• Title/Summary/Keyword: intracellular pH

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Induction of Quinone Reductase , an Anticarcinogenic Marker Enzyme, by Vitamin E in Both Hepalclc7 Cells and Mice

  • Kwon, Chong-Suk;Kim, Jong-Sang
    • Preventive Nutrition and Food Science
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    • v.4 no.2
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    • pp.122-124
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    • 1999
  • Induction of NAD(P)H : (quinone-acceptor) oxidoreductase (QR) which obligatory two electron reduction of quinones and prevents their participation in oxidative cycling and thereby the depletion of intracellular glutathione, has been used as a marker for chemopreventive agents. We postulated that vitamin E, an antioxidant, which induces QR as the gene of QR was reported to contain antioxidant reponsive element in the 5'-flanking region. Vitamin E resulted in significant induction of QR in both hepalclc7 cells and mouse tissues. QR induction was observed; to be maximal at 25uM vitamin E for hepalclc7 cells while it was maximal in the level of 2.5∼5 μmoles vitamin E/㎏ BW for mouse tissues. Thus the cancer-preventive effect of vitamin E may be exerted by it induction of intracellular detoxifying enzymes.

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Anti-oxidative Effect of Piperine from Piper nigrum L. in Caenorhabditis elegans

  • Park, Hyun Mee;Kim, Jun Hyeong;Kim, Dae Keun
    • Natural Product Sciences
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    • v.25 no.3
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    • pp.255-260
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    • 2019
  • Piper nigrum L. (Piperaceae), which is a well-known food seasoning, has been used as a traditional medicine for the treatment of vomiting, abdominal pain, diarrhea and anorexia in Korea, China and Japan. Methanol extract from the fruit of P. nigrum was successively partitioned as n-hexane, methylene chloride, ethyl acetate, n-butanol and $H_2O$ soluble fractions. Among those fractions the ethyl acetate soluble fraction showed the most potent DPPH radical scavenging activity, and piperine was isolated from the ethyl acetate fraction. To know the antioxidant activity of piperine, we tested the activities of superoxide dismutase (SOD) and catalase together with oxidative stress tolerance and intracellular ROS level in Caenorhabditis elegans. To investigate whether piperine-mediated increased stress tolerance was due to regulation of stress-response gene, we quantified SOD-3 expression using transgenic strain including CF1553. Consequently, piperine enhanced SOD and catalase activities of C. elegans, and reduced intracellular ROS accumulation in a dose-dependent manner. Moreover, piperine-treated CF1553 worms exhibited significantly higher SOD-3::GFP intensity.

Effect of pH on the ATP-sensitive $K^+$ Channel in Aortic Smooth Muscle Cells from Rats

  • Kim, Se-Hoon;Kim, Il-Su;Kim, Hoe-Suk;Jeon, Byeong-Hwa;Chang, Seok-Jong
    • The Korean Journal of Physiology and Pharmacology
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    • v.1 no.5
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    • pp.555-563
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    • 1997
  • The effects of pH on $K^+$ currents were investigated in single smooth muscle cells isolated from the thoracic aorta of Wistar-Kyoto rats. Whole-cell $K^+$ currents were recorded in the conventional configuration of the voltage-clamp technique. Pinacidil (10uM) activated the whole-cell current and the pinacidil-activated current was completely inhibited by glibenclamide (10uM) , an inhibitor of ATP-sensitive $K^+$ channel ($K_{ATP}$ channel). Pinacidil-activated current was reversed at near the $K^+$ equilibrium potential. This current was time- and voltage-independent and reduced by elevating intracellular ATP. Pinacidil-activated current was reduced by lowering the external pH. However, alteration of internal pH has controversial effects on pinacidil-activated current. When the single cell was dialyzed with 0.1 mM ATP, alteration of internal pH had no effect on pinacidil-activated $K^+$ current. In the contrast, when the single cell was dialyzed with 3 mM ATP, pinacidil-activated current was increased by lowering internal pH. Our results suggest that $K^+$ channel activated by pinacidil may be $K_{ATP}$ channel and internal $H^+$ may reduce the inhibitory effect of ATP on $K_{ATP}$ channel.

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Mucosal Immunity Related to CD8+ T Lymphocytes in Children with Helicobacter pylori Gastritis

