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Apoptotic Effect of Extract from Artemisia annua Linné by Akt/mTOR/GSK-3β Signal Pathway in Hep3B Human Hepatoma Cells (Hep3B 간암세포에서 개똥쑥추출물로부터 Akt-mTOR-GSK3β 신호경로에 의한 apoptosis 효과)

  • Kim, Eun Ji;Kim, Guen Tae;Kim, Bo Min;Lim, Eun Gyeong;Ha, Sung Ho;Kim, Sang-Yong;Kim, Young Min
    • Journal of Life Science
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    • v.26 no.7
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    • pp.764-771
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    • 2016
  • Extracts from Artemisia annua Linné (AAE) have been known to possess various functions, including anti-bacterial, anti-virus, and anti-oxidant effects. However, the mechanism of those effects of AAE is not well-known. The aim of this study was to analyze the inhibitory effects of AAE on cell proliferation of the human hepatoma cell line (Hep3B) and to examine its effects on apoptosis. Activation by phosphorylation of Akt is cell proliferation through the phosphorylation of TSC2, mTOR, and GSK-3β. We suggested that AAE may exert cancer cell apoptosis through Akt/mTOR/GSK-3β signal pathways and mitochondria-mediated apoptotic proteins. For this, we examined the effects of extracts of AAE on cell proliferation according to treatment concentration. Treatment with AAE not only reduced cell viability, but also resulted in the induced release of lactate dehydrogenase (LDH). These results were determined with a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and a lactate dehydrogenase (LDH) assay. Furthermore, we determined the effects of apoptosis through Hoechst 33342 staining, annexinⅤ-propidium iodide (PI) staining, 5,5′, 6,6′-tetrachloro-1,1′,3,3′-tetraethyl-imidacarbocyanine iodide (JC-1) staining, and Western blotting. Our study showed that the treatment of liver cancer cells with AAE resulted in the inhibition of Akt, TSC2, GSK-3β-phosphorylated, Bcl-2, and pro-caspase 3 and the activation of Bim, Bax, Bak, and cleaved PARP expressions. These results indicate that AAE induced apoptosis by means of a mitochondrial event through the regulate of Akt/mTOR/GSK-3β signaling pathways.

Effects of Non-ionic Surfactant Tween 80 on the in vitro Gas Production, Dry Matter Digestibility, Enzyme Activity and Microbial Growth Rate by Rumen Mixed Microorganisms (비이온성 계면활성제 Tween 80의 첨가가 반추위 혼합 미생물에 의한 in vitro 가스발생량, 건물소화율, 효소활력 및 미생물 성장율에 미치는 영향)

  • Lee, Shin-Ja;Kim, Wan-Young;Moon, Yea-Hwang;Kim, Hyeon-Shup;Kim, Kyoung-Hoon;Ha, Jong-Kyu;Lee, Sung-Sil
    • Journal of Life Science
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    • v.17 no.12
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    • pp.1660-1668
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    • 2007
  • The non-ionic surfactant (NIS) Tween 80 was evaluated for its ability to influence invitro cumulative gas production, dry matter digestibility, cellulolytic enzyme activities, anaerobic microbial growth rates, and adhesion to substrates by mixed rumen microorganisms on rice straw, alfalfa hay, cellulose filter paper and tall fescue hay. The addition of NIS Tween 80 at a level of 0.05% increased significantly (P<0.05) in vitro DM digestibility, cumulative gas production, microbial growth rate and cellulolytic enzyme activity from all of substrates used in this study. In vitro cumulative gas production from the NIS-treated substrates; rice straw, alfalfa hay, filter paper and tall fescue hay was significantly (P<0.05) improved by 274.8, 235.2, 231.1 and 719.5% compared with the control, when substrates were incubated for 48 hr in vitro. The addition of 0.05% NIS Tween 80 to cultures growing on alfalfa hay resulted in a significant increase in CMCase (38.1%), xylanase (121.4%), Avicelase (not changed) and amylase (38.2%) activities after 36 h incubation. These results indicated that the addition of 0.05% Tween 80 could greatly stimulate the release of some kinds of cellulolytic enzymes without decreasing cell growth rate in contrast to trends reported with aerobic microorganism. Our SEM observation showed that NIS Tween. 80 did not influence the microbial adhesion to substrates used in the study. Present data clearly show that improved gas production, DM digestibility and cellulolytic enzyme activity by Tween 80 is not due to increased bacterial adhesion on the substrates.

