• 제목/요약/키워드: toxin components

검색결과 53건 처리시간 0.025초

열분해 질량분석법을 이용한 생물학 물질의 특성 연구 (Characteristics study of biological materials using pyrolysis-mass spectrometry)

  • 최선경
    • 한국군사과학기술학회지
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    • 제7권3호
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    • pp.110-121
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    • 2004
  • Pyrolysis-mass spectrometry, incorporating an in situ thermal hydrolysis and methylation(THM) step, has been used to study biological materials for bacteria, toxin and virus. Newly developed pyrolyzer was used to decompose biological materials, and tetramethylammonium hydroxide(TMAH) was used as a methylation reagent. Chemical ionization(CI) using ethanol and ion trap mass spectrometer(ITMS) were used to ionize and analyze of pyrolysis components, respectively. Analytical characteristics of bacteria (including spore), virus and toxin were analyzed. Also acquisition and interpretation of mass spectra as biomarkers for classification/identification of biological material s were explained.

Expression and Characterization of Helicobacter pylori Adhesin Protein Linked to Cholera Toxin A2/B Subunits in Escherichia coli

  • Kim, Byung-Oh;Shin, Sung-Seup;Yoo, Young-Hyo;Pyo, Shuk-Neung
    • Journal of Microbiology and Biotechnology
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    • 제10권1호
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    • pp.56-62
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    • 2000
  • The hpa gene genetically linked to the ctxa2b gene was cloned into the pTED expression vector, and the constructed pTEDhpa/ctxa2b was transformed into Excherichia coli. The fusion protein, the adhesin fused to the cholera toxin subunit A2B (CTXA2B) subunit, was expressed to high levels as inclusion bodies in E. coli. The expressed protein was partially purified by washing the inclusion bodies with working solution containing 8M Urea and 0.1M DTT. Refolding of denatured fusion protein was carried out in the presence of glutathione redox buffer. The refolded fusion protein was purified by size exclusion chromatography. The expressed fusion protein was verified by SDS-PAGE, western blotting with antibodies to both antigenic components of adhesin and cholera toxin subunit B (CTXB), and its N-terminal amino acid sequence was analyzed. The orderly assembled fusion protein was confirmed by modified Gm1-ganglioside ELISA with Abs to adhesin. The results indicate that the purified fusion protein is an Adhesin/CTXA2B protein containing the H. pylori adhesin and $G_{m1}4-ganglioside binding activity of CTXB and the expressed fusion protein in E. coli could be easily purified by the refolding process, Its molecular weight was 168kDa as estimated by size exclusion chromatography. The Adhesin/CTXA2B protein may be used as a candidate antigen for oral immunization against H. pylori.

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한국산(韓國産) 고등균류(高等菌類)의 성분연구(成分硏究)(제54보)(第54報) -털목이의 독성(毒性) 성분(成分)에 관한 연구(硏究)- (Studies on Constituents of the Higher Fungi of Korea(LIV) -Studies on Toxic Component of Auricularia polytricha-)

  • 김하원;최응칠;김병각
    • 한국균학회지
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    • 제14권4호
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    • pp.265-271
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    • 1986
  • 털목이버섯의 저분자 독성물질로부터 다음과 같은 결론을 얻었다. 털목이버섯 열수추출액의 $LD_{50}$은 복강 및 경구투여시 각각 1.25와 4.18 g/kg이었다. 건조 털목이버섯 100 g에는 650$\sim$700 mouse lethal unit(=MLU)이 함유되어 있었다. 열수총추출액, C분흑 및 최종성분의 1 MLU는 각각 및 28.5, 25.0 및 12.0 mg/mouse이었다. 독성물질의 일반적일 증상은 행동둔화, 체모기립, 전율, 두부진동, 안구돌출, 사지마비, 사망직전의 급격한 다리운동 및 호흡마비 등이었다.

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Comparison of MBA and HPLC Post-column Oxidation Methods for the Quantification of Paralytic Shellfish Poisoning Toxins

  • Yu, Hongsik;Lim, Keun Sik;Song, Ki Cheol;Lee, Ka Jeong;Lee, Mi Ae;Kim, Ji Hoe
    • Fisheries and Aquatic Sciences
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    • 제16권3호
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    • pp.159-164
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    • 2013
  • The mouse bioassay and high performance liquid chromatography (HPLC) post-column oxidation method are different methods of quantifying paralytic shellfish poisoning toxins. In this study, we compared their ability to accurately quantify the toxicity levels in two types of field sample (oysters and mussels) with different toxin profiles for routine regulatory monitoring. A total of 72 samples were analyzed by both methods, 44 of which gave negative results, with readings under the limit of detection of the mouse bioassay ($40{\mu}g/100g$ saxitoxin [STX] eq). In 14 oysters, the major toxin components were gonyautoxin (GTX) 1, -2, -3, -4, -5, decarbamoylgonyautoxin-2 (dcGTX2), and decarbamoylsaxitoxin (dcSTX), while 14 mussels tested positive for dcSTX, GTX2, -3, -4, -5, dcGTX2, neosaxitoxin (NEO), STX, and dcSTX. When the results obtained by both methods were compared in two matrices, a better correlation ($r^2=0.9478$) was obtained for mussels than for oysters ($r^2=0.8244$). Additional studies are therefore needed in oysters to investigate the differences in the results obtained by both methods. Importantly, some samples with toxin levels around the legal limit gave inconsistent results using HPLC-based techniques, which could have a strong economic impact due to enforced harvest area closure. It should therefore be determined if all paralytic shellfish poisoning toxins can be quantified accurately by HPLC, and if the uncertainties of the method lead to doubts regarding regulatory limits.

