• Title/Summary/Keyword: Srb

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Sulforhodamine B Assay to Determine Cytotoxicity of Vibrio vulnificus Against Human Intestinal Cells

  • Lee, Byung-Cheol;Choi, Sang-Ho;Kim, Tae-Sung
    • Journal of Microbiology and Biotechnology
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    • v.14 no.2
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    • pp.350-355
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    • 2004
  • Sulforhodamine B (SRB) assay is a rapid, sensitive, and inexpensive method for measuring cell proliferation and chemosensitivity. However, the lactate dehydrogenase (LDH) release assay is generally used to measure cytototoxicity of infectious microorganisms against host cells. In this study, we investigated the possibility of applying the SRB assay to determine cytotoxicity for infectious microorganisms, and compared the results with those obtained by the LDH release assay. We used Vibrio vulnificus as a model of infectious microorganisms. The SRB assay showed that V vulnificus strongly induced cytotoxic activity against human intestinal cells, Caco-2 and INT-407 cells. The degree of cytotoxicity closely correlated with infection time and number ratios of V. vulnificus to intestinal cells (MOI, multiplicity of infection). Furthermore, cytotoxicity values obtained by SRB assay correlated well with results obtained by the LDH release assay, and both assays gave a linear response with respect to MOI Heat-inactivation of V. vulnificus for 35 min at $60^{\circ}C$ did not induce cytotoxic activity, indicating that viability of V. vulnificus is crucial for cytotoxic activity against intestinal cells. Although both assays are suitable as cytotoxicity endpoints, the SRB assay is recommended for measuring cytotoxicity of infectious microorganisms against host cells because of its significantly lower cost and more stable endpoint than the LDH release assay.

Effect of stabilized rice bran-added high sucrose diet on glucose control in C57BL/6 mice (안정화미강을 첨가한 고서당식이 섭취가 C57BL/6 mice의 혈당조절에 미치는 영향)

  • Lee, Seung-Min;Shin, Mal-Shick;Heo, Young-Ran
    • Journal of Nutrition and Health
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    • v.47 no.3
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    • pp.157-166
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    • 2014
  • Purpose: Rice bran is a byproduct of the hulling of rice and contains a variety of bioactive components. Various studies have reported on the antioxidative, anticancer, immune-enhancing, and hypocholesterolemic effects of rice bran. However, few studies about the physiological activity of stabilized rice bran supplement on dietary intake of sugars is limited. The aim of this study was to investigate the effect of stabilized rice bran supplement on blood glucose in C57BL/6 mice fed a high sucrose diet. Methods: Animals were randomly divided into three groups respectively, and were fed a normal diet (ND group), a high sucrose diet (HSD group) or a high sucrose diet containing 20% stabilized rice bran (HSD-SRB group) for 12 weeks. Results: In the oral glucose tolerance test (OGTT), after seven weeks of feeding on the experimental diets, a significantly lower result was observed for HSD-SRB than for HSD at 30 and 60 minutes after oral administration in glucose solution (2 g/kg body weight). The incremental area under the curve (IAUC) of HSD-SRB was significantly lower than that of HSD. After 12 weeks, fasting blood glucose level of HSD-SRB was significantly lower than that of HSD. No significant difference in the serum insulin level was observed between HSD and HSD-SRB. However, HOMA-IR was significantly decreased in HSD-SRB compared to HSD. In addition, HOMA ${\beta}$-cell was significantly increased in HSD-SRB compared to HSD. Triglyceride in liver of HSD-SRB was significantly lower than that of HSD. Conclusion: Feeding diets containing 20% rice bran improved insulin resistance and insulin secretion by decreasing triglyceride in liver. Thus, rice bran has a positive effect on glycemic control. In addition, the results are expected to be utilized as a basis for human study and development of food products with added rice bran.

Cytotoxicity of Crude Extracts of Rheum uudulatum L. with Human Kidney Epithelial Cell A498 (신장 상피세포주 A498을 이용한 대황(Rheum undulatum L.)추출물의 세포독성)

  • 나명석;진종언;조남철
    • The Korean Journal of Food And Nutrition
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    • v.13 no.5
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    • pp.460-464
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    • 2000
  • We have evaluated cytotoxic effects of four crude extracts of methylene chloride, ethyl acetate, butanol, water layer isolated Rheum undulatum in A498 cell line, human kidney epithelial cells. The cytotoxic evalutation was measured by colorimetric assay using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide(MTT) , neutral red(NR) and sulforhodamine protein B(SRB). These results obtained are as follows : MTT, NR and SRB quantities were significantly decreased in cultured A498 cells treated four crude extracts by increased concentrations. The cell cytotoxic effect of crude extracts of butanol layer was more stronger than others layer. The values of MTT$\sub$50/, NR$\sub$50/, SRB$\sub$50/ of crude extract of butanol layer and were measured both 0.63 mg/ml, 0.65 mg/ml, and 0.68 mg/ml, respectively and the values of water layer were 0.84 mg/ml, 0.82 mg/ml. and 0.80 mg/ml. respectively in cultured A498 cell line.

