• Title/Summary/Keyword: membrane integrity

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Direct and Indirect Membrane Integrity Tests for Monitoring Microbial Removal by Microfiltration (정밀여과(MF)막 미생물 제거율 모니터링을 위한 막 완전성시험)

  • Hong, Seungkwan;Miller, Frank;Taylor, James
    • Journal of Korean Society of Water and Wastewater
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    • v.18 no.6
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    • pp.801-806
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    • 2004
  • The pilot study was conducted to (i) investigate the ability of various membrane integrity monitoring methods to detect changes in membrane integrity during operation, and (ii) determine the impact of membrane damage on microbial removal by microfiltration. Two variations of air pressure hold tests were investigated for direct integrity monitoring: pressure decay (PD) and diffusive air flow (DAF) tests which are most commonly used integrity tests for microfiltration (MF) membranes. Both PD and DAF tests were sensitive enough to detect one damaged fiber out of 66,000 under field operaing conditions. Indirect integrity monitoring such as turbidity and particle counting, however, responded poorly to defects in membrane systems. Microbial challenge study was performed using both new and deliberately damaged membranes, as well as varying the state of fouling of the membrane. This study demonstrated that MF membrane with nominal pore size $0.2{\mu}m$ was capable of removing various pathogens including coliform, spore, and cryptosporidium, at the level required by drinking water regulations, even when high operating pressures were applied. A sharp decrease in average log reduction value (LRV) was observed when one fiber was damaged, emphasizing the importance of membrane integrity in control of microbial contamination.

Effects of Curcumin on Sperm Motility, Viability, Mitochondrial Activity and Plasma Membrane Integrity in Boar Semen

  • Lee, A-Sung;Lee, Sang-Hee;Lee, Seunghyung;Yang, Boo-Keun
    • Biomedical Science Letters
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    • v.23 no.4
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    • pp.406-410
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    • 2017
  • Curcumin is known as a natural antioxidant, decreasing oxidative stress in animal cells. Generally, oxidative stress induces reactive oxygen species in sperm and leads to decreased sperm characteristics in pigs. Therefore, this study investigated the influence of curcumin on sperm motility, viability, mitochondrial activity and plasma membrane integrity in pigs. Curcumin (0, 5 and $10{\mu}M$) was treated in boar semen, which were incubated for 9 hours in $37^{\circ}C$. Then, motility, viability, mitochondrial activity, plasma membrane integrity of sperm was analyzed every 3 hours. In the results, sperm motility was significantly increased by $5{\mu}M$ curcumin after 3 and 9 hours after incubation, and viability was significantly higher in $5{\mu}M$ curcumin treatment at 3 hours (P<0.05). Similarly, sperm mitochondrial activity and plasma membrane integrity were significantly increased by $5{\mu}M$ curcumin at 3, 6 and 9 hours after incubation (P<0.05). There results suggest that curcumin improve sperm characteristics such as motility, viability, mitochondrial activity, and plasma membrane integrity, and may exert a positive effect on sperm fertility in pigs.

Spermatozoa motility, viability, acrosome integrity, mitochondrial membrane potential and plasma membrane integrity in 0.25 mL and 0.5 mL straw after frozen-thawing in Hanwoo bull

  • Kang, Sung-Sik;Kim, Ui-Hyung;Lee, Myung-Suk;Lee, Seok-Dong;Cho, Sang-Rae
    • Journal of Animal Reproduction and Biotechnology
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    • v.35 no.4
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    • pp.307-314
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    • 2020
  • In the present study, we examined the effect of straw size on spermatozoa motility, viability, acrosome integrity, mitochondrial membrane potential, and plasma membrane integrity after freezing-thawing. Hanwoo semen was collected from three bulls and diluted with an animal protein-free extender, divided into two groups, namely, 10 million spermatozoa in 0.25 mL and 20 million spermatozoa in 0.5 mL straw, and cryopreserved. In Experiment 1, the motility and motility parameters of the frozen-thawed spermatozoa were evaluated. After freezing-thawing, the spermatozoa motility parameters fast progressive, straight line velocity, and average path velocity were compared between the 0.25 mL straw and 0.5 mL straw groups. They were 35.2 ± 1.0 and 32.3 ± 0.7%, 34.6 ± 0.7 and 31.8 ± 0.5 μm/s, 51.4 ± 1.3 and 47.1 ± 1.1 μm/s, 0.25 mL straw and 0.5 mL straw groups, respectively. In Experiment 2, the viability, acrosome membrane integrity, and mitochondrial membrane potential of the frozen-thawed spermatozoa were assessed. After freezing-thawing, the percentages of spermatozoa with live, intact acrosomes and high mitochondrial membrane potential were compared between the in 0.25 mL straw and 0.5 mL straw groups. They were 48.0 ± 2.6% and 35.6 ± 2.8% between the 0.25 mL straw and 0.5 mL straw groups. In Experiment 3, the plasma membrane integrity of frozen-thawed spermatozoa was compared. After freezing-thawing, the plasma membrane integrity was higher for the in 0.25 mL straw group than the 0.5 mL straw group. They were 62.0 ± 2.2 and 54.1 ± 1.3% between the 0.25 mL straw and 0.5 mL straw groups. In conclusion, our results suggest that freezing semen in 0.25 mL straw improves the relative motility, viability, and acrosomal, mitochondrial membrane potential, and plasma membrane integrity of Hanwoo bull spermatozoa.

