• KSBB Journal
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• 제16권2호
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• pp.163-169
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• 2001

This study was undertaken to evaluate the effects of green and taste teas on the in-vitro antimicrobial activity and vacuolating toxin titer of Helicobacter pylori. Crude aqueous extracts prepared by adding 2 g of tea leaf or powder to 100 ml of boiling distilled water, and sterilized by passing through a 0.22 $mutextrm{m}$ membrane filter. Green tea, coffee, and ginger tea showed bactericidal activity on H. pylori within 3 hours. Black tea and ssangwha tea also showed bactericidal activity on H. pylori in 24 hours. Arrowroot tea show no bactericidal effect on H. pylori after 48 hours. Two fold diluted green tea and coffee decreased(1/10,000cfu) the growth of H. pylori in 24 hours, but the two fold diluted black tea, ssangwha tea, and ginger tea showed suppression effect upon of(1/10cfu) H. pylori in 24 hours. The two-fold and 10-fold diluted green tea, coffee and two-fold diluted black tea abrogated the vacuolating toxin titer of H. pylori, but the two-fold and 10-fold diluted ginger, ssangwha, ginseng, and arrowroot tea only reduced the vacuolating toxin titer of H.pylori from 1/2 to 1/8. These result suggest that green tea and coffee have effective antibacterial or bactericidal effects on H.pylori, and that they also have a neutralization effect upon the vacuolating toxin of H.pylori.

• 대한화학회지
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• 제59권1호
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• pp.22-30
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• 2015

본 연구에서는 디프테리아 독소가 세포막의 지질에 미치는 영향을 알아보기 위해 HepG2 세포에서 포스포리파제 D와 유리된 지방산(Free fatty acid)의 변화를 살펴보았다. 지질변화는 pH 5.1에서 최고 값을 나타냈으며, 이 pH에서 포스포리파아제 D의 활성을 3.5배 가량, 유리된 지방산의 방출은 5배 정도 증가되었다. 이는 디프테리아 독소가 세포 안으로 들어가는 과정에서 세포막이 교란되어 재배열되었음을 시사한다. 한편 세포막을 무작위로 교란시키는 디지토닌의 영향이 디프테리아 독소의 그것보다 중성 pH에서 4배 이상 상당히 높게 나타난 것으로 미루어 보아 디프테리아 독소의 영향이 상대적으로 선택적인 교란 현상인 것으로 보여진다. 이런 세포막 교란의 연유를 밝히고자 세포막 구멍 형성 저해제인 cibacron blue와 세포막 융합 펩티드를 갖고 있는 hemagglutinin의 영향을 검토하였다. Cibacron blue는 디프테리아 독소에 의한 지질 변화를 50% 정도 저해시켰으며, hemagglutinin에 의한 지질변화는 디프테리아 독소의 그것과 유사함을 관찰 할 수 있었다. 이들 결과들은 디프테리아 독소에 의한 세포막 교란이 구멍형성과 독소의 소수성 펩티드가 세포막에 삽입되는 과정이 서로 연계되어 있음을 암시한다. 그 외 일련의 실험으로 디프테리아 독소가 세포막을 통과하는 과정에서 HepG2 세포의 투과성은 상승시켰으나, 세포의 생존능력은 상당히 높게 유지되었고 DNA 토막내기 같은 세포의 괴사는 일어나지 않았다. 이런 조건하에서 디프테리아 독소는 산성 pH에서 HepG2 세포의 지질의 변화를 가져 온다는 것을 밝힐 수 있었다.

• 한국식품과학회지
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• 제26권5호
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• pp.585-589
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• 1994

시험관 내에서 T-2 toxin이 병아리 비장세포의 blastogenesis에 미치는 영향을 살펴본 결과, B-cell mitogen인 lipopolysaccharide 및 T-cell mitogen인 concanavalin A 자극에 대해 T-2 toxin의 농도가 증가함에 따라 억제정도가 증가하는 경향을 나타내었다. 노출시기를 달리하여 T-2 toxin을 투여한 병아리의 비장세포에서 mitogen 자극에 내한 반응을 안아 본 결과, 부화하기 전과 후에 계속 T-2 toxin에 노출시킨 실험군은 가장 영향을 많이 받은 것으로 나타났고 부화전 혹은 부화후 어느 한 시기에만 T-2 toxin에 노출된 실험군은 비교적 영향을 적게 받는 것으로 나타났다.

