• Title/Summary/Keyword: Membrane integrity

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Alterations in Membrane Transport Function and Cell Viability Induced by ATP Depletion in Primary Cultured Rabbit Renal Proximal Tubular Cells

  • Lee, Sung-Ju;Kwon, Chae-Hwa;Kim, Yong-Keun
    • The Korean Journal of Physiology and Pharmacology
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    • v.13 no.1
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    • pp.15-22
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    • 2009
  • This study was undertaken to elucidate the underlying mechanisms of ATP depletion-induced membrane transport dysfunction and cell death in renal proximal tubular cells. ATP depletion was induced by incubating cells with 2.5 mM potassium cyanide(KCN)/0.1 mM iodoacetic acid(IAA), and membrane transport function and cell viability were evaluated by measuring $Na^+$-dependent phosphate uptake and trypan blue exclusion, respectively. ATP depletion resulted in a decrease in $Na^+$-dependent phosphate uptake and cell viability in a time-dependent manner. ATP depletion inhibited $Na^+$-dependent phosphate uptake in cells, when treated with 2 mM ouabain, a $Na^+$ pump-specific inhibitor, suggesting that ATP depletion impairs membrane transport functional integrity. Alterations in $Na^+$-dependent phosphate uptake and cell viability induced by ATP depletion were prevented by the hydrogen peroxide scavenger such as catalase and the hydroxyl radical scavengers(dimethylthiourea and thiourea), and amino acids(glycine and alanine). ATP depletion caused arachidonic acid release and increased mRNA levels of cytosolic phospholipase $A_2(cPLA_2)$. The ATP depletion-dependent arachidonic acid release was inhibited by $cPLA_2$ specific inhibitor $AACOCF_3$. ATP depletion-induced alterations in $Na^+$-dependent phosphate uptake and cell viability were prevented by $AACOCF_3$. Inhibition of $Na^+$-dependent phosphate uptake by ATP depletion was prevented by antipain and leupetin, serine/cysteine protease inhibitors, whereas ATP depletion-induced cell death was not altered by these agents. These results indicate that ATP depletion-induced alterations in membrane transport function and cell viability are due to reactive oxygen species generation and $cPLA_2$ activation in renal proximal tubular cells. In addition, the present data suggest that serine/cysteine proteases play an important role in membrane transport dysfunction, but not cell death, induced by ATP depletion.

Effect of Ethylene Glycol Concentration and Freezing Speed on Post-thawed Semen Viability and Acrosome Integrity in Korean Jeju Black Bull (제주흑우 동결정액 제조시 Ethylene Glycol의 농도와 예비 동결 조건이 정자의 생존율 및 첨체양상에 미치는 영향)

  • Choi, Sun-Ho;Ko, Min-Hee;Kang, Tae-Young;Cho, Sang-Rae;Park, Yong-Sang;Oh, Shin-Ae
    • Reproductive and Developmental Biology
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    • v.35 no.3
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    • pp.377-383
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    • 2011
  • The objective of this study was to examine effect of ethylene glycol for semen cryopreservation in Korean Jeju Black Bull. The semen was cryopreserved with extenders containing cryoprotectants (7% glycerol and 3%, 5%, 7% ethylene glycol) and packed to 0.5 ml straws. The semen straws were located above 3 cm of liquid nitrogen for 5 min, 5 cm for 10 min and 8 cm for 10 min. And then frozen straw was plunged into $LN_2$. Post-thawed sperm motility, viability and membrane integrity were significantly higher in 5% ethylene glycol ($72.5{\pm}5.00%$, $54.88{\pm}0.66%$ and $46.00{\pm}2.40%$; p<0.05). Motility and viability were similar between 7% glycerol and 5% ethylene glycol. However, the membrane integrity was significantly higher in 5% ethylene glycol ($34.69{\pm}4.64%$ vs $46.00{\pm}2.40%$; p<0.05). The viability and membrane integrity were significantly higher in 5 cm for 10 min and 8 cm for 10 min than 3 cm for 5 min (viability: $55.81{\pm}2.94$, $55.19{\pm}3.34$ vs $47.94{\pm}3.48%$; p<0.05 and membrane integrity: $44.94{\pm}3.51$, $46.06{\pm}2.25$ vs $40.38{\pm}1.03%$; p<0.05). The percentage of capacitated sperm assessed by CTC staining, percentage of F pattern was higher in 7% glycerol, 5% and 7% ethylene glycerol, and AR pattern was significantly higher in 3% ethylene glycol. F pattern was significantly increased in 5 cm for 10 min and 8 cm for 10 min (p<0.05), but AR pattern was significantly increased in 3 cm for 5 min (p<0.05).

