• Title/Summary/Keyword: open-chain

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Cloning and Characterization of Phosphomannomutase/Phosphoglucomutase (pmm/pgm) Gene of Vibrio anguillarum Related to Synthesis of LPS (Lipopolysaccharide 생합성에 관여하는 Vibrio anguillarum의 phosphomannomutase/phosphoglucomutase 유전자 cloning과 특성)

  • Oh, Ryunkyoung;Moon, Soo Young;Cho, Hwa Jin;Jang, Won Je;Kim, Jang-Ho;Lee, Jong Min;Kong, In-Soo
    • Microbiology and Biotechnology Letters
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    • v.44 no.3
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    • pp.355-362
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    • 2016
  • The phosphomannomutase/phosphoglucomutase gene (pmm/pgm) of Vibrio anguillarum (the causative agent of fish vibriosis) was cloned, and the open reading frame corresponded to a protein with 446 amino acids. The pmm/pgm gene showed a significant degree of sequence homology with the previously reported genes from V. mimicus, V. vulnificus, V. splendidus, and V. harveyi, with 92.3%, 91.4%, 89.9%, and 89.9% amino acid identity, respectively. By reverse transcriptase-polymerase chain reaction, we found that the pmm/pgm gene was upregulated under cold stress condition. The PMM/PGM protein is known to catalyze the interconversion between mannose-1-phosphate and mannose-6-phosphate or glucose-1-phosphate and glucose-6-phosphate, which are important intermediates for lipopolysaccharide (LPS) biosynthesis. To confirm the role of PMM/PGM in the LPS biosynthetic pathway, we constructed a knock out mutant by homologous recombination. The respective LPSs were isolated from the V. anguillarum wild-type and mutant strains, and changes were compared by subjecting them to sodium dodecyl sulfate polyacrylamide gel electrophoresis. Based on the different patterns of the LPSs, we expect the pmm/pgm gene to have an important role in LPS biosynthesis. The pmm/pgm-deficient mutant of V. anguillarum will contribute to further studies about the role of LPS in V. anguillarum pathogenesis.

Expression of Monocarboxylate Transporters(MCTs) and Basigin and Estrogen Receptor α(ERα)-Mediated Regulation of MCT1 Expression in The Efferent Ductules of Male Reproductive Tract During Postnatal Development (생후 발달과정동안 남성 생식기의 Efferent Ductules에서 Monocarboxylate Transporters(MCTs)와 Basigin의 발현 양상과 에스트로젠 수용체 α에 의한 MCT1 발현 조절 연구)

  • Lee, Ki-Ho;Choi, I.H.;Chung, C.S.;Chang, J.S.
    • Journal of Animal Science and Technology
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    • v.49 no.3
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    • pp.309-320
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    • 2007
  • The purpose of the present study was to determine if monocarboxylate transporter(MCT) isoforms and Basigin(Bsg) are expressed in the efferent ductules(ED) and if MCT1 expression is under estrogen receptor(ER)α-regulation in the ED of male reproductive tract. The presence of MCT isoforms and Bsg mRNAs was detected by real-time polymerization chain reaction(PCR), and ERα-mediated regulation of MCT1 expression in the ED was indirectly determined by immuno- histochemistry. Current study found differential expression of MCT isoforms(MCT1, 2, 3, 4, and 8) and Bsg mRNAs in rat ED according to postnatal ages. In addition, comparison of MCT1 expression in the ED between wild type and ERα knockout mice at different postnatal ages showed basolateral localization of MCT1 in ciliated cells of the ED and, in part, ERα- mediated regulation of MCT1 expression. It is suggested that MCTs would play a role in regulation of function of the ED.

