• Title/Summary/Keyword: Non-Isolation

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Development of Polyclonal Antibodies to Abdominal and Subcutaneous Adipocytes for Fat-Reduced Hanwoo Beef Production (한우 체지방 감소 쇠고기 생산을 위한 복강 및 피하지방 항체 개발)

  • Choi, Chang-Weon;Kim, Yu-Hyun;Kim, Sang-Jin;Song, Man-Kang;Kwon, Eung-Gi;Oh, Young-Kyoon;Hong, Seong-Koo;Choi, Seong-Ho;Baek, Kyung-Hoon
    • Food Science of Animal Resources
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    • v.28 no.5
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    • pp.651-659
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    • 2008
  • This study aimed to develop polyclonal antibodies to regional inedible adipocytes of Korean native cattle (Hanwoo) and investigate cross-reactivity of the antibodies. Patterns in plasma membrane proteins (PMPs) from abdominal and subcutaneous adipocytes of Hanwoo isolated by collagenase digestion were investigated using SDS-PAGE. As antigens, abdominal and subcutaneous adipocyte PMPs of Hanwoo were injected to sheep 3 times at 3 wk intervals for passive immunization, and non-immunized serum and antisera were collected before and after the injections. Titers of the antisera obtained from sheep and their cross-reactivities with heart, kidney, liver, lung, muscle, and spleen of Hanwoo were determined by ELISA. Isolation and culture of abdominal and subcutaneous adipocytes of Hanwoo were performed for analysing LDH concentration. Based on the SDS-PAGE analysis, specific proteins of PMPs in abdominal and subcutaneous adipocytes appeared despite rather similar patterns between both adipocytes. At the level of 1:1,000 dilution, little antibody reactivity appeared in non-immunized serum whereas the antisera had relatively strong reactivity up to the level of 1:128,000 and 1:64,000 dilution. These findings may indicate that strong antibodies against adipocyte PMPs can be developed using an immunological approach. Extremely low reactivities of abdominal and subcutaneous adipocyte antisera were detected with PMPs of the organs. Both antisera strongly reacted with each adipocyte PMPs and showed statistically (p<0.01) higher cross-reactivities compared with non-immunized serum. In conclusion, these results may indicate that the present polyclonal antibodies against regional inedible adipocyte PMPs are well developed and have safety in cross-reactivities with body organs. Further studies on in vivo cross-reactivity and fat reduction of the antibodies against abdominal and subcutaneous adipocytes PMPs of Hanwoo should be required for inedible fat-reduced high quality beef production.

Evaluation of Selective Media for Isolation of Staphylococcus aureus from Agricultural Products (농산물 중 Staphylococcus aureus의 분리를 위한 선택배지 평가)

  • Kim, Se-Ri;Lee, Seo-Hyun;Seo, Min-Kyoung;Kim, Won-Il;Park, Kyeong-Hun;Yun, Hye-Jeong;Yoon, Yo-Han;Yoo, Soon-Young;Ryu, Kyoung-Yul;Yun, Jong-Chul;Kim, Byung-Seok
    • Journal of Food Hygiene and Safety
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    • v.27 no.2
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    • pp.169-175
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    • 2012
  • Five kinds of selective media, such as mannitol salt agar (MSA), Baird-Parker agar (BPA), Baird-Parker supplemented with rabbit plasma fibrinogen (BPA+RPF), CHROMagar Staphylococcus aureus (CSA), and Petrifilm Staph Express count system (Petrifilm), were compared to recommend the optimum selective media for isolation of Staphylococcus aureus from agricultural products. Seventy four target and non target bacteria were inoculated on five selective media to analyze sensitivity and specificity. In the recovery test of injured S. aureus cells, S. aureus was exposed to acid (1% lactic acid for 10 min), heat ($60^{\circ}C$ for 90s), and cold ($-20^{\circ}C$ for 1h) conditions. And artificially contaminated agricultural products (iceberg lettuce, green pepper, and cherry tomato) was enumerated on five selective media. The sensitivity of BPA+RPF, CSA, Petrifilm, MSA, and BPA were 100%, 100%, 100%, 90.5%, 90.5%, respectively. In addition, the specificity of BPA+RPF, CSA, MSA, BPA and Petrifilm were 100%, 100%, 84.6%, 75.0%, 67.3%, respectively. However, no difference among five selective media was observed in recovery on injured S. aureus cell and enumeration from agricultural products. This results suggest that BPA+RPF and CSA are the optimum media for detection of S. aureus from agricultural products.

