• Title/Summary/Keyword: OPA1

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Analysis genetic diversity of Plasmodiophora brassicae using RFLP and RAPD(oral)

  • Heo, Seung-Hwan;Jang, Chang-Soon;Lee, Hyoun-Kyoung;Lee, Woo-Chung;Jang, Se-Jeong;Kim, Hong-Gi
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.112.1-112
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    • 2003
  • Genetic diversity of Plasmodiophora brassicae from major chinese cabbage cultivating areas in Korea was analyzed by using PCR-RFLP and RAPD. Single spores of P brassicae isolated from galls of club root made induce lesion on chinese cabbage successfully. The PCR-RFLP and RAPD by primers PbITS, URP 3, 6 and OPA 7 revealed that single spore isolates showed various DNA polymorphisms among them unrelated geographic origins. These results indicate that P. brassicae population in Korea showed genetic difference among them. This study could be facilitate to identify genetic characteristics ofP. brassicae based on DNA polymorphisms between single spore isolates and to get basic information which can be used to advanced resistance breeding against club root of chinese cabbage.

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Heptaphylline Induces Apoptosis in Human Colon Adenocarcinoma Cells through Bid and Akt/NF-κB (p65) Pathways

  • Boonyarat, Chantana;Yenjai, Chavi;Vajragupta, Opa;Waiwut, Pornthip
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.23
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    • pp.10483-10487
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    • 2015
  • Heptaphylline derivatives are carbazoles in Clausena harmandiana, a medicinal plant that is utilized for headache, stomach ache, and other treatments of illness. The present study examined the effects of heptaphylline and 7-methoxyheptaphylline on apoptosis of human colon adenocarcinoma cells (HT-29 cell line). Quantification of cell viability was performed using cell proliferation assay (MTT assay) and of protein expression through immunoblotting. The results showed that only heptaphylline, but not 7-methoxyheptaphylline, significantly significantly activated cleaved of caspase-3 and poly (ADP-ribose) polymerase (PARP-1) which resulted in HT-29 cell death. We found that heptaphylline activated BH3 interacting-domain death agonist (Bid) and Bak, proapoptotic proteins. In contrast, it suppressed X-linked inhibitor-of-apoptosis protein (XIAP), Bcl-xL and survivin, inhibitors of apoptosis. In addition, heptaphylline inhibited activation of NF-${\kappa}B$/p65 (rel), a regulator of apoptotic regulating proteins by suppressing the activation of Akt and $IKK{\alpha}$, upstream regulators of p65. The findings suggested that heptaphylline induces apoptosis in human colon adenocarcinoma cells.

Differentiation of Salmonella typhimurium from Gram-negative Intestinal Microbes by Randomly Amplified Polymorphic DNA (RAPD) Fingerprinting

  • Jin, Un-Ho;Chung, Tae-Wook;Kim, June-Ki;Nam, Kyung-Soo;Ha, Sang-Do;Kim, Cheorl-Ho
    • Journal of Microbiology
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    • v.38 no.1
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    • pp.8-10
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    • 2000
  • In order to rapidly identify and differentiate Salmonella typhimurium from the intestinal gram-negative bacteria, randomly amplified polymorphic DNA (RAPD) fingerprinting of Salmonella typhimurium was carried out using random primers designated OPA-13 (5'-CAGCACCCAC-3'), OPB-10 (5'-CGTCTGGGAC-3'), OPB-18 (5'-CCACAGCAGT-3'), and OPJ-10 (5'-AAGCCCGAGG-3'), and its patterns compared with 6 representive intestinal, gram-negative bacterial strains, Vibrio parahaemolyticus, V. vulnificus, Enterobacter cloacae, Escherichia coli O157:H7, Pseudomonas aeruginosa, and Proteus sp., which are often found in foods. S. typhimurium had unique and distinct fingerprinting patterns. RAPD fingerprinting is thus concluded to be a rapid and sensitive method for the identification of S. typhimurium compared to conventional culturing procedures or immunoassays.

