• Title/Summary/Keyword: Techniques

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Multiplex PCR method for environmental monitoring of approved LM cotton events in Korea (국내 승인 LM면화의 자연환경 모니터링을 위한 multiplex PCR 개발)

  • Jo, Beom-Ho;Seol, Min-A;Shin, Su Young;Kim, Il Ryong;Choi, Wonkyun;Eum, Soon-Jae;Song, Hae-Ryong;Lee, Jung Ro
    • Journal of Plant Biotechnology
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    • v.43 no.1
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    • pp.91-98
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    • 2016
  • The growth area of living modified (LM) cotton has steadily increased every year, since its first commercialization in 1996. Development of environmental risk assessment tools and techniques for LM cotton is required for ecosystem safety. We therefore developed multiplex PCR assays for simultaneous detection of two (MON15985, MON531) and four (GHB614, LLCOTTON25, MON88913 and MON1445) LM cotton events approved in Korea, with event specific primer pairs. The PCR reactions were optimized by using event specific primers of six LM cottons at various concentrations. The reactions allows amplification of estimated amplicons of MON15985 (214 bp), MON531 (270 bp), GHB614 (119 bp), LLCOTTON25 (164 bp), MON88913 (276 bp), and MON1445 (389 bp) from multiplex PCR reactions. The multiplex PCR assay developed allowed that two annealing steps (15 cycles at $55^{\circ}C$ and 25 cycles at $60^{\circ}C$) were performed for amplification of distinguished two LM cottons, and only one annealing step (50 cycles at $60^{\circ}C$) was necessary for tetraplex PCR. Primer extension step of all PCR reactions was skipped for time-effective amplification. Our methods suggest that two multiplex PCR assays can be cost-effective and a rapid diagnostic tool for environmental LMO monitoring of six LM cottons.

Clinical Analysis of Recurrent Intussusception and the Pathologic Lead Point in a Single Center (단일 병원에서의 재발성 장중첩증과 병적 선두에 대한 임상적 고찰)

  • Lee, Kun-Song;Park, Yun-Joon
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.12 no.2
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    • pp.163-170
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    • 2009
  • Purpose: Intussusception is the most common cause of intestinal obstruction in young children. Although intussusceptions are easily treated, some intussusceptions with or without a pathologic lead point (PLP) often recur. In this study, we analyzed the clinical characteristics and prognosis of recurrent intussusceptions (RI), the frequency of the PLP, and correlation between RI with PLP. Methods: The medical records of 144 patients, among 590 patients with intussusceptions who had been admitted to the Department of Pediatrics and Pediatric Surgery of Dankook University Hospital between May 1994 and June 2009 were reviewed retrospectively. Results: The overall recurrence rate of intussusceptions in this study was 9.2%. The mean interval between the initial occurrence and the first recurrent attack was 130${\pm}$175 days (range, 12 hours to 3 years). There was no statistically significant difference in the recurrence rate among patients who underwent air, barium, and manual reduction (p=0.131). Eighty-seven cases (92.6%) of RI had a successful reduction by the use of non-operative techniques. A PLP was present in 18 patients (3.0%). The most common PLP was intestinal lymphoid hyperplasia, followed by Meckel's diverticulum, duplication cyst, intestinal polyp, and adenomyoma. The mean number of intussusceptions was 4.7${\pm}$1.9 in 7 patients with PLP, which was significantly higher than (2.4${\pm}$0.9) patients without a PLP (p=0.023). The mean duration of recurrences was 17.4${\pm}$19.8 months (range, 2 days to 72 months). Conclusion: A careful search for a PLP should be performed to prevent recurrence of intussusception, especially when intussusception has recurred more than three times.

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Composite valve graft Replacement of the Aortic Root (Composite valve graft를 이용한 대동맥근부 치환술)

