• Title/Summary/Keyword: yeast cells

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Effect of Chromium Dietary Supplementation on the Immune Response and Some Blood Biochemical Parameters of Transport-stressed Lambs

  • Al-Mufarrej, S.I.;Al-Haidary, I.A.;Al-Kraidees, M.S.;Hussein, M.F.;Metwally, H.M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.5
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    • pp.671-676
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    • 2008
  • Forty-eight Naemi lambs (avg. BW 31.7 kg) were transported by truck for a distance of 1,450 km from Al-Jouf to Riyadh, Saudi Arabia. On arrival day, the lambs were randomly allocated to four groups receiving diets supplemented with 0.0, 0.3, 0.6 and 0.9 ppm organic chromium (Cr). Each group consisted of four separately housed replicates of three lambs each. The animals were fed ad libitum on a grower diet for 84 days. Blood samples were obtained shortly before transportation, upon arrival and at weekly intervals thereafter from all lambs for analysis of plasma and serum. Plasma glucose and serum cortisol, total protein, albumin, urea-N and total cholesterol concentrations were determined. A cursory clinical examination of the lambs, along with rectal temperature, was undertaken at different intervals during the experiment. The lambs were inoculated each with 2 ml i.v. chicken red blood cells (CRBC) on days 0, 21, and 42. Serum total, IgG and IgM antibody titers were determined at weekly intervals post-immunization. An in vivo intradermal hypersensitivity test was carried out on 6 lambs from each group on days 10 and 70. Transportation of the lambs resulted in a significant (p<0.001) elevation of serum cortisol, total protein and albumin levels, as well as increased plasma glucose concentration, with corresponding decrease in total cholesterol, while blood urea-N remained largely unchanged. These constituents returned to normal levels during subsequent weeks, with no significant differences in their concentrations being observed between the Cr-supplemented groups and controls. Rise in rectal temperature after transportation was reduced to a greater extent (p<0.05) in Cr-supplemented versus control lambs. Total, IgG and IgM antibody titers against CRBC rose significantly (p<0.05) during immunizations in all groups, with significantly and linearly higher (p<0.05) total and IgG titers in Cr-supplemented versus control lambs. By contrast, no significant effect due to Cr supplementation was recorded in IgG titers, which increased equally in Cr-fed and control groups. Skin thickness in response to intradermal inoculation of phytohaemagglutinin (PHA) was also significantly (p<0.01) increased as a result of Cr supplementation. These results indicate that dietary Cr supplementation might be useful during stress especially for enhancing immune responses in transport-stressed lambs.

Increased Antioxidative Activities against Oxidative Stress in Saccharomyces cerevisiae KNU5377 (산화 스트레스 대한 Saccharomyces cerevisiae KNU5377의 항산화 활성의 증가)

  • Kim, Il-Sup;Yun, Hae-Sun;Yang, Ji-Young;Lee, Oh-Seok;Park, Heui-Dong;Jin, Ing-Nyol;Yoon, Ho-Sung
    • Journal of Life Science
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    • v.19 no.4
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    • pp.429-435
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    • 2009
  • Oxidative stress is a consequence of an imbalance of the defense system against cellular damage generated by reactive oxygen species (ROSs) such as superoxide anions (menadione; MD). Most organisms have evolved a variety of defense systems to protect cells from adverse conditions. In order to evaluate stress tolerance against oxidative stress generating MD, comparative analyses of antioxidant capacity, or free radical scavenger ability, were performed between S. cerevisiae KNU5377 (KNU5377) and three wild-type S. cerevisiae strains. In a medium containing 0.4 mM MD, the KNU5377 strain showed higher cell viability and antioxidant ability, and contained higher levels of trehalose, superoxide dismutase, thioredoxin system, glucose-6-phosphate dehydrogenase, and some heat shock proteins. The KNU5377 strain also produced a lower level of oxidative stress biomarker than the other three yeast strains. These results indicate that S. cerevisiae KNU5377 has a higher level of tolerance to oxidative stress due to the increased expression of cell rescue proteins and molecules, thus alleviating cellular damage more efficiently than other S. cerevisiae strains.