Analysis of Membrane Integrity and Mitochondrial Activity in Fresh and Cryopreserved Boar Sperm Using Flow Cytometry

  • Park C. S.;Li Z. H.;Sung N. D.;Jin D. I.;Cong P. Q.;Kim E. S.;Song E. S.;Yi Y. J.
    • Reproductive and Developmental Biology
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    • v.29 no.4
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    • pp.253-257
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    • 2005
  • This study was carried out to evaluate the effects of washing medium, breed and washing temperature of fresh and frozen-thawed boar sperm on mitochondrial activity and membrane integrity by flow cytometry. More than $80\%$ of fresh sperm washed with mTLP-PVA medium at $20^{\circ}C$ exhibited an intact membrane and a functional mitochondrion. With frozen-thawed samples, a large number of sperm showed both damaged membrane $(36.4\~46.9\%)$ and nonfunctional mitochondrion $(55.1\~71.1\%)$ in the mTLP-PVA and BTS washing media at $20^{\circ}C$. There were no breed effects of fresh and frozen-thawed sperm on mitochondrial activity and membrane integrity. The percentages of damaged membrane of fresh and frozen sperm, respectively, were higher at $4^{\circ}C$ washing temperature than at $20^{\circ}C$ washing temperature in the mTLP-PVA medium. We found that washing medium and washing temperature of fresh and frozen-thawed boar sperm were important for the analyses of mitochondrial activity and membrane integrity by flow cytometry.

Adaptation of the Hypoosmotic Swelling Test to Evaluate Membrane Integrity of Boar Spermatozoa

  • Jang, Hyun-Yong;Cheong, Hee-Tae;Hwang, Hwan-Sub;Kim, Jong-Taek;Park, Choon-Keun;Lee, Hak-Kyu;Yang, Boo-Keun
    • Reproductive and Developmental Biology
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    • v.31 no.2
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    • pp.121-126
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    • 2007
  • The objective of this study was to establish the optimal conditions for hypoosmotic swelling (HOS) test to assess the functional integrity of the membranes of boar fresh or frozen/thawed spermatozoa. When pooled semen sample was incubated for 30 min at $37^{\circ}C$ with different test solution of varied osmolarity, the highest percentage of HOS positive spermatozoa was observed in a 150 mOsmol fructose/Na-citrate solution (33.6%). Incubation time did not affect significantly the score of HOS positive spermatozoa observed in a 150 mOsmol fructose/Na-citrate solution at $37^{\circ}C$, but the osmolarity affected the score of HOS positive spermatozoa under the same condition above. Fresh semen was significantly better than frozen/thawed semen in semen parameters evaluated such as motility, viability, membrane integrity and lipid peroxidation (p<005). In the relationships of sperm parameters, motility vs viability, motility vs membrane integrity and viability vs membrane integrity were positively correlated ($0.82{\sim}0.94$) but lipid peroxidation vs other estimated factors was negatively correlated ($- 0.90{\sim}- 0.98$). Among the evaluation methods, motility vs Viability, motility vs membrane integrity and lipid peroxidation vs other estimated factors were significantly correlated (p<0.05). These results of this. study indicate that the optimal condition of HOST in boar spermatozoa is a 150 mOsmol fructose/Na-citrate solution for 30 min incubation at $37^{\circ}C$ and HOST can substitute the examination of motility, viability and lipid peroxidation.

