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      • SCOPUSKCI등재

        효소면역측정법을 이용한 Fumonisin의 검출법 개발

        손동화,한성민,임선희,이인원,조선희,강신영,이경애 한국미생물생명공학회 ( 구 한국산업미생물학회 ) 1996 한국미생물·생명공학회지 Vol.24 No.1

        Fumonisin에 대한 효소면역측정법(ELISA)을 개발하기 위하여 특이항체를 생산하고, 분석조건을 확립하여, 인위적으로 오염시킨 옥수수 시료의 분석을 행하였다. Fumonisin B_1 (FB_1)을 cholera toxin (CT)에 결합시킨 FB_1-CT conjugate를 면역원으로 하여 Freund's adjuvant와 함께, 또는 단독으로 두 군의 토끼에 면역하고 특이항체를 생산하였다. 항혈청을 ELISA로 분석한 결과 그 adjuvant와 함께 면역한 경우 항체역가(titer)가 높게 나타났다. 가장 높은 항체역가(1 : 16,000)를 나타낸 항혈청 및 그로부터 정제한 항체를 이용하여 간접법 및 직접법에 의한 경합적 ELISA(ciELISA 및 cdELISA)를 각각 확립하였다. 이 항체의 유사독소와의 교차반응을 ciELISA로 조사하였을 때, FB_3에 대한 교차반응율은 2%로 매우 낮았으나, FB_2에 대한 것은 179%로 다소 높았다. FB_1의 검출한계는 ciELISA에서 0.03ppb, cdELISA에서 0.3 ppb로 각각 나타났다. 실제 곡물시료의 분석에 ELISA 활용가능성을 조사하기 위하여 인위적으로 FB_1을 오염시킨 옥수수를 75% methanol로 추출하고 그 회수율을 측정한 결과, 분석이 불안정하여 회수율이 일정하게 나타나지 않았다. 그러나 방해물질의 제거를 위하여 시료추출액을 strong anion exchange(SAX) cartridge를 거쳐 세척한 다음 분석하였을 때, 3~10 ppm 범위의 FB_1 오염 옥수수에서 평균 34%(CV의 평균, 8.2%)의 안정된 회수율을 보였다. In order to develop enzyme-linked immunosorbent assay (ELISA) for fumonisins, production of specific antibodies, establishement of ELISA conditions, and quantitation of the toxin from spiked corns by ELISA were performed. Fumonisin B_1 (FB_1) conjugated to cholera toxin (CT) with or without Freund's adjuvant was subcutaneously injected into 2 groups of rabbits. When the titer of the antisera produced by each rabbit tested, higher titer was observed in case of the immunization with the adjuvant. By use of the antiserum showing the highest titer (1 : 16,000) and its purified antibodies, competitive indirect and direct ELISA's (ciELISA and cdELISA) were established, respectively. When the cross-reactivity of the antibody against fumonisin analogs was investigate by the ciELISA is was very low against B_3 (2%) but high against fumonisin B_2 (179%). The sensitivity of the ELISAs was also very high, because the detection limit for FB_1 was 0.03 ppb in ciELISA and 0.3 ppb in cdELISA. When the ELISA's were applied to the spiked corns after extraction with 75% methanol, the assay recovery of FB_1 was too unstable to assay. However, when cleanup by strong anion exchange (SAX) cartridge was introduced to remove interfering materials, the mean ELISA recovery of FB_1 from corns spiked to 3~10 ppm was found to be 34.0% and stable (mean of CV, 8.2%).

      • KCI등재

        Inhibitory Effect of Clavicepitaceae on Serotonin Release out of Human Platelets and Human Platelet Aggregation

        박화진,--,--,--,--,--,-- THE KOREAN SOCIETY FOR BIOMEDICAL LABORATORY SCIEN 2004 Journal of biomedical laboratory sciences Vol.10 No.1

        We have investigated the effects of hypha-water extracts (HWE), fruit body-water extracts (FWE) and cordycepin from Cordyceps militaris on serotonin release out of human platelets and human platelet aggregation. HWE and FWE inhibited the release of [^(3)H]-serotonin from human platelet stimulated by thrombin (2 U/ml) or collagen (20 ㎍/ml) in a dose-dependent manner. Furthermore, cordycepin, a major component of Cordyceps militaris, inhibited the human platelet aggregation induced by collagen (10 ㎍/ml) in a dose-dependent manner. These results suggest that cordycepin containing in HWE and FWE may inhibit the serotonin release by suppressing the collagen-induced human platelet aggregation. Accordingly, our data demonstrate that HWE and FWE containing much cordycepin might have antithrombotic and antimigrainous functions.

