• 제목/요약/키워드: pH dependent

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Glutathione Reductase from Oryza sativa Increases Acquired Tolerance to Abiotic Stresses in a Genetically Modified Saccharomyces cerevisiae Strain

  • Kim, Il-Sup;Kim, Young-Saeng;Yoon, Ho-Sung
    • Journal of Microbiology and Biotechnology
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    • 제22권11호
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    • pp.1557-1567
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    • 2012
  • Glutathione reductase (GR, E.C. 1.6.4.2) is an important enzyme that reduces glutathione disulfide (GSSG) to a sulfydryl form (GSH) in the presence of an NADPH-dependent system. This is a critical antioxidant mechanism. Owing to the significance of GR, this enzyme has been examined in a number of animals, plants, and microbes. We performed a study to evaluate the molecular properties of GR (OsGR) from rice (Oryza sativa). To determine whether heterologous expression of OsGR can reduce the deleterious effects of unfavorable abiotic conditions, we constructed a transgenic Saccharomyces cerevisiae strain expressing the GR gene cloned into the yeast expression vector p426GPD. OsGR expression was confirmed by a semiquantitative reverse transcriptase polymerase chain reaction (semiquantitative RT-PCR) assay, Western-blotting, and a test for enzyme activity. OsGR expression increased the ability of the yeast cells to adapt and recover from $H_2O_2$-induced oxidative stress and various stimuli including heat shock and exposure to menadione, heavy metals (iron, zinc, copper, and cadmium), sodium dodecyl sulfate (SDS), ethanol, and sulfuric acid. However, augmented OsGR expression did not affect the yeast fermentation capacity owing to reduction of OsGR by multiple factors produced during the fermentation process. These results suggest that ectopic OsGR expression conferred acquired tolerance by improving cellular homeostasis and resistance against different stresses in the genetically modified yeast strain, but did not affect fermentation ability.

Effect of $Ca^{2+}-channel$ Blockers on Norepinephrine Release in the Rat Hippocampal Slice and Synaptosome

  • Kim, Suk-Won;Jung, Kyu-Yong;Choi, Bong-Kyu
    • The Korean Journal of Physiology and Pharmacology
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    • 제6권2호
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    • pp.87-91
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    • 2002
  • The aim of this study was to investigate the role of $Ca^{2+}-channel$ blockers in norepinephrine (NE) release from rat hippocampus. Slices and synaptosomes were incubated with $[^3H]-NE$ and the releases of the labelled products were evoked by 25 mM KCl stimulation. Nifedipine, diltiazem, nicardipine, flunarizine and pimozide did not affect the evoked and basal release of NE in the slice. But, diltiazem, nicardipine and flunarizine decreased the evoked NE release with a dose-related manner without any change of the basal release from synaptosomes. Also, a large dose of pimozide produced modest decrement of NE release. ${\omega}-conotoxin$ (CTx) GVIA decreased the evoked NE release in a dose-dependent manner without changing the basal release. And ${\omega}-CTxMVIIC$ decreased the evoked NE release in the synaoptosomes without any effect in the slice, but the effect of decrement was far less than that of ${\omega}-CTxGVIA.$ In interaction experiments with ${\omega}-CTxGVIA,\;{\omega}-CTxMVIIC$ slightly potentiated the effect of ${\omega}-CTxGVIA$ on NE release in the slice and synaptosomal preparations. These results suggest that the NE release in the rat hippocampus is mediated mainly by N-type $Ca^{2+}-channels,$ and that other types such as L-, T- and/or P/Q-type $Ca^{2+}-channels$ could also be participate in this process.

Analysis of Cross Sectional Ease Values for Fit Analysis from 3D Body Scan Data Taken in Working Positions

  • Nam, Jin-Hee;Branson, Donna H.;Ashdown, Susan P.;Cao, Huantian;Carnrite, Erica
    • International Journal of Human Ecology
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    • 제12권1호
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    • pp.87-99
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    • 2011
  • Purpose- The purpose of this study was to compare the fit of two prototype liquid cooled vests using a 3D body scanner and accompanying software. The objectives of this study were to obtain quantitative measurements of ease values, and to use these data to evaluate the fit of two cooling vests in active positions and to develop methodological protocol to resolve alignment issues between the scans using software designed for the alignment of 3D objects. Design/methodology/approach- Garment treatments and body positions were two independent variables with three levels each. Quantitative dataset were dependent variables, and were manipulated in 3x3 factorial designs with repeated measures. Scan images from eight subjects were used and ease values were obtained to compare the fit. Two different types of analyses were conducted in order to compare the fit using t-test; those were radial mean distance value analysis and radial distance distribution rate analysis. Findings- Overall prototype II achieved a closer fit than prototype I with both analyses. These were consistent results with findings from a previous study that used a different approach for evaluation. Research limitations/implications- The main findings can be used as practical feedback for prototype modification/selection in the design process, making use of 3D body scanner as an evaluation tool. Originality/value- Methodological protocols that were devised to eliminate potential sources of errors can contribute to application of data from 3D body scanners.

