• Title/Summary/Keyword: cell permeability

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Cytotoxicity of Vibrio vulnificus Cytolysin on Rat Neutrophils

  • Park, Kwang-Hyun;Rho, In-Whan;Park, Byung-Hyun;Kim, Jong-Suk;Kim, Hyung-Rho
    • BMB Reports
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    • v.32 no.3
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    • pp.273-278
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    • 1999
  • Cytolysin produced by Vibrio vulnificus has been known to be lethal to mice by increasing vascular permeability and neutrophil sequestration in the lung. In the present study, a cytotoxic mechanism of V. vulnificus cytolysin on the neutrophil was investigated. Cytolysin rapidly bound to neutrophils and induced cell death, as determined by the trypan blue exclusion test. V. vulnificus cytolysin caused the depletion of cellular ATP without the release of ATP or lactate dehydrogenase. Formation of transmembrane pores was evidenced by the rapid efflux of potassium and 2-deoxy-D-[$^3H$]glucose from cytolysin-treated neutrophils. It was further confirmed by the rapid flow of monovalent ions in the patch clamp of cytolysin-treated neutrophil membrane. The pore formation was accompanied by the oligomerization of cytolysin monomers on the neutrophil membrane as demonstrated by immunoblot, which exhibited a 210 kDa band corresponding to a tetramer of the native cytolysin of $M_r$ 51,000. These findings indicate that V. vulnificus cytolysin rapidly binds to the neutrophil membrane and oligomerizes to form small transmembrane pores, which induce the efflux of potassium and the depletion of cellular ATP leading to cell death without cytolysis.

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Characterization of Nafion/Pt/Polypyrrole Composite Membrane Prepared by Chemical In-situ Polymerization for DMFC (화학적 합성에 의해 제조된 직접 메탄올 연료전지용 나피온/백금/폴리피롤 복합 막의 특성 분석)

  • Park, Ho-Seok;Kim, Yo-Jin;Im, Hun-Suk;Choi, Bong-Gill;Hong, Won-Hi
    • 한국신재생에너지학회:학술대회논문집
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    • 2006.11a
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    • pp.421-424
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    • 2006
  • Nafion/Pt/Polypyrrole composite membranes were fabricated by chemical in-situ polymerization of pyrrole monomers with Pt precursors in Nafion matrix for DMFC. We demonstrated that positively charged pyrrolinum groups of polypyrrole particles were co-interacted with sulfonic groups of Nafion as verified by FT-IR results. Mutual interaction between $Nafion-SO_3^-$ (or negatively charged Pt precursors) and Polypyrrole$-NH_2^+$ influenced the physical properties of pristine Nafion. Thermal property proton conductivity, methanol permeability, and cell performance of pristine and modified Nafion were analyzed for an application of DMFC membrane. Thermal stabilities of sulfonic groups and side chains in Nafion/Pt/polypyrrole composite membranes were higher than those of Nafion due to mutual interaction between sulfonic groups of Nafion and pyrrolinum groups of polypyrrole. Methanol permeabilities of Nafion/Pt/Polypyrrole composite were reduced more proton conductivities with the increase in the content of Pt particles. As a result of that, the enhancement of cell performance by Nafion/Pt/Polypyrole O2 relative to Nafion was more pronounced under the specific experimental condition such as high temperature and more concentrated methanol solution.

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Storage of Bull and Boar Semen: Novel Concepts Derived Using Magnetized Water and Antioxidants

  • Lee, Sang-Hee;Cheong, Hee-Tae;Yang, Boo-Keun;Park, Choon-Keun
    • Reproductive and Developmental Biology
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    • v.38 no.1
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    • pp.1-8
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    • 2014
  • Artificial insemination technique has been contributed immensely for production of livestock worldwide as a critical assisted reproductive technique to preserve and propagate excellent genes in domestic animal industry. In the past decade, methods for semen preservation have been improved mostly in liquid preservation method for boar semen and freezing method for bull semen. Among many factors affecting semen quality during preservation, reactive oxygen species, produced by aerobic respiration in sperm for survival and motility, are unfavorable to sperm physiology. In mammalian cell as well as in the sperm, antioxidant system plays a role in degradation of reactive oxygen species. Magnetized water forms smaller stabilizing water clusters, resulting in high absorption and permeability of the cell for water, implicating its application for semen preservation. Therefore, this review focuses on preservation methods of boar and bull semen with respect to improvement of extender and reduction of reactive oxygen species by using magnetized water and supplementation of antioxidants.