Isolation of Pea and Soybean Nodule Bacteria and Assessment of their Nitrogen-fixing Capacities (완두 및 대두근류균(根瘤菌)의 분리(分離) 및 질소고정능력(窒素固定能力)의 비교(比較))

  • Kim, Seung-Yeol;Choi, Woo-Young
    • Korean Journal of Agricultural Science
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    • v.5 no.2
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    • pp.136-144
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    • 1978
  • A series of experiments over three years was planned for practical application of rhizobia in farms and grass lands in Korea. This is the report for the studies of the first year mainly on the isolation and characterization of rhizobial strains, and on the assessment of their nodulation abilities and nitrogen fixation capacities. 1. Total number of 88 strains for soybean group and 22 st ra ins for pea and vetch group was isolated from nodules which were taken from legumes grown in Daekwanyong, Cheju and various places in Korea. 2. Morphological and cultural characteristics of the strains were studied, and attempts were also made to investigate their antigenic properties and to demonstrate lysogenic strains in these groups. The results were : i) the isolates varied in cultural characteristics on yeast mannitol broth and agar, and in degree of congo reel absorption ; ii) similarities in their antigenic properties were found between/among the strains: G-3/G-9/D216, G-20/G-52 in soybean group; iii) no lysogeny was found in the strains of these groups. 3. Plant infection tests by test tube and bottle method in light room were carried out to elucidate the ability of the strains to nodulate specific legumes and of the capacity of such nodules to fix atmospheric nitrogen. The isolates were grouped into non- invasive, ineffective, or effective to the legumes. Those strains which produced effective nodules, supporting similar level of growth as nitrate control, were: P-3, 4 and 8 in pea and vetch group; G-23, 27, and, D-216 in soybean group.

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Effects of Vitamin E enhanced transgenic soybean cultivation on insect diversity (비타민 E 강화콩 재배가 곤충다양성에 미치는 영향)

  • Oh, Sung-Dug;Suh, SangJae;Park, Soo-Yun;Lee, Kijong;Sohn, Soo-In;Yun, Doh-Won;Chang, Ancheol
    • Korean Journal of Breeding Science
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    • v.49 no.3
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    • pp.129-140
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    • 2017
  • This study was carried out to develop of environmental risk assessments and the biosafety guide for Vitamin E enhanced transgenic soybean at LMO (Living Modified Organism) isolation field. In LMO quarantine area of National Institute of Agricultural Sciences, insect species diversities and population densities on vitamin E enhanced transgenic soybean and non-GM soybeans (Willams 82 and Seoritae) were investigated. A total of 17,717 individuals of 77 species from 8 orders were collected in LMO isolation field. In three type soybeans field, total of 5,250 individuals in Vitamin E enhanced transgenic soybean, 5,510 individuals in Willams 82, and 6,957 individuals in Seoritae were collected, respectively. There was no difference between the population densities of insect pests, natural enemies and other insects on Vitamin E enhanced transgenic soybean and Willams 82, while natural enemies density on Seoritae was higher than on Vitamin E enhanced transgenic soybean, but insect pests density on Vitamin E enhanced transgenic soybean was higher. These results provided the insects diversity for risk assessment survey of Vitamin E enhanced transgenic soybean and suggested that the guideline could be useful to detect LMO crops.

Frequency of Candida Strains Isolated from Candidiasis Patients at A Tertiary Hospital over the Last 10 Years (최근 10년 동안 일개 상급종합병원의 칸디다혈증 환자에서 분리된 칸디다 균종의 빈도)

  • Hwang, Yu-Yean;Kang, On-Kyun;Park, Chang-Eun;Hong, Sung-No;Kim, Young-Kwon;Huh, Hee-Jae;Lee, Nam-Yong
    • Korean Journal of Clinical Laboratory Science
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    • v.54 no.2
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    • pp.110-118
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    • 2022
  • Candidemia is a major cause of nosocomial infections resulting in increased morbidity and mortality. It remains a serious risk in inpatients and increases medical treatment costs. From 2009 to 2018, Candida strains (3,533) isolated from blood culture tests at the S Hospital were analyzed according to the period, year, sex, age, ward, etc. During the entire period, 54,739 of 717,996 blood culture tests showed a positive rate (7.6%) and the Candida isolation rate was 3,533 (6.4%) out of 1,036 patients. Among the Candida isolates, C. albicans was most common (33.8%), followed by C. tropicalis (28.6%), C. glabrata (19.8%), C. parapsilosis (7.8%), and C. krusei (4.0%). In early (2009~2013)/late (2014~2018) isolation, C. tropicalis decreased by 3.8% and C. glabrata increased by 3.4%. After 50 years of age, the higher the separation frequency. C. parapsilosis (31.3%) in 1~10s, C. tropicalis (30.3%) and C. glabrata (27.6%) in 41~50s, and C. tropicalis (28.6%) in 80s are relatively frequent. has been separated C. krusei was isolated in a relatively high proportion from females (60.9%). Therefore, a systematic and continuous nosocomial infection control system should be established for appropriate treatment as per antifungal treatment guidelines. The system should continuously monitor the distribution of Candida species and provide rapid identification results.