Myocardial Function and Metabolic Energetics in Low Flow Ischemia and with $\beta$-Adrenergic Stimulation in Spontaneously Hypertensive Rat Hearts

  • Kang, Young-Hee;Kang, Jung-Sook;Park, Han-Yoon
    • Preventive Nutrition and Food Science
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    • 제6권1호
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    • pp.43-50
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    • 2001
  • The effects of cardiac ischemia-reperfusion and $\beta$-adrenergic stimulation on metabolic function and energetics were investigated in Lan gendorff-perfused spontaneously hypertensive rat (SHR) hearts. Sarcoplasmic reticulum {TEX}$Ca^{2+}${/TEX}-dependent ATPase and cardiac lactate dehydrogenase (LDH) are additionally studied. The perfusion medium (1.0 mM {TEX}$Ca^{2+}${/TEX}) contained 5 mM glucose(+5 U/L insulin) and 2 mM pyruvate as substrates. Global ischemia was induced by reducing perfusion pressure of 100 to 40 cm {TEX}$H_{2}${/TEX}O, followed by 20 min reperfusin. Isoproterenol (ISO, 1$\mu$M) was infused for 10 min. Coronary vascular resistance and myocardial oxygen consumption ({TEX}$MVO_{2}${/TEX}) of SHR were increased in parallel with enhanced venous lactate during ischemia and reperfusion compared to those of Sprague Dawley (SD) hearts. Although ischemia-induced increase in venous lactate and combined adenosine plus inosine was abolished, coronary vasodilation produced in SD during reperfusion. In SHR, depressed reactive hyperemia was associated with a fall in cardiac ATP and CrP/Pi ratio and a rise in intracellular lactate/Pyruvate ratio. On the other hand, ISO produced coronary functional hyperemia and an increase in {TEX}$MVO_{2}${/TEX}. However, these responses were less than those in SHR hearts. The ATPase activity of SHR was attenuated in free {TEX}$Ca^{2+}${/TEX} concentrations used under basal condition and with ISO compared to that of SD. Venous lactate output and cardiac LDH activity were augmented in SHR as influenced by ISO. These results demonstrate that coronary reactive and functional hyperemia was dpressed in SHR, which cold be explained by alterations in the cytosolic phosphorylation potential and the cytosolic redox state manipulated by LDH, and by abnormal free calcium handling.

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리튬이온이차전지용 고밀도 양극활물질의 합성 및 평가 (Synthesized and Characterization of high density cathode materials for Lithium Secondary Batteries)

  • 권용진;최병현;지미정;선양국
    • 한국전기전자재료학회:학술대회논문집
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    • 한국전기전자재료학회 2008년도 추계학술대회 논문집 Vol.21
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    • pp.429-429
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    • 2008
  • Li$[Ni_{1/2}Co_{1/2}]O_2$ powder were synthesized from co-precipitation spherical metal oxide, $[Ni_{1/2}Co_{1/2}](OH)_2$. The preparation of metal hydroxide was significantly dependent on synthetic conditions, such as pH, amount of chelating agent, stirring speed, etc. The optimized condition resulted in $[Ni_{1/2}Co_{1/2}](OH)_2$, of which the particle size distribution was uniform and the particle shape was spherical, as observed by scanning electron microscopy. Calcination of the uniform metal hydroxide with LiOH at higher temperature led to a well-ordered layer-structured Li$[Ni_{1/2}Co_{1/2}]O_2$, as confirmed by X-ray diffraction pattern. Also these materials have ${\alpha}-NaFeO_2$ ($R\bar{3}m$) structure. Due to the homogeneity of the metal hydroxide, $[Ni_{1/2}Co_{1/2}](OH)_2$, the final product, Li$[Ni_{1/2}Co_{1/2}]O_2$, was also significantly uniform, i.e., the average particle size was of about 10 to 15 ${\mu}m$ in diameter and the distribution was relatively narrow. As a result, the corresponding tap-density was also high approximately 2.41 $gcm^{-3}$, of which the value is comparable to that of commercialized $LiCoO_2$.

