• 생명과학회지
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• 제22권8호
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• pp.1092-1098
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• 2012

본 연구에서는 우방자를 70% 에탄올로 추출하여 얻어진 추출물을 n-hexane, methylene chloride, ethyl acetate로 순차용매 분획하였다. 각 분획물에 대해 activity-guided isolation을 수행하여 활성물질의 분리 정제를 실시하였다. 활성을 나타내는 물질은 silica gel chromatography (230 mesh), sephadex LH 20, recrystallization method를 이용하여 분리하였다. 각 화합물의 화학구조는 NMR 스펙트럼 데이터 해석하였고 arctiin, arctigenin, diarctigenin, matairesinol 으로 동정하였다. 이들을 human dermal fibroblast HS68 세포에 처리하여 얻어진 상등액은 ELISA kit를 활용하여 procollagen type I 생합성과 MMP-1 저해활성을 측정하였다. 측정결과 procollagen type I 생합성과 MMP-1 저해활성 결과 모두 arctiin이 가장 우수한 결과를 보였다. 이와 같은 결과를 통해 우방자에서 분리한 리그난 화합물들을 이용하여 주름개선 소재로 개발할 수 있을 것으로 사료 된다.

• 생명과학회지
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• 제19권6호
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• pp.744-750
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• 2009

토양에서 분리한 세포외 다당류 생산균주 PS-12는 형태학적, 생리학적, 화학적 분석에 의하여 Bacillus sp.에 속하는 균주로 동정되었으며, 분리균주 PS-12는 Bacillus sp. PS-12로 명명하였다. Bacillus sp. PS-127가 생산하는 세포외 다당류는 에탄올 침전, cetylpyridinium chloride (CPC) 침전과 gel permeation chromatography를 이용하여 정제하였으며, 정제된 다당류 PS-12의 당조성은 glucose, mannose, galactose와 fucose가 7:3.2:2:1의 몰비로 구성되어있었다. Bacillus sp. PS-12로부터 분리된 다당류 PS-12를 이용하여 면역증강효과를 확인하였다. TNF-${\alpha}$ 및 lL-6 측정은 RAW264.7 대식세포주를 사용하였으며 cytokine 정량을 위하여 ELISA kit를 이용하였다. RAW264.7 세포주에 대한 PS-12의 세포독성을 확인하기 위하여 세포독성이 10% 미만을 나타내는 농도인 2 ${\mu}g$/ml을 PS-12의 최대농도로 측정하였다. PS-12는 ${\mu}g$/ml에서 TNF-${\alpha}$를 정상세포보다 50배 이상 높은 수치로 생산하였다. 또한 lL-6의 생산을 농도 의존적으로 증가시켰다. 이러한 결과로부터 PS-127가 면역세포에 대해 세포독성을 거의 나타내지 않는 농도에서 대식세포로부터 TNF-${\alpha}$와 11-6의 cytokine 생산을 함으로써 면역증강효과를 나타낸다는 것을 확인하였다.

• 한국식품위생안전성학회지
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• 제23권4호
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• pp.343-347
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• 2008

최근 식사대용으로 각광을 받는 선식은 포장 후 더 이상 가열 처리 과정 없이 섭취하는 식품으로 포자를 형성하여 열처리 과정에서 완전히 사멸되지 않으며 생존하는 B. cereus에 의한 위해성이 문제시되고 있다. 따라서 시중에서 유통되고 있는 선식에 대한 B. cereus정량을 통해 오염 수준을 파악한 결과 선식 161점 중 57점 (35.4%)에서 B. cereur가 검출되었고 검출된 시료 57점 중에 100 FU/g 미만은 21점, 100-1000 CFU/g 미만은 33점, 1,000 CFU/g 이상은 3점이 검출되었다. 선식에서 분리한 B. cereus의 생화학적 특성을 통해 독소 형태를 알아본 결과는 enterotoxin 양성이 93%이며 emetic toxin 의심 균주는 5.3%였다. 이러한 B. cereus의 저감화를 위한 방안의 일환으로 선식의 제조 과정에서 B. cereus의 열처리 과정에 대한 사멸 정도를 측정하였다 측정한 결과 tryptic soy broth에 포자를 접종한 결과는 75, 80, 85, 90에서 40.3분, 8.8분, 3.3분, 1.1분이였다. 용질에 따라 수분 활성도가 낮아지면 열에 대한 저항성이 높아지므로 선식에 시료에 대한 D-value를 시험한 결과 $75^{\circ}C$, $80^{\circ}C$, $85^{\circ}C$, $90^{\circ}C$에서 37.1분, 22.5분, 4.9분, 3.1분이였다. Z-value는 tryptic soy broth에서는 $9.8^{\circ}C$이며 선식 시료에서는 $12.8^{\circ}C$였다. 따라서 선식에서 B. cereis에 대한 관리가 요구되어진다.