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An Experimental Study on the Oil Spillage of Damaged Oil Tanker (유조선의 선체손상 시 기름의 해상유출에 대한 실험적 연구)

  • Kim, Ul-Nyeon;Ha, Woo-Il;Choe, Ick-Heung
    • Journal of the Society of Naval Architects of Korea
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    • v.46 no.4
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    • pp.398-408
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    • 2009
  • Crude oil carriers or product oil carriers are confronted with sea pollution due to hull damage from various accidents. To reduce the oil spillage of tankers, IMO(International Maritime Organization) and OPA 90(Oil Pollution Act 1990) adopted the hull structures of double skin type. In this study, oil spillage test of the double skin tanker with 1/100 scaled model was carried out under damaged condition due to collision and grounding accidents. A new structural type of oil tanker was also tested with pipe and valve system arranged in double side and single bottom hulls. Their results were compared with that of conventional type double hull on the view point of ship safety and oil spillage.

De-novo Hybrid Protein Design for Biodegradation of Organophosphate Pesticides

  • Awasthi, Garima;Yadav, Ruchi;Srivastava, Prachi
    • Microbiology and Biotechnology Letters
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    • v.47 no.2
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    • pp.278-288
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    • 2019
  • In the present investigation, we attempted to design a protocol to develop a hybrid protein with better bioremediation capacity. Using in silico approaches, a Hybrid Open Reading Frame (Hybrid ORF) is developed targeting the genes of microorganisms known for degradation of organophosphates. Out of 21 genes identified through BLAST search, 8 structurally similar genes (opdA, opd, opaA, pte RO, pdeA, parC, mpd and phnE) involved in biodegradation were screened. Gene conservational analysis categorizes these organophosphates degrading 8 genes into 4 super families i.e., Metallo-dependent hydrolases, Lactamase B, MPP and TM_PBP2 superfamily. Hybrid protein structure was modeled using multi-template homology modeling (3S07_A; 99%, 1P9E_A; 98%, 2ZO9_B; 33%, 2DXL_A; 33%) by $Schr{\ddot{o}}dinger$ software suit version 10.4.018. Structural verification of protein models was done using Ramachandran plot, it was showing 96.0% residue in the favored region, which was verified using RAMPAGE. The phosphotriesterase protein was showing the highest structural similarity with hybrid protein having raw score 984. The 5 binding sites of hybrid protein were identified through binding site prediction. The docking study shows that hybrid protein potentially interacts with 10 different organophosphates. The study results indicate that the hybrid protein designed has the capability of degrading a wide range of organophosphate compounds.

Drp1 Expression and Phosphorylation in Steroidogenic Corpus Luteum during the Estrous Cycle in Rat Ovaries

  • Park, Ji-Eun;Lee, Seung Gee;Yoo, Young Hyun;Kim, Jong-Min
    • Development and Reproduction
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    • v.26 no.2
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    • pp.71-77
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    • 2022
  • In response to luteinizing hormone (LH), a higher concentration of progesterone (P4) is produced in luteal cells of corpus luteum (CL). Mitochondria are an essential cellular organelle in steroidogenesis. The specific engagement of the concept regarding mitochondrial shaping with early stages of steroidogenesis was suggested in reproductive endocrine cells. Although the specific involvement of GTPase dynamin-related protein 1 (Drp1) with steroidogenesis has been demonstrated in luteal cells of bovine CL in vitro, its actual relationship with ovarian steroidogenesis during the estrous cycle remains unknown. In this study, while Fis1 and Opa1 protein levels did not show significant changes during the estrous cycle, Drp1, Mfn1, and Mfn2 proteins exhibited relatively lower levels at proestrus than at estrus or diestrus. 3β-HSD showed higher levels at proestrus than at estrus or diestrus. In addition, Drp1 phosphorylation (s637) was higher in proestrus than in estrus or diestrus. Immune-positive cells for Drp1, pDrp1 (s637), and 3β-HSD were all localized in the cytoplasm of luteal cells in the CL. The immune-positive cells for 3β-HSD were more frequently seen in the CL at proestrus than at estrus or diestrus. Immunoreactivity for Drp1 in luteal cells at proestrus was weaker than that at estrus or diestrus. However, pDrp1 (s637) immune-positive cells were mostly detected in luteal cells at proestrus. These results imply that steroidogenesis (P4 production) in the CL is closely related to phosphorylation of Drp1 at serine 637. Taken together, this study presents evidence that Drp1 phosphorylation at serine 637 is an important step in steroidogenesis in the CL.