Controlling the Morphology of Polyvinylidene-co-hexafluoropropylene (PVDF-co-HFP) Membranes Via Phase Inversion Method (상전이법을 이용한 P(VDF-co-HFP) 분리막 구조제어)

  • Song, Ye Jin;Kim, Jong Hoo;Kim, Ye Som;Kim, Sang Deuk;Cho, Young Hoon;Park, Ho Sik;Nam, Seung Eun;Park, You In;Son, Eun Ho;Kim, Jeong F.
    • Membrane Journal
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    • v.28 no.3
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    • pp.187-195
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    • 2018
  • In this work, the morphology of polyvinylidene-co-hexafluoropropylene (PVDF-co-HFP) membranes were systemically investigated using phase inversion technique, to target membrane contactor applications. As the presence of macrovoids degrade the mechanical integrity of the membranes and jeopardize the long-term stability of membrane contactor processes (e.g. wetting), a wide range of dope compositions and casting conditions was studied to eliminate the undesired macrovoids. The type of solvent had significant effect on the membrane morphology, and the observed morphology were correlated to the physical properties of the solvent and solvent-polymer interactions. In addition, to fabricate macrovoid-free structure, the effects of different coagulation temperatures, inclusion of additives, and addition of nonsolvents were investigated. Due to the slow crystallization rate of P(VDF-co-HFP) polymer, it was found that obtaining porous membrane without macrovoids is difficult using only nonsolvent-induced phase separation method (NIPS). However, combined other phase inversion methods such as evaporation-induced phase separation (EIPS) and vapor-induced phase separation (VIPS), the desired membrane morphology can be obtained without any macrovoids.

Effect of Seed Priming on the Germination Performance and Membrane Integrity of Tomato(Lycopersicon esculentum Mill.) Seeds (Priming 처리에 의한 토마토 종자의 발아력과 Membrane Integrity에 미치는 영향)

  • Kang, Jum-Soon;Son, Beung-Gu;Ahn, Chong-Kil
    • Journal of Bio-Environment Control
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    • v.12 no.4
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    • pp.221-227
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    • 2003
  • The objective of this research was to determine the effect of seed priming on membrane integrity during poriming and germination. Among the five chemicals, $KNO_3$at 150 mL gave the shortest $T_{50}$ (days required to reach 50% of the final germination percentage). Compared to unprimed, the seeds primed with 150 mL $KNO_3$ at 20$^{\circ}C$ for 4 days had reduced $T_{50}$ values when germinated at 15$^{\circ}C$. These results indicated that seed priming is an effective way for rapid and synchronized germination, especially at low temperature. Changes in conductivity of priming solutions during the 4-days period of priming were highly dependent upon the priming agents. Conductivity of the $KNO_3$ and $K_3PO_4$ solution slowly declined during the first 3 hours ad then increased Amount of amino acids, sugars and proteins exuded from seeds into $KNO_3$ solution were less than those into distilled water and $K_3PO_4$. All the results suggested that the $KNO_3$ priming play a positive role in regulating the permeability of cell membranes.