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Analgesic and antipyretic actions of Muntingia calabura leaves chloroform extract in animal models

  • Zakaria, ZA;Kumar, G Hanan;Zaid, Siti NH Mohd;Ghani, Marwiza A;Hassan, MH;Hazalin, Nurul AMN;Khamis, Mahirah M;Devi, G Rathna;Sulaiman, MR
    • Advances in Traditional Medicine
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    • 제7권1호
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    • pp.34-40
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    • 2007
  • The present study was carried out to elucidate the potential of Muntingia (M.) calabura leaves chloroform extract (MCCE) as antinociceptive, anti-inflammatory and antipyretic agents using various animal models. The dried powdered leaves of M. calabura (20 g) were soaked in chloroform for 72 h and the supernatant obtained was then evaporated to dryness. The crude dried extract (0.912 g), dissolved in dimethyl sulfoxide (1:20; w/v) and considered as a stock solution (100% concentration/strength), was then diluted to the concentrations of 10 and 50% and used together in all experimental models. The MCCE was found to show significant (P < 0.05) antinociceptive and antipyretic activities, but less remarkable anti-inflammatory activity. Only the antinociceptive activity of MCCE measured using the abdominal constriction test and in the first phase of the formalin test occurred in a concentration-dependent manner. The anti-inflammatory activity of 50 and 100% concentrations MCCE was observed only at the range of time interval of 60 - 120 and 60 min, respectively. Based on the results, we conclude that the M. calabura leaves chloroform extract possessed remarkable antinociceptive and antipyretic, but less effective anti-inflammatory, activities and thus justifies the Peruvian folklore claims of its medicinal values.

Ginsenoside Rg3 from Red Ginseng Prevents Damage of Neuronal Cells through the Phosphorylation of the Cell Survival Protein Akt

  • Joo, Seong-Soo;Won, Tae-Joon;Lee, Yong-Jin;Hwang, Kwang-Woo;Lee, Seon-Gu;Yoo, Yeong-Min;Lee, Do-Ik
    • Food Science and Biotechnology
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    • 제15권2호
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    • pp.244-247
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    • 2006
  • Neuronal cell death significantly contributes to neuronal loss in neurological injury and disease. Typically, neuronal loss or destruction upon exposure to neurotoxins, oxidative stress, or DNA damage causes neurodegenerative diseases such as Alzheimer's disease. In this study, we attempted to determine whether ginsenoside Rg3 from red ginseng has a neuroprotective effect via an anti-apoptotic role induced by S-nitroso-N-acetylpenicillamine (SNAP) at the molecular level. We also investigated the antioxidant effect of Rg3 using a metal-catalyzed reaction with $Cu^{2+}/H_2O_2$. Our results showed that Rg3 ($40-100\;{\mu}g/mL$) protected SK-N-MC neuroblastoma cells under cytotoxic conditions and effectively protected DNA from fragmentation. In the signal pathway, caspase-3, and poly (ADP-ribose) polymerase (PARP) were kept at an inactivated status when pretreated with Rg3 in all ranges. In particular, the important upstream p-Akt signal pathway was increased in a dose-dependent manner, which indicates that Rg3 may contribute to cell survival. We also found that oxidative stress can be mitigated by Rg3. Therefore, we have concluded that Rg3 plays a certain role in neurodegenerative pathogenesis via an anti apoptotic, antioxidative effect.

구강세치제에 함유된 SLS(Sodium lauryl Sulfate)가 HaCaT 세포와 NIH-3T3 세포에 미치는 독성 효과 (The effect of the cytotoxicity of sodium lauryl sulfate containing toothpaste on HaCaT and NIH-3T3 cells)

  • 박상례;김영민;최별보라;김지영
    • 한국치위생학회지
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    • 제15권4호
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    • pp.719-725
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    • 2015
  • Objectives: The purpose of this study was to determine the toxic effects of sodium lauryl sulfate(SLS) in human keratinocyte HaCaT cells and mouse fibroblast NIH-3T3 cells. Methods: The effect of sodium lauryl sulfate(SLS) cell viability and proliferation were determined by WST-1 assay and changes shape of nucleus were evaluated by Hoechst staining under fluorescence microscopy. Additionally, observation of cell morphological changes under light microscopy. Results: SLS induced cytotoxicity and a marked apoptosis in both HaCaT and NIH-3T3 cell lines. With the result of the WST-1 assay, SLS induced the cytotoxicity of 0.005% and 0.0075%, 0.01% SLS for 24 h after HaCaT and NIH-3T3 cells in time and dose-dependent manner(p<0.005). SLS inhibited cell growth and caused apoptosis as evidenced by nuclear fragmentation and condensation. Thus, determination of the morphological changes to define apoptosis was visualized using inverted phase contrast microscopy. Conclusions: SLS had toxicity of the human keratinocyte cells and mouse fibroblast cells and this study will provide the basic data for the development of proper SLS concentration in dentifrice.