Characterization of HtrA2-deficient Mouse Embryonic Fibroblast Cells Based on Morphology and Analysis of their Sensitivity in Response to Cell Death Stimuli. (HtrA2 유전자가 결손된 mouse embryonic fibroblast 세포주의 형태학적 특징 및 세포사멸 자극에 대한 감수성 조사)

  • Lee, Sang-Kyu;Nam, Min-Kyung;Kim, Goo-Young;Rhim, Hyang-Shuk
    • Journal of Life Science
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    • v.18 no.4
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    • pp.522-529
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    • 2008
  • High-temperature requirement A2(HtrA2) has been known as a human homologue of bacterial HtrA that has a molecular chaperone function. HtrA2 is mitochondrial serine protease that plays a significant role in regulating the apoptosis; however, the physiological function of HtrA2 still remains elusive. To establish experimental system for the investigation of new insights into the function of HtrA2 in mammalian cells, we first obtained $HtrA2^{+/+}$ and $HtrA2^{-/-}$ MEF cells lines and identified those cells based on the expression pattern and subcellular localization of HtrA2, using immunoblot and biochemical assays. Additionally, we observed that the morphological characteristics of $HtrA2^{-/-}$ MEF cells are different form those of $HtrA2^{+/+}$ MEF cells, showing a rounded shape instead of a typical fibroblast-like shape. Growth rate of $HtrA2^{-/-}$ MEF cells was also 1.4-fold higher than that of $HtrA2^{+/+}$ MEF cells at 36 hours. Furthermore, we verified both MEF cell lines induced caspsase-dependent cell death in response to apoptotic stimuli such as heat shock, staurosporine, and rotenone. The relationship between HtrA2 and heat shock-induced cell death is the first demonstration of the research field of HtrA2. Our study suggests that those MEF cell lines are suitable reagents to further investigate the molecular mechanism by which HtrA2 regulates the balance between cell death and survival.

Selection and Quality Evaluation of Sprout Soybean [Glycine max (L.) Merrill] Variety for Environment-Friendly Cultivation in Southern Paddy Field (남부지역 친환경 논 재배를 위한 나물콩 품종 선발 및 품질 평가)

  • Kim, Young-Jin;Lee, Kwang-Won;Cho, Sang-Kyun;Oh, Young-Jin;Shin, Sang-Ouk;Paik, Chae-Hoon;Kim, Kyong-Ho;Kim, Tae-Soo;Kim, Ki-Jong
    • Korean Journal of Organic Agriculture
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    • v.19 no.3
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    • pp.357-372
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    • 2011
  • We carried out the experiment to select the suitable sprout soybean varieties for environment-friendly cultivation in paddy field of southern part area, compares of excess moisture injury degree and yield ability among 29 sprout soybean varieties. Plant growth of sprout soybean was generally low in beginning and recovered after flowering due to rainfall. In paddy field cultivation, number of pod per individual and number of seed per individual were less in difference than upland cultivation, and maturing date was delayed 5-14 days than upland cultivation in most species. When environment-friendly cultivation, pest injury was not caused major problem for the growth during the vegetative period of soybean due to ground spider as natural enemy to insect pest. However, damage of stink bugs showed severe during grain filling period, and Dawonkong, Anpyeongkong, Dachaekong and Wonhwangkong showed susceptible to sting bug. SMV infection was weak and showed some necrosis symptoms in Sokangkong, but black root rot was not infected at all. Bacterial pustule began to be infected slowly from pod enlargement stage in most species, displayed severe symptoms in Dawonkong, Pungsannamulkong, Seonamkong and Sobaeknamulkong. The symptoms of pod anthracnose, pod blight and purple spot were greatly appeared after flowering. Disease resistance varieties was Paldokong, Kwangankong, Doremikong, Somyeongkong, Pungsannamulkong, Iksa-namulkong, Seonamkong, Sojinkong, Pureunkong, Bosugkong, Namhaekong and Sorokkong. Lodging index showed 3 in Saebyeolkong, and other species displayed slight lodging in 0-3 degree. 100-seed weight is 9.8-17.2g extent and increased 0.1-3.7g than upland cultivation in most species, but decreased in some species. Government purchase standard, species correspond to small-seed-size namulkong (Sizing screen diameter 4.0-5.6 mm) was Dawonkong, Dachaekong, Bosugkong, Seonamkong, Sokangkong, Hannamkong, Somyeongkong and Wonhwangkong. Species which seed yield was higher than Pungsannamulkong (266kg/10a) were Sorokkong, Hannamkong, Bosugkong and Sowonkong. Considering sprout soybean species, disease endurance, insect resistance, lodging resistance, 100-seed weight, yield ability and excess moisture tolerances synthetically, Seonamkong, Hannamkong, Doremikong, Bosugkong, Pungwonkong, Kwangankong, Sowonkong, Dagikong, Paldokong, Eunhakong and Pungsannamulkong were promising for environment-friendly cultivation in paddy field.

