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Optimization of PCR Condition with Conidiospore for Primary Screening of Aspergillus nidulans Transformants (Aspergillus nidulans의 무성포자를 이용한 PCR 조건의 최적화)

  • 박희문;박범찬;박윤희;양소영
    • Korean Journal of Microbiology
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    • v.38 no.2
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    • pp.103-106
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    • 2002
  • Direct PCR from intact fungal cells is not readily suitable to all fungi mainly because of difficulties in rupturing the cell walls. Microwave irradiation has been proven to be useful in fungal DNA extraction protocol. Here we describe a fast template preparation method for PCR amplification from Aspefillus nidulans conidiospores using microwave irradiation. We optimized the duration far microwave irradiation, and the amount of template DNA for PCR. Amplification from samples prepared in this manner was so efficient that we could get PCR products with size enough to identify transformants. We believe that this is a time-saving procedure for screening true transformants of A. nidulans.

HPLC SEPARATION AND QUANTITATIVE DETERMINATION OF GINSENOSIDES FROM PANAX GINSENG, PANAX QUINQUEFOLIUM AND FROM GINSENG DRUG PREPARATIONS

  • Soldati F
    • Proceedings of the Ginseng society Conference
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    • 1980.09a
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    • pp.59-69
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    • 1980
  • A new HPLC-method for separation and quantitative determination of ginsenosides in Panax ginseng, Panax quinquefolium and in pharmaceutical drug preparations is elaborated. A reversed-phase-system with ${\mu}Bondapak\;C_{18}$ column (3.9 mm $I.D.{\times}30\;cm$) using acetonitrile-water (30:70) 2 ml/min and acetonitrile-water (18:82) 4 ml/min is suitable for the base-line separation of $Rb_1,\;Rb_2,\;Rc,\;Rd,\;Rf,\;Rg_2,\;respectively\;Re,\;Rg_1$ in 30 minutes. The ginsenosides are directly detected at 203 nm (without derivatization) with the LC-55 or LC-75 spectrophotometer (Perkin-Elmer) at $100\%$ transmission. Detection limit is 300 ng at a signal-to-noise ratio of 10:1. The ginsenosides-peak identification is carried out with HPTLC (high performance thin layer chromatography), with MIR-IR (multiple internal reflection-IR-spectros-copy) and with FD-MS (field desorption mass spectrometry). The calibration curve of each ginsenoside has a correlation coefficient very near to 1. Relative standard deviation for quantitative determinations depends upon the amount of ginsenosides and is approximately 1\%$ for ginsenoside contents of 1\%$. This method is adaptable for routine analysis in quality control laboratories.

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${\ulcorner}$Aheuichaulyo御醫撮要${\lrcorner}$-Restoration of the lost medical book in ancient Korea I ("어의촬요(御醫撮要)"-실전의서의 복원 I)

  • Ahn, Sang-Woo;Choi, Hwan-Soo
    • Korean Journal of Oriental Medicine
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    • v.4 no.1 s.4
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    • pp.1-25
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    • 1998
  • $\ulcorner$Aheuichaulyo御醫撮要$\lrcorner$ was known to be published by Choi Jong-jun崔宗峻 in Korea高麗 Dynasty(A.D.1226), however its original copy is not available at the present. It was thought that Korean folk medicine and the remedies were recorded in this book, and these were recorded in $\ulcorner$Euibangyoochui醫方類聚$\lrcorner$ published in Chosun 朝鮮 dynasty(1445). $\ulcorner$Euibangyoochui$\lrcorner$ is called a treasure-house of the knowledge of traditional oriental medicine which contains over 50,000 prescriptions and enormous amount of medical information. The restoration of $\ulcorner$Aheuichaulyo$\lrcorner$was possible during the process of establishment of data base work of $\ulcorner$Euibangyoochui$\lrcorner$. $\ulcorner$Aheuichaulyo$\lrcorner$ was restored as 131 chapters mainly from the related contents of $\ulcorner$Euibangyoochui$\lrcorner$ and partially from the contents of $\ulcorner$Hyangyakjipsungbang鄕藥集成方$\lrcorner$ and $\ulcorner$Boonmoononyukyihaebang分門瘟易解方$\lrcorner$ for the supplement.