Magnaporthe oryzae Effector AVR-Pii Helps to Establish Compatibility by Inhibition of the Rice NADP-Malic Enzyme Resulting in Disruption of Oxidative Burst and Host Innate Immunity

  • Singh, Raksha;Dangol, Sarmina;Chen, Yafei;Choi, Jihyun;Cho, Yoon-Seong;Lee, Jea-Eun;Choi, Mi-Ok;Jwa, Nam-Soo
    • Molecules and Cells
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    • v.39 no.5
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    • pp.426-438
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    • 2016
  • Plant disease resistance occurs as a hypersensitive response (HR) at the site of attempted pathogen invasion. This specific event is initiated in response to recognition of pathogen-associated molecular pattern (PAMP) and subsequent PAMP-triggered immunity (PTI) and effector-triggered immunity (ETI). Both PTI and ETI mechanisms are tightly connected with reactive oxygen species (ROS) production and disease resistance that involves distinct biphasic ROS production as one of its pivotal plant immune responses. This unique oxidative burst is strongly dependent on the resistant cultivars because a monophasic ROS burst is a hallmark of the susceptible cultivars. However, the cause of the differential ROS burst remains unknown. In the study here, we revealed the plausible underlying mechanism of the differential ROS burst through functional understanding of the Magnaporthe oryzae (M. oryzae) AVR effector, AVR-Pii. We performed yeast two-hybrid (Y2H) screening using AVR-Pii as bait and isolated rice NADP-malic enzyme2 (Os-NADP-ME2) as the rice target protein. To our surprise, deletion of the rice Os-NADP-ME2 gene in a resistant rice cultivar disrupted innate immunity against the rice blast fungus. Malic enzyme activity and inhibition studies demonstrated that AVR-Pii proteins specifically inhibit in vitro NADP-ME activity. Overall, we demonstrate that rice blast fungus, M. oryzae attenuates the host ROS burst via AVR-Pii-mediated inhibition of Os-NADP-ME2, which is indispensable in ROS metabolism for the innate immunity of rice. This characterization of the regulation of the host oxidative burst will help to elucidate how the products of AVR genes function associated with virulence of the pathogen.

Effect of Combined Use of various Anti-microbial Materials on Brewing of Low Salted Kochujang (저식염 고추장 양조시 항균물질 혼합첨가의 영향)

  • Park, Seon-Ju;Kim, Dong-Han
    • Applied Biological Chemistry
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    • v.50 no.4
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    • pp.287-294
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    • 2007
  • Effect of combined use of anti-microbial materials, such as ethanol, mustard and chitosan, on the quality of low salted kochujang was investigated during fermentation at $20^{\circ}C$ for 12 weeks. Viable cells of yeast increased remarkably during fermentation, but increasing ratio was significantly low in ethanol-mustard added kochujang. Activity of ${\beta}-amylase$ was high in anti-microbial material added kochujang, whereas ${\alpha}-amylase$ and protease activities were low in those groups. Water activity decreased during fermentation with being low in the control kochujang prepared with normal-salt without anti-microbial materials. Hunter L-, a- and b-values of kochujang increased during fermentation, and the degree of increase in total color difference $({\Delta}E)$ was low in ethanol added kochujang. Titratable acidity of kochujang was decreased in anti-microbial materials added group at late aging period, and oxidation-reduction potential was low in the control kochujang. Total sugar and reducing sugar contents of kochujang were high in ethanol-mustard added kochujang. Ethanol contents of kochujang increased at late aging period, with high values in ethanol-chitosan added kochujang. Amino nitrogen content increased during middle of fermentation, and ammonia nitrogen content of kochujang decreased in ethanol-mustard-chitosan added group during fermentation. After 12 weeks fermentation, sensory results showed that ethanol or ethanol-mustard added kochujang were the highest in color and flavor with the highest overall acceptability.

Inactivation of Candida albicans Biofilm by Radachlorin-Mediated Photodynamic Therapy (라다클로린으로 매개된 광역학치료에 의한 백색 캔디다 바이오필름의 비활성)

  • Kwon, Pil Seung
    • Korean Journal of Clinical Laboratory Science
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    • v.47 no.4
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    • pp.273-278
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    • 2015
  • The purpose of this study was to evaluate the in-vitro efficacy of PDT using red light emitting diode (LED) with Radachlorin for biofilm inhibition of clinical Candida albicans isolates. The suspensions containing C. albicans at $9{\times}10^8CFU/mL$ were prepared on yeast nitrogen base containing 5% glucose. The biofilm formation was grown for 3 h after seeding suspensions each 100 ul on a 96-well plate and then supernatant was discarded. Each well was treated with $0.39{\mu}g/mL$ from $50{\mu}g/mL$ concentrations of Radachlorin on adherent biofilm. After a 30-minute incubation, light was irradiated for 30, 60, or 90 minutes using the following light source of wavelength 630 nm LED, at energy densities of 14, 29, and $43J/cm^2$. Afterwards, all supernatant was removed and dried. Adherent cells were stained with safranin O and dried. The cell viability was measured using a microplate reader at 490 nm. Also, a fluorescent signal on C. albicans was observed by saturation of a photosensitizer. In conclusion, a significant inhibition of 72.5% was observed to C. albicans on biofilm at the Radachlorin dose of $50{\mu}g/mL$ with 630 nm LED. The Photosensitizer (Radachlorin) was adequate at 30 minuttes for C. albicans. Overall, the results showed that inhibition of biofilm formation was Radachlorine dose-dependent. The results suggest that PDT, using Radachlorin with 630 nm LED, is able to decrease biofilm formation of C. albicans.