Bandsharing Values and Genetic Distances of Two Wild Shortnecked Clam, Ruditapes philippinarum Populations from the Yellow Sea Assessed by Random Amplified Polymorphic DNAs-Polymerase Chain Reaction

  • Yoon, Jong-Man;Kim, Yong-Ho
    • Journal of Aquaculture
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    • v.17 no.1
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    • pp.12-23
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    • 2004
  • Genomic DNAs were extracted from the muscle of twenty-two specimens of two shortnecked clam, Ruditapes phifippinarum populations collected in Anmyeondo and Seocheon. Genetic differences within and between populations were analysed by random amplified polymorphic DNAs-polymerase chain reaction (RAPD-PCR) using twenty arbitrary decamer primers. Out of 20 primers, 6 generated a total of 1,111 major and minor RAPD bands from individuals of two sites, producing approximately 4.2 average polymorphic bands per primer in individuals from Anmyeondo and ranging in size from less than 50 to larger than 1,500 base pairs (bp). The electrophoretic analysis of RAPD products amplified showed moderate levels of similarity among the different individuals in Seo-cheon population. The average bandsharing values (BS value) of the samples within population from Anmyeondo ranged from 0.155 to 0.684, whereas it was 0.143∼0.782 within population from Seocheon. The average BS value between individuals No. 13 and No. 14 from Seocheon was 0.782 which was higher than that of those from Anmyeondo. The single linkage dendrogram resulted from three primers (OPA-08, -09 and -20), indicating six genetic groupings composed of group 1 (No.4, 8 and 10), group 2 (No. 18), group 3 (No.2, 5 and 7), group 4 (No. 1, 3, 6, 9, 11, 12, 13, 14, 15 and 17), group 5 (16, 19 and 20) and group 6 (No. 21 and 22). In the Seocheon population, the individual No. 18 clustered distinctly from the others of this population. The observed genetic distance between the two populations from Anmyeondo and Seocheon was more than 0.209 (0.247 and 0.275). The shortest genetic distance (0.094) displaying significant molecular differences was between individuals No. 13 and No. 14. Especially, the genetic distance between individuals No. 22 and the remnants among individuals in two geographical populations was highest (0.275). This result illustrated that individual No.22 is distinct from other individuals within two shortnecked populations. The different geographical features of two sites may have caused the genetic diversity in two shortnecked clam populations.

Simultaneous Fluorimetric Determination of On-line Preconcentrated HANs, DCAD and TCAD by Using RPLC with a Postcolumn Derivatization System

  • Jung, Sung-Woon;Choi, Yong-Wook
    • Bulletin of the Korean Chemical Society
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    • v.34 no.6
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    • pp.1783-1790
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    • 2013
  • A simultaneous analytical method has been developed for the fluorimetric determination of haloacetonitriles (HANs) [dichloroacetonitrile (DCAN), trichloroacetonitrile (TCAN), dibromoacetonitrile (DBAN), haloacetamides [dichloroacetamide (DCAD), and trichloroacetamde (TCAD)] in drinking water by using the combined on-line perconcentration/reversed phase liquid chromatography (RPLC)-postcolumn detection system. This on-line perconcentration system was achieved by employing a precolumn packed with a commercial solid phase extraction (SPE) sorbent for the enrichment and purification of the target analytes. The haloacetonitriles and haloacetamides were separated on CN analytical column in a 7.5% methanol-0.02 M phosphate buffered mobile phase at pH 3. The column effluents were reacted with postcolumn reagents of ophthaldialdehyde (OPA) and sulfite ion at pH 11.5, to produce a highly fluorescent isoindole fluorophore, which were measured with a fluorescence detector. Under the optimized conditions for RPLC and the postcolumn derivatization system all of the coefficient of determination of the standard calibration curves for the target analytes were over 0.99 and had a linear range from 5 to 100 ${\mu}g/L$. The detection limits showed 1.6 ${\mu}g/L$ for DCAD, 0.1 ${\mu}g/L$ for TCAD, 0.6 ${\mu}g/L$ for DCAN, 1.6 ${\mu}g/L$ for TCAN and 1 ${\mu}g/L$ for DBAN, and the recoveries were ranged from 64 to 99% except for DCAD with precisions less than 4.9% in distilled water, and from 72(${\pm}4%$) to 116%(${\pm}2%$) in tap water.

