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산성환경에서 Lactobacillus casei YIT9018 의 생존에 관한 연구
김현욱,유광현,이돈성,고준수 ( Hyun Uk Kim,Kwang Hyun You,Don Sung Lee,Jun Soo Goh ) 한국축산학회 1980 한국축산학회지 Vol.22 No.4
Behavior of Lactobacillus casei YIT9018 has been studied in acidic media !MRS and skim milk broth) using three most common acids (acetic, lactic, citric acid) for their survival and the factors affecting their acid tolerance. Survival and death of L. casei YIT9018 were dependent on the concentration and the kinds of acids present in the media. In MRS broth, about 120 mM of lactic acid appeared to be the critical acid concentration for the survival of this useful lactic acid bacterial species. At the same molar concentration of the acids studied, citric acid was the most inhibitory, lactic acid is the next, and acetic acid was the least inhibitory acid to the survival of L. casei YIT9018 in MRS broth. Chemical structure of the acids and pH of acidified media appeared to affect the growth and the survival of this bacterium. Peptone, sucrose, and skim milk powder demonstrated varying degree of the protection for the survival of L.casei YIT9018 in acidified MRS broth. Skim milk was the most protective, peptone was the next, and sucrose was the least protective additive. Synergistic effects were observed when mixtures of acids were used in acidifying the media. Mixture of lactic and citric acid was the most inhibitory and the synergistic inhibition was stronger in MRS broth than in skim milk broth.
박종삼,최가영,유광현,최규서,김진태,김창회,한동현,Park Jong Sam,Choi Ga Young,You Kwang Hyun,Choi Ku Seo,Kim Jin Tea,Kim Chang Hwoe,Han Dong Hyun 대한방사선사협회 2001 대한방사선사협회지 Vol.27 No.2
The two ligaments, the anterior talofibular ligament(ATFL) and the calcaneofibular ligament(CFL), arise from the anteroinferior aspect of the lateral malleolus, and therefore avulsion fragments are superimposed on the lateral malleolus on the srandard rad
YHB6211(recombinant human granulocyte-colony stimulating factor)의 토끼에서 국소자극성에 관한연구
최연식(Yeon Shik Choi),안경규(Kyoung Kyu Ahn),전재현(Jae Hyeon Juhn),이종성(Jong Sung Lee),문병석(Byoung Seok Moon),유광현(Kwang Hyun You) 한국독성학회 1999 Toxicological Research Vol.15 No.2
Local irritation of YHB6211, a recombinant human granulocyte-colony stimulating factor, was assessed in male New Zealand White (NZW) rabbits by dermal and ocular application. In the skin irritation test, 0.5 ml of 0.03% YHB6211 aqueous solution was applied to the back skin of rabbits for 24 hours, and subsequent observation was performed. The solution did not induce any response after the treatment, and the primary irritation index (P.I.I.) of the compound was 0. In the eye irritation test, 0.1 ml of the same solution was instilled into the conjunctiva of the eye. No treatment related reactions were observed at the cornea, iris, and conjunctiva. Based on these results, 0.03% (w/v) YHB6211 solution could be considered as a non-irritating material.
PelB Signal Sequence로 유도된 제조합 인간 상피세포 증식인자 분비 발현 벡터의 제조
박세철,남정현,김정근,권태종,고인영,유광현 한국미생물생명공학회 ( 구 한국산업미생물학회 ) 1996 한국미생물·생명공학회지 Vol.24 No.5
전사개시 단계중에 삽입되는 N말단의 methionine이 제거되고 pelB signal sequence를 삽입하여 인체 epidermal growth factor (hEGF)가 직접 분비발현 되도록 고안된 T7과 tac promoter에 의하여 각각 발현되는 pYHB101과 pYHB2를 제조하였다. 또한 T7 promoter에 의하여 발현되지만 signal sequence를 삽입하지 않은 pYHB1도 제조하여 각각 제조된 재조합 plasmid들의 발현 및 분비효율을 비교하였다. pelB signal sequence hEGF 유전자를 갖도록 제조된 pYHB101을 포함하고 있는 균주가 pYHB1보다 균체 성장속도는 낮았지만 pYHB101을 1 mM IPTG로 5시간 induction 하였을 때 13 mg/l의 rhEGF가 분비발현되어 T7 promoter의 발현 체계를 갖는 경우가 tac promoter보다 높은 발현 효율을 보였다. 이때 pYHB101에서 발현되는 rhEGF의 발현량은 준최적 조건에서 14 mg/l이었다. We have designed nucleotide sequences of hEGF structural gene to eliminate the N-terminal methionine residue incorporated during the translation initiation step, and constructed recombinant human epidermal growth factor (rhEGF) secretion plasmids pYHB101, and pYHB2 in which pelB signal sequence-hEGF gene was expressed under the control of the T7, and tac promoter, respectively. We also constructed pYHB1 vector which contains rhEGF gene controlled by T7 promoter. The transformant with pYHB101 showed relatively slow growth pattern compared to the transformant with pYHB1. However, we observed that the transformant with pYHB101 secreted rhEGF of 13 mg/l significantly after 5 hr induction with 1 mM IPTG and that the T7 promoter was more effective than tac promoter when connected to pelB signal sequence. The amount of rhEGF was 14 mg/l under the sub-optimized condition.
