Biogenetical study on potential regulatory factors involved in expression of region III genes of Escherichia coli K99 adhesion gene cluster

이존화,백병걸,강창원,Lee, John-Hwa,Baek, Byeong-Kirl,Kang, Chang-Won The Korean Society of Veterinary Science 2002 大韓獸醫學會誌 Vol.42 No.4

대장균 K99 섬모의 생합성은 8개로 구성된 K99의 특이 유전자의 발현과 숙주유래 인자에 의해 조절되는 다른 유전자들의 발현에 의존된다. 본 연구에서는 K99섬모 유전자군중 제 3지역 발현에 유전조절자의 관련성 여부를 연구하였다. Gel retardation 분석 방법올 통하여 제3지역의 발현에 관련된 유전조절단위를 함유한 fanF 지역의 단백질 인자가 부착됨을 암시하였다. 이 분석방법을 이용한 결과는 또한 이 단백질 인자가 K99 유전자에서 유래되지 않고 대장균 염색체에서 유래됨을 지적하였다. 이를 보다 더 조사하기 위하여 대장균 염색체에 Tn10 transposon 유전자 변이 실험을 수행하였다. K99 유전자군으로부터 제 1지역과 제2지역의 유전자를 제거시키고, 제 3지역의 유전자인 fanG에 transposon TnlacZ를 삽입한 pTL65-1 plasmid을 제작하였다. 이 pTL65-1는 다시 Tn10으로 염색체가 변이된 대장균에 주입하였다. 3개의 pTL65-1 주입된 Tn10 대장균 변이체 내에서 fanG의 발현이 증가되었다. 이들 변이대장균으로부터 Tn10이 어떤 염색체 유전자 부위를 변이 시켰는지 확인하기 위해서 변이부위 유전자를 cloning하여 염기서열을 분석하였다. 이중 2개의 clone이 동일하였으며 지금까지 알려지지 않은 유전자였다. 이들 2개의 변이체 내에서 fanG의 발현은 대조군과 비교해 약 4.2배 증가 되였다. 결론적으로 이들 2개의 clone으로부터 유래된 인자는 지금까지 알려지지 않은 제 3지역의 억제 조절자임을 나타내었다.

PC12 세포에서 신경전달물질 방출을 저해하는 생리활성물질 FS11052의 탐색

이윤식,이존화,Lee, Yun-Sik,Lee, John Hwa 대한수의학회 2006 大韓獸醫學會誌 Vol.46 No.2

We established an in vitro experimental system using the following procedure. We first introduced tritium-labeled norepinephrine ([$^3$H]-NE) into PC12 cells, The [$^3$H]-NE incorporated into PC12 cells were then stimulated by a high concentration (60 mM) of $K^+$ buffer during 12 minutes. Then, we collected $100{\mu}l$ supernatant and counted the amount of [$^3$H]-NE release from PC12 cells with a scintillation counter. After screening fungal, Streptomyces spp. or bacterial product using this experimental sytem, we obtained FS11052 from Streptomyces spp. which inhibited [$^3$H]-NE release from PC12 cells. FS11052 also inhibits the release of ATP as a neurotransmitter of PC12 cells and rat cortical neurons, The inhibitory effect was seen even when the PC12 cells were treated with low $K^-$ buffer containing ionomycin ($1{\mu}M$) as an ionopore. This result suggests that the inhibitory action of FS11052 on neurotransmitter release appeared after the influx of $Ca^{2+}$.

Brucella abortus 감염 흰쥐에서의 rifaampin과 streptomycin의 치료효과

백병걸,최춘기,임채웅,이존화,김병수,이성일,허진,Baek, Byeong-Kirl,Choi, Chun-ki,Lim, Chae-woong,Lee, John-hwa,Kim, Byeong-Soo,Lee, Sung-il,Hur, Jin,Ibulaimu, Kakoma 대한수의학회 2004 大韓獸醫學會誌 Vol.44 No.3

This study was carried out to investigate the efficacy of rifampin with or without streptomycin in male Sprague-Dawley (SD) rats experimentally inoculated with Brucella abortus. Thirty rats were intraperitoneally inoculated with $1.0{\times}10^9$colony-forming units of B. abortus. They were divided into 3 groups by treatment with antibiotic. 10 rats in Group A were orally administrated with rifampin, 10 rats in Group B with rifampin orally and with streptomycin intramuscularly over 12 weeks starting at 1 week post infection (PI). A placebo recipient in Group C was inoculated with sterile saline without antibiotics. All animals were monitored by tube agglutination test (TAT) and AMOS-PCR to evaluate the efficiency of the antibiotics to B. abortus infection. The antibody titers in Groups A, B and C were 1:400, 1:400 and 1:800 as measured by TAT at the first week PI, respectively. The antibody titer in Group A decreased to 1:100 by the 13th week PI. That in the control group was observed as high antibody titer until 13th weeks PI, but the antibody response in Group B was low(1:50) from the 5th week to the 13th week PI. AMOS-PCR there was evidence of relapse of B. abortus in group A in liver and spleen specimens at the 13th week PI. B. abortus DNA was detected in Group C in liver and spleen specimens from the 1st week to 13th week PI by AMOS-PCR. However AMOS-PCR could not detect any organism in Group B from the 3rd week PI until the end of the study. This study demonstrated that administration of a combination of rifampin and streptomycin was more efficacious than administration of rifampin alone. A significant reduction in antibody titer was observed when a combination of 15 mg/kg/day of rifampin per os and 15 mg/kg/day streptomycin intramuscularly was used in comparison with the antibody of control group.