Fermentation Characteristics of Low Salted Kochujang Prepared with Mixture of Sub-materials (부원료를 혼합 첨가한 저식염 고추장의 발효 특성)

  • Kim, Dong-Han
    • Korean Journal of Food Science and Technology
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    • v.37 no.3
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    • pp.449-455
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    • 2005
  • To reduce saft content of kochujang, various combinations of sub-materials such as ethanol mustard and chitosan were added to kochujang, and their effects on microbial characteristics, enzyme activities, and physicochemical characteristics of kochujang were investigated after 12 weeks of fermentation. Activities of ${\beta}$-amylase and pretense were low in ethanol-mustard-chitosan-added kochujang, whereas no significant difference was observed in ${\alpha$-amylase activity among all groups. Number of viable yeast cells decreased remarkably in mustard-added kochujang during late aging period, and anaerobic bacterial counts decreased in sub-material-added groups. Consistency of kochujang increased by addition of sub-materials, and oxidation-reduction potential was low in chitosan-added group. Mustard-chitosan-added kochujang showed lowest increase in total color difference(${\Dalta}E$) and decrease in water activity. PH of kochujang wns highest in mustard-chitosan-added kochujang, resulting in significantly increased titratable acidity. Addition of sub-material increased reducing sugar contents of kochujang, whereas ethanol production was significantly repressed in mustard-chitosan-added kochujang. Amino nitrogen content was Highest in mustard-chitosan-added kochujang during late aging period, whereas ammonia nitrogen content was lower in ethanol-mustard-added kochujang. Results of sensory evaluation indicated ethanol-mustard-added kochujang was more acceptable than other groups in taste and overall acceptability.

Fermentation of Citrus unshiu Marc. and Functional Characteristics of the Fermented Products (감귤의 발효와 발효산물의 기능적 특성)

  • Moon, Sang-Wook;Kang, Shin-Hae;Jin, Young-Joon;Park, Ji-Gweon;Lee, Young-Don;Lee, Young-Ki;Park, Deok-Bae;Kim, Se-Jae
    • Korean Journal of Food Science and Technology
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    • v.36 no.4
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    • pp.669-676
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    • 2004
  • Functional characteristics of citrus products fermented with lactic acid bacterium and yeast were investigated. Flavonoid composition of fermented citrus extracts increased significantly compared to control, leading to increases of naringenin and hesperetin concentrations. All citrus extracts showed anti-apoptotic effects in HepG2 cells regardless of fermentation, with citrus-fermented products showing greater anti-apoptotic effect and intracellular Reactive Oxygen Species content reduction compared to native citrus extracts. Male Sprague-Dawley rats were orally dosed with native or fermented citrus extracts. Singnificantly higher body weight reductions were observed in higher fermented citrus-dosed (100 mg/kg body weight) group compared to the other groups. Plasma total cholesterol level was slightly, but not significantly, reduced. Fatty liver formation induced by high-fat diet was significantly suppressed in rats administered with fermented citrus extracts. Results suggest fermented citrus extracts have potent anti-apoptotic and anti-oxidative activities in vitro, and inhibitory activity against fatty liver formation by high-fat diet in vivo.

A Specific Role of Ime2, Meiosis-specific Protein Kinase, in the Eary Meiotic Pathway in Saccharomyces cerevisiae (Saccharomyces cerevisiae의 감수분열 특이적 Protein Kinase인 Ime2의 역할)