Construction of Mammalian Cell Expression Vector for pAcGFP-bFLIP(L) Fusion Protein and Its Expression in Follicular Granulosa Cells

  • Yang, Run Jun;Li, Wu Feng;Li, Jun Ya;Zhang, Lu Pei;Gao, Xue;Chen, Jin Bao;Xu, Shang Zhong
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.3
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    • pp.401-409
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    • 2010
  • FLICE inhibitory protein (FLIP) is one of the important anti-apoptotic proteins in the Fas/FasL apoptotic path which has death effect domains, mimicking the pro-domain of procaspase-8. To reveal the intracellular signal transduction molecules involved in the process of follicular development in the bovine ovary, we cloned the c-FLIP(L) gene in bovine ovary tissue with the reverse transcription polymerase chain reaction (RT-PCR), deleted the termination codon in its cDNA, and directionally cloned the amplified c-FLIP(L) gene into eukaryotic expression vector pAcGFP-Nl, including AcGFP, and successfully constructed the fusion protein recombinant plasmid. After identifying by restrictive enzyme BglII/EcoRI and sequencing, pAcGFP-bFLIP(L) was then transfected into follicular granulosa cells, mediated by Lipofectamine 2000, the expression of AcGFP observed and the transcription and expression of c-FLIP(L) detected by RT-PCR and Western blot. The results showed that the cattle c-FLIP(L) was successfully cloned; the pAcGFPbFLIP(L) fusion protein recombinant plasmid was successfuly constructed by introducing a BglII/EcoRI cloning site at the two ends of the c-FLIP(L) open reading frame and inserting a Kozak sequence before the start codon. AcGFP expression was detected as early as 24 h after transfection. The percentage of AcGFP positive cells reached about 65% after 24 h. A 1,483 bp transcription was amplified by RT-PCR, and a 83 kD target protein was detected by Western blot. Construction of the pAcGFP-bFLIP(L) recombinant plasmid should be helpful for further understanding the mechanism of regulation of c-FLIP(L) on bovine oocyte formation and development.

Studies on the Sedative Alkaloids from Zizyphus spinosus Semen

  • Han, Byung-Hoon;Park, Myung-Hwan;Park, Jeong-Hill
    • Korean Journal of Pharmacognosy
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    • v.16 no.4
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    • pp.233-238
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    • 1985
  • A number of sedative alkaloids were isolated from Sanjoin(酸棗仁), the seeds of Zizyphus vulgaris Lamark var. spinosus Bunge (Rhamnaceae) which is an important Chinese medicinal material used to treat insomnia and sometimes to treat sleepiness. Those compounds were designated as Sanjoinine-A, B, C, D, etc. depending on the order of increasing polarity. Sanjoinine-A, $C_{31}H_{42}N_4O_4$, $mp\;249^{\circ}$, $[{\alpha}]^{27}_D-316$, Sanjoinine-B, $C_{30}H_{40}N_4O_4$, $mp\;212{\sim}4^{\circ}$, Sanjoinene, $C_{29}H_{35}N_3O_4$, $mp\;281{\sim}2^{\circ}$, $[{\alpha}]^{22}_D-272$, Sanjoinine-D, $C_{32}H_{46}N_4O_5$, $mp\;256{\sim}8^{\circ}$, $[{\alpha}]^{22}_D-53.6$, Sanjoinine-F, $C_{31}H_{42}N_4O_5$, $mp\;228{\sim}9^{\circ}$, $[{\alpha}]^{22}_D-215$, and $Sanjoinine-G_1,\;C_{31}H_{44}N_4O_5,\;mp\;236{\sim}8^{\circ},\;[{\alpha}]^{22}_D-68.6$, were found as 14-membered cyclic peptide alkaloids, $Sanjoinine-G_2,\;C_{30}H_{42}N_4O_4,\;mp\;182^{\circ},\;[{\alpha}]^{22}_D-79.2$, as being open chain peptide alkaloid, and Sanjoinine-E, $C_{19}H_{21}NO_2$, $mp\;166^{\circ}$, $[{\alpha}]^{20}_D-146.2$, N-Methylasimilobine, $C_{18}H_{19}NO_2$, $mp\;193{\sim}5^{\circ}$, $[{\alpha}]^{20}_D-204$, Sanjoinine-Ia, $C_{18}H_{19}NO_2$, $mp\;155{\sim}7^{\circ}$, $[{\alpha}]^{20}_D-140$, Sanjoinine-Ib, $C_{19}H_{21}NO_4$, $mp\;184^{\circ}$, Sanjoinine-K, $C_{16}H_{19}NO_3$, $mp\;159{\sim}61^{\circ}$, $[{\alpha}]^{20}_D+35$, Caaverine, $C_{17}H_{17}NO_2$, $mp\;204^{\circ}$, $[{\alpha}]^{20}_D-80$, and Zizyphusine, $C_{20}H_{24}NO_4$, $mp\;214{\sim}6^{\circ}$, $[{\alpha}]^{20}_D+317$ as being aporphine alkaloids. The heat treatment of the cyclic peptide alkaloids produced their isomeric products which showed enhanced sedative activity. The chemical structure of the isomeric products will be discussed.