Escherichia coli에서 발현된 재조합 인간 상피세포 증식인자의 정제 및 특성
박세철,유광현 한국미생물생명공학회 ( 구 한국산업미생물학회 ) 1996 한국미생물·생명공학회지 Vol.24 No.4
E. coli BL21(DE3, pYHB101)을 이용한 rhEGF의 최적 생산조건은 25℃, 48시간 배양에서 변형된 MBL 배지에 glucose를 10g /l 첨가한 배지로 배양한 경우이었고 유도물질로 1mM IPTG를 사용하여 2시간 배양 후에 유도 배양하였을 때 최대 rhEGF가 68.7 mg/l가 발현되었다. 배양액으로부터 Amberlite XAD-7, ultrafiltration, DEAE Sepharose column chromatography 정제 과정을 거쳐 rhEGF가 정제되었으며 이때 정제도는 267배이었으며 66.6%의 회수율을 보였다. 정제된 rhEGF는 HPLC 분석 결과 2개의 분획으로 나누어졌으며 N말단 아미노산 서열 분석결과 Asn-Ser-Asp-Ser-Glu-Cys-Pro-Leu-Ser-His로 나타났다. 또한 5'-bromo-2'-deoxy-uridine (BrdU)의 삽입에 의한 DNA 생성능을 조사하였다. 상기의 결과로 E. coli BL21(DE3, pYHB101)에 의하여 생산된 rhEGF는 nhEGF와 동일한 것으로 추정되었다. Recombinant human epidermal growth factor (rhEGF) was produced strain was cultured at 25℃ for 48 hours in the modified MBS medium containing 10 g/l glucose with 1 mM IPTG induction at 2 hours after inoculation. The rhEGF was purified upto 267 folds by Amberlite XAD-7 chromatography, ultrafiltration, and DEAE Sepharose fast fow ion exchange chromatography with an overall yield of 66.6%. The purfied rhEGF was further separated into two fractions by HPLC. The N-terminal amino acid sequence of the second fraction was Asn-Ser-Asp-Ser-Glu-Cys-Pro-Leu-Ser-His. The effect of rhEGF on the DNA synthesis was examined using in vitro biological assay based on the incorporation of 5'-bromo-2'-deoxy-uridine (BrdU). The purified rhEGF shows no difference with natural human epidermal growth factor (nhEGF) in N-terminal amino acids residues and biological activity. From the results, we concluded that rhEGF produced from E. coli harboring the plasmid pYHB101 was apparently the same as nhEGF.
임신부 뇨로부터 정제된 인간 상피세포 증식 인자 유사체의 in vitro bioassay 및 특성
박세철,전재현,남정현,권태종,고인영,유광현 한국미생물생명공학회 ( 구 한국산업미생물학회 ) 1996 한국미생물·생명공학회지 Vol.24 No.4
벤조산흡착, 음이온 교환수지, 단일클론항체을 이용한 immunoaffinity chromatography를 통하여 임신부 뇨로부터 천연의 hEGF를 정제하였다. 정제된 hEGF는 μ Bonda C_18 column을 사용한 HPLC 분석을 통하여 4개의 fraction으로 분리가 가능하였으며 western blot과 double immunodiffusion 실험 결과, 각각의 fraction이 hEGF의 특성을 가진 유사체인 것을 알 수 있었다. 또한 hEGF 표준 물질과의 spiking 및 아미노산 분석 등을 통하여 두 번째 fraction이 nhEGF와 동일한 것으로 확인하였다. nhEGF 및 그 유사체의 생물학적 활성 비교를 위하여 NIH 3T3 세포주에서 5'-Brdu incorporation 측정을 위한 labelling 시간, 혈청 농도의 최적 조건을 결정하였다. NIH 3T3 세포주의 DNA 합성능은 0.2% FCS가 포함된 저혈청 배지에서 hEGF가 0.1~10 ng/ml 농도로 첨가하였을 때 증가하는 경향을 나타냈다. HPLC를 통하여 분리된 두번째 hEGF 유사체가 다른 유사체보다도 생물학적인 활성이 우수하였으며, rhEGF 표준물질과의 spiking 및 아미노산 서열 분석등을 통하여 nhEGF로 밝혀졌다. 임신부의 뇨의 hEGF 유사체 함량중 natural hEGF는 46%이었다. Natural human epidermal growth facto (nhEGF) was purified from pregnant human urine by benzoic acid adsorption, DEAE-Sepharose ion exchange, and immunoaffinity chromatography. The purified nhEGF was further separated into four fractions using Bondapak C_18 HPLC system. Following characterization by Western blot and double immunodiffusion, we found that each fraction corresponds to four derivatives of the nhEGF. For biological analysis of nhEGF, we optimized the labeling time and serum concentration for the incorporation of 5-bromo-2'-deoxy-uridine (BrdU), a non-radioactive alternative for [^3H]-thymidine uptake, into NIH 3T3 cells. The DNA synthesis of NIH 3T3 cells was gradually increased at the nhEGF concentrations between 0.1~10 ng/ml in the Dulbecco's Modified Eagles Medium (DMEM) containing 0.2% Fetal calf serum (FCS). When we assayed the biological activity of four fractions, the activity of the second fraction was superior to that of the others.
김제형,황호근,최두환,유광현,류준형,최석채,김학철 圓光大學校 醫科學硏究所 1995 圓光醫科學 Vol.11 No.1
Biloma has been defined as an extraductular collection of bile within a defined capsular space. Development of an intraabdominal bile collection usually occurs secondary to traumatic or iatrogenic injury including abdominal surgery, percutaneous catheter drainage and transhepatic cholangiogram. Spontaneous bile leakage associated with other primary causes, including sickle cell anemia, gall bladder cancer and hepatocellular carcinoma, has been reported rarely. We present a case of biloma secondary to traumatic injury with liver biopsy, who had right upper guadrant pain, jaundice and yellowish skin discoloration in a patient with cholangiocellular carcinoma. Huge biloma was confirmed by percutaneous transhepatic biliary drainage and drained with pig tail catheter successfully. A case of huge biloma secondary to liver biopsy is presented with brief review of the literatures.