  • Leem, Sun-Hee;Tak, Yon-Soo;Sunwoo, Yang-Il
    • Korean Journal of Microbiology
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    • v.35 no.4
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    • pp.258-265
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    • 1999
  • Entry into meiosis in the yeast Saccharomyces cerevisiae is regulated by two major factors: the cell type MATa/MAT${\alpha}$ and the nutriational state (starvation) of the cell. The two independent regulations act through IME1and IME2 expression to initiate meiosis. IME2 encodes a meiosis-specific protein kinase, and it enabled MATa/MAT${\alpha}$ diploid cells to undergo meiosis and sporulation. The PCR mutagenesis method was applied for the isolation of thermosensitive ime2 mutants. Among sixty two mutants isolated from the phenotype of defective spore formation under the restrictive temperature, three with the most easily observed temperature-sensitive phenotype (ts ${\cdot}$ime2-11, ts ${\cdot}$ime2-12 and ts ${\cdot}$ime2-13) were selected for further study. To understand the detailed functions of IME2, we examined the defects of these mutants in the early meiotic pathway including the premeiotic DNA replication and exhibited decreased level in meiotic recombination. These results suggest that the IME2 gene plays essential role in meiotic recombination pathway as well as premeiotic DNA replication. As the result of the IME2 overexpression in ${\Delta}$mre4. moreover, it was suggested that the IME2 and MRE4 genes act on the same pathway of initiation step in meiotic recombination.

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Identification of Meiotic Recombination Intermediates in Saccharomyces cerevisiae (효모 감수분열과정에서의 유전자 재조합 기전 특이적 DNA 중간체의 구조 변화)

  • Sung, Young Jin;Yoon, Sang Wook;Kim, Keun Pil
    • Korean Journal of Microbiology
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    • v.49 no.1
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    • pp.1-7
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    • 2013
  • During meiosis, genetic recombinants are formed by homologous recombination accompanying with the programmed double-strand breaks (DSBs) and strand exchanges between homologous chromosomes. The mechanism is generated by recombination intermediates such as single-end invasions (SEIs) and double-Holliday junctions (dHJs), and followed by crossover (CO) or non-crossover (NCO) products. Our study was focused on the analysis of meiotic recombination intermediates (DSBs, SEIs, and dHJs) and final recombination products (CO and NCO). We identified these meiotic recombination intermediates using DNA physical analysis under HIS4LEU2 "hot spot" system in budding yeast, Saccharomyces cerevisiae. For DNA physical analysis, when the hot spot locus is recognized by restriction enzyme from synchronous meiotic cells, the fragmented DNA that are forming recombination intermediates can be detected and quantified through Southern hybridization analysis. Our study suggests that this system can analyze the structural change of recombination intermediates during DSB-SEI transition, double-Holiday junctions and crossover/non-crossover products in meiosis.

Transcriptional Regulation of a DNA Repair Gene in Saccharomyces cerevisiae

  • Jang, Yeon-Kyu;Sancar, Gwen-B.;Park, Sang-Dai
    • Proceedings of the Zoological Society Korea Conference
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    • 1998.10b
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    • pp.113-113
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    • 1998
  • In Saccharomyces cerevisiae UV irradiation and a variety of chemical DNA -damaging agents induce the transcription of specific genes, including several involved in DNA repair. One of the best characterized of DNA -damage inducible genes is PHRI, which encodes the apoenzyme for DNA photolyase. Basal-level and damage-induced expression of PHRI require an upstream activation sequence, UASPHRI. Here we report the identification of the UlvIE6 gene of S. cerevisiae as a regulator of UASPHRl activity. Surprisingly, the effect of deletion of UME6 is growth phase dependent. In wild-type cells PHRI is induced in late exponential phase, concomitant with the initiation of glycogen accumulation that precedes the diauxic shift. Deletion of UNIE6 abolishes this induction, decreases the steady-state concentration of photolyase molecules and PHRI mRNA, and increases the UV sensitivity of a rad2 mutant. The results suggest that UM E6 contributes to the regulated expression of a subset of damage-responsive genes in yeast. Furthermore, the upstream repression sequence, URSPHRI, is required for repression and damage-induced expression of PHRl. Here we show identification of YER169W and YDR096W as putative regulators acting through $URS_{PHRI}$. These open reading frames were designated as RPHI (YERl69W) and RPH2 (YDR096W) indicating regulator of PHRI. Simultaneous disruption of both genes showed a synergistic effect, producing a four-fold increase in basal level expression and a similar decrease m the induction ratio following treatment of methyl methanesulfonate(MMS). Mutation of the sequence ($AG_4$) bound by Rphlp rendered the promoter of PHRI insensitive to changes in RPHI or RPH2 status. The data suggest that RPHI and RPH2 act as damage-responsive negative regulators of PHRI. Surprisingly, the sequence bound by Rphlp in vitro is found to be $AG_4$ which is identical to the consensus binding site for the regulators Msn2p and Msn4p involved in stress-induced expression. Deletion of MSN2 and MSN4 has little effect on the induction$.$ ratio following DNA damage. However, all deletions led to a significant decrease in basal-level and induced expression of PHRI. These results imply that MSN2 and MSN4 are positive regulators of P HRI but are not required for DNA damage repression. [Supported by grant from NIH]om NIH]

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