Comparison of Growth, Yield and Quality between Organic Cultivation and Conventional Cultivation in Rice (Oryza sativa L.) Field (벼 유기재배와 관행재배의 생육 및 수량과 품질 비교)

  • Cha, Kwang-Hong;Oh, Hwan-Jung;Park, Ro-Dong;Park, Heung-Gyu;An, Kyu-Nam;Jung, Woo-Jin
    • Korean Journal of Organic Agriculture
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    • v.18 no.2
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    • pp.199-208
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    • 2010
  • To investigate a comparison of rice yield and quality between organic and conventional cultivation, study for occurrence of rice diseases carried out in rice (Oryza sativa L.) field of Noahn and Bannam region. The results obtained as following: 1) Bactericide and insecticide were applied twice and four times at Noahn and Bannam region in conventional cultivation (C.C) of rice field, respectively, it was applied twice at two region in organic cultivation (O.C). Rice strip virus, leaf blast, neck blast, sheath blight, bacterial leaf blight, rice water weevil, rice leaf folder, and plant hoppers were occurred mainly in rice field. Leaf blast, neck blast, and rice leaf folder were occurred highly at O.C compared with C.C. 2) Growth level of rice in clum and panicle length was lower at O.C than C.C. Number of panicles per hill was by 0.6 higher at O.C than C.C. Number of panicles per hill was by 0.6 higher at O.C than C.C in Noahn region. Number of spikelets per panicle, grain filling ratio, brown/rough rice ratio, and weight of 1,000 grains was lower at O.C than C.C. Number of panicles per hill was by 0.8 lower at O.C than C.C in Bannam region. Number of spikelets per panicle and grain filling ratio was higher at O.C than C.C while brown/rough rice ratio was lower at O.C. Total yield percentage of rice in O.C was level of 84% at Noahn region and 94% at Bannam region compared with C.C. 3) Head rice percentage was lower at in O.C than C.C at two region. Protein and palatability (Toyo value) were lower at O.C than C.C in Bannam region causing occurrence of neck blast and rice leaf folder by oversupply of nitrogen fertilizer. Protein, whiteness, and palatability (Toyo value) were adequate level at O.C in Noahn region by lower nitrogen fertilizer.

Feasibility Study of Wetland-pond Systems for Water Quality Improvement and Agricultural Reuse (습지-연못 연계시스템에 의한 수질개선과 농업적 재이용 타당성 분석)

  • Jang, Jae-Ho;Jung, Kwang-Wook;Ham, Jong-Hwa;Yoon, Chun-Gyeong
    • Korean Journal of Ecology and Environment
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    • v.37 no.3 s.108
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    • pp.344-354
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    • 2004
  • A pilot study was performed from September 2000 to April 2004 to examine the feasibility of the wetland-pond system for the agricultural reuse of reclaimed water. The wetland system was a subsurface flow type, with a hydraulic residence time of 3.5 days, and the subsequent pond was 8 $m^3$ in volume (2 m ${\times}$ 2 m ${\times}$ 2 m) and operated with intermittent-discharge and continuous flow types. The wetland system was effective in treating the sewage; median removal efficiencies of $BOD_5$ and TSS were above 70.0%, with mean effluent concentrations of 27.1 and 16.8 mg $L^{-1}$, respectively, for these constituents. However, they did often exceed the effluent water quality standards of 20 mg $L^{-1}$. Removal of T-N and T-P was relatively less effective and mean effluent concentrations were approximately 103.2 and 7.2 mg $L^{-1}$, respectively. The wetland system demonstrated high removal rate (92 ${\sim}$ 90%) of microorganisms, but effluent concentrations were in the range of 300 ${\sim}$ 16,000 MPN 100 $mL^{-1}$ which is still high for agricultural reuse. The subsequent pond system provided further treatment of the wetland effluent, and especially additional microorganisms removal in addition to wetland-pond system could reduce the mean concentration to 1,000 MPN 100 $mL^{-1}$ from about $10^5$ MPN 100 $mL^{-1}$ of wetland influent. Other parameters in the pond system showed seasonal variation, and the upper layer of the pond water column became remarkably clear immediately after ice melt. Overall, the wetland system was found to be adequate for treating sewage with stable removal efficiency, and the subsequent pond was effective for further polishing. This study concerned agricultural reuse of reclaimed water using natural systems. Considering stable performance and effective removal of bacterial indicators as well as other water quality parameters, low maintenance, and cost-effectiveness, wetland- pond system was thought to be an effective and feasible alternative for agricultural reuse of reclaimed water in rural area.

