• Title/Summary/Keyword: Cag

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Highly Efficient Biotransformation of Astragaloside IV to Cycloastragenol by Sugar-Stimulated β-Glucosidase and β-Xylosidase from Dictyoglomus thermophilum

  • Li, Qi;Wu, Tao;Zhao, Linguo;Pei, Jianjun;Wang, Zhenzhong;Xiao, Wei
    • Journal of Microbiology and Biotechnology
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    • v.29 no.12
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    • pp.1882-1893
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    • 2019
  • β-Glucosidases and β-xylosidases are two categories of enzymes that could cleave out non-reducing, terminal β-D-glucosyl and β-D-xylosyl residues with release of D-glucose and D-xylose, respectively. In this paper, two functional β-glucosidase Dth3 and β-xylosidase Xln-DT from Dictyoglomus thermophilum were heterologously expressed in E.coli BL21 (DE3). Dth3 and Xln-DT were relatively stable at 75℃ and were tolerant or even stimulated by glucose and xylose. Dth3 was highly tolerant to glucose with a Ki value of approximately 3 M. Meanwhile, it was not affected by xylose in high concentration. The activity of Xln-DT was stimulated 2.13-fold by 1 M glucose and 1.29-fold by 0.3 M xylose, respectively. Furthermore, the βglucosidase Dth3 and β-xylosidase Xln-DT showed excellent selectivity to cleave the outer C-6 and C-3 sugar moieties of ASI, which established an effective and green method to produce the more pharmacologically active CAG, an exclusive telomerase activator. We measured temperature, pH and dosage of enzyme using a single-factor experiment in ASI biotransformation. After optimization, the optimal reaction conditions were as follows: 75℃, pH 5.5, 1 U of Dth3 and 0.2 U of Xln-DT, respectively. Under the optimized conditions, 1 g/l ASI was transformed into 0.63 g/l CAG with a corresponding molar conversion of 94.5% within 3 h. This is the first report to use the purified thermostable and sugar-tolerant enzymes from Dictyoglomus thermophilum to hydrolyze ASI synergistically, which provides a specific, environment-friendly and cost-effective way to produce CAG.

In Vitro Anti-Helicobacter pylori Activity of Ethanol Extract of Sohamhyoongtang and Coptidis Rhizoma Total Alkaloids (소함흉탕 에탄올 추출물 및 황련 알칼로이드의 헬리코박터 파이로리에 대한 항균활성)

  • Lee, BaWool;Choi, MyungSook;Yim, DongSool;Choi, SungSook
    • Korean Journal of Pharmacognosy
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    • v.45 no.2
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    • pp.168-173
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    • 2014
  • The aim of this study was to evaluate the anti-helicobacter activity of the ethanol extract of Sohamhyoongtang (Coptidis Rhizoma, Pinelliae Tuber and Trichosanthis Semen) and Coptidis Rhizoma total alkaloids, which is one of the components of Sohamhyoongtang. Crude ethanol extract of Sohamhyoongtang (ESHHT) and Coptidis Rhizoma total alkaloids (CRTA) were used for this experiment. Five different types of H. pylori (including H. pylori 26695) were used as test strain. To determine anti-helicobacter activity, minimum inhibitory concentration (MIC) was determined by agar dilution method. The effect of ESHHT and CRTA on the gene expression of H. pylori was investigated by quantitative realtime-PCR (qRT-PCR). MICs of ESHHT against five H. pylori strains were $250{\sim}500{\mu}g/ml$ and MICs of CRTA against five H. pylori strains were $50{\sim}200{\mu}g/ml$. Four representative virulence genes of H. pylori, cagA, ureA, ureB and ureI were tested as target genes for qRT-PCR. According to the qRT-PCR results, both ESHHT and CRTA markedly repressed the expression of cagA gene of H. pylori 26695 (6.91 and 20 folds respecively). These results showed that the ESHHT and CRTA demonstrated antihelicobacter properties, suggesting their potential use in gastritis or duodenal ulcer.

