• Pediatric Infection and Vaccine
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• 제22권2호
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• pp.106-112
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• 2015

목적: 장기간의 항생제 치료는 중이염 어린이 환자의 중이액으로부터 원인균이 배양되는 것을 방해한다. 본 연구는 배양 음성 중이액으로부터 분자적 진단에 의한 신속한 균 검출 가능성 여부를 확인하고자 하였다. 방법: 폐구균 lytA 유전자를 표적으로 하는 PCR과 LAMP로 민감도와 특이도를 비교 결정하고, 임상중이액에서의 폐구균 검출에 적용하였다. 결과: PCR 기법에 의한 폐구균 검출 최소한계는 약 $10^4$ 집락형성단위(CFU)이고, LAMP의 검출 최소한계는 10 CFU에서 결정되었다. 한편 두 가지 검사법 모두 Haemophilus influenzae 와 Moraxella catarrhalis 에 대해 $10^6$ CFU 이상에서도 DNA를 증폭하지 않았다. 22개의 배양음성 중이액 중에서 12개 검체가 LAMP-양성(54.5%, 12/22)으로 확인되었고, 이들 12개 LAMP-양성 검체 중, 3개의 검체만이 PCR-양성으로 확인되었다(25%, 3/12). 본 연구의 결과는 LAMP 기법의 폐구균 검출 해상력이 PCR 기법에 비교하여 4배 이상 높음을 보여준다(P<0.01). 결론: lytA -특이 LAMP 기법은, 중이액 내의 타 병원균과는 교차반응 없이 10 CFU 폐구균의 DNA를 검출할 수 있는 고해상 기술로서, 중이액 폐구균 검출 및 폐구균 백신의 보급에 따른 백신 효과 평가에 적용이 기대된다.

• 농업과학연구
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• 제12권2호
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• pp.199-205
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• 1985

1. 벼 종자(種子)의 변색정도(變色程度)에 따른 균류(菌類)의 검출(檢出)을 조사(調査)한 결과(結果) 변색(變色)이 심한 종자(種子)에서 Alternaria tenuis, Cladosporium cladosporioides, Curvularia lunata, Fusarium semitectum, Gerlachia oryzae, Phoma sp. Trichoconiella padwickii 등이 높은 비율(比率)로 검출(檢出)되었고 특히 반점종자(斑點種子)에서는 Helminthosporium 균(菌)이 높은 비율(比率)로 검출(檢出)되었다. 2. 벼 종자상(種子上)에서 진균(眞菌)의 검출(檢出)에 미치는 광선(光線)의 영향(影響)을 조사(調査)한 결과(結果) Alternaria tenuis, Helminthosporium oryaze, Trichoconiella padwickii, Fusarium moniliforme, Fusarium semitectum 등(等)은 암흑구(暗黑區)에서 보다 광선조사구(光線照射區)에서 검출율(檢出率)이 높았고 Phoma sp. Curvularia lunata, Epicoccum purpurascens, Verticillium sp.는 광선조사구(光線照射區)나 암흑구(暗黑區)나 거의 차이가 없었으며 Gerlachia oryzae는 암흑구(暗黑區)에서 오히려 검출율(檢出率)이 높았다. 3. 벼 종자상(種子上)의 배양온도(培養溫度)가 균류(菌類)의 검출(檢出)에 미치는 영향(影響)을 조사(調査)하였던 바 Alternaria tenuis, Cladosporium cladosporioides는 저온구$(15^{\circ}C)$에서 검출율(檢出率)이 높았고 Phoma sp.는 고온구$(30^{\circ}C)$에서 검출율(檢出率)이 높았다. 4. 검출(檢出)된 각(各) 균류(菌類)가 종자(種子) 변색(變色)및 수량(收量)에 미치는 영향(影響)을 조사(調査)하였던 바 공시(供試)한 7게(個)의 균류(菌類) 모두가 종자(種子) 변색(變色)에 관여(關與)하여 인공접종(人工接種)한 이삭에서의 천립중(千粒重)은 대조구의 천립중(千粒重)에 비해 모두 낮았고 특히 공시(供試)한 7개(個)의 균(菌)을 혼합접종(混合接種)한 이삭의 천립중(千粒重)과 등숙비율(登熟比率)은 대조구보다 저하(低下)됨을 알 수 있었다.