Effect of Lactic acid bacteria and Enzyme Supplementation on Fermentative Patterns of Ensiling Silages, Their In vitro Ruminal Fermentation, and Digestibility (젖산균과 효소제 처리에 의한 동계사료작물 발효성상, In vitro 반추위 발효 및 소화율에 미치는 영향 연구)

  • Lee, A-Leum;Shin, Su-Jin;Yang, Jinho;Cho, Sangbuem;Choi, Nag-Jin
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.36 no.1
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    • pp.7-14
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    • 2016
  • The objective of this study was to determine the effect of bacterial inoculation (Lactobacillus plantarum or combo inoculant mixed with Lactobacillus plantarum and Lactobacillus buchneri) and addition of fibrolytic enzyme on chemical compositions and fermentation characteristics of whole crop barley (WCB) and triticale (TRT) silage, their ruminal in vitro fermentation, and digestibility. In TRT silage, enzyme addition significantly (p<0.01) decreased NDF content compared to no enzyme addition treatment. Organic acids such as lactate and acetate contents in WCB and TRT silages were significantly (p<0.01) higher compared to those in the control. Particularly, lactate content was the highest in L. plantarum treatment. Fibrolytic enzyme treatment on both silages had relatively higher lactic acid bacteria content, while mold content was lower in both treatments compared to that in the control. In vitro dry matter digestibility was generally improved in WCB silages. It was higher (p<0.01) in TRT with mixed treatment of L. plantarum, L. buchneri, and enzyme compared to others. In vitro ruminal acetate production was relatively higher in treatments with both enzyme and inoculant additions compared to that in the control. Therefore, the quality of silage and rumen fermentation could be improved by inoculants (L. plantarum and L. buchneri) regardless whether whole crop barley (WCB) or triticale (TRT) silage was used. Although it was found that fibrolytic enzyme addition to both silages had various quality and rumen fermentation values, further study is needed.

Microbiological Evaluation of Raw Vegetables (비가열 섭취 채소류의 미생물 오염도 조사)

  • Jung, Seung-Hye;Hur, Myung-Je;Ju, Jeong-Hwa;Kim, Kyung-Ae;Oh, Sung-Suck;Go, Jong-Myoung;Kim, Yong-Hee;Im, Jeong-Soo
    • Journal of Food Hygiene and Safety
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    • v.21 no.4
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    • pp.250-257
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    • 2006
  • The purpose of this study is to evaluate microbiological contamination of leafy vegetables. Total aerobic bacteria and coliforms were monitored to get the contamination levels and Staphylococcus aureus, Bacillus cereus, Clostridium perfringens, Escherichia coli, Escherichia coli O157:H7, Salmonella spp., Vibrio parahaemolyticus, Listeria monocytogenes, Yersinia enterocolitica, Campylobacter jejuni to detect pathogens with risk of foodpoisoning from fresh vegetables. The colony count of total aerobes and coliforms was also performed to determine the efficacy of washing with tab water by common consumers. 124 samples which are divided into 8 kinds of vegetables - Sesame leaf, Dropwort, Chinese cabbage, Korean leek, Lettuce, Crown daisy, Pimpinella brachycarpa, Chicory were sampled in 2 wholesale markets in Incheon. Mean counts of total aerobic bacteria for individual vegetables ranged from $2.2\times10^6\;CFU/g\;to\;6.0\times10^7\;CFU/g$ and total coliforms were from $4.1\times10^5\;CFU/g\;to\;9.8\times10^6\;CFU/g$. Both show the peaks in summer on this study from March to September. Decrease rates after washing with tab water averaged 81.0% and 82.5% in total aerobic bacteria and coliform counts respectively. Staphylococcus aureus was isolated 8.1%, Bacillus cereus 14.5%, Clostridium perfringens 5.6%, Escherichia coli 18.5%. 11 samples showed overlapped bacterial contamination. For respective vegetables Staphylococcus aureus isolated from 0.0% to 22.2%, Bacillus cereus from 0.0% to 29.4%, Clostridium perfringens from 0.0% to 23.1 %, Escherichia. coli from 0.0% to 35.0%. Escherichia coli O157:H7, Salmonella spp., Vibrio parahaemolyticus, Listeria monocytogenes, Yersinia enterocolitica, Campylobacter jejuni were not isolated. This study is expected to be available as the reference for the basal data of pathogens in fresh vegetables.

