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이혜신,최진혁,손태권,이희준,배성진,서지혜,박지현,류수형,이단비,장명국,유은실,정영화,김규원 생화학분자생물학회 2018 Experimental and molecular medicine Vol.50 No.-
Liver fibrosis can be reversed by removing its causative injuries; however, the molecular mechanisms mediating the resolution of liver fibrogenesis are poorly understood. We investigate the role of a scaffold protein, A-Kinase Anchoring Protein 12 (AKAP12), during liver fibrosis onset, and resolution. Biliary fibrogenesis and fibrosis resolution was induced in wild-type (WT) or AKAP12-deficient C57BL/6 mice through different feeding regimens with 0.1% 3,5- diethoxycarbonyl-1,4-dihydrocollidine (DDC)-containing chow. AKAP12 expression in portal fibroblasts (PFs) and liver sinusoidal endothelial cells (LSECs) gradually decreased as fibrosis progressed but was restored after cessation of the fibrotic challenge. Histological analysis of human liver specimens with varying degrees of fibrosis of different etiologies revealed that AKAP12 expression diminishes in hepatic fibrosis from its early stages onward. AKAP12 KO mice displayed reduced fibrosis resolution in a DDC-induced biliary fibrosis model, which was accompanied by impaired normalization of myofibroblasts and capillarized sinusoids. RNA sequencing of the liver transcriptome revealed that genes related to ECM accumulation and vascular remodeling were mostly elevated in AKAP12 KO samples. Gene ontology (GO) and bioinformatic pathway analyses identified that the differentially expressed genes were significantly enriched in GO categories and pathways, such as the adenosine 3′,5′-cyclic monophosphate (cAMP) pathway. Knockdown of the AKAP12 gene in cultured primary PFs revealed that AKAP12 inhibited PF activation in association with the adenosine 3′,5′-cyclic monophosphate (cAMP) pathway. Moreover, AKAP12 knockdown in LSECs led to enhanced angiogenesis, endothelin-1 expression and alterations in laminin composition. Collectively, this study demonstrates that AKAP12-mediated regulation of PFs and LSECs has a central role in resolving hepatic fibrosis.
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항혈청 기반 진단 스트립을 이용한 과수 화상병 현장진단
허광일,신두산,손수형,오창식,박덕환,이영기,차재순 한국식물병리학회 2017 식물병연구 Vol.23 No.4
Recently fire blight occurred in the Republic of Korea and eradication program for the disease has been executed since then. Specificity and detection sensitivity of the 2 antibody-based diagnostic strips to Korean isolates of Erwinia amylovora (Ea) and their application for on-site diagnosis were evaluated in this study. Ea AgriStrip, a commercial diagnostic kit, and EB strip, developed in this study, reacted positively to the all tested Korean Ea strains and also to most of Erwinia pyrifoliae (Ep) strains causing black shoot blight. They reacted negatively to all Pusedomonas syringae pv. syringae (Pss) strains that cause shoot blight on apple. Detection sensitivity was similar between the 2 strips. For on-site diagnosis, the two strips reacted positively only to the extractions of the fire-blighted samples on all fire blight occurred orchards except one orchard at which onsite diagnosis was carried out at winter time. In addition, they reacted positively to the black-shoot blighted extractions from the black shoot blight occurred apple orchard. These results suggest that both EB strip and Ea AgriStrip would be useful for on-site diagnosis of fire blight in Korea.
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신두산,허광일,손수형,오창식,이영기,차재순 한국식물병리학회 2018 Plant Pathology Journal Vol.34 No.3
Fast and accurate diagnosis is needed to eradicate and manage economically important and invasive diseases like fire blight. Loop-mediated isothermal amplification (LAMP) is known as the best on-site diagnostic, because it is fast, highly specific to a target, and less sensitive to inhibitors in samples. In this study, LAMP assay that gives more consistent results for on-site diagnosis of fire blight than the previous developed LAMP assays was developed. Primers for new LAMP assay (named as DS-LAMP) were designed from a histidine-tRNA ligase gene (EAMY_RS32025) of E. amylovora CFBP1430 genome. The DS-LAMP amplified DNA (positive detection) only from genomic DNA of E. amylovora strains, not from either E. pyrifoliae (causing black shoot blight) or from Pseudomonas syringae pv. syringae (causing shoot blight on apple trees). The detection limit of DSLAMP was 10 cells per LAMP reaction, equivalent to 104 cells per ml of the sample extract. DS-LAMP successfully diagnosed the pathogens on four fire-blight infected apple and pear orchards. In addition, it could distinguish black shoot blight from fire blight. The Bühlmann-LAMP, developed previously for on-site diagnosis of fire blight, did not give consistent results for specificity to E. amylovora and on-site diagnosis; it gave positive reactions to three strains of E. pyrifoliae and two strains of P. syringae pv. syringae. It also, gave positive reactions to some healthy sample extracts. DSLAMP, developed in this study, would give more accurate on-site diagnosis of fire blight, especially in the Republic of Korea, where fire blight and black shoot blight coexist.
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Xanthomonas euvesicatoria Causes Bacterial Spot Disease on Pepper Plant in Korea
Min-Seong Kyeon,손수형,노영희,김용언,이혁인,차재순 한국식물병리학회 2016 Plant Pathology Journal Vol.32 No.5
In 2004, bacterial spot-causing xanthomonads (BSX)were reclassified into 4 species—Xanthomonas euvesicatoria, X. vesicatoria , X. perforans , and X. gardneri . Bacterial spot disease on pepper plant inKorea is known to be caused by both X. axonopodispv. vesicatoria and X. vesicatoria . Here, we reidentifiedthe pathogen causing bacterial spots on pepperplant based on the new classification. Accordingly, 72pathogenic isolates were obtained from the lesions onpepper plants at 42 different locations. All isolates werenegative for pectolytic activity. Five isolates were positivefor amylolytic activity. All of the Korean pepperisolates had a 32 kDa-protein unique to X. euvesicatoriaand had the same band pattern of the rpoB gene asthat of X. euvesicatoria and X. perforans as indicatedby PCR-restriction fragment length polymorphismanalysis. A phylogenetic tree of 16S rDNA sequencesshowed that all of the Korean pepper plant isolates fitinto the same group as did all the reference strains ofX. euvesicatoria and X. perforans . A phylogenetic treeof the nucleotide sequences of 3 housekeeping genes—gapA , gyrB , and lepA showed that all of the Koreanpepper plant isolates fit into the same group as did allof the references strains of X. euvesicatoria . Based onthe phenotypic and genotypic characteristics, we identifiedthe pathogen as X. euvesicatoria . Neither X. vesicatoria, the known pathogen of pepper bacterial spot,nor X. perforans , the known pathogen of tomato plant,was isolated. Thus, we suggest that the pathogen causingbacterial spot disease of pepper plants in Korea is X. euvesicatoria .
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