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Factors Affecting the Turnover Intention of Dental Hygienists: Emotional Labor, Job Satisfaction, and Social Support

  • Noh, Sun-Mi;Lim, Hee-Jung;Kim, Myoung-Hee;Im, Ae-Jung;Lim, Do-Seon
    • Journal of dental hygiene science
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    • v.18 no.5
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    • pp.271-279
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    • 2018
  • This study aims to contribute to managing the human resource of dental hygienists and qualitatively enhancing dental medical services by examining factors that affect their turnover intention. Therefore, it attempted to examine their degrees of emotional labor, job satisfaction, and social support and the effects of each variable on turnover intention. This study administered self-reporting questionnaires to dental hygienists working in dental hospitals, dental clinics, and general or university hospitals in Seoul Metropolis and Gyeonggi-do by conducting convenience sampling, from May 18, 2017 to August 4, 2017. Among a total of 224 copies that were distributed, a total of 223 copies, excluding 1 copy with poor responses, were used in the data analysis. The research tools comprised 12 questions on general characteristics, 24 questions on emotional labor (4-point Likert scale), 16 questions on job satisfaction (5-point Likert scale), 8 questions on social support (4-point Likert scale), and 4 questions on turnover (5-point Likert scale). The scores of dental hygienists were as follows: emotional labor, 2.49 out of 4; job satisfaction, 3.14 out of 5; social support, 3.04 out of 4; and turnover intention, 3.07 out of 5. Their turnover intention has a positive correlation with emotional labor, but a negative correlation with job satisfaction and social support. It was found that some factors that significantly influence turnover intention included the amount of overtime work and job satisfaction. Thus, dental medical institutes should search for measures, including improvement of the working environment, to reduce the amount of overtime work and enhance job satisfaction.

Detection of Salmonella in Milk by Polymerase Chain Reaction

  • Park, Weon-Sang
    • Journal of Food Hygiene and Safety
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    • v.15 no.3
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    • pp.262-266
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    • 2000
  • The polymerase chain reaction was used to selectively detect sequences within the fimbrial antigen of Salmonella enteritidis. Sterile milk was artificially inoculated with known amount of S. enteritidis and then DNA was extracted with guanidine thiocyanate/phenol/chloroform, followed by PCR. A detection limit of as few as 100 colony forming unit (cfu) per 0.5 ml milk was obtained with this method. For the whole procedure, it took only 5 h. A semi-quantitative polymerase chain reaction assay which allows an estimation of colony forming unit of S. enteritidis was developed. Known amount of standard plasmid pGem-4Z-Sef B(-) containing cloned S. enteritidis fimbrial antigen gene was co-amplified with Salmonella genomic DNA isolated from artificially inoculated milk. The same set of primers were used for the amplification and the products were cleaved with Bam HI. The concentration of the target DNA could be estimated by comparing the intensity of the two bands after electrophoresis. The PCR-based protocol described in this paper provides a rapid, simple, and sensitive method for detecting S. enteritidis in milk.