• 식물병연구
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• 제17권3호
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• pp.369-375
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• 2011

Fusarium verticillioides(완전세대: Gibberella moniliformis)는 Gibberellea fujikuroi 종 복합체에 속하는 식물병원균으로서 옥수수의 줄기와 이삭에 썩음병을 일으킬 뿐 아니라 인축에 중독증을 일으키는 fumonisin 곰팡이 독소를 생산한다. 본 연구의 목적은 옥수수에 주로 발생하는 fumonisin 생성가능 G. fujikuori 종 복합체 소속 Fusarium 곰팡이 중 F. verticillioides와 그 외 F. proliferatum, F. fujikuori 등을 서로 구별할 수 있는 종 특이적 PCR primer 조합을 개발하는 것이다. RNA polymerase II beta subunit 유전자(RPB2)의 염기서열로부터 제작된 특이 primer 조합(RVERT1와 RVERT2)은 우리나라 옥수수에서 분리한 잠재적인 fumonisin 생성 G. fujikuori 종 복합체 균주 중 오직 F. verticillioides로부터 208 bp 크기의 단일 DNA 절편을 증폭하였다. 한편 F. verticillioides를 포함한 모든 조사균주는 fumonisin 생합성에 필수적인 FUM1 유전자를 포함하고 있었다. 개발된 특이 primer 조합의 검출한계는 분석 곰팡이 DNA 0.125 pg/${\mu}l$ 수준이었다. 한편, 같은 primer 조합으로 Fusarium spp.에 오염된 옥수수 시료의 게놈 DNA로부터 F. verticillioides 특이 DNA 절편이 증폭되었다. 이와 같은 결과를 종합할 때, 본 연구에서 개발된 primer 조합은 여러 곡물 시료에 오염되어 있는 F. verticillioides 균주의 검출과 종 동정에 유용하게 사용될 것이다.

• Food Science and Biotechnology
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• 제17권2호
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• pp.262-266
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• 2008

This study was conducted to provide a vitamin $B_{12}$ database for the representative Korean food items and to assess the dietary intake assessment of vitamin $B_{12}$ for Koreans. The vitamin $B_{12}$ content of 106 foods had been determined by high performance liquid chromatography (HPLC) using column switching method. Rich sources of vitamin $B_{12}$ were meats, milk, and egg ($0.3-3.4\;{\mu}g/100g$). Vegetables and fruits contained vitamin $B_{12}$ below limit of detection (LOD). The major food sources for vitamin $B_{12}$ intake were milk (72.0%), meats (22.3%), egg (3.6%), and fishes & shellfishes (2.1%). Mean vitamin $B_{12}$ intake of Koreans was $3.16\;{\mu}g$/person/day. The proportion of population with intake below estimate average requirement (EAR) and above recommended intake (RI) of vitamin $B_{12}$ was 60.7 and 36.5%, respectively. The vitamin $B_{12}$ intake level of young children with 1-2 years which was 834.6% of RI while the intake level of the older adults 50 years and older was only 70.0% of RI. Also, there were regional differences between urban and rural area. The population with intake below RI was larger than that with intake above RI in Korea.

• 한국임상수의학회지
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• 제8권2호
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• pp.147-152
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• 1991

습도가 높고 환기가 불량하며 비위생적인 우리에서 같이 사육한 두 마리의 어린 염소에서 발생한 진균성피부염의 일차원인이 Trichophyton mentagrophytes이었음을 보고하였다. 두 환축의 진단은 피부소파 시료에 KOH를 사용하여 균요소를 검출하고 3$0^{\circ}C$의 진균배지에서 순수배양하여 균을 분리하여 확인하였다. 한 마리는 안면에서 나머지 한 마리는 목과 귀에서 병변이 발견되었다. 같은 우리에서 사육 중이던 두 마리의 성축에서는 병변이 발견되지 않았다. 더욱이 염소의 주인과 주인가족들에서도 병변이 발견되지 않았다. 두 분리균주를 개량된 sunflower seed medium을 이용하여 유전적 교배실험을 한 결과 둘 다(-) mating type에 속했다. Hair performance test결과 두 균주는 모두 각질용해작용을 나타내었다. 공중위생학적인 중요성과 화학요법에 대해서도 고찰하였다. 감염경로에 대해서는 명확히 밝힐 수 없었다.

