispinatus, Labidocera rotunda and Paracalanus parvus s. l. in the polyhaline zone. Their density was fastly reduced in the other zones. In particular, the oligohaline species migrated and aggregated into deeper water during ebb tides in order to retain their populations, while the same tendency was weaker for polyhaline species, suggesting that evolutionary traits primarily control population retention behaviors in estuarine environments.

Insect-killing fungi have high potential for controlling agriculturally harmful pests. However, their pathogenicity is slow and this is one reason for their poor acceptance as a fungal insecticide. The expression of bumblebee, Bombus ignitus, venom serine protease (VSP) by Beauveria bassiana ERL1170 induced melanization of yellow spotted longicorn beetles, Psacothea hilaris as an over-reactive immune response, and caused substantially earlier mortality in beet armyworm, Spodopetra exigua larvae when compared to the wild type. No fungal outgrowth or sporulation was observed on the melanized insects, thus suggesting a self-restriction of the dispersal of the genetically modified fungus in the environment. The research is the first use of a multi-functional bumblebee VSP to significantly increase the speed of fungal pathogenicity, while minimizing the dispersal of the fungal transformant in the environment

ORF78 (ac78) of Autographa californica multiple nucleopolyhedrovirus (AcMNPV) is a baculovirus core gene of unknown function. To determine the role of ac78 in baculovirus life cycle, an ac78-deleted mutant AcMNPV, Ac78KO, was constructed. Quantitative PCR analysis revealed that ac78 is a late gene in the viral life cycle. After transfection into Spodoptera frugiperda cells, Ac78KO produced a single-cell infection phenotype indicating that no infectious budded viruses (BVs) were produced. The defection in BV production was also confirmed by both viral titration and Western blot. However, viral DNA replication is unaffected. Analysis of BV and occlusion derived virus (ODV) revealed that AC78 is associated with both forms of the virions and is a structural protein located to viral envelope. Electron microscopy showed that ac78 also plays an important role in embedding of ODV into occlusion body. This study therefore demonstrates that AC78 is a late virion associated protein and is essential for the viral life cycle.

ORF11 (ac11) of Autographa californica multiple nucleopolyhedrovirus (AcMNPV) is a highly conserved gene of unknown function. To determine the role of ac11 in baculovirus life cycle, an ac11-knockout mutant AcMNPV, Ac11KO, was constructed. qPCR analysis revealed that ac11 is an early gene in the life cycle. After transfection into Spodoptera frugiperda cells, Ac11KO produced a single cell infection phenotype indicating that no infectious budded viruses (BVs) were produced. The defection in BV production was confirmed by both viral titration and Western blot. However, viral DNA replication is unaffected. Electron microscopy showed that ac11 is required for nucleocapsids envelopment to form ODV and their subsequent embedding into OB. This study therefore demonstrates that ac11 is an early gene which is essential for the viral life cycle.

Worldwide studies on Apis cerana variation for biogeography and genetic diversity depended largely on a 86~93 bp-long mitochondrial non-coding region (internal spacer region) located between tRNALeu and COII (named as NC2), possibly due to higher variability among available markers. In order to incorporate the A. cerana occurring in South Korea into world extensive data, we also sequenced the NC2 from 118 A. cerana samples collected over nine Korean localities and 66 A. cerana samples over seven Asian localities, such as China, Vietnam, and Thailand. These data were combined with preexisting world data to scrutinize genetic relationships of A. cerana in South Korea to outside distributional range. Sequencing of 184 samples provided a total of ten haplotypes: five from Korea, six from China, one from Vietnam, and two from Thailand. Among them eight were new, whereas two were previously reported ones. Phylogenetic analysis of A. cerana NC2 haplotypes so far found including ours has confirmed the presence of four major groups of A. cerana (Asian mainland group, Sundaland group, Palawan group, and Luzon-Mindahnao group) and all haplotypes found in this study also were included in the Asian mainland group. In order to find further variable regions that can be used as sequence-based marker several mitochondrial non-coding regions and nuclear intron regions are in the middle of testing.

