• Title/Summary/Keyword: 면역독소

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Survey on Fusarium Mycotoxin Contamination in Oat, Sorghum, Adlay, and Proso Millet during the Harvest Season in Korea (귀리, 수수, 율무, 기장의 수확기에 발생하는 Fusarium 곰팡이독소 오염도 조사)

  • Lee, Mi Jeong;Wee, Chi-Do;Ham, Hyenheui;Choi, Jung-Hye;Baek, Ji Sun;Lim, Soo Bin;Lee, Theresa;Kim, Jeom-Soon;Jang, Ja Yeong
    • Journal of Food Hygiene and Safety
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    • v.35 no.1
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    • pp.13-22
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    • 2020
  • A total of 244 cereal samples (oat, sorghum, adlay, and proso millet) were collected from fields to examine the contamination of Fusarium mycotoxins in cereals during harvest season in 2017 and 2018. The contamination levels of deoxynivalenol (DON), nivalenol (NIV), and zearalenone (ZEA) were analyzed individually by using the immunoaffinity column clean-up method with ultra performance liquid chromatography, and fumonisins (FUM) were analyzed by using the QuEChERS method with liquid chromatography-mass spectrometry. Highest level of NIV contamination (120.0-3277.0 mg/kg) was observed in oat samples among the analyzed cereals. In the adlay samples, DON contamination was the highest (maximum level 730.0 ㎍/kg). The proso millet samples had a high frequency of detection of NIV and ZEA (61.5% and 57.9%, respectively), but the levels were low (average detection level of NIV, 75.6 ㎍/kg, for ZEA, 21.5 ㎍/kg). Among the cereal samples, sorghum had the highest contamination frequency of DON, ZEA, and FUM, and the co-occurrence of Fusarium mycotoxin was 70.0%, which was higher than the average of 29.9%. In order to safely manage Fusarium mycotoxin levels in cereals, continuous research on the development of contamination prevention technologies together with monitoring of mycotoxin contamination is needed.

Effects of Chitosan on the Production of Th1 and Th2 Cytokines in Mice (키토산이 Th1과 Th2 사이토카인 생성에 미치는 효과)

  • Kim, Kwang-Hyuk
    • Journal of Life Science
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    • v.19 no.3
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    • pp.411-416
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    • 2009
  • Chitosan is derived from chitin by a process of controlled deacetylation. In the present study, we investigated the effects of chitosan on the production of cytokines such as interleukin-2 (IL-2), interferon-$\gamma$ (IFN-$\gamma$), interleukin-4 (IL-4), and interleukin-10 (IL-10) in mice. The culture supernatants of splenocytes exposed with chitosan alone or chitosan plus cell stimulants, lipopolysaccharide (LPS), concanavalin A (Con A), and phytohemagglutinin-P (PHA-P) were harvested to assay IL-2, IFN-$\gamma$, IL-4, and IL-10 production. IL-2, IFN-$\gamma$, and IL-4 from splenocytes exposed to chitosan showed a greater increase compared to the PBS control group. IL-2 and IFN-$\gamma$ levels in the culture supernatants from splenocytes exposed to LPS+chitosan were higher than those of the groups exposed to LPS alone. IL-4 and IL-10 levels in the culture supernatants from splenocytes exposed to LPS+chitosan were lower than those of the groups exposed to LPS only. These findings demonstrate that chitosan upregulates the immune responses by Th1 cytokines (IL-2 and IFN-$\gamma$) and downregulates those by Th2 cytokines (IL-4 and IL-10) in LPS-associated immunity. These results show the potential of its usefulness for balancing the Th1/Th2 immune response, if more research results were accumulated.

Application of Fluorescence Polarization Immunoassay for the Screening of Ochratoxin A in Unpolished Rice (현미에서의 오크라톡신 A의 검색을 위한 형광편광면역분석법의 응용)

  • Park, Jung-Hyun;Chung, Duck-Hwa;Lee, In-Seon
    • Journal of Life Science
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    • v.16 no.6
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    • pp.1006-1013
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    • 2006
  • To High Throughput Screening (HTS), a homogeneous fluorescence polarization immunoassay (FPIA) was developed for the quantitative determination of ochratoxin A(OTA) using a $Victor^3$ (PerkinElmer). The homologous tracer, fluorescein-labelled OTA-EDF were synthesized and a specific OTA antibody has been used in the development of the method. It allowed the determination of OTA in the concentration range 0.5-200 ng/ml, with the detection limit of 0.3 ng/ml. The method developed was highly specific and reproducible. OTA spikes in unpolished rice extracts were determinable by FPIA with good recovery. For naturally contaminated unpolished rice samples some disagreement was observed between the results obtained by FPIA and HPLC, which could be related to the a little matrix effect observed for FPIA. Further research is needed to validate the procedure. On the basis of these initial results, this FPIA appears to meet the performance criteria for OTA screening of food samples without a complicated clean-up.