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The Safety of Brucella abortus strain RB51 in pregnant cattle (부루세라 RB51 백신의 임신소에 대한 안전성)

  • Kim, Jong-man;Woo, Sung-ryong;Lee, Ji-youn;Jung, Suk-chan;Jean, Young-hwa;Kim, Jae-hoon;Kweon, Chang-hee;Yoon, Young-dhuk;Yoo, Han-sang
    • Korean Journal of Veterinary Research
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    • v.43 no.4
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    • pp.667-675
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    • 2003
  • The safety of Brucella abortus strain RB51(SRB51) was investigated in dairy cows and Korean native cattle of 4~7th month of gestation. From experimentally inoculated cattle, 18 of 25 (72.0%) dairy cows, and 3 of 10 (30.0%) Korean native cattle were aborted or delivered premature fetus. There were no significant differences in the incidence of abortion depending on the inoculation route (intramuscular and subcutaneous) and dosages ($1{\times}10^9$, $2.8{\times}10^9$, and $4.0{\times}10^9$ CFU). The antibodies to the SRB51 were measured by a dot blot enzyme-linked immunosorbent assay. The highest titers to SRB51 were detected between 5~7 weeks after inoculation and the specitic antibody could be detected up to 28 weeks after inoculation. The SRB51 was isolated from amnio-allantoic fluid, bronchial lymph node, mammary gland, and supramammary lymph node in 5 of 25 dairy cows during 4 weeks after either abortion or delivery. Although SRB51 was isolated from 4 of 24 aborted fetus or normally delivered calves at parturition time, it was not isolated during 4 weeks afterward. Eleven of twentyfive dairy cows showed the endometritis and/or necrosis until 6 weeks after delivery, no lesions were seen at 8 weeks after delivery and uterus from control dairy cows. The results of present study revealed that SRB51 might induce the clinical signs of brucellosis in the pregnant cattle at 4~7th month of gestation.

Trichloroethylene Removal Using Sulfate Reducing Bacteria and Ferric Iron (황환원균과 3가철을 이용한 Trichloroethylene의 제거에 관한 연구)

  • Hwang, Ki-Chul;Min, Jee-Eun;Park, In-Sun;Park, Jae-Woo
    • Journal of Soil and Groundwater Environment
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    • v.13 no.1
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    • pp.24-31
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    • 2008
  • Sulfate reducing bacteria (SRB) is universally distributed in the sediment, especially in marine environment. SRB reduce sulfate as electron acceptor to hydrogen sulfide in anaerobic condition. Hydrogen sulfide is reducing agent enhancing the reduction of the organic and inorganic compounds. With SRB, therefore, the degradability of organic contaminants is expected to be enhanced. Ferrous iron reduced from the ferric iron which is mainly present in sediment also renders chlorinated organic compounds to be reduced state. The objectives of this study are: 1) to investigate the reduction of TCE by hydrogen sulfide generated by tht growth of SRB, 2) to estimate the reduction of TCE by ferrous iron generated due to oxidation of hydrogen sulfide, and 3) to illuminate the interaction between SRB and ferrous iron. Mixed bacteria was cultivated from the sludge of the sewage treatment plant. Increasing hydrogen sulfide and decreasing sulfate confirmed the existence of SRB in mixed culture. Although hydrogen sulfide lonely could reduce TCE, the concentration of hydrogen sulfide produced by SRB was not sufficient to reduce TCE directly. With hematite as ferric iron, hydrogen sulfide produced by SRB was consumed to reduce ferric ion to ferrous ion and ferrous iron produced by hydrogen sulfide oxidation decreased the concentration of TCE. Tests with seawater confirmed that the activity of SRB was dependent on the carbon source concentration.

Comparative Analysis of Stress Distribution in Composite Resin Brackets with Metal Slot of Permanent Maxillary Central Incisor Using the Finite Element Method: A Pilot Study

  • Im, Jae-Jung;Song, Jae-Joon;Kim, Nan-Hee;Heo, Jin-Young;Jung, Gyu-Un;Hwang, Kyung-Gyun;Park, Chang-Joo
    • Journal of Korean Dental Science
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    • v.4 no.2
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    • pp.46-51
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    • 2011
  • Purpose: For aesthetic reasons, composite resin brackets are widely used. However, related studies are rare. This pilot study sought to compare the stress distributions in two commercially available composite resin brackets with metal slot. Materials and Methods: Two commercially available resin brackets -- full-metal slot resin bracket (fSRB) and partial-metal slot resin bracket (pSRB) with straight wire appliance dimension of $0.022{\times}0.028$ in -- were selected. In each bracket, 3-dimensional finite element models were constructed, and stress level was evaluated using finite element analysis. By loading the tipping force and torsion moment, which are similar to those applied by the stainless steel rectangular wire ($0.019{\times}0.025$ -in), stress distributions were calculated, and von Mises stress values were obtained. Results: In pSRB and fSRB, the stress value of the torque moment was much higher than that of the tipping force. The pSRB showed higher stress value than fSRB in both tipping force and torque moment because of the difference in size and configuration of the metal frame inserted into the slot. More stress was also found to be concentrated on the slot area than the wing area in fSRB. Conclusion: The slot form of fSRB was found to be more resistant to the stress of tipping and torque than the slot form of pSRB. In addition, the slot areas -- rather than the wing areas -- of the bracket showed breakage susceptibility. Therefore, resistance to the torque moment on the slot area should be considered in bracket design.