QTL Analysis of Soybean Seed Weight Using RAPD and SSR Markers

  • Chung, Jong-Il;Ko, Mi-Suk;Kang, Jin-Ho
    • Plant Resources
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    • v.3 no.3
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    • pp.184-193
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    • 2000
  • Soybean [Glycine max (L.) Merr.] seed weight is a important trait in cultivar development. Objective of this study was to identify and confirm quantitative trait loci (QTLs) for seed weight variation in the F2 and F2:3 generations. QTLs for seed weight were identified in F2 and F2:3 generations using interval mapping (MapMaker/QTL) and single-factor analysis of variance (ANOVA). In the F2 plant generation (i.e., F3 seed), three markers, OPL9a, OPM7a, and OPAC12 were significantly (P<0.01) associated with seed weight QTLs. In the F2:3 plant row generation (i.e., F4 seed), five markers, OPA9a, OPG19, OPL9b, OPP11, and Sat_085 were significantly (P<0.01) associated with seed weight QTLs. Two markers, OPL9a and OPL9b were significantly (P<0.05) associated with seed weight QTLs in both generations. Two QTLs on USDA soybean linkage group C1 and R were identified in both F2 and F2:3 generations using interval mapping. The linkage group C1 QTL explained 16% of the variation in seed weight in both generations, and the linkage group R QTL explained 39% and 41% of the variation for F2 and F2:3 generation, respectively. The linkage group C2 QTL identified in F2:3 generation explained 14.9% of variation. Linkage groups C1, C2 and R had previously been identified as harbouring seed size QTLs. The consistency of QTLs across generations and populations indicates that marker-assisted selection is possible in a soybean breeding program.

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N-oleoyl-D-erythro-sphingosine-based Analysis of Ceramide by High Performance Liquid Chromatography and Its Application to Determination in Diverse Biological Samples

  • Lee, Youn-Sun;Choi, Heon-Kyo;Yoo, Jae-Myung;Choi, Kyong-Mi;Lee, Yong-Moon;Oh, Sei-Kwan;Kim, Tack-Joong;Yun, Yeo-Pyo;Hong, Jin-Tae;Okino, Nozomu;Ito, Makoto;Yoo, Hwan-Soo
    • Molecular & Cellular Toxicology
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    • v.3 no.4
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    • pp.273-281
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    • 2007
  • Ceramide is involved in cell death as a lipid mediator of stress responses. In this study, we developed an improved method of ceramide quantification based on added synthetic ceramide and thin layer chromatography (TLC) separation, and applied to biological samples. Lipids were extracted from samples spiked with N-oleoyl-D-erythro-sphingosine ($C_{17}$ ceramide) as an internal standard. Ceramide was resolved by TLC, complexed with fatty-acidfree bovine serum albumin (BSA), and deacylated by ceramidase (CDase). The released sphingosine was derivatized with o-phthalaldehyde (OPA) and measured by high performance liquid chromatography (HPLC). The limit of detection for ceramide was about 1-2 pmol and the lower limit of quantification was 5 pmol. Ceramide recovery was approximately 86-93%. Ceramide concentrations were determined in biological samples including cultured cells, mouse tissues, and mouse and human plasma. TLC separation of ceramide provides HPLC chromatogram with a clean background without any interfering peaks and the enhanced solubility of ceramide by BSAceramide complex leads to the increased deacylation of ceramide. The use of an internal standard for the determination of ceramide concentration in these samples provides an accurate and reproducible analytical method, and this method can be applicable to diverse biological samples.