An Enzyme-Linked Immunosorbent Assay for $Aflatoxin\;M_1$ in Cow's Milk without a Cleanup Procedure (희석에 의한 우유 중 $Aflatoxin\;M_1$의 효소면역측정법)

  • Shon, Dong-Hwa;Lim, Sun-Hee;Lee, Yin-Won
    • Korean Journal of Food Science and Technology
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    • v.28 no.6
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    • pp.1184-1187
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    • 1996
  • A simple and rapid detection system for $aflatoxin\;M_1\;(AFM_1)$ in cow's milk by an enzyme-linked immunosorbent assay (ELISA) was developed. Specific antibodies against $AFM_1$, conjugated to bovine serum albumin $(AFM_1-BSA)$ were raised in rabbits and purified. The cross-reactivities of the antibodies against aflatoxin analogs were less than 29.9%. When a competitive direct ELISA (cdELISA) for $AFM_1$, established by use of the antibodies was applied to the spike test of $AFM_1$ onto uncontaminated cow's milk, the assay recovery was unstable unless cow's milk was diluted to 40% (2:3) with phosphate buffered saline (PBS). In that condition of sample dilution, the mean ELISA recovery of $AFM_1$, from the cow's milk was 113% (coefficient of variation (CV) of each recovery percentage, 8.2%) in the range of $0.3{\sim}3.0\;ppb$. These results showed that the ELISA system could be a convenient tool to monitor the contamination of AFM1 more than 0.5 ppb in cow's milk (FDA allowance limit) easily.

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리보솜 불활성화 단백질의 삼차원 구조 규명과 그 구조의 신약개발에의 응용

  • 서세원
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1994.04a
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    • pp.264-264
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    • 1994
  • 본 연구는 보리 씨앗에 존재하는 리보솜 불활성화 단백질(RIP) 의 삼차원 구조를 X-선 결정학 방법을 이용하여 밝히고, 그 결과로 분자 차원에서 기능을 이해하는 것을 목적으로 하고있다. 리보솜불활성화 단백질은 N-glucosidase 반응을 통하여 단백질 합성을 저해하기 때문에 세포를 죽일 수 있다. 따라서 암세포만을 특정적으로 인식하는 다른 물질과 결합시키면 암세포만을 특정적으로 죽일 수 있는 면역독소로 이용될 수 있다. 또, 최근에는 항바이러스의 작용을 함이 밝혀져 많은 연구가 진행되고 있다. 단백질 삼차원 구조 규명을 위해서는 여러가지 단계가 있는데 지난 번 과제까지 성공적으로 리보솜 불활성화 단백질의 대량 분리와 X-선 결정학의 필수 요건 좋은 결정을 길렀고, 이번에는 구조 해석을 위해 꼭 해결해야하는 위상문제를 극복하기 위하여 여러가지 실험을 진행하였다. 우선, 비슷한 구조인 피마자씨에서 분리한 Ricin의 A-체인과 미국자리공 잎에서 분리한 Pokeweed antiviral protein의 삼차원 분자좌표를 이용하여 분자치환법으로 위상문제 해결을 시도하였다. Ricin 의 A-체인을 이용하였을 때 분자의 위치가 정확히 찾아지지 않았고, 다른 모델인 Pokeweed antiviral protein을 이용하여 X-PLOR 프로그램내의 PC refinement법으로 분자치환을 시도하였다. Euler각도로 (187.37, 22.5, 311.94) 의 회전해 (Rotation solution) 를 가지고 있었고, 이러한 해에 맞추어서 분자를 돌려둔 후, 이동해 (Transaltion solution) 을 구해서 그 위치 (Orientation) 로 분자를 이동하였다. 이 때 R값은 53.9 % (8.0 - 3.5$\AA$) 이였고, 부분좌표 (Fractional coorcdinate) 에서는 0.102, 0.000, 0.261 이고, 직교좌표 (Orthogonal coorclinate) 에서는 4.616, 0.000, 13.167 의 결과를 얻었다.