Most Probable Number 방법을 이용하여 측정한 중랑천 하상토양의 혐기성 세균의 수와 수질과의 상관 관계

  • Park, Doo-Hyun;Kim, Byung-Hong;Lim, Si-Keun;Choi, Young-Hyo
    • Microbiology and Biotechnology Letters
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    • v.24 no.3
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    • pp.364-370
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    • 1996
  • Sediments collected from the Jungnang-cheon and its tributaries were used to enumerate anaerobic bacteria by most probable number (MPN) methods. A simple method was developed to detect ferrous ion in the culture fluid in order to count the number of iron ion reducers, and sulfate-reducing bacteria (SRB) and methanogens were detected by the presence of FeS precipitate in the culture or methane in the head space, respectively. The numbers of iron reducer was in the range of 10$^{7}$ - 10$^{8}$ /g in the sediment of the stream containing higher organic content than the tributaries. The sediments of tributaries were analyzed to contain iron reducers less than 10$^{7}$ cells/g. With one exception the numbers of SRB and methanogens were less than 10$^{3}$ cells/g in the sediment. From these results it is concluded that organics in the sediment support the growth of iron reducers, which out-compete SRB and methanogens.

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Microbiologically Induced Corrosion of Three Tubular Materials

  • Mukadam, S.;Al-Hashem, A.
    • Corrosion Science and Technology
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    • v.14 no.6
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    • pp.267-272
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    • 2015
  • The performance of three tubular materials (C-90, L-80, and N-80) was evaluated in a synthetic brine inoculated with sulfate-reducing bacteria (SRB) in the absence and presence of biocides. A flow loop was used in the evaluation of the three alloys. Morphological examination of the alloy surfaces after exposure to SRB and after biocide treatment was performed by scanning electron microscopy (SEM) to determine the nature of any localized corrosion. The SE images of the coupon samples showed a marked difference between the biocide-treated and untreated samples. Small pits were observed on the ultrasonically cleaned surfaces of the three alloys after exposure to SRB. The biocide treatment reduced the number of SRB on the surfaces of the alloys. Results indicated that C-90 and L-80 alloys exhibited better MIC resistance than N-80 under the conditions used in this study.

Inhibitory effects of Saiko-Ka-Ryukotsu-Borei-To on the migration and proliferation of vascular smooth muscle cell

  • Chung, Hwa-Jin;Ikuro Maruyama;Tadato Tani;Lee, Sang-Kook
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2003.11a
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    • pp.100-100
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    • 2003
  • We have reported that oral administration of Saiko-Ka-Ryukotsu-Borei-To (SRB), a traditional Chinese formulation, inhibited the intimal thickening in carotid artery after balloon injury in cholesterol-fed rats. To elucidate its mechanism, the effects of SRB on migration and proliferation of vascular smooth muscle cell (VSMC) were examined in vivo and in vitro. We have reported that oral administration of Saiko-ka-Ryukotsu-Borei-To (SRB), a traditional Chinese formulation, inhibited the intimal thickening in carotid artery after balloon injury in cholesterol-fed rats. To elucidate its mechanism, the effects of SRB on migration and proliferation of vascular smooth muscle cell (VSMC) were examined in vivo and in vitro.

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전남지역 생활하수처리장과 농공단지폐수처리장 방류수에 대한 세포독성 평가 - XTT, SRB in vitro bio-assay를 이용한 -

  • Lee, Moon-Hee;Han, Sang-Guk
    • Proceedings of the Korean Environmental Sciences Society Conference
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    • 2005.11a
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    • pp.185-188
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    • 2005
  • XTT assay와 SRB assay를 이용하여 Hep3B와 L929 세포에서 생활하수처리장과 농공단지폐수처리장 방류수의 세포독성을 알아본 결과 다음과 같은 결론을 얻었다. 1) XTT, SRB 실험법에 적용하여 대표적인 발암성 물질인 Benzo(a)Pyrene의 세포독성을 평가 할 수 있었다. Benzo(a)Pyrene 30uM농도는 Hep 3B와 L929 세포에서 약 50%의 세포독성을 나타내었다. 2) 단백질합성과 세포호흡의 활성도로 세포독성을 평가할 수 있는 실험법인 XTT와 SRB 실험법에 적용하여 방류수를 추출 농축한 실제 시료에 대한 세포독성을 평가할 수 있었다. 3) 생활하수처리장의 방류수에 비해 다종 고농도의 화학물질로 오염되어있는 농공단지폐수처리장 방류수에서의 독성발현이 더 높았다. 4) 생활하수처리장 방류수와 농공단지폐수처리장 방류수 중 오염도가 가장 심각한 4지점의 화학물질 분석 결과와 in vitro bio-assay에 의한 세포독성 결과가 일치하였다.

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