Evaluation of Efficacy and Development of Predictive Reduction Models for Escherichia coli and Staphylococcus aureus on Food Contact Surfaces as a Function of Concentration and Contact Time of Chlorine Dioxide (대장균과 황색포도상구균에 대한 이산화염소의 살균소독력 평가 및 살균예측모델 개발)

  • Yoon, So-Jeong;Park, Shin Young;Kim, Yong-Soo;Ha, Sang-Do
    • Journal of Food Hygiene and Safety
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    • v.32 no.6
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    • pp.507-512
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    • 2017
  • There has been increasing concern regarding misuse of disinfectants and sanitizers such as ethanol, sodium hypochlorite, and hydrogen peroxide for food contact surfaces in the food industry. Examining the efficacy of the concentration of currently used disinfectants and sanitizers is urgently required in the Korean society. This study aimed to develop predictive reduction models for Escherichia coli and Staphylococcus aureus in suspension, as a function of $ClO_2$ (chlorine dioxide) and contact time using response surface methodology. E. coli ATCC 10536 and S. aureus ATCC 6538 (initial inoculum, 8-9 log CFU/mL) in tryptic soy broth were treated with different concentrations of $ClO_2$ (5, 20, and 35 ppm) for different contact times (1, 3, and 5 min) following a central composite design. The polynomial reduction models for $ClO_2$ on E. coli and S. aureus were developed under the clean condition. E. coli reduction by 35 ppm $ClO_2$ for 1, 3, and 5 min was 2.49, 2.70, and 3.65 log CFU/mL, respectively. Also, S. aureus reduction by 35 ppm $ClO_2$ for 1, 3, and 5 min was 4.59, 5.25, and 5.81 log CFU/mL, respectively. The predictive response polynomial models developed were $R=0.43231-0.056492^*X_1-0.097771^*X_2+9.24167E-003^*X_1^*X_2+3.06333E-003^*X_1{^2}$ ($R^2=0.98$) on E. coli and $R=1.10542-0.20896^*X_1-0.046062^*X_2+8.30000E-003^*X_1^*X_2+8.73300E-003^*X_1{^2}$ ($R^2=0.99$) on S. aureus, where R was the bacterial reduction (log CFU/mL), $X_1$ was the concentration and $X_2$ was the contact time. Our predictive reduction models should be validated in developing the optimal concentration and contact time of $ClO_2$ for inhibiting E. coli and S. aureus on food contact surfaces.

Measurement of Lactoferrin, IgA, IgG1, IgG2, Antibacterial Activity, and Lactic Acid Bacterial Growth in Holstein Colostrum (Holstein 초유 중 Lactoferrin, IgA, IgG1, IgG2 정량과 미생물의 성장에 미치는 영향)

  • Renchinthand, Gereltuya;Bae, Hyoung-Churl;Nam, Myoung-Soo
    • Food Science of Animal Resources
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    • v.27 no.4
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    • pp.522-530
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    • 2007
  • This experiment was carried out to measure the content of lactoferrin, IgA, $IgG_1,\;IgG_2$, in Holstein colostrum, and to test the effect of it's colostrum on the antibacterial activity to pathogenic bacteria and the growth stimulation of lactic acid bacteria. Colostrum was collected at the first, second, and third day after parturition in summer and winter season. The levels of lactoferrin, IgA, $IgG_1,\;and\;IgG_2$ in Holstein cow colostrum were 0.30 mg/mL, 0.37 mg/mL, 4.00 mg/mL, 0.37 mg/mL, respectively, on the first day of the summer season whereas they were 1.16 mg/mL, 2.60 mg/mL, 13.35 mg/mL, 1.30 mg/mL on the first day of the winter season, postpartum. Heat treated ($65^{\circ}C$ for 30 min) or non-treated colostrum showed antibacterial activity toward Escherichia coli. The growth of commercial mixed strains (Bifidobacterium longum, Lactobacillus acidophilus, Streptococcus themophilus), L. acidophilus, L. casei, L. bulgaricus, and L. lactis subsp. cremoris were improved in first, second and third day colostrum compared to normal milk. Commercial miked strains (B. longum, L. acidophilus S. themophilus) lowered the pH to 4.97-5.22 and 4.89 while increasing the titratable acidity to 0.75-0.88% and 0.70% in colostrum and normal milk, respectively. However, L. casei, L. bulgaricus, L. lactis subsp. cremoris lowered the pH to 5.96-6.47 and 6.5-6.8 while increasing the titratable acidity to 0.29-0.48% and 0.20-0.25% in colostrum and normal milk, respectively.