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Immunogenicity of the recombinant Pasteurella multocida toxin for development of subunit vaccine against swine atrophic rhinitis (돼지 위축성 비염 단위 백신 개발을 위한 재조합 파스튜렐라 독소 단백질의 면역원성 검정)

  • Lee, Jeongmin
    • Korean Journal of Veterinary Research
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    • v.47 no.1
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    • pp.59-65
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    • 2007
  • Pasteurella multocida Pasteurella multocida toxin (PMT) is a causal pathogenin atrophic rhinitis in pigs. To investigate the protective immunity and vaccination effect of recombinantPMT, the gene for PMT was isolated from the infective P. multocida D:4. The 2.3 kb XhoI/PstI fragment(PMT2.3) of PMT gene was expressed in E. coli BL21 (DE3) using the induced expression vector system.The recombinant protein of PMT2.3 having molecular weight of 84 kDa was purified by Ni-afinitycolumn chromatography. The PMT2.3 raised slightly less anti-PMT antibody titer than formalin-killedwhole cel, however, it showed more protective imunity against P. multocida D:4 infection in vaccinationand chalenge.

Immunological Analysis of Endotoxin Proteins Produced by Bacillus thuringiensis serovar. kurstaki HD1 and HA73 (Bacillus thuringiensis serovar. kurstaki HD1과 HD 73이 생산하는 내독소 단백질의 면역학적 분석)

  • 오상수;이영종;김창규;구본성;김종배;이형환
    • Microbiology and Biotechnology Letters
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    • v.16 no.2
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    • pp.168-173
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    • 1988
  • Immunological analysis between endotoxin proteins produced by Bacillus thuringiensis serovar. kurstaki HD1 and HD73 have been investigated by using polyclonal antibodies. The antisera against the endotoxin proteins were prepared from rabbits injected with the endotoxin protein antigens. When about 2mg/$m\ell$ of the antigens were injected for 7 times, the titers were highest. The stability of the antigens was reduced to about 50% after 9 days incubation at 4$^{\circ}C$. The sensitibity of endotoxin protein from B. thuringiensis HD1 and HD73 by indirect ELISA was 50ng/$m\ell$ and 400ng/$m\ell$, respectively. The cross reaction of antiserum appeared that anti-HD1 partialy reacted with crystal protein but anti-HD73 reacted with HD1 endotoxin about 100%.

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Simultaneous Analysis and Survey for Contamination of Nivalenol, Deoxynivalenol, T-2 toxin and Zearalenone in Feed (사료 중 Nivalenol, Deoxynivalenol, T-2 foxin과 Zearalenone의 동시분석과 오염도조사)

  • Kim, Dong-Ho;Kim, Hyun-Jung;Jang, Han-Sub;Kim, Yeong-Min;Choi, Heng-Bo;Ahn, Jong-Sung
    • Journal of Food Hygiene and Safety
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    • v.26 no.1
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    • pp.1-11
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    • 2011
  • Nivalenol (NIV), deoxynivalenol (DON), T-2 toxin (T-2) and zearalenone (ZEN) are mycotoxins produced by some Fusarium species known to be very frequently contaminated in feed. The study for simultaneous analysis and contamination survey in animal feed carried out. All mycotoxins were analysed by using high performance liquid chromatography tandem mass with internal standard. The limits of detection (LOD) were $2.0\;{\mu}g/kg$, $1.0\;{\mu}g/kg$, $1.0\;{\mu}g/kg$ and $0.1\;{\mu}g/kg$ for NIV, DON, T-2 and ZEN, respectively. Two hundred and thirty nine samples of feed were collected. The average concentration of DON was $212.3\;{\mu}g/kg$, $207.8\;{\mu}g/kg$ and $812.1\;{\mu}g/kg$ in chicken, pig and cattle feed, respectively. The average concentration of ZEN was $31.2\;{\mu}g/kg$, $35.6\;{\mu}g/kg$ and $147.2\;{\mu}g/kg$ for them, respectively. Especially, the levels of contamination for DON and ZEN were higher than those of NIV or T-2. And, the levels of contamination for four Fusarium mycotoxins in cattle feed appeared higher than those of pig and chicken feed. It was investigated that the high level of mycotoxin contamination in cattle feed was caused by com gluten feed of ingredients for feed, mainly.