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Incidence of Viral Diseases on Red Pepper in Yeongyang-Gun, Gyeongbuk Provice (경북 영양지역 노지고추의 바이러스병 발생양상)

  • Kwon, Oh-Hun;Lee, Joong-Hwan;Jang, Kil-Su;Kim, Dong-Jin;Kim, Chan-Yong;Jeon, Su-Gyeong;Kwon, Jung-Bae;Jung, Hee-Young
    • Research in Plant Disease
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    • v.23 no.3
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    • pp.234-240
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    • 2017
  • Incidence of virus diseases in red pepper of open field in Yeongyang-Gun, Gyeongbuk Province was investigated by reverse transcription-polymerase chain reaction in 2012-2016. The infection rate of Cucumber mosaic virus (CMV), Broad bean wilt virus 2 (BBWV2), Pepper mottle virus, Potato virus Y, Pepper mild mottle virus and Tomato spotted wilt virus was 46.1%, 41.5%, 2.0%, 2.0%, 4.4% and 0.1% respectively. Incidence rate of single and mixed infection was 31.2% and 62.6%. Most of single infections were CMV and BBWV2. Among mixed infections, the incidence rate of CMV+BBWV2 mixed infection was the highest as 49.3% and most of mixed infections of triplex and tetraplex included CMV+BBWV2 mixed infection. CMV single infection caused mosaic, chlorosis, yellowing and vein necrosis and BBWV2 single infection induced cholosis and mosaic. CMV+BBWV2 mixed infection caused severe mosaic with chlorosis or malformation.

Clinical Analysis of Posterior Thoracic and Endoscopic Surgical Approach for Essential Hyperhidrosis (본태성 다한증의 후흉추 접근법 및 내시경수술의 임상고찰)