• 한국식품위생안전성학회지
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• 제28권2호
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• pp.181-187
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• 2013

전분의 사용원료를 확인하는 방법은 전분입자의 크기 또는 형태 등으로 분류하는 이화학적인 방법이 연구되었으나 원료별 또는 동일한 원료라도 품종에 따른 차이점으로 인하여 명확하게 확인하기 어려운 단점이 있어 유전자분석법을 시도하였다. 시료는 고구마 전분, 감자 전분, 옥수수 전분 및 타피오카 전분 등 총 11종을 사용하였으며, 유전자추출은 DNeasy plant mini 키트, magnetic DNA purification system 및 CTAB 방법으로 하였으며 추출유전자의 증폭을 위하여 WGA 키트로 처리하였다. 그리고 고구마, 감자, 옥수수 및 타피오카 검출을 위한 유전자 부위는 SSR (simple sequence repeat, ib-286-F/ib-286-R), 자당합성효소(potato sucrose synthase, Pss 01n-5'/Pss 01n-3'), 전분합성효소(starch synthase, SSllb 3-5'/SSllb 3-3') 및 SSR (SSRY26-F/SSRY26-R)를 각각 사용하였다. 그 결과 대부분의 경우 WGA를 처리한 경우에는 사용원료의 확인이 가능하였다.

• 한국농림기상학회지
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• 제20권2호
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• pp.205-213
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• 2018

작물 모델의 개발과 검증에 사용되는 생체중 자료는 파괴적 샘플링을 통해 얻어져 왔다. 파괴적 샘플링이 가지는 단점을 보완하기 위해 저가형 3D 센서인 Kinect 센서와 무료 공개 소프트웨어들을 사용하여 생체중을 추정하는 기법을 개발하였다. 특히, 많은 작물모델들이 개발되어 있지 않은 배추를 대상으로 입체이미지를 생성하여, 그로부터 얻어진 부피와 생체중 추정치의 신뢰도를 분석하고자 하였다. 크기가 다른 배추 결구 부위를 스캔하기 위해 Kinect 센서와, Microsoft가 무상으로 제공하는 Software Development Kit 내 Kinect Fusion Explorer 프로그램을 사용하였다. 개별 배추의 입체이미지를 생성하기 위해 3D 그래픽 편집 소프트웨어인 Meshlab을 활용하여 배경과 불필요한 물체를 수동으로 제거하였다. 또한, 불완전한 입체모델로부터 생체중 추정을 위해 3D 프린터 소프트웨어인 Makerbot Desktop 을 사용하여 배추를 생성하기 위해 필요한 플라스틱 필라멘트 소모량을 추정하였다. 입체모델 편집 프로그램인 Blender를 사용하여 부피를 추정하였을 때, 실제 부피에 비해 17.6%에서 2160.6% 범위의 상당한 오차가 있었다. 반면, 필라멘트 소요량은 실제 배추 생체중 변이의 98.7%를 설명할 수 있었다. 또한, 이들의 상관관계는 5% 수준에서 유의하였다. 이러한 결과들은 직접적인 부피 계산 절차를 제외하더라도 간편하게 생체중을 추정할 수 있음을 확인하였다. Kinect 센서를 사용하여 배추의 생체중 추정이 가능하다는 것이 확인 되었으나, 기존의 고가형 3D 센서에 비해 낮은 해상도와 주간에 활용이 어려운 점이 있다. 그럼에도 불구하고, 배추 생육 모델의 시계열적 검증 자료를 Kinect 센서를 이용하여 간편하고 신속하게 획득할 수 있어 모델의 불확도를 감소하는 데에 기여할 수 있을 것으로 판단된다. 따라서, 후속 연구에서 보다 저렴한 가격대의 3D 센서들을 대상으로 야외 및 주간조건애서 작물의 생체중 측정 가능성에 대해 검토하고 작물 모형 개발 및 개선을 위한 기술개발이 이루어져야 할 것으로 사료된다.