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Effect of antibiotic resistant factors in effluent of wastewater treatment plant on stream (하수처리장 방류수에 존재하는 항생제 내성인자가 하천에 미치는 영향)

  • Jang, Yejin;Yoo, Yong-Jae;Sul, Woo Jun;Cha, Chang-Jun;Rhee, Ok-Jae;Chae, Jong-Chan
    • Korean Journal of Microbiology
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    • v.53 no.4
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    • pp.316-319
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    • 2017
  • The antibiotic resistant genes (ARG) and mobile genetic elements (MGE) were investigated with the effluent of waste-water treatment plant (WWTP), and river waters of upstream and downstream in order to elucidate the effect of effluent on antibiotic resistance in a natural river. Total numbers of 134~183 of ARG and MGE were detected and the abundance of ARG and MGE was 0.063~0.422 copies per one of 16S rRNA gene in three water samples. Effluent sample contained the highest amount of the total number and abundance of ARG and MGE whereas total viable cells were observed in the lowest amount among the three samples. This indicated that the genes were originated from cells died during the wastewater treatment process. In addition, the co-relationship of abundance between ARG and MGE suggested that acquired resistance was a prevalent mechanism among the antibiotic-resistant bacteria existing in WWTP.

In vitro ruminal fermentation of fenugreek (Trigonella foenum-graecum L.) produced less methane than that of alfalfa (Medicago sativa)

  • Niu, Huaxin;Xu, Zhongjun;Yang, Hee Eun;McAllister, Tim A;Acharya, Surya;Wang, Yuxi
    • Animal Bioscience
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    • v.34 no.4
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    • pp.584-593
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    • 2021
  • Objective: The objective of this study was to compare fenugreek (FG) with alfalfa (Alf) in ruminal fermentation and methane (CH4) production in vitro. Methods: Whole-plant FG harvested at 11- and 15-wk and Alf harvested at early and mid-bloom maturities, alone or as 50:50 mixture of FG and Alf at the respective maturity, were assessed in a series of 48-h in vitro batch culture incubations. Total fermentation gas and methane gas production, dry matter (DM) disappearance, volatile fatty acids, microbial protein and 16S RNA gene copy numbers of total bacteria and methanogens were determined. Results: Compared to early bloom Alf, FG harvested at 11-wk exhibited higher (p<0.05) in vitro DM and neutral detergent fibre disappearance, but this difference was not observed between the mid-bloom Alf and 15-wk FG. Regardless plant maturity, in vitro ruminal fermentation of FG produced less (p<0.001) CH4 either on DM incubated or on DM disappeared basis than that of Alf during 48-h incubation. In vitro ruminal fermentation of FG yielded similar amount of total volatile fatty acids with higher (p<0.05) propionate percentage as compared to fermentation of Alf irrespective of plant maturity. Microbial protein synthesis was greater (p<0.001) with 11-wk FG than early bloom Alf as substrate and 16S RNA gene copies of total bacteria was higher (p<0.01) with 15-wk FG than mid-bloom Alf as substrate. Compared to mid-bloom Alf, 15-wk FG had lower (p<0.05 to 0.001) amount of 16S RNA methanogen gene copies in the whole culture during 48-h incubation. Conclusion: In comparison to Alf, FG emerges as a high quality forage that can not only improve rumen fermentation in vitro, but can also remarkably mitigate CH4 emissions likely due to being rich in saponins.

Genetic Transformation of Chrysanthemum with Cold Regulated Gene (BN115) (저온저항성 유전자를 이용한 국화 형질전환)

  • Han, Soo-Gon;Choi, In-Young;Kang, Chan-Ho;Ko, Bok-Rai;Choi, Joung-Sik;Lee, Wang-Hyu
    • Journal of Plant Biotechnology
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    • v.33 no.1
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    • pp.19-25
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    • 2006
  • With the use of Agrobacterium and gene-gun, cold regulated gene (BN115) has been injected in Chrysanthemum leaf disc and transgenic plants have been produced successfully on the selection media containing phytohormone. To determine the presence of the transferred cold regulated gene (BN115) in the transgenic Chrysanthemum, PCR-amplification indicated the presence of that gene. Real-Time PCR for confirmation of the putative transgenic plants was established. The copy number of cold regulated gene (BN115) is extrapolated on the basis of a standard curve. Serial dilutions of known number of gene copies were in triplicates. In this diagram, PCR cycles are plotted against the fluorescence intensity. The cycle at which the fluorescence reaches a threshold cycle is inversely proportional to the starting amount of target DNA.