  • 백만종;나찬영;김웅한;오삼세;김수철
    • Journal of Chest Surgery
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    • v.35 no.2
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    • pp.102-112
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    • 2002
  • This study was undertaken to analyze the outcome of composite valve graftreplacement(CVGR) for the treatment of aneurysms of the ascending aorta involving the aortic root. Material and Method: Between April 1995 and June 2001, 56 patients had replacement of the ascending aorta and aortic root with a composite graft valve and were reviewed retrospectively. Aortic regurgitation was present in 50 patients(89%), Marfan's syndrome in 18 patients(32%), and bicuspid aortic valve in 7(12.5%). The indications for operation were annuloaortic ectasia(AAE) in 30 patients(53.6%), aortic dissection in 13(23.2%), aneurysms of the ascending aorta involving aortic root in 11(19.6%), and aortitis in 2(3.6%). Cardiogenic shock due to the aortic rupture was present in 2 patients. Nine patients(16%) had previous operations on the ascending aorta or open heart surgery. The operative techniques used for CVGR were the aortic button technique in 51 patients(91%), the modified Cabrol technique in 4, and the classic Bentall technique in 1. The concomitant procedures were aortic arch replacement in 24 patients(43%), coronary artery bypass graft in 8(14.3%), mitral valve repair in 2, redo mitral valve replacement in 1, and the others in 7 The mean time of circulatory arrest, total bypass, and aortic crossclamp were 21$\pm$14 minutes, 186$\pm$68 minutes, and 132$\pm$42 minutes, respectively. Result: Early mortality was 1.8%(1/56). The postoperative complications were left ventricular dysfunction in 16 patients(28.6%), reoperation for bleeding in 7(12.5%), pericardial effusion in 2, and the others in 7. Fifty-three patients out of 55 hospital survivors were followed up for a mean of 23.2 $\pm$ 18.7 months(1-75 months). There were two late deaths(3.8%) including one death due to the traumatic cerebral hemorrhage, and CVGR-related late mortality was 1.9%. The 1- and 6-year actuarial survival was 98.1$\pm$1.9% and 93.2$\pm$5.1%, respectively. Two patients required reoperation for complication of CYGR(3.8%) and two other patients required subsequent operations for dissection of the remaining thoracoabdominal aorta. The 1- and 6-year actuarial freedom from reoperation was 97.8$\pm$2.0% and 65.3$\pm$26.7%, respectively.

Effect of Various Organic Materials on Weed Control in Environment-friendly Rice Paddy Fields (벼 친환경재배에서 다양한 유기자원별 잡초방제효과)

  • Kwon, Oh-Do;Park, Heung-Gyu;An, Kyu-Nam;Lee, Yeen;Shin, Seo-Ho;Shin, Gil-Ho;Shin, Hae-Ryoung;Kuk, Yong-In
    • Korean Journal of Weed Science
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    • v.30 no.3
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    • pp.272-281
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    • 2010
  • The objective of this research was to find out the weed management techniques in environment-friendly rice paddy fields through the study on herbicidal effects and problems of various organic materials. This experiment was conducted under different conditions of weed species and weed densities in environment-friendly rice paddy fields. There was no difference in weedy efficacy on golden apple snail (GAS), paper mulching (PM), and machine weeding (MW) between low and high weed densities. However, the effect of weed control in rice bran (RB) and effective microorganism (EM) + molasses was higher in high weed density than in low weed density. In general, the effect of weed control as affected by various organic materials was in the order of GAS (97-100%) > PM (93-98%) > RB (15-80%) > EM (7-31%). GAS provided excellent control of all weed species tested except for Persicaria hydropiper. PM gave acceptable control of the weed species except for Echinochloa crus-galli, Ludwigia prostrata, and Eleocharis kuroguwai. However, MW gave fair control (70% biomass reduction) of all weed species tested. BR followed by MW or EM followed by MW treatments had similar effect on weed control compared to each treatment alone. However, BR followed by GAS or EM followed by GAS provided 100% control of weed species tested. The level of rice foliar injury caused by various organic materials was in the order of GAS and MW (10-20%) > RB (10-15%) > PM and EM (5-7%). Typical symptoms of organic materials are wilting, inhibition of growth, missing hill, and reduction of tiller. Cost for weed control of GAS, RB, EM, and PM were 2.1, 3.1, 2.3, and 13.2 times higher than that of the herbicide. These data indicate that GAS was the best method for weed management in environmentfriendly rice paddy fields. Further study is required to elucidate the mechanisms underlying the rice injury as affected by GAS.