• Pediatric Infection and Vaccine
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• 제3권2호
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• pp.185-193
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• 1996

Purpose : The incidence of measles was gradually decreased since 1963 when measles vaccination was firstly developed and liscenced in the world. But, recently the outbreaks of measles in infants and school children have been reported despite of wide spread use of measles vaccination. This study was performed to evaluate the efficacy of measles vaccination and the necessity of revaccination in Korean infants and children. Methods : 168 subjects of mothers and neonates, infants and children were enrolled in this study during the periods of 10 months from March to December in 1995. Measles specific IgG in the sera of mothers and children was measured using EIA kit (Sigma Co., MO, USA). Antibody titer of over or equal to 110 AU/ml was considered positive. Results : The results obtained were as follows. 1) Values of measles specific IgG in the sera of mother and neonate were 82.9 AU/ml and 89.3 AU/ml respectively and were rapidly decreased within 6 month after birth. Positive antibody levels (${\geq}$ 110 AU/ml) were observed in only 25 % of neonates. 2) In vaccinated children, values of measles specific IgG were 117.4 AU/ml in 9~15 month group, 76.9 AU/ml in 3~6 year group and 79.5 AU/ml in 10~15 year group after either one or two times of measles vaccination. Positive antibody levels in vaccinated children were observed in 57.7% of 9~15 month group, 38.4% of 3~6 year group and 34.7% of 10~15 year group. Conclusion : These results suggest that primary measles vaccination before 6 months of age can be considered and revaccination of measles should be recommended before 3~6 years of age. Further studies will be needed to clarify the reasons of high proportion of primary measles vaccination failure and to established the appropriate schedule of measles vaccination in korean infants and children.

• Archives of Plastic Surgery
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• 제42권1호
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• pp.11-19
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• 2015

Background Wound healing is an interaction of a complex signaling cascade of cellular events, including inflammation, proliferation, and maturation. $K^+$ channels modulate the mitogen-activated protein kinase (MAPK) signaling pathway. Here, we investigated whether $K^+$ channel-activated MAPK signaling directs collagen synthesis and angiogenesis in wound healing. Methods The human skin fibroblast HS27 cell line was used to examine cell viability and collagen synthesis after potassium chloride (KCl) treatment by Cell Counting Kit-8 (CCK-8) and western blotting. To investigate whether $K^+$ ion channels function upstream of MAPK signaling, thus affecting collagen synthesis and angiogenesis, we examined alteration of MAPK expression after treatment with KCl (channel inhibitor), NS1619 (channel activator), or kinase inhibitors. To research the effect of KCl on angiogenesis, angiogenesis-related proteins such as thrombospondin 1 (TSP1), anti-angiogenic factor, basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF), pro-angiogenic factor were assayed by western blot. Results The viability of HS27 cells was not affected by 25 mM KCl. Collagen synthesis increased dependent on time and concentration of KCl exposure. The phosphorylations of MAPK proteins such as extracellular-signal-regulated kinase (ERK) and p38 increased about 2.5-3 fold in the KCl treatment cells and were inhibited by treatment of NS1619. TSP1 expression increased by 100%, bFGF expression decreased by 40%, and there is no significant differences in the VEGF level by KCl treatment, TSP1 was inhibited by NS1619 or kinase inhibitors. Conclusions Our results suggest that KCl may function as a therapeutic agent for wound healing in the skin through MAPK signaling mediated by the $K^+$ ion channel.

• Parasites, Hosts and Diseases
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• 제52권3호
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• pp.305-310
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• 2014

Ascidian soft tunic syndrome (AsSTS) caused by Azumiobodo hoyamushi (A. hoyamushi) is a serious aquaculture problem that results in mass mortality of ascidians. Accordingly, the early and accurate detection of A. hoyamushi would contribute substantially to disease management and prevention of transmission. Recently, the loop-mediated isothermal amplification (LAMP) method was adopted for clinical diagnosis of a range of infectious diseases. Here, the authors describe a rapid and efficient LAMP-based method targeting the 18S rDNA gene for detection of A. hoyamushi using ascidian DNA for the diagnosis of AsSTS. A. hoyamushi LAMP assay amplified the DNA of 0.01 parasites per reaction and detected A. hoyamushi in 10 ng of ascidian DNA. To validate A. hoyamushi 18S rDNA LAMP assays, AsSTS-suspected and non-diseased ascidians were examined by microscopy, PCR, and by using the LAMP assay. When PCR was used as a gold standard, the LAMP assay showed good agreement in terms of sensitivity, positive predictive value (PPV), and negative predictive value (NPV). In the present study, a LAMP assay based on directly heat-treated samples was found to be as efficient as DNA extraction using a commercial kit for detecting A. hoyamushi. Taken together, this study shows the devised A. hoyamushi LAMP assay could be used to diagnose AsSTS in a straightforward, sensitive, and specific manner, that it could be used for forecasting, surveillance, and quarantine of AsSTS.