• Journal of Microbiology and Biotechnology
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• 제19권1호
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• pp.108-112
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• 2009

A simplified method for the purification of cholera toxin was developed. The 569B strain of Vibrio cholerae, a recognized hyper-producer of cholera toxin, was propagated in a bioreactor under conditions that promote the production of the toxin. The toxin was separated from the bacterial cells using 0.2-${\mu}m$ crossflow microfiltration, the clarified toxin was passed through the membrane into the permeate, and the bacterial cells were retained in the retentate. The 0.2-${\mu}m$ permeate was then concentrated 3-fold and diafiltered against 10 mM phosphate buffer, pH 7.6, using 30-kDa crossflow ultrafiltration. The concentrated toxin was loaded onto a cation exchange column, the toxin was bound to the column, and most of the impurities were passed unimpeded through the column. The toxin was eluted with a salt gradient of phosphate buffer, pH 7.0, containing 1.0 M NaCl. The peak containing the toxin was assayed for cholera toxin and protein and the purity was determined to be 92%. The toxin peak had a low endotoxin level of $3.1\;EU/{\mu}g$ of toxin. The purified toxin was used to prepare antiserum against whole toxin, which was used in a $G_{M1}$ ganglioside-binding ELISA to determine residual levels of toxin in an oral inactivated whole-cell cholera vaccine. The $G_{M1}$ ganglioside-binding ELISA was shown to be very sensitive and capable of detecting as little as 1 ng/ml of cholera toxin.

• 미생물학회지
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• 제36권2호
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• pp.91-96
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• 2000

불와전 균류인 Fusarium s^g pp.를 이용하여 여러 가지 배양조건과 생리적 영향에 따른 균주의 성장 및 T-2 toxin의 생성에 관하여 고찰하였다. T-2 toxin 의 검출방법은 thin layer chromatography (TCL) 법과 미생물학적 검출방법을 사용하였다. 고체 배지의 경우 횐옥수수 가루(Quaker사 제품)베지에서 다른 곡물보다 많은 양의 T-2 toxin이 생성되었으며,비교적 깨끗한 T-2 toxin이 정제되었다. 이 경우 배지 100g당 약 700 mg의 T-2 toxin이 생성되었으며, 그중 약 30%정도가 깨끗한 결정으로 정제되었다. 고온(20-$25^{\circ}C$)에서는 생장은 많았으나, T-2 toxin의 생성은 적었으며, 저온(10-$15^{\circ}C$)에서는 비교적 생장이 적었지만, T-2 toxin의 생성이 많았고, 젖당, 글리세롤, 솔비톨의 경우는 적었다. 유일 탄소원으로 구연산과 초산은 이용하지 못하였으며, 녹발의 경우 생장은 많았으나 T-2 toxin의 생성양은 적었다. 질소원의 경우 $NaNO_2$를 제외하고는 $(NH_4)_2NO_4$, $NH_4Cl_3$, $NH_4NO_3$, $KNO_3$ 를 거의 동일하게 이용하였다. 초기 pH값에 생성과 균주의 성장은 pH4.0-5.0일 경우 최적을 나타냈으며 ph6.0이상에서는 성장도 저하되고, T-2 toxin생성도 적었다. 회전속도에 따른 T-2 toxin 생성과 균주의 성장을 보면 회전속도가 속돠 증가함에 따라 균주의 생장과 T-2 toxin 생성량이 모두 증가하였다. $15^{\circ}C$에서 7일간 배양 후, $25^{\circ}C$로 옮겨 7일간 배양하여, toxin의 생성을 보면, $15^{\circ}C$에 7일간 배양했을 때보다 T-2 toxin양이 적었다. 이는 생성되었던 T-2 toxin이 분해되었음을 보여주는 것이다. 이상의 결과를 볼 때 T-2 toxin 대사 경로는 온도에 의한 효소 억제 또는 효소 유지 시스템에 의해 조절되는 것이라고 생각할 수 있다.