Quantitative analysis of periodontal pathogens present in the saliva of geriatric subjects

  • Shet, Uttom K.;Oh, Hee-Kyun;Kim, Hye-Jeong;Chung, Hyun-Ju;Kim, Young-Joon;Kim, Ok-Su;Choi, Hong-Ran;Kim, Ok-Jun;Lim, Hoi-Jeong;Lee, Seok-Woo
    • Journal of Periodontal and Implant Science
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    • v.43 no.4
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    • pp.183-190
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    • 2013
  • Purpose: At present, information regarding periodontal disease in geriatric patients is scarce. The purpose of this study was to quantify the periodontal pathogens present in the saliva of Korean geriatric patients and assess the relationship between the bacterial levels and the periodontal condition. Methods: Six putative periodontal pathogens were quantified by using a real-time polymerase chain reaction assay in geriatric patient groups (>60 years) with mild chronic periodontitis (MCP), moderate chronic periodontitis (MoCP), and severe chronic periodontitis (SCP). The copy numbers of Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola, Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum, and Prevotella intermedia were measured. Results: It was found that the bacterial copy numbers increased as the severity of the disease increased from MCP to SCP, except for P. intermedia. For P. intermedia, it was found that samples in the MCP group yielded the largest amount. It was also found that the quantities of P. gingivalis, T. forsythia, and T. denticola, the so-called "red complex" bacteria, were lower than those of F. nucleatum, A. actinomycetemcomitans, and P. intermedia in all of the samples. Conclusions: Collectively, the results of this study suggest that the levels of P. gingivalis, T. forsythia, F. nucleatum, and T. denticola present in saliva are associated with the severity of periodontal disease in geriatric patients.

Effects of Tillage on Organic Matters and Microbial Communities in Organically Cultivated Corn Field Soils (유기농 옥수수밭에서 경운이 토양 유기물 함량 및 미생물군집에 미치는 영향)

  • Ahn, Dalrae;An, Nan-Hee;Kim, Da-Hye;Han, Byeong-Hak;You, Jaehong;Park, InCheol;Ahn, Jae-Hyung
    • Korean Journal of Environmental Agriculture
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    • v.39 no.1
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    • pp.65-74
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    • 2020
  • BACKGROUND: Soil carbon sequestration has been investigated for a long time because of its potential to mitigate the greenhouse effect. No- or reduced tillage, crop rotations, or cover crops have been investigated and practiced to sequester carbon in soils but the roles of soil biota, particularly microorganisms, have been mostly ignored although they affect the amount and stability of soil organic matters. METHODS AND RESULTS: In this study we analyzed the organic matter and microbial community in organically cultivated corn field soils where no-tillage (NT) or conventional tillage (CT) had been practiced for about three years. The amounts of organic matter and recalcitrant carbon pool were 18.3 g/kg dry soil and 4.1 g C/kg dry soil, respectively in NT soils, while they were 12.4 and 2.5, respectively in CT soils. The amounts of RNA and DNA, and the copy numbers of bacterial 16S rRNA genes and fungal ITS sequences were higher in NT soils than in CT soils. No-tillage treatment increased the diversities of soil bacterial and fungal communities and clearly shifted the bacterial and fungal community structures. In NT soils the relative abundances of bacterial phyla known as copiotrophs, Betaproteobacteria and Bacteroidetes, increased while those known as oligotrophs, Acidobacteria and Verrucomicrobia, decreased compared to CT soils. The relative abundance of a fungal phylum, Glomeromycota, whose members are known as arbuscular mycorrhizal fungi, was about two time higher in NT soils than in CT soils, suggesting that the higher amount of organic matter in NT soils is related to its abundance. CONCLUSION: This study shows that no-tillage treatment greatly affects soil microbial abundance and community structure, which may affect the amount and stability of soil organic matter.