• Title/Summary/Keyword: isolation tube

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Isolation and Characterization of Mitochondrial DNA from Arehicityars and Metacitrus (Archicitrus와 Metacitrus로부터 Mitochondrial DNA의 분리 및 특성)

  • 이숙영;박민희
    • The Korean Journal of Food And Nutrition
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    • v.8 no.4
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    • pp.307-317
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    • 1995
  • The purity of mtDNAs isolated from Archicitrus and Metacitrus leaves was higher in percoll density gradient centrifugation than differential and sucrose density gradient centrifugation. The most clear mtDNAs were obtained from mitochondria included in the Interface band of between 21% and 45% under isomotic, low viscosity conditions in the three step discontinuous percoll density gradient centrifugation. DNase treatment to the crude mitochondrlal suspension still more increased purity of mtDNA by the effective removal of the nuclear and chloroplast DNA and mtDNAs were appeared as a single band at middle position of tube by EtBr /cscl density gradient centrifugation. Agarose gel electrophoresis of mtDNAs resolved a single, broad band containing high molecular weight DNAs In all preparation. Yield of mtDNAs was about 110 and 2 ug Per 2009 in mature and immature leaves respectively. The mtDNA fragment patterns showed by EcoR I treatment were indistinguishable with respect to nom bet and position of bands in Archicitrus and Metacitrus. In the pattern of Hind E restriction, the Metacitrus displayed the unique band between 5.0 and 4.0kb, in addition to four fragments about 5.0, 2.4, 2.15, and 2.0kb, respectively, different from Archicitrus. Also the pattern of total mtDNAs fragment by the treatment of Pst I showed that the distinguishable fragment pat tern was not appeared in Archicitrus(C. iyo Tanaka), but about 6.0, 5.5, 5.0 and 2.Bkb fragments were appeared only in Metacitrus(C. junos Sieb). Therefore it was indicated that two species in intra-subgenus were identical each other, whereas considerable difference was revealed for inter-subgenus.

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Production of Polyclonal Antibody against $\alpha$-Fetoprotein and Polyclonal Antibody-Based Competitive Enzyme-Linked Immunosorbent Assay for $\alpha$-Fetoprotein (인간 $\alpha$-fetoprotein (AFP)에 대한 폴리클로날 항체의 생산 및 $\alpha$-fetoprotein 측정용 효소면역분석법 (competitive ELISA)의 개발)

  • Michung Yoon
    • Biomedical Science Letters
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    • v.3 no.2
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    • pp.115-123
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    • 1997
  • $\alpha$-Fetoprotein (AFP) has been a useful marker in screening and/or monitoring patients with hepatocellular carcinoma, gonadal germ cell tumor, gastric carcinoma and neural tube defects. In the present study, it was attempted to produce anti-human AFP polyclonal antibodies and to develop a competitive enzyme-linked immunosorbent assay (ELISA) for the measurement of AFP in human plasma and amniotic fluid. AFP was isolated from amniotic fluid using an isolation procedure consisting of affinity chromatography and preparative polyacrylamide gel electrophoresis. The antibody directed against AFP was raised in rabbits. Double immunodiffusion and Western blotting methods showed that the antiserum was highly specific, reacting with only AFP-containing samples. Standard curve was obtained by using purified AFP and specific antiserum. The assay sensitivity was 5ng/ml and the working range was 5~l,000ng/ml. The within-assay and between-assay coefficient of variance (CV) was 4.5% and 8.5%, respectively. These results indicate that the assay is valuable for the measurement of AFP and found to be simple, reproducible, and accurate.

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Measurement of Ochratoxin A and Isolation of the fungi producing Ochratoxin A from Korean traditional fermented soybean foodstuffs (전통 대두발효식품(醱酵食品)중에 존재하는 Ochratoxin A 생산균(菌) 분리(分離)와 Ochratoxin A량 측정)

  • Kang, Sung-Chul;Lee, Sang-Sun;Shin, Hyun-Kil;Kim, Joung-Bae
    • The Korean Journal of Mycology
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    • v.19 no.2
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    • pp.148-155
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    • 1991
  • Fungal ochratoxin A wes extracted and measured from the Korean traditional and fermented soybean foodstuffs (12 samples of Maeju, 28 samples of Dwangjang and 28 samples of Kangjang) collected from the whole nation wide regions. The various fungi were isolated from the foodstuffs and they were also examined whether the isolates produce ochratoxin A (OA) under the artificial conditions or not. Determinations of OA produced by the fungi were done with the antibody-attached CIA method, which was accurate or sensitive at the range of 20 pg per sample with a ninty percent recovery. Out of the 222 fungal isolates, 39 isolates produced the OA under the artificial conditions, and were identified as species of Aspergillus, Penicillium, Paecilom­yces or other genera. The OA detected in all soybean foodstuffs was presumed to be originated from the first fermentation step of maeju.