  • Cheon, Hyo Cheol;Kim, Jae Hyoo;Lee, Jung Kil;Kim, Tae Sun;Jung, Shin;Kim, Soo Han;Kang, Sam Suk;Lee, Je Hyuk
    • Journal of Korean Neurosurgical Society
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    • v.30 no.8
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    • pp.992-997
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    • 2001
  • Objectives : Essential hyperhidrosis is a common condition characterized by excessive body sweating. Excessive sweating beyond what is necessary to maintain normal body temperature need not be considered pathological unless it interferes with one's occupation and/or life-style. The existing non-operative therapeutic options seldom give sufficient relief or show a transient effect. In this regard, the thoracic sympathectomy may provide a definitive cure. In the past, surgical procedures were highly invasive and caused significant morbidity, but the minimally invasive thoracoscopic procedure provided detailed visualization of sympathetic ganglia and is associated with minimally postoperative morbidity. Nowadays, thoracoscopic transthoracic sympathectomy is accepted as the treatment of choice for essential hyperhidrosis. In palmar hyperhidrosis, however, the level of sympathetic chain to be blocked has been somewhat obscure. It is assumed that the incidence of compensatory hyperhidrosis may closely related to the extent of thoracic sympathectomy. Material & Methods : To compare the results of posterior midline approach with endoscopic sympathectomy, and the results of T2 with T2, 3 sympathectomy or sympathicotomy, we retrospectively studied 62 patients treated for palmar hyperhidrosis between September 1993 and May 2000. We reviewed medical records and recently interviewed the patients by telephone calls. Results : The treatment effect of T2 sympathectomy is no different from T2, 3 sympathectomy. But, the incidence of compensatory hyperhidrosis is less in the T2 sympathectomy group than the T2, 3 sympathectomy group. Conclusion : Thoracoscopic sympathectomy is considered a simple, safe, and effective method for treating palmar hyperhidrosis, with a shorter operation time, fewer hospital days, and a better cosmetic result, as compared with the open approaches. However, sympathicotomy seems to provide the advantages of a limited extent of denervation and the resultant decrease of compensatory hyperhidrosis compared to sympathectomy.

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Cloning of Coat Protein Gene from Korean Isolate Potato Leafroll Virus (PLRV) and Introduction into Potato (Solanum tuberosum) (한국 분리주 감자 잎말림 바이러스 (PLRV) 외피 단백질 유전자의 클로닝 및 감자 내 도입)

  • Seo Hyo-Won;Yi Jung-Yoon;Park Young-Eun;Cho Ji-Hong;Hahm Young-Il;Cho Hyun-Mook
    • Journal of Plant Biotechnology
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    • v.32 no.4
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    • pp.243-250
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    • 2005
  • The coat protein gene (AF296280) of the Korean isolate Potato leafroll virus (PLRV) was cloned and the open reading frame (627 bp) was transformed into potato (Solanum tuberosum cv. Superior). Out of seventeen individual transgenic lines, five lines were identified to confer resistance to PLRV through the five generation's selection program in the greenhouse as well as isolated trial field. Successful introduction and genetic stability of coat protein gene in the genome of potato were confirmed by polymerase chain reaction (PCR), Southern blot hybridization and northern blot hybridization. Some of the transgenic lines were highly resistant to PLRV but did not show any resistance to less homologous Potato virus Y (PVY). Our results suggest that the resistance to PLRV is due to homology dependent gene silencing by sense strand coat protein gene. In addition, the results of field test through five generations showed that there were no significant differences comparing to nontransgenic potatoes in the morphological aspect of shoot as well as tuber, Ho remarkable differences were also observed in the major agronomic characters and yields except for the resistance to PLRV.

Association between HSP70 Genotypes and Oocytes Development on In vitro Maturation/Fertilization in Pig

  • Wee, M.S.;Park, C.K.;Cho, S.R.;Lee, S.S.;Yeon, S.H.;Kim, C.D.;Cho, C.Y.;Choi, S.H.;Sang, B.D.;Son, D.S.;Li, Z.D.;Jin, H.J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.10
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    • pp.1404-1410
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    • 2008
  • This study was performed to clarify whether the variation of stress related heat shock protein 70 (HSP70) (GenBank X68213) gene was associated with the nuclear morphological change of in vitro maturation and in vitro capacitation in oocytes of pig ovaries obtained at the slaughterhouse. The nucleic acid substitution of C to G at the 483rd position was found out in HSP70 K1 (290-512) from X68213. The ovaries were categorized into CC, CG, and GG genotypes using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) (BsiHKA I). After the second in vitro maturation of immature fresh oocytes, the relation of nuclear morphological change in oocytes with the genotype of HSP70 K1 gene was such that the MII ratios of the genotype GG and CG (46.93% and 42.20%, respectively) were significantly higher than that of the CC genotype (10.71%) (p<0.05). With respect to in vitro maturation of frozen-thawed oocytes by an open pulled straw (OPS) method, the percentage of oocytes matured to MII stage of the CG genotype showed a higher trend than CC and GG genotypes. After the in vitro maturation of immature fresh oocytes and frozen-thawed oocytes by the OPS method, the relation of the pronuclei change in oocytes matured in vitro with HSP70 genotype was assessed, and the result showed that the enlarged sperm heads (ESH) of matured fresh oocytes and frozen-thawed oocytes were 80.0% and 60.0% in the CC genotype, respectively. The CC genotype group had a significantly higher rate of ESH than the CG and the GG genotype group (p<0.05). The ratios of polyspermic invasion were not different among HSP70 of the three genotypes. It was considered that the rate of in vitro maturation of fertilized oocytes was expected to differ according to genotype of the stress related gene.