To enhance the production efficiency of foreign protein in baculovirus expression system, the effects of polyhedrin fragments were investigated by fusion expression them with the enhanced green fluorescence protein (EGFP). Recombinant viruses were generated to express EGFP fused with polyhedrin fragments based on the minimal region for self-assembly and the KRKK nuclear localization signal (NLS). The increase of EGFP production by fusion expressions was confirmed through protein and fluorescence intensity analyses. The importance of nuclear localization for enhanced production of EGFP was shown by the mutation of the NLS within the fused polyhedrin fragment. Among the fusion expressed protein in cytoplasm, the most hyper-expression was observed in the fusion of amino acids 32 to 59 of polyhedrin. Polyhedrin fragment fusion expression with classical swine fever virus E2 protein also resulted hyper-enhanced expression of E2 protein. However, the fusion expression of porcine circovirus ORF2 with polyhedrin fragment did not show significant enhance of ORF2 production. These results suggested that the enhancement of foreign protein production when fused with polyhedrin is caused by the enhanced stability of expressed protein.

Hyphantria cunea is a fall webworm is considered an agricultural pest. It is a major pest of many board-leaved trees. H. cunea nucleopolyhedrovirus (HcNPV) and H. cunea granulovirus (HcGV) were isolated from the fall webworm cadavers in Korea. To better understand HcNPV and HcGV, their genomic sequences were determined, analyzed and compared to two viruses together. The entire nucleotide sequence of the HcNPV genome was fully sequenced using 454 pyrosequencing. The genome of the HcNPV was 131,302 bp with a 45 % G+C content. Computer assisted analysis predicted 146 open reading frames (ORFs) of 50 or more amino acids that showed minimal overlap. Further more, when the phylogenetic relationship was analyzed, HcNPV was closely related to Orgyia pseudotsugata MNPV (OpMNPV) which belong to Group I NPV. The HcGV genome was 114,557 bp with a 39% G+C content and contained 130 putative ORFs of 50 or more amino acids. When phylogenetic relationships were analyzed, HcGV was closely related to Xestia c-nigrum granulovirus, which belong to the Type-II GV. HcNPV shares 48 ORFs with HcGV. The most significant difference between HcNPV and HcGV is fgf gene. HcNPV contains one fgf gene, whereas HcGV contains three fgf genes. The presence of fgf reduces the time and efficient systemic infection it takes the virus to kill its host. The difference of fgf number from HcNPV and HcGV suggested that different affect for the speed of systemic infection.

Cu2ZnSn(S,Se)4 material is receiving an increased amount of attention for solar cell applications as an absorber layer because it consists of inexpensive and abundant materials (Zn and Sn) instead of the expensive and rare materials (In and Ga) in Cu(In,Ga)Se2 solar cells. We were able to achieve a cell conversion efficiency to 4.7% by the selenization of a stacked metal precursor with the Cu/(Zn + Sn)/Mo/glass structure. However, the selenization of the metal precursor results in large voids at the absorber/Mo interface because metals diffuse out through the top CZTSe layer. To avoid the voids at the absorber/Mo interface, binary selenide compounds of ZnSe and SnSe2 were employed as a precursor instead of Zn and Sn metals. It was found that the precursor with Cu/SnSe2/ZnSe stack provided a uniform film with larger grains compared to that with Cu2Se/SnSe2/ZnSe stack. Also, voids were not observed at the Cu2ZnSnSe4/Mo interface. A severe loss of Sn was observed after a high-temperature annealing process, suggesting that selenization in this case should be performed in a closed system with a uniform temperature in a SnSe2 environment. However, in the experiments, Cu top-layer stack had more of an effect on reducing Sn loss compared to Cu2Se top-layer stack.