Eimeria tenella : 抗콕시듐제 投藥 中斷에 따른 닭 콕시듐증 誘發

Byeong Kirl BAEK(白秉杰),Jin Ho KIM(金鎭浩),John Hwa LEE(李存華) 한국예방수의학회 1988 예방수의학회지 Vol.12 No.2

In order to odbserve the pattern fo coccidiosis of 4 weeks chickens which were administrated with 0.01 % amprolium and 0.01% nitrofurazone in water as an inticocidial drug, were inoculated with 1000 eimeria tenella orally during administration of anticoccidial drug and reinoculated with it at the second day after anticoccideal drug withdrawn. And to determine the resistance against reinoculation with E.tenella, OPG, lesion score and microscopical observation of cecum in chicken were compared chronically the difference betwiin before withdrawal of anticoccidial drug with after withdrawal of anticoccideal drug to chickens inoculated with E. tenella. The results were summarized as follfws: 1. The typical coccidiosis in the nonmedicated chickens were shown up not only the destruction of epitherial cell layer and lamina propia of cecum by the procedure of developing oocysts, but also the signs of inflammation of cecum microscopically from the 7th day through the 10th day and after reinoculation.The signs of coccidiosis are almostly same. OPG in chickens inoculated with E. tenella were 4400-26090 at the 7th day through the 10th day. But after reinoculation, it was decreased to 150-5930. 2. The coccidiosis of chickens which were administrated 0.01% amprolium was not shown up, However withdrawing of anticoccidial drug, chickens were reinoculated with E. tenella, have brought the typical coccidiosis in cecum accompanied with the sloughing of epitherial cell layer and lamina propian and a great number of oocyt in faecal examinatoin(++++). OPG in the reinoculatoin chickens were 1520-22800 at the 7th day through the 14th day. 3. The slight pattern of coccidiosis(++) was shown up in the chickens which were administrated with 0.01% nitrofurazone and also are same pattern in the reinoculated chinkens withdrawn nitrofurazone. and OPG in the chickens administrated and in the chickens withdrawn nitrofurazone are apparently less than the OPG of nonmedicated chickens and reinoculated chickens after wethdrawing amprolium.

肝蛭症의 酵素免疫學的 診斷을 위한 抗原分劃

李宰求(Jae Ku Rhee),白秉杰(Byeong Kirl Baek),李存華(John Hwa Lee) 대한기생충학열대의학회 1986 The Korean Journal of Parasitology Vol.24 No.2

Succinic acid inhibits the uptake of Brucella abortus 544 into RAW 264.7 cells and promotes survival of B. abortus in ICR mice

Tran Xuan Ngoc Huy,Trang Thi Nguyen,Alisha Wehdnesday Bernardo Reyes,김희진(Heejin Kim),민원기(WonGi Min),이후장(Hu Jang Lee),이존화(John Hwa Lee),김석(Suk Kim) 한국예방수의학회 2021 예방수의학회지 Vol.45 No.4

This study aims to investigate the effects of exogenous succinic acid (SCA) on Brucella (B.) abortus infection in macrophage RAW 264.7 cells and ICR mice. Firstly, the in vitro experiment was conducted by MTT cytotoxicity and bacterial internalization assay to evaluate the uptake of B. abortus into macrophage cells. Two non-cytotoxic concentrations of SCA demonstrated attenuated invasion of Brucella into macrophages at 30 and 45 min post- infection (pi). Secondly, ICR mice were treated with SCA and infected with B. abortus. On day-14 pi, spleen and blood serum were collected to evaluate the bacterial burden and total spleen weight as well as the production of cytokine/chemokine, respectively. The results showed that SCA treatment promoted bacterial growth and reduced the total spleen weight in mice. Furthermore, SCA treatment increased the level of IL-10 cytokine in the sera, while dampening the production of MCP-1 chemokine compared to the control. The results of bacterial load in spleen and spleen weight together with cytokine/chemokine production profile in the sera indicated that SCA induced the host anti-inflammatory response which is beneficial for the survival of Brucella. Therefore, these findings suggest that SCA contributed to host immunity against Brucella infection and the emerging potential topic-immunometabolism should be invested for further investigations.