Clinical Significance of Measuring Levels of CEA, CA19-9 in Peritoneal Washing Fluid in Patients with Gastric Cancer (위암 환자에서 복강 세척액의 CEA, CA19-9 측정의 임상적 의의)

  • Shim, Gyu-Beom;Park, Ji-Hun;Koo, Tea-Young;Min, Hyun-Sik
    • Journal of Gastric Cancer
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    • v.6 no.3
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    • pp.125-131
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    • 2006
  • Purpose: Free cancer cells exfoliated from cancer-invaded serosa contribute to peritoneal dissemination, the most frequent pattern of recurrence in patients with gastric cancer. To detect free cancer cells, CEA and CA19-9 were introduced as the markers of gastric cancer, and many methods, such as cytology, immunoassay, and reverse transcription polymerase chain reaction (RT-PCR), exist for detecting them. The aim of this study is to define the clinical significance of using immunoassay to measure the levels of CEA and CA19-9 in the peritoneal washings in patients with gastric cancer. Materials and Methods: The peritoneal washing fluids were obtained from 130 patients with gastric cancer who received a curative gastrectomy, palliative gastrectomy or open and closure. The pCEA and pCA19-9 levels were measured by using immunoassay and cytology. The results were compared with the clinicopathological data. Results: The pCEA and pCA19-9 levels were correlated with tumor invasion, lymph-node metastasis, and stage (P<0.05). Conclusion: A correlation was found between elevated pCEA and pCA19-9 levels measured by immunoassay and the TNM stage. Therefore, a combined pCEA and pCA19-9 assay could be a sensitive detector of peritoneal dissemination, as well as a predictor of postoperative prognosis. pCEA and pCA19-9 may also determine the adjuvant management strategy.

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A Study on Anti-collision Algorithm in Gen2 Protocol Based RFID Systems (Gen2기반 RFID시스템에서의 충돌방지 알고리즘에 관한 연구)

  • Quan Cheng-Hao;Mo Hee-Sook;Choi Gil-Young;Pyo Cheol-Sig;Chae Jong-Suk
    • The Journal of Korean Institute of Communications and Information Sciences
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    • v.31 no.6B
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    • pp.561-571
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    • 2006
  • RFID technology, as one contact-less identification technology with using electronic tag, is the field where its multiple researches have been studied. Especially, in the field of supply chain management, when it is necessary to identify a lot of objects by real-time, performance of anti-collision algorithm for multiple tag identification has an effect on performance of entire system. GEN2 is global, open, interoperability protocol for UHF RFID system, the active researches and developments are processed about it. In this paper, we introduce an anti-collision algorithm based on Slotted-ALOHA for GEN2 protocol, focus on estimating number of tags and setting number of slots and review the previous works. We propose new scheme that can estimate number of tags efficiently, and define system efficiency newly when it's used in setting number of slots. We also present result of compare and analysis with previous scheme through simulation. The proposed scheme of estimating number of tags is easy to implement, number of tags which is less than 4 times of number of slots can estimate tags efficiently, we also proved that identification efficiency based on weighted-slot distinguished with definition of previous slot efficiency can improve system efficiency.