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Epidemiological study for infection route of brucellosis in a infected dairy farms (Brucella 감영농장에서 감염경로의 역학적 연구)

  • 윤여백;김영진;김추철;노영선;권미순;김철민;임채웅
    • Korean Journal of Veterinary Service
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    • v.27 no.2
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    • pp.159-164
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    • 2004
  • A dairy farm that has been suffered continuously(more than 2 years) from brucellosis in Korea in spite of repeated legal test-and-slaughter was investigated the main source of infection in the farm. All cattle(22 milking cows, 44 heifers, 60 calves, 8 bull), dogs(3 mixed breed), feces from wild birds(3 samples), drinking water(3 sites), and soil in the paddocks(14 sites) inside the farm were examined with serological and/or bacteriological methods including specific DNA detection with PCR method. Brucella spp in the milk and blood were detected in 12/22 and 5/22 milking cows, respectively, although all of them were negative with conventional tube agglutination test. The number of serologically positive heifer was 15(15/44), but the isolation of Brucella spp was succeeded in the only 11(11/15) of them. Brucella were detected in vagina 1(1/11) and nasal(3/12) excretion in serologically positive heifers. All the three dogs were serologically positive, and Brucella spp were isolated from their blood. However, Brucella spp were not detected in the drinking water, soil in the paddocks, nor the feces of wild birds. The results suggest that milking cow secrete Brucella spp through milk, genital tract and nasal cavity, which are the major source of infection in this farm, The main infection route of Brucella spp is contact to contact with Brucella spp excreting animals rather than environmental contamination. The animals, living together with infected cow such as dogs, are the readily susceptible and are required to be examined for Brucella spp.

A Study on SNP of IL10 in Cerebral Infarction Patients

  • Jung, Tae-Young;Choi, Sung-Hun;Kim, Kyung-Woon;Lee, Yoon-Kyung;Lim, Seong-Chul;Lee, Kyung-Min;Seo, Jung-Chul
    • Journal of Acupuncture Research
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    • v.23 no.2
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    • pp.173-179
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    • 2006
  • Objectives : In this study, we investigated the SNP (single-nucleotide polymorphism) of IL10 in patients with stroke. The present study was undertaken to see if specific genotypic and allelic variations are associated with stroke in the Korean population. Methods : Blood samples from all subjects were obtained for DNA extraction and collected in EDTA tube. Genomic DNA was extracted using DNA isolation kit for Mammalian Blood (Boehringer Mannheim, IN, USA). The extracted DNA was amplified by polymerase chain reaction (PCR). Pyrosequencing was performed according to manufacturer's standard protocol. Results : There was no statistically significant genotypic distribution difference between control and stroke group. The frequencies of A/A homozygotes and A/C heterozygotes among control subjects were 91 (87.5%) and 13 (12.5%). The frequencies of A/A and A/C among the stroke patients were 85 (89.5%) and 10 (10.5%). There was not statistically significant allelic frequency difference between control and stroke group. The allelic frequency of A and C was 195 (93.8%) and 13 (6.2%) among the control subjects and 180 (94.7%) and 10 (5.3%) in stroke patients, respectively. Conclusion : The cytokine IL10 may not be pathogenetic factors in stroke. But further studies including different cytokine gene can be a useful for predicting stroke. Establishment of more systemic approach and high quality of prospective cohorts will be necessary for the good prediction of genetic markers.

