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김창열 ( Chang-yul Kim ),송은섭 ( Eun-seob Song ),신소정 ( So-jung Shin ),조지훈 ( Ji-hun Jo ),( Ravi Gautam ),이재희 ( Jae-hee Lee ),양수정 ( Su-jeong Yang ),김형아 ( Hyoung-ah Kim ),허용 ( Yong Heo ) 대구가톨릭대학교 자연과학연구소 2019 자연과학연구논문집 Vol.17 No.1
Endotoxin in dust generated at agricultural environments is known to contribute to occurrence of farmer’s lung. Iimmunological markers related with respiratory allergy was evaluated for bovine husbandry workers, and compared with the age-sex matched non - agricultural subjects. Peripheral bloods were collected from 6 workers at 3 bovine farms, and 14 control subjects. Peripheral mononuclear cells were stimulated with phorbol 12-myristate 13-acetate and ionomycin for 72 hours. Level of various cytokines produced at culture supernatants was determined using commercially available ELISA kit. Plasma levels of IgE and IgG subclass were determined. Concentration of inhalable and respirable particulate matter in indoor of the bovine farms was evaluated, and endotoxin level in the dust was measured by Limulus Amebocyte Lysate Kinetic QCL method. Level of endotoxin in the inhalable dust was approximately 154 EU/m<sup>3</sup>, and 9 EU/m<sup>3</sup> in the respirable dust. Plasma IgG4 level, a humoral indicator for allergic propensity, was higher in the milk cow farmers tan the control. Ratio (interferon - γ vs interleukin-4 or intrleukin-13), indicating immunologic skewedness against allergic reactivities, was lower at the milk cow workers compared with those of control subjects. Even though sufficient number of bovine husbandry workers and farms were not investigated, this study suggests in general that immunological function of milk cow farm workers exposed to high level of endotoxin could be modulated toward allergic reactivities.
한국형 인공피부조직을 이용한 탐색적 피부과민성 대체시험법 개발
박정은 ( Jung Eun Park ),( Ravi Gautam ),김창열 ( Chang Yul Kim ),장원희 ( Won Hee Jang ),석승혁 ( Seung Hyeok Seok ),허용 ( Yong Heo ) 한국동물실험대체법학회 2015 동물실험대체법학회지 Vol.9 No.1
Keraskin™-VM, reconstructed human epidermis, was previously investigated for its applicability in development of skin irritation alternative test methods. Considering its expression of various inflammatory mediators, Keraskin™-VM was further evaluated whether it could be useful for development of skin sensitization alternative test method. Keraskin™-VM was incubated 24h in the presence of various skin sensitizing or non-sensitizing test substances followed by collection of culture supernatants. Level of interleukin(IL)-18, a representative pro-inflammatory cytokine, in the supernatants was determined. All the skin sensitizing substances resulted in significantly higher IL-18 levels than the their vehicle control leading to maximum 64-fold, 28-fold, and 85-fold induction of IL-18 expression from 2,4-dintrochlorobenzene, eugenol, and citral, respectively. Among three non-sensitizing test substances (sodium lauryl sulfate, lactic acid, and glycerol), the maximum fold induction of IL-18 was 16 from 0.01% sodium lauryl sulfate. Cell viability was not substantially affected on the magnitude of IL-18 fold induction. Overall, our results suggest the potential usage of Keraskin™-VM for skin sensitization alternative test method.
연구논문 : 한국형 인공피부조직을 이용한 탐색적 피부과민성 대체시험법 개발
박정은 ( Jung Eun Park ),( Ravi Gautam ),김창열 ( Chang Yul Kim ),장원희 ( Won Hee Jang ),석승혁 ( Seung Hyeok Seok ),허용 ( Yong Heo ) 한국동물실험대체법학회 2015 동물실험대체법학회지 Vol.9 No.1
Keraskin™-VM, reconstructed human epidermis, was previously investigated for its applicability in development of skin irritation alternative test methods. Considering its expression of various inflammatory mediators, Keraskin™-VM was further evaluated whether it could be useful for development of skin sensitization alternative test method. Keraskin™-VM was incubated 24h in the presence of various skin sensitizing or non-sensitizing test substances followed by collection of culture supernatants. Level of interleukin(IL)-18, a representative pro-inflammatory cytokine, in the supernatants was determined. All the skin sensitizing substances resulted in significantly higher IL-18 levels than the their vehicle control leading to maximum 64-fold, 28-fold, and 85-fold induction of IL-18 expression from 2,4-dintrochlorobenzene, eugenol, and citral, respectively. Among three non-sensitizing test substances (sodium lauryl sulfate, lactic acid, and glycerol), the maximum fold induction of IL-18 was 16 from 0.01% sodium lauryl sulfate. Cell viability was not substantially affected on the magnitude of IL-18 fold induction. Overall, our results suggest the potential usage of Keraskin™-VM for skin sensitization alternative test method.
Candida kefyr β- galactosedase 의 고정화에 관한 연구
이용규,정기철,김창열 한국낙농학회 1989 韓國酪農學會誌 Vol.11 No.3
Candids kefyr CBS 834의 β-galactosidase를 cellulose triacetate에 고정화하였다. 효소를 고정화하기 위한 최적조건은 100㎖ methylene chloride에 10g의 cellulose triagetate를 용해한 액에 효소액 15%을 첨가하여 만든 후, 200 strokes/min으로 진탕반응했을 때 얻어졌다. 또한 섬유의 절단길이는 효소활성에 큰 영향을 주지 않았다. 고정화효소의 최적온도와 pH는 각각 50℃와 7.0이었고, 활성화에너지는 8,539 cal/mole이었다. ONPG를 기질로 하였을 경우 고정화 효소의 K_m과 V_(max)은 각각 7.4×10^(-5)M과 66.7 unit/㎎ protein이었고, 유당에 대한 K_m과 V_(max)은 3.3×10^(-4)M과 29.4 unit/㎎ protein이었다. 고정화 효소를 4℃에서 30일간 저장후 잔존활성과 3회 재사용후 잔존활성을 조사할 결과 각각 95%와 94%을 유지하였다. 고정화 효소 1g(190 unit)을 50㎖의 탈지유와 5% 유당용액에 첨가한 후 30℃에서 10시간 반응시켰을 때 각각 10%와 15%의 유당분해율을 보여주었다. β-Galactosidase from Candida kefyr CBS 834 was immobilized in the cellulose triacetate. The optimal condition for the immobilized enzyme was obtained by 15% of the enzyme solution added to 100㎖ of methylene chloride and 10g of cellulose triacetate. The length of fiber had hardly any an effect on the enzyme activity and the optimal condition for reciprocating water bath shaker was 200 strokes per minute. The optimal temperature and pH for the immobilized enzyme were 50℃ and 7.0, respectively. The activation energy for the immobilized enzyme was 8,539 cal/mole. The Km and Vmax of the immobilized enzyme, on ONPG were 7.4 × 10^(-5)M and 66.7 unit/㎎ protein, whereas those on lactose were 3.3 × 10^(-4)M and 29.4 unit/㎎ protein, respectively. The rate of remained enzyme activity for the immobilized enzyme stored at 4℃ for 30 days was 95%, and when reused for three times was 94%. When the skim milk (4.8% lactose) and 5% lactose solution of 50㎖ were reacted with the immobilized enzyme (190 unit) for 10hrs at 30℃, 10% of the lactose in skim milk was hydrolyzed, whereas 15% of the lactose in 5% lactose solution was hydrolyzed.