• 대한수의학회지
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• 제37권4호
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• pp.793-807
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• 1997

We tried to develop detection system of porcine reproductive and respiratory syndrome virus(PRRSV) by in situ hybridization(ISH) in the piglets experimentally infected with KPRRS-2, the Korean isolate(12 piglets) or Mn-1b, the American isolate(4 piglets), and in the natural infection suspected 6 piglets. Twelve 30-days-old piglets(two pigs per each inoculated group) were inoculated by nasal instillation of KPRRS-2 virus(total dose $10^{4.5}TCID_{50}$), Six piglets(one pig per each group) were induced contact infection with inoculated piglets, during the experiment, and two piglets were used as control. Inoculated or contacted piglets were euthanized at 1, 3, 5, 7, 14 and 21 days postinoculation(DPI). The respiratory signs such as coughing and nasal discharge were observed on day 3 DPI, and ear cyanosis were on day 5 DPI, including contacted piglets. Through the necropsy, purple discolorization of dorsal part of lung, and hypertrophy of local lymph nodes were observed. The histopathological lesions of lung were interstitial pneumonia characterized by type 2 pneumocyte hyperplasia. We prepared the probe for ISH by RNA isolation from KPRRS-2, RT-PCR, and biotin labeling. We performed the ISH within only 1~2 hours using $Microprobe^{TM}$ capillary action system. As the results, the strong red specific positive signals, means PRRSV distribution, was mainly observed in the cytoplasm of alveolar macrophages. And also signals were detected in some type 2 pneumocytes and bronchiolar epithelium of lung, myocardium, liver, kidney, tonsil, spleen, gastrointestinal mucosa, testis and lymph nodes.

• Asian Pacific Journal of Cancer Prevention
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• 제15권4호
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• pp.1851-1857
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• 2014

Objective: The main purpose of this work was to investigate the effect of berberine hydrochloride (BH) on the proliferation, apoptosis, migration, and invasion of CNE-1 nasopharyngeal carcinoma cells. Our results shed light on the functional components of traditional Chinese herbs for potential use in modern medicine. Methods: The CNE-1 cell line was treated with different concentrations of BH and effects on cell viability and proliferation were evaluated using the Cell Counting Kit-8 (CCK-8) assay. Anti-migratory and anti-invasive actions of BH were investigated using wound healing assays and the Millicell Hanging cell culture insert system, respectively. Expression of the epithelial-mesenchymal transition (EMT)-related gene twist (Twist) was analyzed by real-time PCR and Western blotting. Apoptosis was estimated with an annexin-V fluorescein (FITC) apoptosis detection kit, as well as with reference to levels of activated caspase-3 of CNE-1 cells before and after treatment with BH utilizing fluorescence spectroscopy. Results: BH was capable of reducing proliferation and viability of CNE-1 cells in a dose- and time-dependent manner, also demonstrating anti-migratory and anti-invasive capacities which correlated with reduction in expression of Twist. Finally, BH was able to induce significant amounts of apoptosis in CNE-1 cells, as demonstrated by an increase in the activity of caspase-3 and in annexin-V staining following treatment. Conclusion: BH extracted from rhizoma coptidis demonstrated an ability to block proliferation, induce apoptosis, and impair the migration and invasion of the CNE-1 cell line Considering these properties, our results suggest that BH could be an important compound for consideration in the treatment of nasopharyngeal carcinoma.

