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Chinese Hamster Lung Cell을 이용한 in vitro 소핵시험의 세포질 최적화 연구
백민경 ( Min Kyoung Paik ),김아름누리 ( Areumnuri Kim ),신혜림 ( Hye Rim Shin ),전경미 ( Kyongmi Chon ),박경훈 ( Kyung-hun Park ),류지혁 ( Byeong Chul Moon ),문병철 한국환경농학회 2018 한국환경농학회지 Vol.37 No.3
In vitro 소핵시험(vitMNT)은 유전독성의 유망한 대체시험법 중 하나로, OECD에서 TG로 채택되어 화학물질의 등록에 사용되고 있다. 본 시험에서는 CHL cell을 사용한 vitMN test에서 소핵을 판별하기 위한 최적화된 세포질 조건을 찾고자 하였으며, 양성대조물질로 MMC와 Col을 사용하고 세포 염색을 위해 giemza 용액을 사용하였다. 시험결과, 세포현탁을 위해 사용되는 고정액의 acetic acid의 농도는 1%로 하는 것이 band의 두께와 세포질의 퍼짐성 측면에서 적당하였다. 또한 세포의 깨짐을 최소화하는 최종 고정액의 적하시간은 현탁 후 1~4시간이었다. 이러한 결과는 vitMNT에서 소핵관찰의 신속성, 용이성 및 정확성을 확보하는데 도움이 될 것이다. BACKGROUND: in vitro micronucleus test (vitMNT) is one of the promising alternative testing methods in genotoxicity test and was adopted as OECD test guideline for chemical registration. This study was conducted to optimize the cytoplasm conditions in vitMNT using Chinese hamster lung (CHL) cell. METHODS AND RESULTS: In this study cytokinesisblock micronucleus test was conducted. Mitomycin C and colchicine were used as positive control chemicals and were treated for three hours (short time) or twenty-four hours (long time). Giemsa solution was used for cell staining. For optimization of vitMNT, the final fixative was prepared as five concentrations (0%, 1%, 3%, 5%, and 25%) of acetic acid in methanol, and treatment times of the final fixative were varied under four conditions (immediately, one hour, four hours, and one day). CONCLUSION: Acetic acid at 1% in methanol as the final fixative was most adequate to preserve the cytoplasm around the nucleus in the interphase cells. Also, fixative treatment time of cell suspension for one to four hours may minimize the cell rupture. These results can be helpful for getting an accurate result promptly due to clear visual distinction to score micronucleus in vitMNT using giemsa solution.
백민경 ( Min Kyoung Paik ),임정택 ( Jeong Taek Im ),전경미 ( Kyongmi Chon ),박경훈 ( Kyung-hun Park ),최용수 ( Yong-soo Choi ),이명렬 ( Myeong-lyeol Lee ),배철한 ( Chul-han Bae ),김진호 ( Jin-ho Kim ),문병철 ( Byeong Chul Moon 한국환경농학회 2016 한국환경농학회지 Vol.35 No.4
BACKGROUND: Recently, the widespread distribution of pesticides in the hive has been of concern about pesticide exposure on honeybee (Apis mellifera L.) health. Larval toxicity was adapted to assess the synergistic and antagonistic interaction of cumulative mortality to the honeybee larvae of the four most common pesticides detected in pollen. METHODS AND RESULTS: Acetamiprid(3.0 μl/L), chlorothalonil (803.0 μl/L), coumaphos (128.0 μl/L), and tau-fluvalinate (123.0 μl/L) were tested in combination; binary, ternary and four component mixture. Larvae were exposed to four pesticides mixed in diet at the average levels detected in pollen. As a result, synthetic toxicity was observed in the binary mixture of acetamiprid with coumaphos. The binary and ternary component mixtures of tested pesticides have mostly demonstrated additive effect in larval bees. The significant antagonistic effects were found in four parings of mixtures including chlorothalonil added to acetamiprid/tau-fluvalinate or acetamiprid/coumaphos/tau-fluvalinate, and tau-fluvalinate added to acetamiprid/chlorothalonil or acetamiprid/coumaphos/chlorothalonil. CONCLUSION: Interactions between combinations of four pesticides showed mostly additive or antagonistic effects in larval bees. Therefore, predicting the larval mortality of pesticides mixtures on the basis of the results of single pesticide may actually overestimate the risk. We suggest that pesticide mixture in pollen be evaluated by adding their toxicity together for complete data on interactions.