      • SCOPUSKCI등재

        효소면역측정법에 의한 우유중의 Aflatoxin M1 분석

        손동화,임선희,이인원 한국미생물생명공학회 ( 구 한국산업미생물학회 ) 1996 한국미생물·생명공학회지 Vol.24 No.5

        국내산 우유중 aflatoxin M_1 (AFM_1)의 오염현황을 조사하기 위하여, 효소면역측정법(ELISA)을 개발하고 이를 이용한 독소의 정량을 행하였다. 항체의 생산을 위하여 AFM_1 bovine serum albumin 결합체를 Freund's adjuvant와 함께 토끼에 피하주사하여 면역하였다. 가장 높은 항체역가를 보인 항혈청과 AFB_1-horseradish peroxidase 결합체를 이용하여 직접법에 의한 경합적 효소면역측정법(competitive direct ELISA ; cdELISA)의 조건을 확립하였으며, 그 검출한계는 0.003ppb로 나타났다. 또한 유사독소에 대한 교차반응율은 aflatoxin M_1, M_2, B_1, _2, G_1, G_2, B_2a, and G_2a 에 대하여 각각 100, 29.9, 25.0, 2.7, 13.0, 0.65, 0, 및 0%이었다. 다음으로, AFM_1을 인위적으로 오염시킨 우유를 C_18 cartridge로 세척후 cdELISA로 분석하였을 때, 0.01~1 ppb(10~1,000 ppt)의 범위에서 그 회수율의 평균은 104%(CV의 평균, 6.4%)로 나타났다. 시유 및 목장우유를 대상으로 AFM_1의 오염현황을 cdELISA로 조사하였을 때, 평균 오염도는 80.4±55.0 ppt (n=64; range, 5,6~280 ppt)이었다. For a survey of the occurrence of aflatoxin M_1 (AFM_1) in domestic cow's milk, we developed an enzyme-linked immunosorbent assay (ELISA) system, and quantitated the toxin in cow's milk. In order to produce specific antibodies AFM_1 conjugated to bovine serum albumin (AFM_1-BSA) and Freund's adjuvant were immunized subcutaneously to rabbits. By use of the antiserum showing the highest titer and AFB_1-HRP conjugate, we established a competitive direct ELISA (cdELISA) for AFM_1 whose detection limit was 0.003 ppb. The cross-reactivities of the antiserum against aflatoxin M_1, M_2, B_1, _2, G_1, G_2, B_2a, and G_2a were 100, 29.9, 25.0, 2.7, 13.0, 0.65, 0, and 0%, respectively. When the cdELISA was applied to the cow's milk spiked with AFM_1 and followed by cleanup with C_18 cartridge, the mean recovery of the assay was 104% (mean of CV, 6.4%) in the final concentration of 0.01~1 ppb (10~1,000 ppt). When cow's milk samples gathered from markets and farms were assayed by the cdELISA, the mean concentration and SD of AFM_1 was 80.4±55.0 ppt (n=64; range, 5,6~280 ppt).

      • KCI등재후보

        골절단을 이용한 급속 치아이동 후 치수 및 치주조직 변화

        강경화,김은철,이선경,임채웅,마쓰다 기꾸,태기출,김상철 대한치과교정학회 2004 대한치과교정학회지 Vol.34 No.2

        본 연구는 골절단을 이용한 치아-치조골 분절의 급속 치아이동 후 치수, 치주인대 및 치조골의 변화를 유성견에서 평가하고자 하였다. 하악 제4소구치의 근심, 원심, 치근단 부위에서 피질골을 절단하여 치아-골 분절을 형성하고 하악 제3소구치를 발거하면서 협측, 설측 부위의 피질골을 삭제하였다. 1주 휴지기 부여에 따라 휴지기군과 비휴지기군으로 나누어 치아의존형 견인장치로 6일 동안 견인하고 강화기를 거친 후 0주, 1주, 2주, 4주, 6주, 8주에 희생시켜 치수, 치주인대 및 치조골의 조직 변화를 임상적, 방사선적, 조직학적 및 면역화학적으로 평가하여 다음과 같은 결과를 얻었다. 1. 치아이동 양과 강화기 동안의 조직 치유 양상에서 휴지기 유무에 따른 차이는 없었다. 2. 견인측에서 골형성은 강화기 8주까지 계속되었는데 강화기 1-2주에 가장 활발하였으며 6-8주간의 변화는 적었다. 3. 비휴지기군의 강화기 1주에서 치조골 흡수 및 파골세포 출현, 염증세포 침윤이 가장 많았으며, 특징적으로 파상아세포가 압박측의 치주인대와 치수 내에 나타났다. 4. TGF-β는 치조골의 골기질 및 골모세포, 파골세포, 치수 내 파상아세포에서 강양성 발현을 보인 반면 치수, 백악모세포, 무세포성 백악질에서 경미한 양성 발현을 나타내는 부위 특이성이 있었다. 5. TGF-β는 견인측 치주인대의 치조골에 인접한 혈관 및 치주인대세포, 골모세포에서 강화기 초기 1-2주에 주로 발현되었으며 6주 이후에는 발현이 크게 감소하였다. The movement of tooth-bone segments by osteotomy can simultaneously shift tooth and surrounding alveolar bone in a relatively short period. The purpose of this study was to evaluate the tissue changes in pulp, periodontal ligament, and alveolar bone in rapid tooth bone movement with osteotomy. The mandibular 3rd premolar of a dog was extracted and cortical bones of the buccal and lingual area were eliminated, and then cortical bones around the mesial and distal area of root, and below the root apex of the mandibular 4th premolar were osteotomized. After a one week latency period, a tooth-borne distraction device was activated for 6 days. And pulp, periodontal ligament and alveolar bone were evaluated clinically, radiologically, histologically and immunohistochemically at 0, 1, 2, 4, 6, 8 weeks of the consolidation period and conclusions were reached as follows. 1. Latency period didn't affect total amount of tooth movement and healing process of tissue during consolidation period. 2. Bone formation continued through 8 weeks of consolidation in distracted side, with a high peak at 1-2 weeks, and the lowest at 6-8 weeks of consolidation. 3. At 1 week of consolidation, alveolar bone resorption, osteoclast appearance and inflammatory cell infiltration were the most active, and dentinoclasts characteristically appeared on the pulp and pressure side of the periodontal ligament. 4. The expression of TGF-β was area-specific, as it was strong-positive at bone matrix, osteoblast, osteoclast of alveolar bone, and dentinoclast inside pulp, but weak in pulp, cementoblast and acellular cementum. 5. The expression of TGF-β was generally observed at the initial 1-2 weeks of consolidation at vessels, periodontal ligament cells, and osteoblast near alveolar bone on the distraction side of the periodontal ligament, and was significantly decreased after 6 weeks of consolidation.