Isolation and Characterization of Cellulolytic Anaerobic Fungi from the Guts of the Hanwoo Cattle and the Korean Native Goat (한우 및 산양의 장내 섬유소 분해 혐기 곰팡이의 분리 및 특성 구명)

  • Kim, C.H.;Lee, S.S.
    • Journal of Animal Science and Technology
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    • v.45 no.6
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    • pp.1019-1030
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    • 2003
  • The study was conducted to isolate and identify highly fibrolytic anaerobic fungi from the guts of a Hanwoo steer and a Korean native goat, and then investigate the characterization of cellulolytic activity of an anaerobic fungus. Twenty-one anaerobic fungal colonies were isolated in the study, in which 16 colonies were isolated from the rumen contents of the Hanwoo steer and 5 colonies from the duodenal fluids of the Korean native goat. Four anaerobic fungi were selected based on higher cellulolytic enzyme activities to identify under a optical microscope. NLRI-M003 and -T004 belong to Neocallimastix genus and NLRI-M014 belongs to Piromyces genus based on the morphology of their thallus, sporangia, rhizoid and the number of flagella. NLRI-M001 appeared to be an unknown strain of anaerobic fungi due to its different morphology from existing types of anaerobic fungi, though the morpholgoy is similar to Orpinomyces sp. Supplementation of 2% anaerobic fungal culture(NLRI-M003) in rumen-mixed microorganisms increased in vitro DM degradability of rice straw and filter paper up to 4 and 11%, respectively, compared with non-supplementation(control). CMCase and xylanase activities in in vitro culture were also higher in 2% fungal supplementation than controls in both rice straw and filter paper substrates.

A Recombinant $Plasmodium$ $vivax$ Apical Membrane Antigen-1 to Detect Human Infection in Iran

  • Haghi, Afsaneh Motevalli;Khoramizade, Mohammad Reza;Nateghpour, Mehdi;Mohebali, Mehdi;Edrissian, Gholam Hossein;Eshraghian, Mohammad Reza;Sepehrizadeh, Zargham
    • Parasites, Hosts and Diseases
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    • v.50 no.1
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    • pp.15-21
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    • 2012
  • In Iran, $Plasmodium$ $vivax$ is responsible for more than 80% of the infected cases of malaria per year. Control interventions for vivax malaria in humans rely mainly on developed diagnostic methods. Recombinant $P.$ $vivax$ apical membrane antigen-1 (rPvAMA-1) has been reported to achieve designing rapid, sensitive, and specific molecular diagnosis. This study aimed to perform isolation and expression of a rPvAMA-1, derived from Iranian patients residing in an endemic area. Then, the diagnostic efficiency of the characterized Iranian PvAMA-1 was assessed using an indirect ELISA method. For this purpose, a partial region of AMA-1 gene was amplified, cloned, and expressed in pET32a plasmid. The recombinant $His-tagged$ protein was purified and used to coat the ELISA plate. Antibody detection was assessed by indirect ELISA using rPvAMA-1. The validity of the ELISA method for detection of anti-$P.$ $vivax$ antibodies in the field was compared to light microscopy on 84 confirmed $P.$ $vivax$ patients and compared to 84 non-$P.$ $vivax$ infected individuals. The ELISA cut-off value was calculated as the mean+2SD of OD values of the people living in malaria endemic areas from a south part of Iran. We found a cut-off point of OD=0.311 that showed the best correlation between the sera confirmed with $P.$ $vivax$ infection and healthy control sera. A sensitivity of 81.0% and specificity of 84.5% were found at this cut off titer. A good degree of statistical agreement was found between ELISA using rPvAMA-1 and light microscopy (0.827) by Kappa analysis.