The Characterization of the Increase of Membrane Conductance after Depolarization in Single Rat Adrenal Chromaffin Cells

  • Lim, Won-Il;Kim, Sang-Jeong;Kim, Jun
    • The Korean Journal of Physiology and Pharmacology
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    • v.2 no.1
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    • pp.95-100
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    • 1998
  • The conductance change evoked by step depolarization was studied in primarily cultured rat adrenal chromaffin cells using patch-clamp and capacitance measurement techniques. When we applied a depolarizing pulse to a chromaffin cell, the inward calcium current was followed by an outward current and depolarization-induced exocytosis was accompanied by an increase in conductance trace. The slow inward tail current which has the same time course as the conductance change was observed in current recording. The activation of slow tail current was calcium-dependent. Reversal potentials agreed with Nernst equation assuming relative permeability of $Cs^+\;to\;K^+$ is 0.095. The outward current and tail current were blocked by apamin (200 nM) and d-tubocurarine (2 mM). The conductance change was blocked by apamin and did not affect membrane capacitance recording. We confirmed that conductance change after depolarization comes from the activation of the SK channel and can be blocked by application of the SK channel blockers. Consequently, it is necessary to consider blocking of the SK channel during membrane capacitance recording.

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Changes in Liver and Serum Guanase Activity following Administration of Carbon Tetrachloride in Rats (四鹽化炭素에 依한 肝損傷時 Guanase 活性 變動)

  • Kang, Hoe Yang;Yoon, Chong Guk
    • Journal of Environmental Health Sciences
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    • v.12 no.1
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    • pp.25-38
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    • 1986
  • This paper was intented to charily the cause of an increase of serum guanase activity in rats following injection of $CCI_4$. The cause of increasing serum guanase was focused on the change of guanase activities in both serum and liver, and additionally, these results were compared with the previously known alanine aminotransferase (ALT). Concomitantly the microscopic investigation on the histologic changes, and the determination of lipid peroxides of liver were combined in this experiment for a correlation to observe that the activity of guanase would be effected by the various degree of hepatic injury induced by $CCI_4$. The serum levels of guanase were increased about 2 fold in the fatty change stage (3-12 days), 5.2 fold representing the peak value in necrosis stage (21days), 4.5 fold in early cirrhosis stage (48 days), and 2 fold in severe cirrhosis stage (92 days). These changes of serum guanase activity showed similar patterns to those of ALT activity and lipid peroxides in liver cell. The changes of liver guanase activities showed an increase, whereas ALT activities in liver were markedly decreased. It is likely that the increase of serum guanase activity is based on the excess leaking of guanase into blood by the result of accelerated enzyme synthesis in liver cell of $CCI_4$ intoxicated rats. In addition, the possibility could not be ruled out, however, that the increase of serum guanase activity would be caused by the alteration of membrane permeability.

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Development of Inhibitors of $\beta$-Amyloid Plaque Formation

  • Kim, Dong-Jin
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2006.04a
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    • pp.123-135
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    • 2006
  • Alzheimer's disease (AD) is the most common form of dementia in the aging population and is clinically characterized by a progressive loss of cognitive abilities. Pathologically, it is defined by the appearance of senile plaques - extracellular insoluble, congophilic protein aggregates composed of amyloid $\beta$ (A$\beta$) and neurofibrillary tangles (NFTs) - inyracellular lesions consisting of paired helical filaments from hyperphosphorylated cytoskeletal tau protein as described by Alois Alzheimer a century ago. These hallmarks still serve as the major criteria for a definite diagnosis of the disease. Consequently, one of the key strategy for drug development in this disease area focuses on reducing the concentration of cerebral A$\beta$ plaque by using substances that inhibit A$\beta$ fibril formation. We focused on developing inhibitors by synthesizing several kinds of aromatic molecules. The synthetic compounds were initially screened to evaluate the effective compound by tioflavin T fluorescence assay. The selected effective compounds were tested cytotoxicity and protective effect from A$\beta$-induced neuronal toxicity by cell based MTT assay with HT22 hippocampal neurons. The BBB permeability on effectors was also tested in in vitro co-culture model(HUVEC/C6 cell line). The behavior test wea carried out in mutant APP/PS1 transgenic mouse model of Alzheimer's disease. And inhibition of A$\beta$ fibril formation by the effective compound was monitored with transmitted electron microscopic images.