• 한국수정란이식학회:학술대회논문집
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• 한국수정란이식학회 2002년도 국제심포지엄
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• pp.73-73
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• 2002

Nuclear transfer (NT) techniques have advanced in the last years, and cloned animals have been produced by using somatic cells in several species including pig. However, it is difficult that the nuclear transfer porcine embryos development to blastocyst stage overcoming the cell block in vitro. Abnormal segregation of chromosomes in nuclear transferred embryos on genome activation stage bring about embryo degeneration, abnormal blastocyst, delayed and low embryo development. Thus, we are evaluated that the correlations of the frequency of embryo developmental rates and chromosome aberration in NT and In viかo fertilization (IVF) derived embryo. We are used for ear-skin-fibroblast cell in NT. If only karyotyping of embryonic cells are chromosomally abnormal, they may difficultly remain undetected. Then, we evaluate the chromosome aberrations, fluorescent in situ hybridization (FISH) with porcine chromosome 1 submetacentric specific DNA probe were excuted. In normal diploid cell nucleus, two hybridization signal was detected. In contrast, abnormal cell figured one or three over signals. The developmental rates of NT and IVF embryos were 55% vs 63%, 32% vs 33% and 13% vs 17% in 2 cell, 8 cell and blastocyst, respectively. When looking at the types of chromosome aberration, the detection of aneuploidy at Day 3 on the embryo culture. The percentage of chromosome aneuploidy of NT and IVF at 4-cell stage 40.0%, 31.3%, respectively. This result indicate that chromosomal abnormalities are associated with low developmental rate in porcine NT embryo. It is also suggest that abnormal porcine embryos produced by NT associated with lower implantation rate, increase abortion rate and production of abnormal fetuses.

• Asian-Australasian Journal of Animal Sciences
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• 제15권9호
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• pp.1237-1243
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• 2002

We have developed a reliable and noninvasive method for swine genotyping of single locus nuclear gene with aged single hair follicles delivered by general mail. The method is based on booster and nested PCR amplification with step-wise increase of primers and dNTPs concentrations followed by restriction endonuclease digestion. To establish this method, the ryanodine receptor (RYR 1) locus which is an economically important trait in swine industry was employed for genotyping experiment. The 3-step PCR amplication method is much less dependent on the quantity and quality of template DNA and produces enough amplification product for the detection on the ethidium bromide-stained gel such as RFLP analysis. A total of 120 pigs were subjected to the RYR 1 genotyping analysis using three-step PCR method which amplified enough quantity of PCR products from the aged single hair follicles for RFLP analysis and genotyping results were identical to the results of the corresponding ethanol-fixed skeletal muscle tissue. This approach will be a great help for porcine breeders and investigators in genotyping of swine. They can receive genotyping results later by simply plucking single hairs of their pigs at farm and sending them in general mail to the diagnostic laboratory which eliminates the inconveniences to collect ear tissue or blood cells from pigs, or the investigator's need for travel to farms in order to collect fresh hair samples.

• 한국동물위생학회지
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• 제36권2호
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• pp.105-110
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• 2013

Bovine viral diarrhea virus (BVDV) is very important disease in domestic and wild ruminants and has a world wide distribution. Cattle persistently infected with BVDV (BVDV-PI) are the primary reservoir for BVDV infection in Korean native cattle herds. The prevalence of cattle persistently infected with BVDV (BVD-PI) was determined using 4,260 heads from 29 Korean native cattle farms at 8 districts from 2011 to 2012. The sera and ear nothches were collected for each sample. We surveyed BVD-PI cattle using antibody ELISA and antigen capture ELISA for detection of antibody and antigen respectively. Three thousand seventy-six cattle (72.2%) were positive for BVDV antibody and a total of 27 BVD-PI cattle were found in 12 farms. 11 cattle (40.7%) out of the total 27 BVDV-PI cattle were six months old or under. The positive rate of BVDV antibody (83.2%) from 12 farms with BVD-PI cattle was higher than the positive rate of BVDV antibody (63.6%) from 17 farms without BVD-PI cattle.