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Study on the Development of Home Care Nursing Intervention Protocol for Stroke Patients (뇌졸중 환자의 가정간호중재 프로토콜 개발)

  • Yoo Ji-Soo
    • Journal of Korean Academy of Fundamentals of Nursing
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    • v.7 no.1
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    • pp.122-136
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    • 2000
  • Stroke patient needs rehabilitation after receiving an acute treatment in a hospital. When stroke patient gets involved in an early discharge program, home care nurse plays a pivotal role to make them to gain a full strength and to come back to his/her prior life before he/she is sick. In spite of the importance of home care nursing intervention protocol for home care nurses to perform home care nursing autonomously, home care nursing intervention protocol for stroke patient is rarely developed. Therefore this study was conducted to develop home care nursing protocol that is applicable for stroke patients in home care nursing area. 41 home care nursing charts for stroke patients registered in home care nursing agencies from December 1st 1994 to August 31st 1999 at Y hospitals in Seoul and Won-Ju city were analyzed. 44 home care nurses who were having over three years' experience on stroke patients were participated in this study as a user validity validation group. The results of this study are as follows. 1. 28 nursing diagnoses were selected on the basis of evaluation of nursing diagnoses of stroke patients presented in a previous literature and case studies on home care nursing. 2. 17 nursing diagnoses were classified through the frequency analysis of home care nursing charts for 41 stroke patients who had received home care nursing. The order of sequence was like these: impaired skin integrity, risk for infection, nutritional deficit, impaired physical mobility, constipation, knowledge deficit, ineffective airway clearance, anxiety in family members, risk for aspiration, self care deficit, altered urinary elimination, ineffective individual coping, social isolation, risk for injury, self-esteem disturbance, impaired verbal communication, fatigue of family caregiver. 3. Based on validation on expert and user validities, 44 nursing interventions which were above ICV=.80 were chosen. 4. Nursing intervention protocols which showed above ICV=.90 were developed and were like these; pressure ulcer care, position change, preventive care for circulatory dysfunction, tube care : catheter, vital sign monitor, constipation/impaction management, artificial airway management, suction of airway secretion, environmental management : safety, and fall prevention.

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Isolation and Characterization of Cold-Adapted PGPB and Their Effect on Plant Growth Promotion

  • Li, Mingyuan;Wang, Jilian;Yao, Tuo;Wang, Zhenlong;Zhang, Huirong;Li, Changning
    • Journal of Microbiology and Biotechnology
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    • v.31 no.9
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    • pp.1218-1230
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    • 2021
  • Cold-adapted plant growth-promoting bacteria (PGPB) with multiple functions are an important resource for microbial fertilizers with low-temperature application. In this study, culturable cold-adapted PGPB strains with nitrogen fixation and phosphorus solubilization abilities were isolated. They were screened from root and rhizosphere of four dominant grass species in nondegraded alpine grasslands of the Qilian Mountains, China. Their other growth-promoting characteristics, including secretion of indole-3-acetic acid (IAA), production of siderophores and ACC deaminase, and antifungal activity, were further studied by qualitative and quantitative methods. In addition, whether the PGPB strains could still exert plant growth-promoting activity at 4℃ was verified. The results showed that 67 isolates could maintain one or more growth-promoting traits at 4℃, and these isolates were defined as cold-adapted PGPB. They were divided into 8 genera by 16S rRNA gene sequencing and phylogenetic analysis, of which Pseudomonas (64.2%) and Serratia (13.4%) were the common dominant genera, and a few specific genera varied among the plant species. A test-tube culture showed that inoculation of Elymus nutans seedlings with cold-adapted PGPB possessing different functional characteristics had a significant growth-promoting effect under controlled low-temperature conditions, including the development of the roots and aboveground parts. Pearson correlation analysis revealed that different growth-promoting characteristics made different contributions to the development of the roots and aboveground parts. These cold-adapted PGPB can be used as excellent strain resources suitable for the near-natural restoration of degraded alpine grasslands or agriculture stock production in cold areas.