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LIGHT DEPENDENT CHANNELS AND EXCHANGER IN THE INTERNAL LIMITING MEMBRANE OF VERTEBRATE EYE

  • Hyuk Jung;Kim, You-Young
    • Journal of Photoscience
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    • v.6 no.2
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    • pp.77-83
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    • 1999
  • Calcium has a variety of functions in neuron and muscle cells and blood clotting, especially in the visual system where dark adapted rods cotransport with Na$\^$+/ into the cell. An influx of Ca$\^$++/ flows out of the cell through the Na$\^$+/-Ca$\^$++/ exchanger. By using a modified Using chamber in order to bring in vivo environment close, we have known that Ca$\^$++/ blocks the activity of guanylate cyclase, in consequence, having an effect on the amplitude of electroretinogram (ERG). We have measured the Ca$\^$++/, K$\^$+/, and Na$\^$+/ concentration in dark and light adapted bullfrog's (Rana catesbeiana) vitreous humor. The calcium concentration of the light adapted bullfrog's vitreous humor was higher than that of the dark adapted bullfrog's vitreous humor This means that ion activity between the photoreceptor and vitreous humor side is light dependent and we have found that a Ca$\^$++/ channel and Ca$\^$++/K$\^$+/ exchanger exist in the vitreous humor side. Taken together permeability of Ca$\^$++/, K$\^$+/ and K$\^$+/ ion internal limiting membrane faced in the vitreous humor side has light-dependent activity during the illumination.

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Ionic Cluster Mimic Membranes Using Ionized Cyclodextrin

  • Won Jong-Ok;Yoo Ji-Young;Kang Moon-Sung;Kang Yong-Soo
    • Macromolecular Research
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    • v.14 no.4
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    • pp.449-455
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    • 2006
  • Ionic cluster mimic, polymer electrolyte membranes were prepared using polymer composites of crosslinked poly(vinyl alcohol) (PVA) with sulfated-${\beta}$-cyclodextrins (${\beta}-CDSO_3H$) or phosphated-${\beta}$-cyclodextrins (${\beta}-CDPO(OH)_2$). When Nafion, developed for a fuel cell using low temperature, polymer electrolyte membranes, is used in a direct methanol fuel cell, it has a methanol crossover problem. The ionic inverted micellar structure formed by micro-segregation in Nafion, known as ionic cluster, is distorted in methanol aqueous solution, resulting in the significant transport of methanol through the membrane. While the ionic structure formed by the ionic sites in either ${\beta}-CDSO_3H$ or ${\beta}-CDPO(OH)_2$ in this composite membrane is maintained in methanol solution, it is expected to reduce methanol transport. Proton conductivity was found to increase in PVA membranes upon addition of ionized cyclodextrins. Methanol permeability through the PVA composite membrane containing cyclodextrins was lower than that of Nafion. It is thus concluded that the structure and fixation of ionic clusters are significant barriers to methanol crossover in direct methanol fuel cells.

In vitro Alternatives to Skin Irritation Test

  • Shin, Dae-Sup;Kim, Dai-Byung;Ryu, Seung-Rel;Lee, Sun-Hee;Koh, Jae-Sook;Park, Won-Sae;Kim, Pu-Young
    • Biomolecules & Therapeutics
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    • v.3 no.3
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    • pp.242-244
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    • 1995
  • In vitro cell culture system has been proposed as a promising alternative model to in vivo skin irritation test. These studies were performed to screen the cytotoxicity effects of surfactants using normal human skin fibroblasts. Cell membrane integrity assessed by the leakage of lactate dehydrogenase (LDH) and mitochondrial integrity by MTT [3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromides reduction test were affected in a dose dependent manner. The irritation potential of surfactants to human skin patch test, and the changes of capillary permeability by rabbit intradermal safety test were assessed as in vivo methods. Our results suggest that LDH leakage assay and MTT reduction test using cultured human fibroblasts could be predictive for the irritancy of various surfactants in human, and LDH assay is superior correlated with in vivo test (r=0.886) to MTT test with in vivotest (r=0.757).

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Degradation of Polymer Electrolyte Membrane under Low Current/Low Humidity Conditions (저전류/저가습 조건에서 고분자전해질 막 열화)

  • Kim, Tae-Hee;Lee, Jung-Hun;Lee, Ho;Lim, Tae-Won;Park, Kwon-Pil
    • Transactions of the Korean hydrogen and new energy society
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    • v.18 no.2
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    • pp.157-163
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    • 2007
  • During PEMFC operation, low current and low humidity conditions accelerate the degradation of perfluorosulfonic acid membrane. But, there have been no studies that clearly explain why these conditions accelerate the membrane degradation. In this study, the hydrogen permeability through the membrane, I-V polarization of MEA, fluoride emission rate(FER) in effluent water were measured during cell operation under low current densities and low relative humidity(RH). The experimental results were evaluated with oxygen radical mechanism the most commonly known for membrane degradation. It seems that low RH of anode is a good condition for $H{\cdot}$ radical formation on the Pt catalyst and the low current condition accelerates the $H{\cdot}$ to form $HO_2{\cdot}$ radical attacking the polymer membrane.