Initial tissue response of biodegradable membrane in rat subcutaneous model (백서 피하층에서 흡수성 차단막의 초기 조직 반응)

  • Lim, Hyun-Chang;Chae, Gyung-Joon;Jung, Ui-Won;Kim, Chang-Sung;Lee, Yong-Keun;Cho, Kyoo-Sung;Kim, Chong-Kwan;Choi, Seong-Ho
    • Journal of Periodontal and Implant Science
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    • v.37 no.4
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    • pp.839-848
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    • 2007
  • Purpose: Various kinds of biodegradable membranes are currently used in dental clinics. And the frequency and the necessity of their usage are increasing due to their numerous advantages. Therefore it is important to understand the difference of various membranes and histological reaction against implanted membranes. Materials and Methods: Biodegradable membranes of $Biogide^{(R)}$, $Resolute^{(R)}$, and $Tutodent^{(R)}$ were cut into small pieces by $1.0{\times}0.5cm$. The membranes were implanted 1.5cm apart from each other under the epithelium on the skull of 18 Sprague Dawley rats. The animals were sacrificed at 3, 7, and 14 days after surgical procedure. The specimens were examined by histological analysis. Results: 1. Early period after implantation of the membranes showed connective tissues surrounding membranes and there were a few inflammatory cells present. 2. In $Biogide^{(R)}$ and $Tutodent^{(R)}$ specimens, inflammatory cells and surrounding tissues were shown to infiltrate from outside with slight density difference inside. In $Resolute^{(R)}$ specimens, membranes were fragmented. Inflammatory cells and connective tissues were also observed inside. 3. In $Resolute^{(R)}$ specimen, giant cells were present which implicates that foreign body reaction has occurred. 4. $Biogide^{(R)}$ had lower integrity than other membranes and is not enough to be used alone in defect area. However, $Resolute^{(R)}$ had superior firmness than others. $Tutodent^{(R)}$ had middle level of integrity. Conclusion: This experimental model enabled to observe early inflammatory reactions and morphological changes of materials and can be used to develop and evaluate the efficacy of biodegradable membranes. Duplication of standardized human oral environment will be required in future experiments.

Effect of Adding Taurine, Hypotaurine and Trehalose as Antioxidants to a Tris-based Egg Yolk Extender on Korean Jeju Black Bull Sperm Quality Following Cryopreservation (제주흑우 동결정액 제조에 있어 난황 Tris 희석제에 항산화제로서 Taurine, Hypotaurine 그리고 Trehalose의 첨가가 동결 융해 후 정자의 성상에 미치는 영향)

  • Oh, Shin-Ae;Ko, Min-Hee;Kang, Tae-Young;Choi, Sun-Ho;Ko, Moon-Suck;Chung, Young-Ho;Cho, Won-Mo
    • Journal of Animal Science and Technology
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    • v.54 no.4
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    • pp.283-290
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    • 2012
  • Cryopreservation induces sublethal damage to the spermatozoa, which leads to their reduced fertile life. The objective of this study was to investigate the effect of taurine, hypotaurine and trehalose as antioxidants on the function of the freezing-thawed sperm in Korean Jeju Black Bull. The semen was cryopreserved with tris egg yolk extendercontaining 7% glycerol and treated with 20mM taurine, hypotaurine and trehalose. Frozen-thawed sperms were evaluated for sperm motility, viability, membrane integrity, acrosome integrity and sperm penetration ability. The results were compared to semen cryopreserved in tris egg yolk extender containing 7% glycerol only as control. Frozen-thawed semen evaluation clearlyindicated that the addition of taurine or hypotaurine significantly improved (p<0.05) the motility and viability compared to control spermatozoa. Moreover, in membrane integrity, swollen sperm ratio was significantly increased (p<0.05) in taurine, hypotaurine or trehalose compared to control. In sperm acrosome integrity, F pattern ratio was increased (p<0.05) in hypotaurine among treatments, and AR pattern was significantly lowered (p<0.05) in taurine, hypotaurine and trehalose. In assessed sperm fertilizing ability, taurine, hypotaurine or trehalose significantly improved (p<0.05) the ratio of pronucleus formation and SFI. Finally, compared with the control, addition of taurine, hypotaurine or trehalose as an antioxidant to the freezing extender showed more positive effects on the frozen-thawed spermatozoa. It is concluded that the addition of taurine, hypotaurine, or trehalose to the freezing extender could reduce cryodamage of the Korean Jeju Black Bull spermatozoa.