• 대한약리학회지
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• 제31권1호
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• pp.115-122
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• 1995

C5a 또는 PMA에 의하여 활성화된 호중구에서의 superoxide와 HOCl 생성에 나타내는 staurosporine, genistein과 pertussis toxin의 효과를 관찰하였다. C5a에 의한 superoxide과 $H_2O_2$의 생성은 staurosporine, genistein과 pertussis toxin에 의하여 억제되었다. PMA의 자극효과는 staurosporine에 의하여 억제되었으나 pertussis toxin에 의하여 영향을 받지 않았으며, 한편 이는 genistein에 의하여 더 촉진되었다. Staurosporine, genistein은 sodium fluoride에 의한 superoxide 생성을 억제 하였으나 pertussis toxin은 영향을 나타내지 않았다. PMA에 의한 $H_2O_2$의 생성은 staurosporine에 의하여 억제되었으나 pertussis toxin은 영향을 나타내지 않았다. Genistein은 PMA에 의한 $H_2O_2$의 생성에 자극효과를 나타내지 않았다. Staurosporine과 pertussis toxin은 C5a 또는 PMA에 의한 HOCl 생성을 억제하였으나, 이에 반하여 genistein은 자극하였다. C5a와 PMA에 의한 myeloperoxidase 유리는 genistein에 의하여 억제되었나, pertussis toxin의 효과는 나타나지 않았다. Staurosporine은 유리에 대한 PMA의 자극효과에 영향을 주지 않았다. Myeloperoxidase 활성은 genistein에 의하여 현저하게 증가되었으나 staurosporine과 pertussis toxin의 영향은 받지 않았다. 이상의 결과는 호중구의 respiratory burst가 protein kinase C와 protein tyrosine kinase에 의하여 조절된다고 제시한다. Protein kinase C의 직접적인 자극에 따른 superoxide 생성은 protein tyrosine kinase의 영향을 역으로 받을 것으로 추정된다. Genistein은 아마도 myeloperoxidase를 활성화하여 HOCl 생성을 촉진할 것으로 시사된다.

• 생약학회지
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• 제16권3호
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• pp.129-135
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• 1985

In order to search for potential antidotes for T-2 toxin poisoning, seven Chinese herbal drug extracts and five natural constituents were tested on mice intoxicated with T-2 toxin. When extracts of Panax ginseng and Atractylodes japonica (500 mg/kg) were administered p.o. once 3 hrs before and once 1 hr after T-2 toxin treatment, a 30% complete survival rate was noted. In case of Paeonia albiflora var. typica, a 30% complete survival rate was also produced at a dose of 250 mg/kg. Other extracts, Glycyrrhiza uralensis, Scutellaria baicalensis, Rehmannia glutinosa and Plantago asiatica exhibited no significant protection from the T-2 toxin poisoning. A nucleoside, thymidine showed protective activity against T-2 toxin toxicity and it produced a 40% complete survival rate when administered i.p. once 0.5 hr after T-2 toxin treatment. Other natural constituents, aucubin, vitamin C and E, and lipoic acid did not show any significant protective activities.

• Journal of Acupuncture Research
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• 제33권2호
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• pp.1-9
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• 2016

Objectives : I investigated whether snake venom can synergistically strengthen the cytotoxic effects of NK-92 cells, and enhance the inhibition of the growth of lung cancer cells including NCI-H358 through the induction of death receptor dependent extrinsic apoptosis. Methods : Snake venom toxin inhibited cell growth of NCI-H358 Cells and exerted non influence on NK-92 cell viability. Moreover, when they were co-cultured with NK cells and concomitantly treated with $4{\mu}g/m{\ell}$ of snake venom toxin, more influence was exerted on the inhibition of growth of NCI-H358 cells than BV or NK cell co-culture alone. Results : The expression of Fas, TNFR2 and DR3 and in NCI-H358 lung cancer cells was significantly increased by co-culture of NK-92 cells and treatment of $4{\mu}g/m{\ell}$ of snake venom toxin, compared to co-culture of NK-92 cells alone. Coincidentally, Bax, caspase-3 and caspase-8 - expressions of pro-apoptotic proteins in the extrinsic apoptosis pathway, demonstrated significant increase. However, in anti-apoptotic NF-${\kappa}B$ activities, activity of the signal molecule was significantly decreased by co-culture of NK-92 cells and treatment of $4{\mu}g/m{\ell}$ of snake venom toxin, compared to co-culture of NK-92 cells or snake venom toxin treated by NCIH358 alone. Meanwhile, in terms of NO generation, there is a significant increase, in co-culture of NK-92 cells with NCI-H358 cells as well as the co-culture of NK-92 cells and concomitant treatment of $4{\mu}g/m{\ell}$ of snake venom toxin. However, no synergistic increase of NO generation was shown in co-culture of NK-92 cells and treatment of $4{\mu}g/m{\ell}$ of snake venom toxin, compared to co-culture of NK-92 cells with NCI-H358 cells. Conclusion : Consequently, this data provides that snake venom toxin could be useful candidate compounds to suppress lung cancer growth along with the cytotoxic effect of NK-92 cells through extrinsic apoptosis.