Dose-dependent Effects of Bee Venom Acupuncture on MPTP-induced Mouse Model of Parkinson's Disease (MPTP로 유발된 파킨슨병 Mouse 모델에 대한 봉약침의 농도의존적 효과)

  • Jun, Hyung-Joon;Kim, Yong-Suk
    • Journal of Acupuncture Research
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    • v.27 no.5
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    • pp.59-68
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    • 2010
  • 목적 : 최근 한의학에서 널리 사용되며, 신경계 질환에도 응용되고 있는 봉약침의 농도의존적 효과를 알아보기 위하여, 대표적인 신경 퇴행성 질환인 파킨슨병의 동물모델을 통해 세포보호효과와 세포사멸 및 신경염증 기전을 관찰하였다. 방법 : C57BL/6 mice에 신경독소인 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine(MPTP)를 4번 복강내 주입하여 중뇌의 흑질 도파민 신경세포를 파괴하여 Parkinson 질병동물 모델을 만든 후, 2개의 군에는 마지막 MPTP 투여 2시간 후에 1차, 그 후로 48시간이 지날 때마다 양측 신수에 각각 0.06mg/kg 농도와 0.6mg/kg 농도의 봉약침을 시행하여 총 4회 시술한 후, 도파민 세포를 측정하는 TH 면역조직 화학법을 통해 세포의 보존 정도를 관찰하고, 세포사멸과 관련된 양상을 확인하기 위하여 Caspase 3, 신경염증과 관련된 양상을 확인하기 위하여 iNOS의 발현여부를 면역 조직화학법을 이용하여 관찰하였다. 결과 : 관찰결과 MPTP 투여 후 MPTP 투여군의 흑질의 도파민 세포 수는 감소하였으나 0.6mg/kg 봉약침을 투여한 경우에는 유의성 있게 세포 수가 유지되었다. Caspase-3와 iNOS 발현억제 실험에서 0.6mg/kg 봉약침군은 MPTP 투여군과 0.06mg/kg의 봉약침군과 비교하여 Caspase-3, iNOS 발현을 유의하게 억제하였다. 결론 : 봉약침은 MPTP 투여로 인한 신경세포 손상에 대하여 농도에 따라 세포사멸 기전과 신경염증 기전을 억제함으로 신경세포를 보호하는 것으로 추정되며, 추후 적절한 경혈점 및 최적의 봉약침 농도를 찾는데 지속적인 연구가 필요할 것이다.

Inactivation of Pathogenic Escherichia coli Using Crude Extract of Immunized Silkworm (면역유도누에 추출물을 이용한 병원성 대장균의 불활성화)

  • Park, Jong Woo;Jeong, Chan Young;Lee, Chang Hoon;Kang, Sang Kuk;Ju, Wan-Taek;Kim, Seong-Wan;Kim, Nam-Suk;Kim, Kee Young
    • Journal of Life Science
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    • v.31 no.8
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    • pp.755-760
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    • 2021
  • Swine diarrhea is a livestock disease that causes huge economic losses to pig farms. In general, diarrhea occurs because of the proliferation of pathogenic Escherichia coli (E. coli). The toxins produced by the proliferated E. coli cause edema in pigs. Although the proliferation of these coliforms can be prevented by using a vaccine, the vaccines containing chemically produced dead bacteria are not very effective, making it difficult to control the proliferation of E. coli. Therefore, there is a need to develop new, more effective vaccines. In this study, we prepared killed F4+ and F18ab+ E. coli, which induce diarrhea and edema in pigs, using the extracts of immune-induced silkworms containing antimicrobial peptides and examined their availability as a killed-bacteria vaccine. First, the antimicrobial activity analysis of the prepared immune-induced silkworm extract was conducted using the radial diffusion assay. The results showed high activity against both F4+ and F18ab+ E. coli. The production efficiency of E. coli dead cells was determined using the colony-counting method. The concentration of the E. coli dead cells was the highest (50 mg/ml) when treated at 4℃. In addition, the analysis of the prepared dead cells using a transmission electron microscope confirmed that E. coli leaked out of the cytoplasm and the cell membrane remained intact. Therefore, F4+ and F18ab+ E. coli produced using immune-induced silkworms extract are considered to be highly available as bacterial ghost vaccines that can help prevent swine diarrhea and the resulting edema.