Toxic Components of Auricularia polytricha

  • Kim, Ha-Won
    • Archives of Pharmacal Research
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    • 제16권1호
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    • pp.36-42
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    • 1993
  • To find biologically active components of the higher fungi of Korea, the carpophores of Auricularia polytricha, a well-known edible mushroom, were extracted with 0.14 M NaCl solution. The extract was successively fractionated by adding ammonium sulfate at various concentrations, and the respective precipitates were separated by centrifugation, then dialyzed and freeze-dried. When a does of 60 mg/kg of each was injected i.p. into ICR mice, the fraction which precipitated at 20% ammonium sulfate showed the highest toxicity, killing seven out of seven mice within two days. The fraction obtained at 40% ammonium sulfate showed the second highest toxicity. The two fractions were named auritoxin I and II after the genus name. However, they Nere shown to have nearly identical composition by physicochemical and 6.8% protein. The polysaccharide moiety was found to have 12.3% $\alpha$-linkage and 87.7% $\beta$-linkage and to be a heteromannoglucan consisting of 45.1% glucose, 435 mannose and 11.0% xylose. The protein moiety contained ten amino adids. The molecular weight of the toxin was $1.5\times10^6$ dalton by Sepharose CL-4B gel filtration. The modian lethal doses of auritoxin in mice were 56.4, 157.2 and 454.6 mg/kg by i.p., s.c. and p.o.administrations, respectively. The signs of intrxication were convulsion during the first 30 minutes after the injection, coma or sleeping within an hour, termor, lacrimation, nasal bleeding congestion, and death in 24 hours. Smong the various organs, the spleen was found to be enlarged remarkably. Human platelet aggregation was inhibited by the addition of auritoxin. The activity of malic dehydrogenase in vitro was inhibited by the toxin.

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Purification and Anti-pathogenic Properties of Immunoglobulin Concentrates from Porcine Blood

  • Jung, Tae-Hwan;Choi, Jae-Hwan;Koh, Kyung-Chul;Jeon, Woo-Min;Han, Kyoung-Sik
    • 한국축산식품학회지
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    • 제37권5호
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    • pp.743-751
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    • 2017
  • During slaughtering, animal blood is typically discarded, resulting in water pollution. However, this discarded blood has valuable components, such as immunoglobulin (Ig). Although several studies have been conducted to develop methods for effective recycling of slaughterhouse blood, they have not been commercially utilized in Korea. Here, we extracted an Ig-rich fraction from porcine blood that was then subjected to various in vitro tests, including pathogen growth inhibition, antigenic cross-reactivity, and anti-toxin activity. The porcine immunoglobulin concentrate (PIC) was effectively purified by eliminating other components, such as albumin, and consisted of approximately $63.2{\pm}2.9%$ IgG and $7.2{\pm}0.4%$ IgM on a protein basis. The results showed that it significantly suppressed the growth of pathogenic bacteria, and bound to all tested pathogens, including both gram-positive and gram-negative species, although the degree of activity differed according to strain. The PIC bound to two types of lipopolysaccharide (LPS) obtained from Escherichia coli O111:B4 and Salmonella enterica serotype typhimurium in a concentration-dependent manner. In addition, the PIC restored the proliferation activity of the lymphoblast K-562 cells when co-incubated with pathogenic LPS. These results confirm that the PIC prepared in this study is a potentially valuable functional food material or diet supplement as an alternative to antibiotics that can protect animals from pathogenic bacteria.

생쥐 대식세포의 활성시 신호전달에 미치는 인삼성분들의 영향 (The Effects of Ginseng Components on the Signal Transduction in the Activation of Murine Macrophages)

  • 신은경;박한우
    • Journal of Ginseng Research
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    • 제20권2호
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    • pp.159-167
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    • 1996
  • To study the effects of ginseng saponin components on the signal transduction in the ac tivation of murine macrophages, phagocytosis and Intracellular calcium concentration of peritoneal exuded mouse macrophages were examined. The phagocytosis was increased significantly after treatment with total saponin, diol-saponin, $Rg_1$ and $Rg_2$, but triol-saponin was unable to increase phagocytosis. The phagocytosis were increased when H7, a PKC inhibitor, was pretreated and increased significantly by saponin fractions except total saponin. Pertussis toxin, which inactivates G-protein, decreased the phagocytosis. But the phagocytosis was restored to the control level by saponin fractions and the phagocytosis was increased significantly by $Rg_2$ and $Rg_2$. The triol saponin increased phagocytosis approximately by 2-fold as compared with the TMB-8 treated group. Peritoneal exuded macrophages displayed a prominent rise in cytosolic calcium following treatment with triol-saponin, $Rg_1$, $Rg_2$ and $Rg_2$. Incubation of macrophages with PT resulted in an inhibition of cytosolic calcium mobilization, but increased cytosolic calcium mobilization with saponin fraction.