A Study on Establishment of Basic Safety and Essential Performance Criteria of Mobile Computed Tomography (이동형 전산화단층촬영장치의 기본 안전 및 필수 성능 기준을 마련하기 위한 연구)

  • Kim, Eun Hye;Park, Hye Min;Kim, Jung Min
    • Journal of radiological science and technology
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    • v.44 no.3
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    • pp.261-267
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    • 2021
  • As the number of Coronavirus Disease-19 (COVID-19) patients increases in a global pandemic situation, the usefulness of mobile computed tomography (CT) is gaining attention. Currently, mobile CT follows the basic safety and essential performance evaluation criteria of whole-body or limited-view X-ray CT in order to obtain device approval and evaluation in the Republic of Korea. Unlike stationary CT, mobile CT is not operated in shielded areas but rather areas such as intensive care units, operating rooms, or isolation rooms. Therefore, it requires a different basic safety and essential performance evaluation standard than stationary CT. In this study, four derived basic safety evaluation criteria related to electrical, mechanical, and radiation safety were included (dose indication test, protection against stray radiation, safety measures against excessive X-rays, half-value layer measurement); and seven essential performance evaluation criteria were included (tube voltage accuracy, mAs accuracy, radiation dose reproducibility, CT number of water, noise, uniformity, and spatial resolution); total eleven basic safety and essential performance evaluation criteria were selected. This study aims to establish appropriate basic safety and essential performance evaluation criteria for simultaneously obtaining images with diagnostic value and reducing the exposure of nearby patients, medical staff, and radiologic technologists during the use of mobile CT.

Protoplast Regeneration, Reversion and Isolation of Auxotrophic Mutants in Flammulina velutipes (팽이버섯 (Flammulina velutipes)원형질체(原形質體)의 재생(再生), 환원(還元) 및 영양요구성(營養要求性) 균주선발(菌株選拔))

  • Shin, Gwan Chull;Park, Jong Seung;Yoo, Young Bok;Yeo, Un Hyung
    • Korean Journal of Agricultural Science
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    • v.15 no.1
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    • pp.15-22
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    • 1988
  • Factors affecting the regeneration, reversion of protoplasts from mycelium of F. velutipes were investigated and the selection of auxotrophic mutants from protoplasts of F. velutipes was performed. PDP medium stabilized with 0.6M sucrose was suitable for the regeneration of protoplasts, and regeneration frequency was 0.47-1.32. The regeneration frequency of protoplasts was increased when nutrients were added to the regeneration medium. Especially, yeast extract was the most effective to regeneration of protoplasts. Regeneration pattern of protoplasts was formation of germ tubes from bud-like cells. 13-18% of monokaryotic strains was appeared from reverted protoplasts. Five of auxotrophic mutants were isolated from strains showed survival frequency of 1.9-16.

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Comparison among the Sperm Preparation Methods on the Human Spermatozoa (인간정자에 있어서 정자처리법의 비교)

  • Pang, Myung-Geol;Chung, Ku-Min;Shin, Chang-Jae;Kim, Jung-Gu;Moon, Shin-Yong;Chang, Yoon-Seok;Lee, Jin-Yong;Lee, Sang-Hoon;Chung, Yung-Chai;Kim, Chang-Keun
    • Clinical and Experimental Reproductive Medicine
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    • v.20 no.2
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    • pp.107-115
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    • 1993
  • Procedures to separate motile. normal & motile and acrosome-reacted sperm with high efficiency have clinical application in Assisted Reproductive Technology in terms of increasing the probability of fertilization by a normal sperm and subsequent normal embryonic development. This study evaluated the effects of 10 sperm preparation techniques [Swim-up from a washed pellet (SU). Swim-up from semen (SO). Continuous Percoll Gradients I (PIC). Discontinuous Percoll Gradients I (PID). Continuous Percoll Gradients II(P II C). Discontinuous Percoll Gradients II(P II D), SpermPrep (SFC). Wang's tube (WT). Albumin Gradients (AG), Low temperature capacitation (LTC)] on motility (%), normal morphology (%), motile sperm recovery rate(%). morphologically normal & motile sperm recovery rate (%), true acrosome reaction (%) and fertilizing ability. A P II D proved to be an effective means of separating morphologically normal & motile sperm. Our results indicated the P II D has advantages as compared with other methods in terms of recovery rate. enhancement of motility and normal morphology. And a LTC seems to be an effective means of enhancing the true acrosome reaction and fertilizing ability. These results suggest that the combined method of LTC and P II D for separation of morphologically normal & motile sperm and acrosome reacted sperm may be a useful procedure for intrauterine insemination and in vitro fertilization in the management of male factor infertility as well as for isolation of subpopulation of sperm for basic research.

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