Artificial insemination (AI) has been performed widely in swine industry using fresh liquid sperm instead of frozen type of sperm. However fresh sperm are not able to preserve more than three days with optimal motility and other sperm parameters for the successful fertilization, since in vitro stored sperm has an oxidative stress that resulted increase of abnormality and acrosome reation. To overcome these major problems, novel preservative formulation is needed to neutralize the oxidative stress and to provide suitable physiological environment for sperm in in vitro. In this study, naturally derived substances such as Poncirus trifoliate (Trifoliate orange), Garcinia mangostana (Mangosteen), pig placenta and testis extracts were tested as sperm preservative agents. Placenta extracts (PE), trifoliate orange extracts (TOE), testes extracts (TE) and mangosteen extracts (ME) were applied to analyze specific parameters for sperm motion characteristics individually and combinatorial. Each individual extract treatment can accelerate the sperm motility but noticeably TOE, TE and ME treatments exhibited the considerable and significant preservation of sperm motility. PE, TE and ME showed a significant (p<0.05) increase in ALH after one week. Further we evaluated the five different combinations of these extracts on sperm motility and its motion characteristics. Surprisingly even after one week ME, TOE and TE combination significantly preserved the sperm motility about 75%. It is noteworthy that unlike individual extract treatment, combination of ME, TOE and TE simultaneously protect the sperm motility and its motion characteristics. Taken together these data conclude that addition of ME, TOE and TE can be effective for preservation of pig sperm.

This study aims to examine the effects of taping of the ankle joint on the static and dynamic balance and gait ability of stroke patients. Twenty-six stroke patients receiving physical therapy at a hospital located in Gyeonggi-do were divided equally into a group that had taping in physical therapy and an ordinary physical therapy group. They exercised for 30 minutes each, 3 times per week for 8 weeks from June to August 2011. Romberg’s eye open and eye closed tests, limits of stability(LOS), forward and back test, timed up and go test(TUG) and 10-meter gait velocity test were performed to evaluate static balance, dynamic balance, and gait ability, respectively, prior to and 8 weeks after the intervention. Differences within each group in relation to the lapse of time were compared by a paired t-test. Differences between the two groups were compared by an independent t-test. Regarding comparison of differences within each group, all tests resulted in significant changes in both groups after the intervention (p<.05). Comparison of differences between the two groups showed that taping in the physical therapy group had significantly better test results than the ordinary physical therapy group in all measured items(p<.05). The after effects of ankle taping on stroke patients are more efficient and effective than ordinary physical therapy alone in improving balance and gait ability.

This study was conducted to evaluate the effect of the exercise on elderly balance ability by using hippotherapy and therapeutic ball exercise. 10 patients were assigned to the hippotherapy group and they got with 30 minutes of hippotherapy. Another 10 elderly were assigned to the therapeutic ball group and they got with 30 minutes of therapeutic ball exercise. All procedures were repeated 5 times a week for the total of four weeks. To investigate the participants balancing abilities, the Time“ Up & Go”(TUG) and One Leg Stand Test(OLST) were evaluated. The results of study were significant differences between pre-test and post-test of TUG and OLST(p<.05), and there were no significant differences between hippotherapy and therapeutic ball exercise(p>.05). The conclusion showed that both the hippotherapy and the therapeutic ball exercises were effective on elderly balancing ability. Consequently, it would be better to practice therapeutic ball than hippotherapy for elderly exercise because the more economical and there is less restriction of space than the hippotherapy.

Insect-killing (entomopathogenic) fungi have high potential for controlling agriculturally harmful pests. However, their pathogenicity is slow and this is one reason for their poor acceptance as a fungal insecticide. The expression of bumblebee, Bombus ignitus, venom serine protease (VSP) by Beauveria bassiana (ERL1170) induced melanization of yellow spotted longicorn beetles (Psacothea hilaris) as an over-reactive immune response, and caused substantially earlier mortality in beet armyworm (Spodopetra exigua) larvae when compared to the wild type. No fungal outgrowth or sporulation was observed on the melanized insects, thus suggesting a self-restriction of the dispersal of the genetically modified fungus in the environment. The fungal transformant also shows mammal fibrinolytic activity, by which the transformant can be used pharmaceutically. The research is the first use of a multi-functional bumblebee VSP to significantly increase the speed of fungal pathogenicity, while minimizing the dispersal of the fungal transformant in the environment.