      • KCI등재
      • 옥수수 뿌리에서 전처리 방법에 따른 굴중성 반응의 효과

        김순영,임선화,조명제 안동대학교 기초과학연구소 2000 基礎科學 硏究論文集 Vol.11 No.1

        This research were focused on the methods of the pretreatment of chemicals in the study of gravitropic response. The filter paper method which was wrapped the whole roots showed the similar response to the control. According to the methods, we examined the effect of 10(-6) MIAA. The treatment of IAA inhibited the gravitropic response in all the type of pretreatment. Specially, the filter paper method showed the significant effect in the primary roots of maize.

      • Anti-inflammatory and antiobesity effects of mulberry leaf and fruit extract on high fat diet-induced obesity.

        Lim, Hyun Hwa,Lee, Sung Ok,Kim, Sun Yeou,Yang, Soo Jin,Lim, Yunsook The Society 2013 Experimental biology and medicine Vol.238 No.10

        <P>The purpose of this study was to investigate the anti-inflammatory and antiobesity effect of combinational mulberry leaf extract (MLE) and mulberry fruit extract (MFE) in a high-fat (HF) diet-induced obese mice. Mice were fed a control diet or a HF diet for nine weeks. After obesity was induced, the mice were administered with single MLE at low dose (133 mg/kg/day, LMLE) and high dose (333 mg/kg/day, HMLE) or combinational MLE and MFE (MLFE) at low dose (133 mg MLE and 67 mg MFE/kg/day, LMLFE) and high dose (333 mg MLE and 167 mg MFE/kg/day, HMLFE) by stomach gavage for 12 weeks. The mulberry leaf and fruit extract treatment for 12 weeks did not show liver toxicity. The single MLE and combinational MLFE treatments significantly decreased plasma triglyceride, liver lipid peroxidation levels and adipocyte size and improved hepatic steatosis as compared with the HF group. The combinational MLFE treatment significantly decreased body weight gain, fasting plasma glucose and insulin, and homeostasis model assessment of insulin resistance. HMLFE treatment significantly improved glucose control during intraperitoneal glucose tolerance test compared with the HF group. Moreover, HMLFE treatment reduced protein levels of oxidative stress markers (manganese superoxide dismutase) and inflammatory markers (monocyte chemoattractant protein-1, inducible nitric oxide synthase, C-reactive protein, tumour necrosis factor-α and interleukin-1) in liver and adipose tissue. Taken together, combinational MLFE treatment has potential antiobesity and antidiabetic effects through modulation of obesity-induced inflammation and oxidative stress in HF diet-induced obesity.</P>

      • SCOPUSKCI등재

        Brassinosteroids Affect Ethylene Production in the Primary Roots of Maize(Zea mays L.)

        Lim, Sun Hwa,Chang, Soo Chul,Lee, June Seung,Kim, Seong-Ki,Kim, Soon Young 한국식물학회 2002 Journal of Plant Biology Vol.45 No.3

        To better understand the physiological roles of brassinosteroids (BRs) in the primary roots of maize, we examined their effect on ethylene production. Exogenously applied brassinolide (BL; 10^-9 to 10^-7M) incrementally increased the level ethylene in a dose-dependent manner. This BL-induced production was enhanced in the presence of IAA, thereby implying a synergistic effect between BR and IAA. At 10^-7 M BL, the level of free ACC was increased, but that of conjugated ACC was diminished. Moreover, greater concentrations of BL proportionally increased ACC oxidase activity. In contrast, higher levels of IAA increased the endogenous content of conjugated ACC as well as ACC synthase activity. Based on these results, we conclude that BR activates ethylene production mainly via ACC oxidase, and interacts with IAA to produce ethylene. However, the functional site for ethylene production is different for each hormone.

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