Porcine tissue factor promoter induces specific expression of target gene in response to human serum in porcine aortic endothelial cells

  • No, Jin Gu;Lee, Haesun;Lee, Na Hyun;Sun, Wu-Sheng;Hwang, In-Sul;Lee, Minguk;Oh, Keon Bong
    • Korean Journal of Agricultural Science
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    • v.48 no.2
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    • pp.353-365
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    • 2021
  • Previously, we reported that tissue factor (Tf) was included in the list of differentially expressed genes as an upregulated gene in a rejected porcine heart after xenotransplantation into monkey. In this study, we analyzed that expression of Tf in aortic endothelial cells (pAEC) isolated from alpha 1,3-galactosyltransferase knockout pig in response to allogeneic porcine serum and xenogeneic human serum. The consequence was significant upregulation of Tf expression by responding to human serum compared with porcine serum. To analyze the function of Tf gene as a promoter, we constructed reporter vectors for expression of luciferase linked to 1,246 and 787 base pairs of porcine Tf (pTF1246 and pTF787), and 535 base pairs of human TF (hTF535) sequences including putative promoter regions and AP-1 biding site at the 5' end. The reporter vectors were transfected into pAEC including cytomegalovirus enhancer/chicken β-actin (CAG)-luciferase vector as a control. Luciferase assay showed that all of the promoters were insufficient to express luciferase compared with CAG promoter in basic culture conditions. Notably, pTF1246, pTF787, and hTF535 led to a significant increase of luciferase expression in response to human serum compared with porcine serum while no change of CAG. pTF1246 and pTF787 showed higher expression than hTF535. Taken together, our findings suggest that pTF1246 and pTF787 promoters could mediate target gene expression specifically at xenogeneic stress condition.

Expression of lewis antigen in gastric mucosa of children with Helicobacter pylori infection (Helicobacter pylori에 감염된 소아의 위점막에서 Lewis 항원의 발현)

  • Chung, Ju-Young;Lim, Seong Jig;Han, Tae Hee
    • Pediatric Infection and Vaccine
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    • v.14 no.1
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    • pp.97-103
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    • 2007
  • Purpose : Lewis antigen has been known to have a role in the attachment of H. pylori to the gastric mucosa, but its expression pattern in children with H. pylori infection is still unclear. The recently described blood group antigen-binding adhesin BabA is known to mediate adherence of H. pylori to Lewis B receptors on gastric epithelium. We investigated the expression of Lewis antigen in gastric mucosa of Korean children with H. pylori infection. Methods : The expression of Lewis A ($Le^a$), B ($Le^b$), X ($Le^x$), and Y ($Le^y$) was evaluated by immunohistochemistry in H. pylori positive biopsy specimens from 35 children (antral gastritis in 30, peptic ulcer in 5) and in H. pylori negative specimens from 19 children. PCR assays for cagA and babA2 gene of H. pylori were performed. Results : We confirmed the expression of $Le^a$ in 60%, $Le^b$ in 97%, $Le^x$ in 100%, and $Le^y$ in 100% of the superficial epithelium of the 35 H. pylori positive children. In H. pylori negative patients, $Le^a$, $Le^b$, $Le^x$, and $Le^y$ expression was 52%, 100%, 89%, and 100%, respectively. The cagA gene was detected in 65% and babA2 gene in 25% of 35 patients. No differences in neutrophil activity and chronic inflammation were found according to the presence of cagA and babA2 genes in H. pylori. Conclusion : $Le^b$, $Le^x$ and $Le^y$ antigen were highly expressed in gastric mucosa of Korean children, but they were not associated with the status of H. pylori infection and the positivity of babA2 gene. Further studies for other mucosal receptors and toxins are needed to define the immune responses to H. pylori infection in gastric mucosa of Korean children.