Study on Measurement of Carcinogen-DNA adducts in exfoliated urothelial cells among workers by 32P-postlabelling methods (근로자의 뇨중 상피세포에서 32P-postlabeling에 의한 발암물질의 DNA adducts측정방법에 대한 연구)

  • Lee, Jin Heon;Roh, Jaehoon;Talaska, Glenn
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.10 no.1
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    • pp.1-17
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    • 2000
  • Carcinogen-DNA adduct analysis has potential for biomonitoring the earliest effects of exposure to many chemical carcinogens. They are the covalent reaction products of electrophiles and nucleophilic sites on DNA and the initial damage to DNA induced by many carcinogens. So many researchers begin to use them as biomarker for monitoring the earliest exposure of carcinogens and develop the effective analytical techniques about them. Randerath, Gupta and coworkers(1981, 1982) has also developed a $^{32}P$-postlabelling method as one among them. A major project for biomonitoring workers with carcinogen-DNA adducts is to develop non-invasive samples instead of tissues of target organs such as baldder and lung. This study use the exfoliated urothelial cells in urine for examine benzidine-DNA adducts. The content of exfoliated urothelial cells is not enough to significantly measure DNA content with spectrophotometer, and require the another way. So firstly washing the collected cells with PBS and 70% ethanol and centrifuge them for removing the crystals in urine, which block the isolation of DNA adducts. And then, measure the total nucleotide after $^{32}P$-postlabelling for calculating RAL. $[{\gamma}-^{32}P]ATP$ using for $^{32}P$-postlabelling, can synthesize with $[^{32}P]H_3PO_4$, and reagent and enzyme mixture (RM, EM), which is very economic in case of requiring a lot of them. Chromatography was composed of two steps. First step was to separate adduct ones from unadducted nucleotide, and secondary step was separate each adduct, which were performed with 4 kinds of solvents and different directions on TLC. With this procedure, we measure the DNA adducts in exfoliated urothelial cells of workers who were employed in benzidine and benzidine-dye company. RAL of adducts were $89.0{\times}10^7$ and $57.0{\times}10^7$ in them. In conclusion, we can significantly measure the DNA adduct in exfoliated urothelial cells by using the above $^{32}P$-postlabelling procedures, and use them to be biomonitoring workers who exposed carcinogens.

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Isolation of Bacillus amyloliquefaciens ATC6 Producing Acidic Cellulase (산성 Cellulase를 분비하는 Bacillus amyloliquefaciens ATC6의 분리)

  • Lee, Se-Hyung;Chae, Jong-Pyo;Kim, Min-Jeong;Kang, Dae-Kyung
    • Journal of Animal Science and Technology
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    • v.52 no.1
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    • pp.65-70
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    • 2010
  • A bacterium producing acidic cellulase was isolated from pig feces. The isolate, ATC6 strain, was found to be Gram-positive, non-motile, catalase-positive, and spore-forming stain. Under an electron microscope, the cells were observed to be rod-shaped. The isolate was identified as Bacillus amyloliquefaciens ATC6 on the basis of morphological and biochemical properties as well as 16S rRNA gene sequences. Optimum pH and temperature for the cellulase activity of the culture supernatant of B. amyloliquefaciens ATC6 were found to be pH 4.5 and $55^{\circ}C$, respectively. More than 80% of its maximum activity was maintained at pH 4.0. The cellulase activity was maintained at temperatures ranging from 35 to $55^{\circ}C$ after 2 h incubation at pH 4.5, whereas it's activity decreased rapidly at $65^{\circ}C$.

Isolation and Characterization of Oxygen-tolerant Mutant of Bifidobacterium longum. (Bifidobacterium longum 산소변이주의 분리와 변이주의 산소내성)

  • 안준배;김광엽;박종현
    • Microbiology and Biotechnology Letters
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    • v.26 no.6
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    • pp.476-482
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    • 1998
  • Growth sensitivity of bifidobacteria on oxygen hindered their industrial applications so that it was necessary to select oxygen-tolerant strains. Studies on their responses to oxygen might facilitate the effective utilization of bifidobacteria in industry. Oxygen-tolerant strain of Bifidobacterium longum JI-1 was able to remove 3% dissolved oxygen within 10 min whilst oxygen-sensitive strain of B. adolescentis, slime non-former, was not. The ability to remove environmental oxygen seemed to be related to the oxygen-tolerance of bifidobacteria. Mutant B. longum ADJ-1 was induced from the B. longum JI-1 under microaerobic atmosphere. There were no differences in sugar utilization pattern, NADH oxidative enzymes and cellular fatty acid compositions between them. The maximal cell density of the mutant was a little bit reduced to 81% of that of the mother strain. However, the mutant formed thick slime layer around its cell. The layer visualized with confocal scanning laser microscopy from the mutant was 6 ${\mu}{\textrm}{m}$ in diameter but that from the mother strain was only 3 ${\mu}{\textrm}{m}$. Therefore, the improved tolerances of the mutant might come from the slime layer, indicating the increase of the layer might be one of oxygen tolerance mechanisms for bifidobacteria.

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