• Asian-Australasian Journal of Animal Sciences
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• 제15권7호
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• pp.1051-1056
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• 2002

In vitro binding efficacy of esterified glucomannan (E-GM) (0.1%) on aflatoxin B1 (AF) (300 ppb), ochratoxin A (OA) (2 ppm) and T-2 toxin (T-2) (3 ppm), when present alone or in combination, was evaluated in toxin-contaminated feed at pH 4.5 and 6.5. Esterified glucomannan showed significantly (p<0.01) higher binding with AF (81.6%), whereas those recorded with T-2 (27.8%) and OA (25.6%) were moderate. Binding of each toxin decreased as the number of toxins in feed increased. pH of medium showed no effect on mycotoxin binding ability of E-GM. A $2{\times}2{\times}2{\times}2$ factorial experiment of 5 week duration was conducted to study the effects of two dietary levels each of AF (0 and 300 ppb), OA (0 and 2 ppm), T-2 (0 and 3 ppm ) and E-GM (0 and 0.1%) on the immune competence of a total of 960 day-old commercial broilers. Reductions in size of thymus (by AF and T-2) and bursa (by AF) and antibody titers against Newcastle disease and Infectious Bursal disease (by all the toxins) were noted. Additive and antagonistic interactions were seen among the toxins on certain parameters. Esterified glucomannan significantly (p<0.01) improved antibody titers and weights of bursa ofFabricius and thymus indicating its counteracting efficacy against immunosuppression in mycotoxicosis of multiple origin.

• Journal of Microbiology and Biotechnology
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• 제18권8호
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• pp.1475-1481
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• 2008

B3 antibody specifically binds the $Lewis^Y$-related carbohydrate antigen of many carcinomas, and it is used as a model antibody in this study. In a previous study, the Fab fragment of the antibody was fused to a 38 kDa truncated form of Pseudomonas exotoxin A, PE38, to make Fab-PE38, where PE38 is fused to the Fd fragment of the Fab domain. This parent monomer molecule, Fab-PE38, had no cysteine in the hinge region, and it could not make a disulfide bond to form a disulfide bond bridged homodimer. In this study, we constructed three different kinds of divalent Fab-toxin fusion homodimers where the toxin is fused to the light chain of Fab, $(Fab-PE38fl)_2$. In addition to the PE38 toxin fused to the light chain, these three molecules have different hinge sequences hi, h2, and h3 making Fabh1-, Fabh2-, and Fabh3-PE38fl monomers, respectively. These hinges contain only one cysteine on different positions of the hinge sequence. The disulfide bond between the hinge region of two monomers forms homodimers $(Fabh1-PE38fl)_2$, $(Fabh2-PE38fl)_2$, and $(Fabh3-PE38fl)_2$. The refolding yields of these dimers were 5-16-fold higher than a previously constructed dimer where the PE38 was fused to the Fd fragment $(Fabh2-PE38)_2$ [8]. Our data suggest that the steric repulsion between the two PE38s in $(Fabh1-PE38)_2$ during disulfide bridge formation is relieved by fusing it at the end of the light chain. The best cytotoxicity value of these dimers showed about 2.5-fold higher on an MCF7 cell line than that of the monovalent reference molecule in ng/ml scale, which is 15-fold higher in pM scale.