Characterization of Insulin-like Growth Factor (IGF)-1, IGF- Binding Proteins and Proteolytic Activity of IGFBP-3 in Normal and Osteoathritis Synovial Fluid of Bovine

Jin, Song Jun,Lee, John Hwa,Kang, Chang Won 한국수의공중보건학회 2001 예방수의학회지 Vol.25 No.1

한우의 골 관절염에서 IGF-I/IGFBPs axis의 변동을 연구하기 위하여 활액 내 IGF-I의 농도, IGFBPs의 patterns 및 IGF-I과 IGFBP-3의 protease activity를 정상 한우 혈청과 비교 분석하여 다음과 같은 결과를 얻었다. 한우 골관절염 활액 내 총 IGF-I의 농도는 정상 활액 보다 증가하였고 free IGF-I의 농도는 정상 활액 보다 감소하였다.(n = 30, p < 0.05). 한우 골 관절염 활액 내 IGF-I proteolytic activity는 정상 활액 보다 활성화되었다. Western ligand blotting의한 IGFBPs의 partterns에서 정상 활액과 골 관절염 활액에서 IGFBP-3, -2, -1 및 -4을 관찰하였다. 이 중 IGFBP-2는 골 관절염 활액에서 정상 활액보다 증가하였으며, Western immunoblotting에서의 IGFBP-3는 골 관절염 활액에서 정상 활액보다 증가하였다. 정상 활액 내 IGFBP-3 proteolytic activity는 정상 혈청과 골 관절염 활액보다 활성이 증가하였으며, 이는 EDTA에 의하여 차단되었다. 정상 활액과 골 관절염 활액 내 IGF-I의 150 kDa과 50 kDa carrier proteins는 유사하였다. 따라서 한우 골 관절염 활액 내 IGF-I/IGFBPs axis 변화는 골 관절염에 관여할 것으로 사료된다.

Eimeria Tenella : 항콕시듐제 投藥 中斷에 따른 닭 콕시듐증 誘發 Pattern of Coccidiosis in Chicken Withdrawing of Anticoccidial Drug

白秉杰,金鎭浩,李存華 한국수의공중보건학회 1988 예방수의학회지 Vol.12 No.2

Antibody Responses in Goats Vaccinated with a New Epsilon Toxoid Vaccine of Clostridium perfringens

Rahmam, Mohammad Siddiqur,Rahmam, Mohammad Mostafizur,Baek, Byeong Kirl,Lee, John Hwa 한국수의공중보건학회 2001 예방수의학회지 Vol.25 No.1

산양의 Clostridium균에 의한 장성중독증의 예방을 목적으로, Clostridium perfrigens의 독소로 제작된 새로운 epsilon toxoid를 예방백신으로서의 타당성을 증명하기 위해 이 toxoid에 대한 산양 내 항체생산능을 측정하였다. 본 실험에 사용된 산양은 총 20마리로서 이 toxoid 접종실험군 16마리와 대조군 4마리로 구분하였으며, 항체생산능 측정은 3주간 3회 실시하였다. 이 toxoid를 접종한 실험군에서는 접종 후 21일째 최고의 중화항체가를 보인 반면에 접종을 하지 않은 대조군에서는 항체가를 감지할 수 없었다. 실험군에서 접종 후 14일과 21일째 항체가 상승은 뚜렷한 유의성을 보였으며 (P<0.01), 이들 항체가는 또 높은 방어치(Protective level)를 나타냈다. 이 결과를 토대로 C. perfrigens로 제작된 새로운 이 epsilon toxoid는 산양의 Clostridium균에 의한 장성중독증을 효과적으로 예방할수 있을 것으로 사료된다.

열대열 말라리아에 대한 상피목 및 죽과의 항 말라리아의 효과

박현,김명수,전병훈,이존화,Takaya, Yoshiaki,Wataya, Yusuke,김혜숙 대한동의병리학회 2003 동의생리병리학회지 Vol.17 No.3

Among extracts prepared from Alstonia scholaris, Phyllostachys pubescens and Bambusa veitchii, methanol fraction of Alstonia scholaris was found to have antiplasmodial effect by inhibiting growth of the chloroquine-resistant Plasmodium falciparum strain FCR-3 with less than 14 ㎍/ml of EC50 values. Methanol fraction 2 of Alstonia scholaris revealed the strongest antiplasmodial effect with 40 ㎍/ml of EC50 value. Especially, this fraction showed higher than 3-folds selective toxicity on a Plasmodium as the EC value was 116 ㎍/ml on the host FM3A cell. This is the first report on which an extract compound from Alstonia scholaris showed antimalarial effect.