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Comparison of Moxifloxacin Monotherapy versus Cephalosporin-Azithromycin Combination Therapies for the Treatment of Community Acquired Pneumonia (원외획득폐렴 환자 치료에서 Moxifloxacin 단독요법과 Cephalosporin-Azithromycin 병용요법의 비교)

  • Cheong, Eun-Jin;Lee, Suk-Hyang
    • Korean Journal of Clinical Pharmacy
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    • v.15 no.2
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    • pp.75-81
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    • 2005
  • Community acquired pneumonia (CAP) remains a prevalent and potentially life threatening illness. American Thoracic Society and Infectious Disease Society America recommend combination therapies with ${\beta}-lactam$ plus a macrolide or a fluoroquinolone monotherapy for the empirical treatment of CAP. The aim of this study was to compare moxifloxacin monotherapy with cephalosporin plus azithromycin combination therapies. From January 2004 to March 2005, 18 patients in the moxifloxacin group(MG) and 21 patients in the cefuroxime or ceftriaxone plus azithromycin group(CAG) with CAP were retrospectively reviewed with regard to clinical, laboratory and microbiological data. Each patient was stratified into mild (risk class I-II), moderate (risk class III) and severe (risk class VI, V) group according to and PSI (Pneumonia Severity Index) score. Each group was compared for microbiological eradication, clinical assessment, the length of hospital stay. As results, Total 39 patients with CAP were reviewed. The appropriateness of admission was 83.3% in MC vs. 76.2% in CAC. The mean length of the hospital day was for 8.31 days vs. 7.39 days, days switching parenteral to oral antibiotics in 5.19 days vs. 5.28 days, clinical improvement in 2.43 days vs. 2.61 days in MG vs. CAC. Radiological improvement required 3.75 days vs 3.63 days in MG vs. CAG and bacteriological eradication rate at discharge was the same in the both groups. Mortality rate was 11.1% (2 of 18) vs 14.3% (3 of 21) in MG vs. CAG (p=0.77). Drug cost of the mean 5 hospital days requiring parenteral antibiotics was the most inexpensive in moxifloxacin group for the 147,045 won, and ceftriaxone 1g-azithromycin group for the 170,285 won, cefuroxime bid-azithromycin group for the 207,800 won, ceftriaxone 2g-azithromycin group far the 220,570 won, cefuroxime tid-azithromycin group for the 251,700 won. There was no significant statistical difference in clinical, bacterial, radiological cure and hospital days, and switch to oral days. In conclusion, that i.v. moxifloxacin monotherapy was as effective as azithromycin plus cefuroxime or ceftriaxone combination therapies fur the treatment of CAP. In drug cost analysis, moxifloxacin is less expensive than CAG.

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CAgM, USDA and the National Drought Policy Commission Associated with WAMIS (농업기상웹서버관련 농업기상위원회, 농무성 및 한발정책위원회 현황)

  • Motha, Raymond P.
    • Korean Journal of Agricultural and Forest Meteorology
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    • v.6 no.2
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    • pp.140-147
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    • 2004
  • Agrometeorological information is essential in many agricultural decisions if it reaches the user in a timely and appropriate manner. Agriculture is the backbone to local, regional, and global economic development. Thus, strengthening agrometeorological application to diverse agricultural sectors will benefit economic development. This paper discusses three distinct organizational minions that all share the same need for improved information technology. The World Meteorological Organization's (WMOs) Commission for Agricultural Meteorology (CAgM) has global responsibility for improved agrometeorological services of Members to aid agricultural production and to conserve natural resources. The United States Department of Agriculture, World Agricultural Outlook Board, publishes monthly World Agricultural Supply and Demand Estimates, considered to be a benchmark for both government and industry in production and trade decisions. The National Drought Policy Commission (NDPC), created by an act of the United States Congress, formulated a national drought policy based on preparedness rather than on crisis management. All three organizations recognize the need for IT applications in agricultural meteorology and have been active in implementing this technology. The development of information technology offers new means of dissemination of agrometeorological products. World Agrometeorological Information Service (WAMIS) has taken advantage of the global Internet application to offer WMO Members a dedicated web server to host agrometeorological bulletins and training modules.

