Five phaP family genes and one phaR gene have been identified in the genome of Burkholderia gut symbiont. PhaP proteins function as surface proteins of polyhydroxyalkanoate (PHA) granules, and PhaR protein acts as a negative regulator of PhaP biosynthesis. To address the biological roles of four phaP family genes (phaP1, phaP2, phaP3, and phaP4) and the phaR gene during insect-gut symbiont interaction, these Burkholderia mutants were fed to the second instar nymph. The ΔphaR mutant decreased the colonization ability in the host midgut compared to wild-type Burkholderia cells and negatively affected the host insect’s fitness compared with wild-type infected host. These results demonstrate that PhaR plays an important role in the biosynthesis of PHA granules and it is significantly related to the colonization of the Burkholderia gut symbiont in the host insects’ midgut

목 적: 안경과 콘택트렌즈 구매장소에 따라 안경사의 직무신뢰도 및 안경과 콘택트렌즈 가격의 만족도에 대한 의견 차이가 있는지에 대하여 알아보고자 하였다. 방 법: 2015년 5월에서 6월 두 달 동안 대구·경북, 부산·경남지역의 안경광학과 재학생과 고객을 대상 으로 안경과 콘택트렌즈 구매 장소, 안경사의 판매 및 검안에 대한 신뢰도 및 안경과 콘택트렌즈 구매가격 만족도에 대한 설문을 실시하였다. 결 과: 직무신뢰도에서 검안은 남자의 평균값이 가장 높았고(3.89±0.88), 가격만족도에서는 모두 여자 의 평균값이 높았다(3.18±0.72). 구매장소에 따른 상관관계에서는 비 프랜차이즈의 검안신뢰도가 가장 높 았다(3.99±0.91). 결 론: 구매장소에 따른 직무신뢰도와 가격만족도에 대하여 일부 항목에서 유의한 차이가 나타났고, 이 결과를 바탕으로 직무신뢰도와 가격만족도를 높이는 자료로 이용될 수 있을 것이라고 생각한다.

Using a partial D-loop sequence of mtDNA, a comprehensive molecular evolutionary analysis was performed of the maternal lineages of the Jeju native pigs(JNPs) that presented in Jeju Island. A total of 100 DNA sequences from Asian domestic pigs(ADP), European domestic pigs(EDP), Asian wild boars(AWB), European wild boars(EWB), and JNPs were used for the molecular evolutionary analyses including phylogeny and network analyses. The most fitted model for the phylogenetic analysis was determined using hierarchical likelihood-ratio tests conducted by MrModeltest implemented in the PAUP package. A consensus tree was established from 5,000,000 iterations using the MR BAYS program. Three recognizable JNP groups–one(JNPE) in the European cluster and two(JNPA1 and JNPA2) in the Asian cluster–were detected in the estimated phylogenetic tree and network. The maternal lineage of JNPE was the most adjacent to that of EWB and a clear haplogroup sharing an identical haplotype(hap16) among 15 individuals of JNPE and 2 individuals of EWB was detected.

The purpose of this study is to evaluate the quality characteristics of blanched Capsella Bursa-pastoris with quick freezing, and thawing methods. Blanched Capsella bursa-pastoris was packed by vacuum packing (VP) and air containing packing (AP). After then, they were frozen at -20, -40, and 60℃ (immersion freezing). Then, they were stored at -20℃ during 1 week. They were thawed with normal (room temperature), running water and high frequency method. The pH and color of Capsella bursa-pastoris were not different before and after freezing. Likewise, total bacterial counts were shown without variation among them. The amount of drip from thawed Capsella bursa-pastoris was generally more in VP than AP. The cutting intensity of immersion freezing wes higher than that of other freezing methods (P<0.05). Therefore, we selected AP, immersion freezing and high frequency thawing methods for the high quality according to above results