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한국산(韓國産) 고등(高等) 균류(菌類)의 성분(威分) 연구(硏究)(제48보)(第48報) -노란다발버섯의 독(毒) 성분(成分) 네마톡신- (Studies on Constituents of the Higher Fungi of Korea(XLVIII) -Nematoxin of Naematoloma fasciculare-)

  • 김병각;심미자
    • 한국균학회지
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    • 제12권3호
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    • pp.117-119
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    • 1984
  • 한국에 야생하는 독버섯의 하나인 노란다발버섯을 물로 가열 추출한 후 추출액에 3배의 에탄올을 첨가하여 고분자 물질을 침전시켰으며 이를 다시 투석하여 정제 하였다. 이것을 냉동 건조하여 분말을 얻었으며 60mg/kg 용량을 백서의 복강내에 주사 하였던 바, 6일 후 부터 사망하기 시작하여 7일째에는 7마리중 4마리가 절명하였다. 이 유독 성분을 "네마톡신"이라 명명하고자 한다.

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가열조리가 복어 추출물 함질소화합물의 조성에 미치는 영향 (Changes of the Composition of Nitrogenous Compounds in Globefish Meat Extracts by the Cooking Method)

  • 양영;한영실;변재형
    • 한국식품조리과학회지
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    • 제6권2호
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    • pp.85-96
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    • 1990
  • Globefish, Fugu xanthopterus, known to have a severe toxin, is one of the favorite food in Korea and Japan when the toxic part is removed. In this paper, the effect of cooking on the composition of nitrogenous components in the extractives from globefish cooked investigated and the changes of the taste compounds originated from the nitrogenous components in the extractives were discussed. When the sample fish was analysed for general composition, drip amount and pH by the different methods of thawing, the method effective method was the running water thawing. Total nitrogen content in raw globefish and the frozen globefish was not different, and the nitrogen content was increased with the heat treatment. It seemed that the nitrogen content was higher in the extract from the boiled globefish than that of the steamed globefish. Taurine, lysine, glycine and alanine were occupied about 70% of the total free amino acids. Total free amino acid content was higher in the extracts from the frozen sample than those from the raw sample. The amount of free amino acids was increased when the globefish soup cooked under the direct-heat cooking than in the microwave oven-heat cooking. Among nucleotides in the extracts from the thawed and cooked fishes, IMP and inosine contents were increased, and the both components were decreased with the heating time and by the heating method. Tne content of total creatinine-nitrogen were 50% of the total nitrogen content of the extracts, but the concentration of glycinebetaine, TMA and TMAO were only a few amounts. It could be concluded that total creatinine components, including free amino acids such as taurine, lysine, glycine and alanine, and IMP might be the important components contributing to the taste of the cooked globefish.

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탄저균 pagA 유전자의 분자적 다양성 (Molecular Diversity of pagA Gene from Baciilus anthracis)

  • 김성주;조기승;최영길;채영규
    • 미생물학회지
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    • 제37권1호
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    • pp.49-55
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    • 2001
  • 탄저(anthrax)는 그람양성이고 포자형성 세균인 탄저균(Bacillus anthracis)으로부터 발명되어진다. 탄저독소는 세가지 요소로 구성되어 있으며, 방어항원(PA)은 숙주세포 표면에서 탄저 독소단백질 및 이종단백질을 세포질 내로 이동시키는 역할을 한다. 본 연구에서는 PA의 분자적 다양성과 국내에서 탄저균의 진화를 이해하고 확인하기 위해 국내외에서 발견된 탄저균 4 균주와 기존에 보고된 탄저균 26 균주로부터 2,294 bp의 PA유전자(pagA)의 DNA 염기서열을 분식하였다. 탄저균 30 균주으로부터 PA유전자의 염기서열을 비교 분식한 결과, 8개 부위에서 돌연변이를 확인 하였다. 돌연변이가 일어난 부위에 따라서 탄저균을 10종류의 PA 유전자형과 4 종류의 PA 표현형으로 구분하였다. 한국 경주에서 분류된 B. anthracis BAK는 600번째 아미노산 alanine이 valine으로 바뀌어서 B. anthracis ATCC 14185 보다 LF와 PA의 결합 위치를 근접하게 하였다. 탄저균의 Pag의 염기서열을 통한 계통분석학적인 분석 결과는 염색체상에서의 분류와 일치하여 탄저균사이에서 pXO1 플라스미드의 수평적인 이동은 없는 것으로 사료된다.

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