Anti-inflammatory Effects of Purpurogallin Carboxylic Acid, An Oxidation Product of Gallic Acid in Fermented Tea (발효차중의 미량 성분인 gallic acid 산화물 purpurogallin carboxylic acid의 항염증 효과)

  • Jhoo, Jin-Woo
    • Korean Journal of Food Science and Technology
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    • v.40 no.6
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    • pp.707-711
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    • 2008
  • The principal objective of the current study was to isolate a purpurogallin derivative as an oxidation product from gallic acid, in an effort to assess the anti-inflammatory effects of this compound. Purpurogallin derivative is known to be the one of the oxidation products of gallic acid. This compound has been identified as a minor chemical component in fermented tea products. It has been previously demonstrated that theaflavins, the oxidation products of catechins found in fermented tea products, exert profound antioxidant and anti-inflammatory effects. However, the biological activities of a minor chemical component in fermented teas have yet to be evaluated. Purpurogallin carboxylic acid (PCA) was identified as a major oxidation product of gallic acid from a peroxidase/hydrogen peroxide oxidation model system. The identity of the PCA was verified by $^{1}H$ NMR, $^{13}C$ NMR and MS techniques. PCA treatment significantly suppressed the generation of pro-inflammatory mediators including nitric oxide and IL-6 in lipopolysaccharide (LPS)-stimulated RAW264.7 murine macrophages. According to the nitrite assay, PCA 100, 75, and $50{\mu}g/mL$ treatment dose-dependently inhibited NO production by 57.6, 41.5, and 21.8%, respectively, in LPS-stimulated RAW264.7 murine macrophage cells. Moreover, IL-6 production was inhibited to a significant degree with PCA treatment of 100 and $75{\mu}g/mL$ at 43.1 and 23.9%, respectively. PCA treatment also significantly suppressed $PGE_2$ production at levels of 100 and $75{\mu}g/mL$. These results showed that PCA exerts inhibitory effects on the production of inflammatory mediators.

Growth Characteristics and Productivity of Spring Sowing Time and Mixed Sowing of Winter Annual Forage Crops (동계 사료작물 봄 파종 시기와 혼파에 따른 생육특성 및 수량 평가)

  • Nam, Cheol Hwan;Kim, Ki Soo;Park, Man Ho;Yun, An A;Park, Jong Ho;Han, Ouk Kyu;Kim, Won Ho;Sun, Sang Soo
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.39 no.4
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    • pp.207-215
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    • 2019
  • This study was conducted to establish spring sowing techniques in preparation for the impacts of climate change on sowing time and wintering rates of winter forage crops such as barley, oat and IRG. Oat showed the highest yield in 2017 which had relatively dry climate condition. And when sowing in late Febrnary 2017 yielding 9,408kg/ha were obtained, yielding 4,011kg/ha more than IRG's sown in the same period. In 2018 which had relatively wet climate condition, four barley species decreased in the production from the previous year. Oat also had decreased by 70% from 9,408kg/ha to 2,851kg/ha. On the other hand, IRG maintained the production in the mid-5,000kg/ha range. It was also found that IRG had the least variability due to external influences regardless of seeding period for 2 years. Mixed sowing with IRG and oat in 50:50 ratio was the highest dry matter, 6,584kg/ha, and IRG was 18.5% and Oat was 2.3 times higher than single planting.

Isolation and Structure Identification of Photosensitizer from Perilla frutescens Leaves Which Induces Apoptosis in U937 (들깻잎(Perilla frutescens)으로부터 U937 세포에 apoptosis를 유도하는 광과민성 물질의 분리 및 구조동정)

  • Ha, Jun Young;Kim, Mi Kyeong;Lee, Jun Young;Choi, Eun Bi;Hong, Chang Oh;Lee, Byong Won;Bae, Chang Hwan;Kim, Keun Ki
    • Journal of Life Science
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    • v.25 no.1
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    • pp.53-61
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    • 2015
  • In this study, we tried to separate the photosensitizer that induces apoptosis of leukemia cells (U937) from perilla leaves. Perilla leaves (Perilla frutescens Britt var. japonica Hara) are a popular vegetable in Korea, being rich in vitamins (A and E), GABA, and minerals. Dried perilla leaves were extracted with methanol to separate the photosensitizer by various chromatographic techniques. The structure of the isolated compound (PL9443) was identified by 1D-NMR, 2D-NMR, and FAB-mass spectroscopy. Absorbance of the UV-Vis spectrum was highest at 410 nm and was confirmed by the 330, 410, and 668 nm. PL9443 compound was determined to be pheophorbide, an ethyl ester having a molecular weight of 620. It was identified as a derivative compound of pheophorbide structure when magnesium comes away from a porphyrin ring. Observation of morphological changes in U937 cells following cell death induced by treated PL9443 compound revealed representative phenomena of apoptosis only in light irradiation conditions (apoptotic body, vesicle formation). Results from examining the cytotoxicity of PL9443 substance against U937 cells showed that inhibition rates of the cell growth were 99.9% with the concentration of 0.32 nM PL9443. Also, the caspase-3/7 activity was 99% against U937 cells with the concentration of 0.08 nM of PL9443 substance. The result of the electrophoresis was that a DNA ladder was formed by the PL9443. The PL9443 compound is a promising lead compound as a photosensitizer for photodynamic therapy of cancer.