  • Da Hee Yang;Ha Young Lee;Woohyuk Choi;Chang-Lim Hyun;Ki Soo Kang
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.27 no.1
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    • pp.26-36
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    • 2024
  • Purpose: We investigated the role of CD8+T cells as host immune factors in pediatric patients with Helicobacter pylori gastritis. Methods: Gastric mucosal tissue and blood samples were collected from 39 children, including 11 children with H. pylori infection and 28 children as controls. Anti-CD8 and anti-T-bet antibodies were used for immunohistochemistry of the gastric mucosa. For the cell surface and intracellular staining, peripheral blood mononuclear cells were stained with anti-IL7Rα, anti-CX3CR1, anti-CD8, anti-T-bet, and anti-IFN-γ antibodies. Cytokines of sera such as tumor necrosis factor alpha (TNF-α) and CX3CL1 were analyzed using enzyme- linked immunosorbent assay (ELISA). Results: In the immunohistochemistry of gastric mucosa, the frequency of CD8+ and T-bet+ T cells cells was higher in the H. pylori-positive group than in the control group (26.9± 7.8% vs. 16.9±3.3%, p<0.001; 5.0±2.5% vs. 2.2±0.7%, p=0.001). Between the control and H. pylori-positive groups, the frequency of IL-7RαlowCX3CR1+ CD8+ and T-bet+ INF-γ+ CD8+ T cells were not significantly different between surface and intracellular staining, respectively (40.4±24.0% vs. 38.2±17.8%, p=0.914; 40.4±24.0% vs. 38.2±17.8%, p=0.914). In the ELISA, no significant differences in TNF-α and CX3CL1 concentrations were observed between the control and H. pylori-positive groups (34.3±12.1 pg/mL vs. 47.0±22.6 pg/mL, p=0.114/0.5± 0.1 pg/mL vs. 0.5±0.1 pg/mL, p=0.188). Conclusion: CD8+ T and Th1 cells, which secrete IFN-γ, might play important roles in the mucosal immunity of the stomach in children with H. pylori infection.

Antioxidative and Neuroprotective Effects of Extract and Fractions from Adenophora triphylla (잔대 추출물과 이들 분획물들의 항산화 및 뇌신경세포 보호 효과)

  • Chung, Mi Ja;Lee, Sanghyun;Park, Yong Il;Kwon, Ki Han
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.11
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    • pp.1580-1588
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    • 2016
  • The 70% ethanol extract from Adenophora triphylla showed a strong antioxidant effect and reduced cytotoxicity of $H_2O_2$ in SK-N-SH cells. The chloroform fraction from A. triphylla extract (AT-CH) among the six fractions showed strong DPPH radical and intracellular reactive oxygen species (ROS) scavenger effects and the highest protective effect against $H_2O_2$-induced SK-N-SH cell death. Bioactivity compounds were purified from AT-CH, and the chemical structures of the compounds were determined as ${\beta}$-sitosterol and daucosterol on the basis of $^1H-NMR$, $^{13}C-NMR$, and EI mass spectra. ${\beta}$-Sitosterol and daucosterol also had protective effects against oxidative stress in SK-N-SH cells. Phospho-p38 MAPK levels were elevated by $H_2O_2$ but were inhibited by treatment with AT-CH and phytosterols (${\beta}$-sitosterol and daucosterol) isolated from AT-CH. These results suggest that AT-CH has brain neuroprotective effects against oxidative stress ($H_2O_2$) by inhibiting activation of p38 pathways and scavenging intracellular ROS.

Enzymatic production of Fructo-oligosaccharides from Sucrose (자당으로부터 프럭토올리고당의 효소적 생산 연구)

  • 신형태;백순용;이수원;서동상;권석태;김종남;임유범;이재흥
    • KSBB Journal
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    • v.17 no.6
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    • pp.555-559
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    • 2002
  • Three different strains of Aureobasidium pullulans were grown in batch cultures to compare their abilities of enzyme production. It was found that specific enzyme activity was the highest with strain ATCC 9348 and the enzyme production was closely coupled to growth. Studies on morphology during the growth of A. pullulans revealed that mycelia cells were dominant at the initial stages of growth. However, yeast-like cells and chlamydospores were dominant in the latter stages of batch culture. The pattern of morphological changes during the growth period was not affected by pH. However, it appears that the ratio of intra- to extracellular enzyme activity tended to increase with fermentation time irrespective of the pH employed, suggesting that the secretion efficiency of intracellular enzyme to broth likely depends on cell morphology Using molasses as a cheap source of sucrose, enzymatic production of fructo-oligosaccharides as a feed additive with A. pullulans cells could be achieved successfully at 55$\^{C}$ and pH 5.5.

Histological and Histochemical Studies on the Mucous Secreting Cells of the Gastric Mucosae according to the Development of Frog, Rana nigromaculata (개구리(Rana nigromaculata) 발생에 따른 위점막 점액분비세포의 조직학적 및 조직화학적 연구)