가축분뇨 유래 퇴비 및 농경지 중 축산용 항생제의 잔류 및 위해성 평가
백민경 ( Min-kyoung Paik ),류송희 ( Song-hee Ryu ),김성철 ( Sung-chul Kim ),홍영규 ( Young-kyu Hong ),김진욱 ( Jin-wook Kim ),김정규 ( Jeong-gyu Kim ),권오경 ( Oh-kyung Kwon ) 한국응용생명화학회(구 한국농화학회) 2021 Journal of Applied Biological Chemistry (J. Appl. Vol.64 No.2
축산용 항생제는 투여된 양의 일부만이 체내에서 사용되며 나머지는 분뇨로 배출되며 이를 활용한 퇴비를 농경지에 살포함으로써 농업환경에 유입되어 2차 오염 등을 초래하고 있다. 따라서, 농업환경 중 항생제 관리기준 설정 등 사후 관리 기술이 필요하다. 본 연구는 국내 사용빈도가 높은 것으로 알려진 tetracycline 및 sulfonamide 계열 등의 항생제를 대상으로 매체별 잔류량을 비교하고 퇴비 시용 전·후 농경지 토양 중 잔류 항생제의 위해성을 평가하기 위하여 수행되었다. Buffer 및 SPE를 사용한 전처리 방법은 ppb 수준에서 70% 이상의 회수율을 나타냈으며, 검출한계(LOD)의 범위는 퇴비와 토양에서 각각 0.13-0.46 μg/kg과 0.05-0.25 μg/kg이었다. 잔류 항생제 분석결과 퇴비 중 tetracycline 계열 항생제의 잔류 농도는 5.38-196.0 μg/kg, sulfonamide 계열은 below the detection of limit (BDL)-259.0 μg/kg 수준으로 검출되었다. 농경지 토양의 경우 각각 0.30-53.3 μg/kg, BDL-4.16 μg/kg의 잔류 수준을 나타냈으며 토양분배계수(Kd) 값이 높은 tetracycline 계열 항생제의 잔류 농도가 sulfonamide 계열보다 높았다. 퇴비 시용 전후의 농경지 토양의 항생제에 대한 인체위해도는 항생제 종류에 따른 차이가 있었으나, 전체 HQ가 1 이하에서 안전하다는 기준에 의하면 조사된 항생제 5종 모두 인체 위해성이 매우 낮았으며 시용전·후의 영향이 전체 위해도에 미치는 비율을 고려하면, 퇴비 시용이 토양의 항생제에 대한 인체위해성에 미치는 영향은 미비한 것으로 판단되었다. While veterinary antibiotics are used only in a part of the dose administered, the rest are excreted as urine or feces. Residual antibiotics enter the adjacent agricultural environments by spraying manure-based composts on farmlands and lead to secondary pollution. Therefore, it is necessary to develop the technique for post management such as regulatory levels of antibiotics in the agricultural environments. This study was conducted to compare by different matrices the amount of residual antibiotics such as tetracyclines and sulfonamides, which are known to be frequently used in Korea and to practice risk assessment by different antibiotics in soils before and after application of composts. Pre-treatment with modified typical method using buffer and solid phase extraction showed the recovery of composts and soils was more than 70% at ppb level and the limits of detection were 0.13-0.46 and 0.05-0.25 μg/kg, respectively. Analysis of manure-based composts revealed concentrations from 5.38 to 196.0 μg/kg for tetracyclines, from below the detection of limit (BDL) to 259.0 μg/kg for sulfonamides. In case of agricultural soils, residual concentrations of selected veterinary antibiotics were ranged 0.30-53.3 μg/kg, BDL-4.16 μg/kg respectively and the concentration level of tetracyclines, which had higher soil distribution coefficient (Kd) values, was higher than that of sulfonamides. There was a difference in human risk assessment by different antibiotics in soil before and after application of composts. But, it was indicated that detection values of all of 5 antibiotics were very safe on the basis that Hazard Quotient was safe below 1.
박경훈 ( Kyung Hun Park ),윤현주 ( Hyun Joo Yoon ),한범석 ( Beom Seok Han ),이제봉 ( Je Bong Lee ),정미혜 ( Mi Hye Jeong ),조남준 ( Nam Jun Cho ),엄애선 ( Ae Son Om ),백민경 ( Min Kyoung Paik ) 한국환경농학회 2014 한국환경농학회지 Vol.33 No.4
BACKGROUND: Azadirachta indica Extract(AIE) containing azadirachtin as active ingredient have been used worldwide as environment-friendly organic material having pest control properties. However, the extracts prepared with different solvent and from different plant site is very diverse and have different toxicity. METHODS AND RESULTS: In this study, the four week repeated oral dose toxicity test of aqueous AIE in Sprague-Dawley rats was carried out to investigate the toxic effect of liver, main toxicity target organ of AIE. The male and female rats were divided into 4 groups, respectively; control(0 g/Kg bw), low-dose group(0.5 g/Kg bw), middle-dose(1.0 g/Kg bw) and high-dose group(2.0 g/Kg bw). As a results, relative liver weight increased with dose-dependent of AIE(p<0.05). Serum LDH in all AIE-treated groups were significantly lower than the control in male rats(p<0.05). However, serum GOT andGPT were significantly increased in all male AIE-treated groups in male rats(p<0.05) and, in particular, increase of serum GPT in dose-dependent manner raise the possibility of liver damage. Even through serum GLU was increased significantly in high-dose group in male rats compared to control, there were no significant differences of urinary GLU among all groups(p<0.05). In addition, histopathological examination of the liver did not reveal any lesions in all AIE-treated groups. CONCLUSION: In conclusion, 4 weeks of the repeated oral administration of AIE 2.0 g/Kg to rats has resulted no toxic response in liver. Therefore, AIE was no indicated to have any toxic effect in the SD rats, when it was orally administrated below the dosage 2.0 g/Kg/day for 4weeks.