Spinocerebellar ataxia 7 (SCA7) (척수소뇌성 운동실조증 제7형)

  • Seon-Yong, Jeong;Seok-Hun, Jang;Hyon-J., Kim
    • Journal of Genetic Medicine
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    • v.4 no.1
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    • pp.22-37
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    • 2007
  • The autosomal dominant spinocerebellar ataxias (SCAs) are a group of neurodegenerative diseases, clinically and genetically heterogeneous, characterized by degeneration of spinocerebellar pathways with variable involvement of other neural systems. At present, 27 distinct genetic forms of SCAs are known: SCA1-8, SCA10-21, SCA23, SCA25-28, DRPLA (dentatorubral-pallidoluysian atrophy), and 16q-liked ADCA (autosomal dominant cerebellar ataxia). Epidemiological data about the prevalence of SCAs are restricted to a few studies of isolated geographical regions, and most do not reflect the real occurrence of the disease. In general a prevalence of about 0.3-2 cases per 100,000 people is assumed. As SCA are highly heterogeneous, the prevalence of specific subtypes varies between different ethnic and continental populations. Most recent data suggest that SCA3 is the commonest subtype worldwide; SCA1, SCA2, SCA6, SCA7, and SCA8 have a prevalence of over 2%, and the remaining SCAs are thought to be rare (prevalence <1%). In this review, we highlight and discuss the SCA7. The hallmark of SCA7 is the association of hereditary ataxia and visual loss caused by pigmentary macular degeneration. Visual failure is progressive, bilateral and symmetrical, and leads irreversibly to blindness. This association represents a distinct disease entity classified as autosomal dominant cerebellar ataxia (ADCA) type II by Harding. The disease affectsprimarily the cerebellum and the retina by the moderate to severe neuronal loss and gliosis, but also many other central nervous system structures as the disease progresses. SCA7 is caused by expansion of an unstable trinucleotide CAG repeat in the ATXN7 gene encoding a polyglutamine (polyQ) tract in the corresponding protein, ataxin-7. Normal ATXN7 alleles contain 4-35 CAG repeats, whereas pathological alleles contain from 36->450 CAG repeats. Immunoblott analysis demonstrated that ataxin-7 is widely expressed but that expression levels vary among tissues. Instability of expanded repeats is more pronounced in SCA7 than in other SCA subtypes and can cause substantial lowering of age at onset in successive generations termed ‘anticipation’ so that children may become diseased even before their parents develop symptoms. The strong anticipation in SCA7 and the rarity of contractions should have led to its extinction within a few generations. There is no specific drug therapy for this neurodegenerative disorder. Currently, therapy remains purely symptomatic. Cellular models and SCA7 transgenic mice have been generated which constitute valuable resources for studying the disease mechanism. Understanding the pathogenetic mechanisms of neurodegeneration in SCAs should lead to the identification of potential therapeutic targets and ultimately facilitate drug discovery. Here we summarize the clinical, pathological, and genetic aspects of SCA7, and review the current understanding of the pathogenesis of this disorder. Further, we also review the potential therapeutic strategies that are currently being explored in polyglutamine diseases.

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Midterm Patency after Off-Pump Coronary Artery Bypass Grafting (심폐바이패스없이 심박동상태에서 시행한 관상동맥우회술후 중기 개통율)

  • Lee, Cheul;Chang, Woo-Ik;Lim, Cheong;Kim, Ki-Bong;Chae, In-Ho;Oh, Byung-Hee;Lee, Myoung-Mook;Park, Young-Bae
    • Journal of Chest Surgery
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    • v.34 no.8
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    • pp.583-590
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    • 2001
  • The aim of this study was to compare one-year graft patency after coronary artery bypass grafting without cardiopulmonary bypass(OPCAB) with that of conventional CABG and that of on-pump beating CABG, and to demonstrate any differences in patency of various conduits among the three groups. Material and Method: We analyzed the results of OPCAB cases(group I; n=122) compared with those of conventional CABG cases(group II; n=65) and those of on-pump beating CABG cases(group III; n=19). In group I, coronary angiography(CAG) was performed immediately postoperatively and 1 year after surgery. In group II and III, CAG was performed 1 year after surgery. Graft patency was graded as grade A(excellent), grade B(fair), or grade O(occluded). Result: The average number of distal anastomoses in groups I, II, and II were 3.1$\pm$1.1, 3.7$\pm$0.9, and 3.6$\pm$0.9, respectively. In group I, postoperative CAG was performed in 92%(112/122) of patients before discharge. The patency rate(grade A+B) was 96.4$(162/168) for arterial grafts, and 85.6%(160/187) for saphenous vein grafts(SVG). One-year follow-up CAG was performed in 74%(90/122) of patients. The patency rate was 97.8%(132/135) for arterial grafts, and 67.9%(106/156) for SVG. In group II, one-year follow-up CAG was performed in 65%(42/65) of patients. The patency rate(grade A+B) was 93.5%(43/46) for arterial grafts, and 86.8%(33/38) for SVG. Conclusion: Our results demonstrated that the patency rate of SVG after OPCAB was significantly lower than that of arterial grafts in the early postoperative CAG(p<0.001), and was also significantly lower than those of SVG of group II(p<0.001) and group III(p<0.01) in the postoperative one-year CAG, although there was no significant difference is one-year patency of arterial grafts among the three groups. Our data suggest that a specific perioperative anticoagulant therapy may be advisable in patients undergoing OPCAB with SVG.