Development of Broad-range and Specific 16S rRNA PCR for Use in Routine Diagnostic Clinical Microbiology (임상미생물 검출을 위한 광대한 범위와 특이도를 가지는 16S rRNA PCR법 개발)

  • Kim, Hyun-Chul;Kim, Yun-Tae;Kim, Hyogyeong;Lee, Sanghoo;Lee, Kyoung-Ryul;Kim, Young-Jin
    • Journal of Life Science
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    • v.24 no.4
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    • pp.361-369
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    • 2014
  • Broad-range and specific 16S rRNA gene PCR is used for detection and identification of bacterial pathogens in clinical specimens from patients with a high suspicion for infection. We describe the development of a broad-range and specific PCR primer, based on bacterial 16S rRNA, for use in routine diagnostic clinical microbiology services. The primers were designed by using conservative regions of 16S rRNA sequences from 10 strains. Ninety-eight clinical strains were isolated from clinical patient specimens. A total of 98 strains of bacteria were identified by phenotypic methods; PCR with newly designed primers and universal primers. All purified PCR products were sequenced using both forward and reverse primers on an automated DNA analyzer. In this study, we evaluated the usefulness of the newly designed primers and the universal primers for the detection of bacteria, and both these techniques were compared with phenotypic methods for bacteria detection. When we also tested 98 strains of clinical isolates with newly designed primers, about 778 bp DNA fragments were amplified and identified from all strains. Of the 98 strains, 94 strains (95.9%) correspond in comparison with phenotypic methods. The newly designed primers showed that the identities of 98 (100%) strains were the same as those obtained by universal PCR primers. The overall agreement between the newly designed primers and universal primers was 100%. The primer set was designed for rapid, accurate, and cheap identification of bacterial pathogens. We think the newly designed primer set is useful for the identification of pathogenic bacteria.

Introduction of Denitrification Method for Nitrogen and Oxygen Stable Isotopes (δ15N-NO3 and δ18O-NO3) in Nitrate and Case Study for Tracing Nitrogen Source (탈질미생물을 이용한 질산성 질소의 산소 및 질소 동위원소 분석법 소개)

  • Lim, Bo-La;Kim, Min-Seob;Yoon, Suk-Hee;Park, Jaeseon;Park, Hyunwoo;Chung, Hyen-Mi;Choi, Jong-Woo
    • Korean Journal of Ecology and Environment
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    • v.50 no.4
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    • pp.459-469
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    • 2017
  • Nitrogen (N) loading from domestic, agricultural and industrial sources can lead to excessive growth of macrophytes or phytoplankton in aquatic environment. Many studies have used stable isotope ratios to identify anthropogenic nitrogen in aquatic systems as a useful method for studying nitrogen cycle. In this study to evaluate the precision and accuracy of denitrification bacteria method (Pseudomonas chlororaphis ssp. Aureofaciens ($ATCC^{(R)}$ 13985)), three reference (IAEA-NO-3 (Potassium nitrate $KNO_3$), USGS34 (Potassium nitrate $KNO_3$), USGS35 (Sodium nitrate $KNO_3$)) were analyzed 5 times repeatedly. Measured the ${\delta}^{15}N-NO_3$ and ${\delta}^{18}O-NO_3$ values of IAEA-NO-3, USGS 34 and USGS35 were ${\delta}^{15}N:4.7{\pm}0.1$${\delta}^{18}O:25.6{\pm}0.5$‰, ${\delta}^{15}N:-1.8{\pm}0.1$${\delta}^{18}O:-27.8{\pm}0.4$‰, and ${\delta}^{15}N:2.7{\pm}0.2$${\delta}^{18}O:57.5{\pm}0.7$‰, respectively, which are within recommended values of analytical uncertainties. Also, we investigated isotope values of potential nitrogen source (soil, synthetic fertilizer and organic-animal manures) and temporal patterns of ${\delta}^{15}N-NO_3$ and ${\delta}^{18}O-NO_3$ values in river samples during from May to December. ${\delta}^{15}N-NO_3$ and ${\delta}^{18}O-NO_3$ values are enriched in December suggesting that organic-animal manures should be one of the main N sources in those areas. The current study clarifies the reliability of denitrification bacteria method and the usefulness of stable isotopic techniques to trace the anthropogenic nitrogen source in freshwater ecosystem.