Zygosaccharomyces rouxii Combats Salt Stress by Maintaining Cell Membrane Structure and Functionality

  • Wang, Dingkang;Zhang, Min;Huang, Jun;Zhou, Rongqing;Jin, Yao;Wu, Chongde
    • Journal of Microbiology and Biotechnology
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    • v.30 no.1
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    • pp.62-70
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    • 2020
  • Zygosaccharomyces rouxii is an important yeast that is required in the food fermentation process due to its high salt tolerance. In this study, the responses and resistance strategies of Z. rouxii against salt stress were investigated by performing physiological analysis at membrane level. The results showed that under salt stress, cell integrity was destroyed, and the cell wall was ruptured, which was accompanied by intracellular substance spillover. With an increase of salt concentrations, intracellular Na+ content increased slightly, whereas intracellular K+ content decreased significantly, which caused the increase of the intracellular Na+/K+ ratio. In addition, in response to salt stress, the activity of Na+/K+-ATPase increased from 0.54 to 2.14 μmol/mg protein, and the ergosterol content increased to 2.42-fold to maintain membrane stability. Analysis of cell membrane fluidity and fatty acid composition showed that cell membrane fluidity decreased and unsaturated fatty acid proportions increased, leading to a 101.21% rise in the unsaturated/saturated fatty acid ratio. The results presented in this study offer guidance in understanding the salt tolerance mechanism of Z. rouxii, and in developing new strategies to increase the industrial utilization of this species under salt stress.

Blue Light Effect on the Fatty Acid Composition of Membrane Lipid of Plant Leaves (식물 세포막의 지방산 조성에 미치는 고강도 청색광선의 효과)

  • Jung, Bo-Kung;Kim, Chang-Sook;Jung, Jin
    • Korean Journal of Environmental Agriculture
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    • v.11 no.3
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    • pp.261-268
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    • 1992
  • The membranes of mitochondria and chloroplasts contain a number of pigments that can act as endogenous sensitizers to produce activated oxygen species, most efficiently in blue light, which, in turn, attack functional targets in membranes. Therefore, intense blue light from the sun can exert various adverse effects on the functional and structural integrity of the membranes: one of the biochemical events of these negative effects could be the oxidative degradation of the unsaturated fatty acid constituents of membrane polar lipid. It may be assumed that as a strategy to avoid the light induced fatty acid degradation in membranes plant cells, responding to high intensity blue light, change the fatty acid compositions of membrane lipid in such that more-unsaturated fatty acid constituents are replaced by lessunsaturated fatty acid constituents. The results obtained in the present study, most importantly the measurements of double bond index of membrane polar lipid in concert with other measurements such as light quaility-dependent membrane peroxidation and the activities of membrane-bound proteins, seem to support this assumption.

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Morphologic Changes of Anchoring Fibers in Vocal Polyps (성대용종에서 부착섬유의 형태학적 변화)

  • 정광윤;최종욱
    • Korean Journal of Bronchoesophagology
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    • v.1 no.1
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    • pp.64-68
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    • 1995
  • Vocal folds injury from vocal abuse is important topics of phonosurgery. Recent advances in diagnostic equipment, phonosurgery and speech analysis equipment have provided a lot of Information about fine movement of the vocal folds. However, predicting the reaction of the vocal folds to phonatory trauma remains difficult. The vocal folds need to withstand great vibratory and shearing stress and anchoring fibers of basement membrane Bone play a role in maintaining structural integrity of histologically different epidermis and superficial layer of lamina propria(cover of vocal folds). The purpose of this study is to demonstrate the changes of anchoring fibers in vocal polyp using transmission electron microscope. Various defects were observed : a irregular thickening of basement membrane, a near absence of normal anchoring fiber, a lot of electron dense material in superficial layer of lamina propria, a destruction of hemidesmosome and many vesicles carrying electron dense material In basal keratinocyte. These observations were suggestive of a hyperactivity of basal keratinocyte of vocal folds epithelium in response to vibratory stress.

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