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Internet-based RAMINS II as a Future Communication Framework for AgroMeteorological Information in Asia (아시아 지역 농업기상정보 공유를 위한 인터넷기반 기상정보 연동시스템)

  • Byong-Lyol Lee;G. Ali Kamali;Wang Shili
    • Korean Journal of Agricultural and Forest Meteorology
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    • v.4 no.2
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    • pp.127-132
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    • 2002
  • All the countries in RA II (Asia Region in WMO) welcome the establishment of a Web site dedicated to agricultural meteorology, because it is believed that the best way to improve and speed up the flow of information is the use of the Internet and the establishment of a Web site. In providing recommendation for the promotion and improvement of the AgroMeteorological service in RA II, a couple of key suggestions were proposed: (a) Exchanges of data and AgroMeteorological knowledge between member countries and between RAs, (b) Exchanges of experts between member countries as a necessary way to share the knowledge, and (c) Joint research between member countries to solve common problems in AgroMeteorological affairs. In order to meet the above requirements for RA II, an AgroMeteorological information network will be the most critical and dynamic aspect in sustainable agriculture in this region. In addition, the establishment of a Core AgroMeteorological station, recommended by CAgM of WMO, will require its own information sharing systems for communication among member countries. Inevitable use of information technologies (IT) such as information networks, databases, simulation models, GIS, and RS for regional impact assessment of environmental change on AgroEcosystem will be enforced. Thus, the regional Internet-based Agrometeorological information network has been in place since 1999, though all contributions to it have been volunteered by individuals, institutes, universities, etc.

Identification of AFLP Marker Linked to a SCN Resistant Gene in Soybean

  • Ko, Mi-Suk;Kim, Myung-Sik;Han, Soung-Jin;Chung, Jong-Il;Kang, Jin-Ho
    • Plant Resources
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    • v.5 no.3
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    • pp.169-175
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    • 2002
  • The soybean cyst nematode (Heterodera glycines Inchinoe; SCN) is a devastating pest of soybean and is responsible for significant losses in yield. The use of resistant cultivars is the effective method to reduce or eliminate SCN damage. The objective of this research is to identify AFLP markers linked to the SCN resistant genes. Bulked genomic DNA was made from resistant and susceptible genotypes to SCN and a total of 19 primer combinations were used. About 31 fragments were detected per primer combination. The banding patterns were readily distinguished in resistant and susceptible bulked genotypes. Polymorphic fragments were detected between resistant and susceptible bulked genotypes in the primer combination of CGT/GGC, CAG/GTG and CTC/GAG. In primer combinations of CGT/GGC and CAG/GTG, bulked resistant genotype produced a polymorphic bands. However, in primer of CTC/GAG, bulked susceptible genotype produced a polymorphic fragments. Three AFLP markers identified as a polymorphic fragments between bulked genomic DNA were mapped in 85 F2 population. Among them, only two markers, CGT/GGC and CTC/GAG, was linked and was mapped. Broad application of AFLP marker would be possible for improving resistant cultivars to SCN.

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