Performance of Drip Irrigation System in Banana Cultuivation - Data Envelopment Analysis Approach

  • Kumar, K. Nirmal Ravi;Kumar, M. Suresh
    • Agribusiness and Information Management
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    • v.8 no.1
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    • pp.17-26
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    • 2016
  • India is largest producer of banana in the world producing 29.72 million tonnes from an area of 0.803 million ha with a productivity of 35.7 MT ha-1 and accounted for 15.48 and 27.01 per cent of the world's area and production respectively (www.nhb.gov.in). In India, Tamil Nadu leads other states both in terms of area and production followed by Maharashtra, Gujarat and Andhra Pradesh. In Rayalaseema region of Andhra Pradesh, Kurnool district had special reputation in the cultivation of banana in an area of 5765 hectares with an annual production of 2.01 lakh tonnes in the year 2012-13 and hence, it was purposively chosen for the study. On $23^{rd}$ November 2003, the Government of Andhra Pradesh has commenced a comprehensive project called 'Andhra Pradesh Micro Irrigation Project (APMIP)', first of its kind in the world so as to promote water use efficiency. APMIP is offering 100 per cent of subsidy in case of SC, ST and 90 per cent in case of other categories of farmers up to 5.0 acres of land. In case of acreage between 5-10 acres, 70 per cent subsidy and acreage above 10, 50 per cent of subsidy is given to the farmer beneficiaries. The sampling frame consists of Kurnool district, two mandals, four villages and 180 sample farmers comprising of 60 farmers each from Marginal (<1ha), Small (1-2ha) and Other (>2ha) categories. A well structured pre-tested schedule was employed to collect the requisite information pertaining to the performance of drip irrigation among the sample farmers and Data Envelopment Analysis (DEA) model was employed to analyze the performance of drip irrigation in banana farms. The performance of drip irrigation was assessed based on the parameters like: Land Development Works (LDW), Fertigation costs (FC), Volume of water supplied (VWS), Annual maintenance costs of drip irrigation (AMC), Economic Status of the farmer (ES), Crop Productivity (CP) etc. The first four parameters are considered as inputs and last two as outputs for DEA modelling purposes. The findings revealed that, the number of farms operating at CRS are more in number in other farms (46.66%) followed by marginal (45%) and small farms (28.33%). Similarly, regarding the number of farmers operating at VRS, the other farms are again more in number with 61.66 per cent followed by marginal (53.33%) and small farms (35%). With reference to scale efficiency, marginal farms dominate the scenario with 57 per cent followed by others (55%) and small farms (50%). At pooled level, 26.11 per cent of the farms are being operated at CRS with an average technical efficiency score of 0.6138 i.e., 47 out of 180 farms. Nearly 40 per cent of the farmers at pooled level are being operated at VRS with an average technical efficiency score of 0.7241. As regards to scale efficiency, nearly 52 per cent of the farmers (94 out of 180 farmers) at pooled level, either performed at the optimum scale or were close to the optimum scale (farms having scale efficiency values equal to or more than 0.90). Majority of the farms (39.44%) are operating at IRS and only 29 per cent of the farmers are operating at DRS. This signifies that, more resources should be provided to these farms operating at IRS and the same should be decreased towards the farms operating at DRS. Nearly 32 per cent of the farms are operating at CRS indicating efficient utilization of resources. Log linear regression model was used to analyze the major determinants of input use efficiency in banana farms. The input variables considered under DEA model were again considered as influential factors for the CRS obtained for the three categories of farmers. Volume of water supplied ($X_1$) and fertigation cost ($X_2$) are the major determinants of banana farms across all the farmer categories and even at pooled level. In view of their positive influence on the CRS, it is essential to strengthen modern irrigation infrastructure like drip irrigation and offer more fertilizer subsidies to the farmer to enhance the crop production on cost-effective basis in Kurnool district of Andhra Pradesh, India. This study further suggests that, the present era of Information Technology will help the irrigation management in the context of generating new techniques, extension, adoption and information. It will also guide the farmers in irrigation scheduling and quantifying the irrigation water requirements in accordance with the water availability in a particular season. So, it is high time for the Government of India to pay adequate attention towards the applications of 'Information and Communication Technology (ICT) and its applications in irrigation water management' for facilitating the deployment of Decision Supports Systems (DSSs) at various levels of planning and management of water resources in the country.