• 대한수의학회지
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• 제32권4호
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• pp.641-647
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• 1992

This study was conducted to determine the serum antibodies against toxoplasma in the artificially infected dogs, pet and street dogs by latex agglutination (LA) and indirect fluorescent antibody (IFA) test. LA test was carried out with commercial Toxo-MT kit (Eiken chemical Co.) and IFA test was carried out with rabbit-anti-dog IgG labelled with FITC (Cappel Co.) and toxo-antigen slides prepared in laboratory. The results obtained were as follows ; 1. Antibodies to Toxoplasma gondii in the artificially infected dogs were detected firstly at the Day 8 in IFA and Day 9 in LA test after inoculation. Positive antibody reactions by these tests were declined gradually afterward but maintained up to 12 weeks. 2. In LA test serum antibody titers in 310 test sera were shown as 10 cases(32%) in 1 : 32.5(1.0%) in 1 : 64, 4(1.3%) in 1 : 128 and 2(0.7%) in 1 : 256. In IFA test serum antibody titers 310 test sera were shown as 17 cases(5.5%) in 1 : 64, 8(2.6%) in 1 : 128 and 5(1.6%) in 1 : 256. 3. In the total of 310 sera from pet and street dogs toxoplasma antibody positive rates were 21 cases (6.8%) by LA and 30 cases (9.7%) by IFA test and the positive detection rates between these two groups by LA and IFA test were not significant(p<0.05). 4. In the total of 115 sera from pet dogs toxoplasma antibody positive rates were 12 cases(10.4%) by LA and 15(13.0%) by IFA test. And in the 195 street dogs the positive rates were 9 cases(4.6%) by LA and 15(7.7%) by IFA test. Also, the positive detection rates between these two groups by LA and IFA test were not significant(p<0.05). 5. Agreement of reactivity between LA and IFA test for 310 sera was 91.3% in total of 283 cases consisting of 12 cases(3.9%) of both LA and IFA positive and 271 cases(87.4%) of LA and IFA negative. 6. LA test was almostly equivalent to the IFA test in producibility and proved to be a simple tool for the screening of toxoplasma antibody in laboratory.

• 대한수의학회지
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• 제49권2호
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• pp.149-155
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• 2009

The objective of this study was to analyze data from the planned national serological monitoring program for Aujeszky's disease (AD) using a simulation model to evaluate probable outcomes expected in the sample derived from the simulated herds at predefined within-herd prevalence and herd prevalence. Additionally, prevalence at animal- and herd-level estimated by the stochastic simulation model based on the distributions of the proportion of infected herds and test-positive animals was compared with those of data from a national serological survey in 2006, in which 106,762 fattening pigs from 5,325 herds were tested for AD using a commercial ELISA kit. A fixed value of 95% was used for test sensitivity, and the specificity was modeled with a minimum, most likely and maximum of 95, 97 and 99%, respectively. The within-herd prevalence and herd prevalence was modeled using Pert and Triang distributions, respectively with a minimum, most likely and maximum point values. In all calculations, population size of 1,000 was used due to lack of representative information. The mean number of infected herds and true test-positives was estimated to be 27 herds (median = 25; 95% percentile 44) and 214 pigs (median = 196; 95% percentile 423), respectively. When testing 20 pigs (mean of 2006 survey) in each herd, there was a 3.3% probability that the potential for false-positive reactions due to less than 100% specificity of the ELISA test would be detected. It was found that the model showed prevalence of 0.21% (99% percentile 0.50%) and 0.5% (99% percentile 0.99%) at animal- and herd-level, respectively. These rates were much similar to data from the 2006 survey (0.62% versus 0.83%). The overall mean herd-level sensitivity of the 2006 survey for fattening pigs was 99.9%, with only a 0.2% probability of failing to detect at least one infected herd.