  • Kim, Han-Hwa;Noh, Yong-Tai;Chung, Young-Wha
    • The Korean Journal of Zoology
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    • v.19 no.1
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    • pp.25-32
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    • 1976
  • To observe the changes of mucosubstances of the mucous secreting cells, stomach tissues in frog tadpoles at each stage of metamorphosis were fixed in 10% buffered formalin at $4^{\circ}C$, embedded in paraffin, sectioned to 4 $\mu$m thickness and stained with periodic acid-Schiff(PAS) and alcian blue (AB) of pH 2.5 and pH 1.0. The results obtained were as follows: 1. The reactivities of the surface mucous cells, which exhibited strong PAS-positivity and weak alcianophilia at both pH 2.5 and 1.0, were not changed in metamorphosis stages and the intracellular contents of neutral mucosubstances in the surface mucous cells increased significantly in XXIV and XXV stages of metamorphosis. 2. In the foveolar mucous cells, which appear after metamorphosis XXI, the staining reactivities to PAS, AB of pH 2.5 and 1.0 were the same as that of surface mucous cells during metamorphosis and the alcianophilia were stronger at pH 1.0 than at pH 2.5. 3. THe mucous neck cells, which appear after metamorphosis XXIV, exhibited a strong PAS-positive reaction and weak alcianophilia at metamorphosis XXIV but at metamorphosis XXV weak reactivity to PAS and strong alcianophilia at pH 1.0.

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EFFECTS OF PROLONGED EXPOSURE TO THE SUN ON BODY WATER TURNOVER AND VOLUME OF THE BLOOD IN SWAMP BUFFALOES

  • Chaiyabutr, N.;Buranakarl, C.;Loypetjra, P.;Chanpongsang, S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.3 no.1
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    • pp.47-52
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    • 1990
  • During prolonged exposure to the sun for 8 h each day for 10 days in which the highest ambient temperature around 14:00 h was $39^{\circ}C$, buffaloes exposed to the sun without shade increased the turnover of body water by 35% and 76% on day 5 and day 10 of exposure respectively. The total body water markedly decreased on day five and this amount was maintained thereafter. Plasma and blood volumes did not change significantly on day five but markedly decreased on day 10. Packed cell volume significantly decreased on day five and day 10 of the exposure period. The reduction of packed cell volume on day 10 coincided with the decrease in total plasma water. On day 10 of the exposure, an increase in the rate of liquid flow from the rumen was noted. It is concluded that on the fifth day of exposure, the increase in the evaporative cooling process was attributed to initial mobilization of water from the intracellular compartment. The reduction of both plasma and cell volumes occurring from day five to day 10 indicated a loss of body water from both intracellular and extracellular compartments.

Effects of Tea Constituents on Intracellular Level of the Major Tea Catechin, (-)-Epigallocatechin-3-gallate

  • Hong, Jun-Gil;Yang, Chung-S.
    • Food Science and Biotechnology
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    • v.16 no.1
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    • pp.167-170
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    • 2007
  • (-)-Epigallocatechin-3-gallate (EGCG), a mai or tea catechin has been shown to have many interesting biological activities. In the present study, we studied the effects of green tea catechins, EGCG metabolites, and black tea theaflavins on accumulation of EGCG in HT-29 human colon cells. Intracellular levels of [$^3H$]-EGCG were not changed significantly in the presence of other tea catechins including (-)-epicatechin, (-)-epigallocatechin, and (-)-epicatechin-3-gallate. EGCG methyl metabolites and EGCG 4"-glucuronide did not affect cellular levels of [$^3H$]-EGCG. Black tea theaflavins and theasinensin A (TsA), an EGCG oxidative dimer, however, significantly decreased cellular accumulation of EGCG in HT-29 cells by 31-56%. This decrease was more pronounced when cells were incubated in the presence of theaflavin-3',3"-digallate (TFdiG) or TsA. When EGCG was added separately from TFdiG or TsA, the accumulation of EGCG in HT-29 cells was also significantly decreased regardless of when TFdiG or TsA was added during the uptake study (p<0.01). The results suggest that theaflavins and TsA may interrupt EGCG absorption through the gastrointestinal epithelium.

Structural characterization of As-MIF and hJAB1 during the inhibition of cell-cycle regulation

  • Park, Young-Hoon;Jeong, Suk;Ha, Ki-Tae;Yu, Hak Sun;Jang, Se Bok
    • BMB Reports
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    • v.50 no.5
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    • pp.269-274
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    • 2017
  • The biological activities of macrophage migration inhibitory factor (MIF) might be mediated through a classical receptor-mediated or non-classical endocytic pathway. JAB1 (C-Jun activation domain-binding protein-1) promotes the degradation of the tumor suppressor, p53, and the cyclin-dependent kinase inhibitor, p27. When MIF and JAB1 are bound to each other in various intracellular sites, MIF inhibits the positive regulatory effects of JAB1 on the activity of AP-1. The intestinal parasite, Anisakis simplex, has an immunomodulatory effect. The molecular mechanism of action of As-MIF and human JAB1 are poorly understood. In this study, As-MIF and hJAB1 were expressed and purified with high solubility. The structure of As-MIF and hJAB1 interaction was modeled by homology modeling based on the structure of Ace-MIF. This study provides evidence indicating that the MIF domain of As-MIF interacts directly with the MPN domain of hJAB1, and four structure-based mutants of As-MIF and hJAB1 disrupt the As-MIF-hJAB1 interaction.