Differentiation of Pleurotus eryngii is laborious and time-consuming tasks especially in mycelial status. For development of a method for differentiation of P. eryngii cultivars, simple sequence repeats (SSR) from whole genomic DNA sequence analysis was used for genotyping and two multiplex-SSR primer sets were developed. These SSR primer sets were employed to distinguish 12 cultivars and strains. Five polymorphic markers were selected based on the genotypes. PCR with the each primer produced one to four distinct bands ranging in size from 200 to 300 bp. Polymorphism information content (PIC) values of the five markers were in range of 0.6627 to 0.6848 with an average of 0.6775. Unweighted pair-group method with arithmetic mean clustering analysis based on genetic distances using five SSR markers classified 12 cultivars into 2 clusters. Cluster I and II comprised of 4 and 8 cultivars, respectively. Two multiplex sets, Multi-1 (SSR312 and SSR366) and Multi-2 (SSR178 and SSR277) completely discriminated 12 cultivar and strains with 21 allele with a PIC value of 0.9090. These results might be useful to provide an efficient method for the identification of P. eryngii cultivars with separate PCR reactions. (This work was supported by a grant from the Gold Seed Project [Supported by a grant from the IPET (213003-04-3-WTI11), MIFAFF, Republic of Korea.]

Peptidoglycan recognition proteins (PGRPs) are family of innate immune molecules that recognize bacterial peptidoglycan. PGRP-LE, a member of the PGRP family, selectively binds to diaminopimelic acid (DAP)-type peptidoglycan to activate both the immune deficiency (IMD) and proPhenoloxidase (proPO) pathways in insects. A PGRP-LE-dependent induction of autophagy to control Listeria monocytogenes has also been reported. We identified and partially characterized a novel PGRP-LE homologue, from Tenebrio molitor and analyzed its functional role in the survival of the insect against infection by a DAP-type PGN containing intracellular pathogen, L. monocytogenes. The cDNA is comprised of an open reading frame (ORF) of 990 bp and encodes a polypeptide of 329 residues. TmPGRP-LE contains one PGRP domain, but lacks critical residues for amidase activity. Quantitative RT-PCR analysis showed a broad constitutive expression of the transcript at various stages of development spanning from larva to adult. RNAi mediated knockdown of the transcripts followed by a challenge with L. monocytogenes showed a significant reduction in survival rate of the larvae, suggesting a putative role of TmPGRP-LE in sensing and control of L. monocytogenes infections in T. molitor. These results implicate PGRP-LE as a defense protein necessary for survival of T. molitor against infection by L. monocytogenes.

CD63, a member of tetraspanin membrane protein family, plays pivotal role in cell growth, motility, signal transduction, host-pathogen interactions and cancer. In this work, the cDNA encoding CD63 homologue (TmCD63) was cloned from larvae of coleopteran beetle, Tenebrio molitor. The cDNA is comprised of an open reading frame of 705 bp, encoding putative protein of 235 amino acid residues. In silico analysis shows that the protein has four putative transmembrane domains and one large extracellular loop. The characteristic ‘Cys-Cys-Gly’ motif and ‘Cys188’ residues are highly conserved in the large extracellular loop. Phylogenetic analysis of TmCD63 revealed that they belong to the insect cluster with 50-56% identity. Analysis of spatial expression patterns demonstrated that TmCD63 mRNA is mainly expressed in gut and Malphigian tubules of larvae and the testis of the adult. Developmental expression patterns of CD63 mRNA showed that TmCD63 transcripts are detected in late larval, pupal and adult stages. Interestingly, TmCD63 transcript was upregulated the maximum 4.5 fold in response to DAP-type peptidoglycan during the first 6 h, although other immune elicitors also made significant increase in the transcript level at later time-points. These results suggest that CD63 might contribute to T. molitor immune response against various microbial pathogens.