고지방 식이로 유도된 비만 생쥐에서 쑥부쟁이 에탄올 추출물의 항비만 효과 (Anti-obesity Effect of Aster Yomena Ethanol Extract in High Fat Diet-induced Obese Mice)

  • 이호재;김현식;서상완
    • 동의생리병리학회지
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    • 제31권6호
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    • pp.348-355
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    • 2017
  • Aster yomena (AY) have been used as a traditional medicine to treat cough, bronchial asthma, and insect bites in Korea. In this study, we evaluated the inhibition of adipogenesis in 3T3-L1 cells and in high-fat diet (HFD)-induced obese mice by AY ethanol extract. Lipid accumulation measurement indicates that AY markedly inhibited adipogenesis in a dose-dependent manner. qRT-PCR results demonstrated that the mRNA expression of adipogenic transcription factors such as peroxisome proliferator-activated receptor-${\gamma}$ ($PPAR-{\gamma}$) in 3T3-L1 cells were significantly down-regulated by AY treatment. And inhibited the expression of FAS, a protein responsible for lipid synthesis, transport and storage. Oral administration of AY (100, 250, and 500 mg/kg, P.O/daily for 4 weeks) was conducted in high-fat diet induced obese mice and C57BL/6 mice. AY was orally administered for 4 weeks to extract liver and epididymal fat, and hematoxylin and eosin staining(H&E staining) was observed. Observation showed that the fat concentration of liver tissue tended to decrease dose-dependently and decreased significantly at 500 mg/kg concentration. The AY-administered group of HFD-induced mice had a lower body weight gain, along with decreased triglycerides and total cholesterol compared with the control mice, however, the HDL-cholesterol/total cholesterol ratio was increased. These results indicate that AY exhibits anti-obesity effects in obese mice by decreasing in serum lipid levels and lipogenesis related gene.

Novel Dioxygenases, HIF-α Specific Prolyl-hydroxylase and Asparanginyl-hydroxylase: O2 Switch for Cell Survival

  • Park, Hyun-Sung
    • Toxicological Research
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    • 제24권2호
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    • pp.101-107
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    • 2008
  • Studies on hypoxia-signaling pathways have revealed novel Fe(II) and $\alpha$-ketoglutarate-dependent dioxygenases that hydroxylate prolyl or asparaginyl residues of a transactivator, Hypoxia-Inducible $Factor-\alpha(HIF-\alpha)$ protein. The recognition of these unprecedented dioxygenases has led to open a new paradigm that the hydroxylation mediates an instant post-translational modification of a protein in response to the changes in cellular concentrations of oxygen, reducing agents, or $\alpha$-ketoglutarate. Activity of $HIF-\alpha$ is repressed by two hydroxylases. One is $HIF-\alpha$ specific prolyl-hydroxylases, referred as prolyl-hydroxylase domain(PHD). The other is $HIF-\alpha$ specific asparaginyl-hydroxylase, referred as factor-inhibiting HIF-1(FIH-1). The facts (i) that many dioxygenases commonly use molecular oxygen and reducing agents during detoxification of xenobiotics, (ii) that detoxification reaction produces radicals and reactive oxygen species, and (iii) that activities of both PHD and FIH-1 are regulated by the changes in the balance between oxygen species and reducing agents, imply the possibility that the activity of $HIF-\alpha$ can be increased during detoxification process. The importance of $HIF-\alpha$ in cancer and ischemic diseases has been emphasized since its target genes mediate various hypoxic responses including angiogenesis, erythropoiesis, glycolysis, pH balance, metastasis, invasion and cell survival. Therefore, activators of PHDs and FIH-1 can be potential anticancer drugs which could reduce the activity of HIF, whereas inhibitors, for preventing ischemic diseases. This review highlights these novel dioxygenases, PHDs and FIH-1 as specific target against not only cancers but also ischemic diseases.