Apolipophorin III (apoLp-III) is a well-known hemolymph protein having a functional role in lipid transport and immune response of insects. We cloned full-length cDNA encoding putative apoLp-III from larvae of the coleopteran beetle, Tenebrio molitor (TmapoLp-III), by identification of clones corresponding to the partial sequence of TmapoLp-III, subsequently followed with full length sequencing by a clone-by-clone primer walking method. The complete cDNA consists of 890 nucleotides, including an ORF encoding 196 amino acid residues. Excluding a putative signal peptide of the first 20 amino acid residues, the 176-residue mature apoLp-III has a calculated molecular mass of 19,146 Da. Genomic sequence analysis with respect to its cDNA showed that TmapoLp-III was organized into four exons interrupted by three introns. Several immune-related transcription factor binding sites were discovered in the putative 5’-flanking region. BLAST and phylogenetic analysis reveals that TmapoLp-III has high sequence identity (88%) with Tribolium castaneum apoLp-III but shares little sequence homologies (<26%) with other apoLp-IIIs. Homology modeling of Tm apoLp-III shows a bundle of five amphipathic helices, including a short helix 3’. The ‘helix-short helix-helix’ motif was predicted to be implicated in lipid binding interactions, through reversible conformational changes and accommodating the hydrophobic residues to the exterior for stability. Highest level of TmapoLp-III mRNA was detected at late pupal stages, albeit it is expressed in the larval and adult stages at lower levels. The tissue specific expression of the transcripts showed significantly higher numbers in larval fat body and adult integument. In addition, TmapoLp-III mRNA was found to be highly up-regulated in late stages of L. monocytogenes or E. coli challenge. These results indicate that TmapoLp-III may play an important role in innate immune responses against bacterial pathogens in T. molitor.

We have identified novel ricin-type (R-type) lectin by sequencing of random clones from cDNA library of the coleopteran beetle, T.molitor. The cDNA sequence is comprised of 495 bp encoding a protein of 164 amino acid residues and shows 49% identity with galectin of Tribolium castaneum. Bioinformatics analysis shows that the amino acid residues from 35 to 162 belong to ricin-type β-trefoil structure. The transcript was significantly upregulated after early hours of injection with peptidoglycans derived from Gram (+) and Gram (-) bacteria, beta-1, 3 glucan from fungi and an intracellular pathogen, L. monocytogenes suggesting putative function in innate immunity.

In the process of revising the tachinid genus Linnaemya Robineau-Desvoidy in Korea, we have discovered two species for the first time in Korea. They closely resemble each other and need to be identified with caution. We here provide detailed redescriptions and illustrations with their diagnostic characters indicated. L. atriventris can be distinguished from L. hirtipennis by the combination of the following characteristics: 1) abdomen black in ground color; 2) wing vein R1 without setulose; 3) hypandrium without secondary posterior lobe; 4) male with antero-basal 1/3 of flagellomere I distinctly swollen; and 5) postgonite very weakly curved dorsally.

The present case study highlights the effects of a novel Comprehensive Hand Repetitive Intensive Strengthening Training (CHRIST) on morphological changes and associated upper extremity (UE) muscle strength and motor performance in a child with spastic quadriplegic cerebral palsy (CP). The Child, a 10-year-old girl with spastic quadriplegic CP, was treated with CHRIST for 60 minutes a day, five times a week, for 5 weeks. The CHRIST was designed to improve motor function and strength. Clinical tests including the modified Wolf Test, Jebsen-Taylor Hand Function Test, and Pediatric Motor Activity Log questionnaire were used to determine motor function. Ultrasound imaging was performed to determine the changes in the cross-section area (CSA) of the extensor carpi radialis (ECR) and triceps brachii (TRI). Muscle strength was measured with a dynamometer at pretest, and post-test, and 3-month follow-up. Ultrasound imaging data showed that the CSAs of both ECR and TRI muscles were enhanced as a function of the intervention. These changes were associated with muscle strength and motor performance and their effects remained even at a 3-month follow-up test. Our results suggest that the CHRIST was effective at treating muscle atrophy, weakness and motor dysfunction in a child with spastic quadriplegic CP.