Effects of Turine and Vitamin E on Sperm Viability, Membrane Integrity and Mitochondrial Activity damaged by Bromopropane in Fresh Boar Semen

  • Lee, Seunghyung;Park, Hee-Woo;Cheong, Hee-Tae;Park, Choon-Keun;Yang, Boo-Keun
    • Journal of Embryo Transfer
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    • v.31 no.1
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    • pp.13-17
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    • 2016
  • The purpose of this study was to examine the effects of taurine and vitamin E on sperm characteristics damaged by bromopropane (BP) in pig. We evaluated toxicity of BP on viability, membrane integrity and mitochondrial activity of spermatozoa. 1-BP (0, 2.5, 5.0, 10, and $50{\mu}M$), 2-BP (0, 2.5, 5.0, 10, and $50{\mu}M$), taurine (0, 5.0, 10, and $25{\mu}M$) and vitamin E (0, 50, 100, and $200{\mu}M$) were treated in fresh boar semen for 6 h. 10 and $50{\mu}M$ of 1-BP and 2-BP inhibited sperm viability, membrane integrity and mitochondrial activity in fresh boar semen (P<0.05). $25{\mu}M$ of taurine increased sperm viability and membrane integrity (P<0.05), $100{\mu}M$ of vitamin E enhanced viability and mitochondrial activity of sperm (P<0.05). Finally, $10{\mu}M$ of 1-BP and 2-BP was co-treated with taurine ($25{\mu}M$) and vitamin E ($100{\mu}M$) in the fresh boar semen. The co-treated samples did affected viability, membrane integrity and mitochondrial activity of sperm. In conclusion, taurine and vitamin E can improve and maintain sperm quality in fresh boar semen.

Myricetin Disturbs the Cell Wall Integrity and Increases the Membrane Permeability of Candida albicans

  • Lee, Heung-Shick;Kim, Younhee
    • Journal of Microbiology and Biotechnology
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    • v.32 no.1
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    • pp.37-45
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    • 2022
  • The fungal cell wall and membrane are the principal targets of antifungals. Herein, we report that myricetin exerts antifungal activity against Candida albicans by damaging the cell wall integrity and notably enhancing the membrane permeability. In the presence of sorbitol, an osmotic protectant, the minimum inhibitory concentration (MIC) of myricetin against C. albicans increased from 20 to 40 and 80 ㎍/ml in 24 and 72 h, respectively, demonstrating that myricetin disturbs the cell wall integrity of C. albicans. Fluorescence microscopic images showed the presence of propidium iodide-stained C. albicans cells, indicating the myricetin-induced initial damage of the cell membrane. The effects of myricetin on the membrane permeability of C. albicans cells were assessed using crystal violet-uptake and intracellular material-leakage assays. The percentage uptakes of crystal violet for myricetin-treated C. albicans cells at 1×, 2×, and 4× the MIC of myricetin were 36.5, 60.6, and 79.4%, respectively, while those for DMSO-treated C. albicans cells were 28.2, 28.9, and 29.7%, respectively. Additionally, myricetin-treated C. albicans cells showed notable DNA and protein leakage, compared with the DMSO-treated controls. Furthermore, treatment of C. albicans cells with 1× the MIC of myricetin showed a 17.2 and 28.0% reduction in the binding of the lipophilic probes diphenylhexatriene and Nile red, respectively, indicating that myricetin alters the lipid components or order in the C. albicans cell membrane, leading to increased membrane permeability. Therefore, these data will provide insights into the pharmacological worth of myricetin as a prospective antifungal for treating C. albicans infections.

The Cryoprotective Effect on Frozen-thawed Boar Semen of Egg Yolk Low Density Lipoproteins

  • Hu, Jian-hong;Li, Qing-Wang;Li, Gang;Chen, Xiao-Yu;Hai-Yang, Hai-Yang;Zhang, Shu-Shan;Wang, Li-Qiang
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.4
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    • pp.486-494
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    • 2006
  • In order to protect the spermatozoa against cold shock, hen egg yolk is widely used as a cryoprotective agent in semen freezing extenders for domestic animals. The protective action of yolk is largely presumed to be due to low density lipoproteins (LDL). The effects of LDL on sperm quality of bull and northern pike (Esox lucius) after freezing-thawing have been reported, but no study has been made to evaluate the effect of LDL on boar sperm motility and other characteristics. The experiment was carried out to investigate the effect of LDL on the freezing of boar sperm in 0.25 ml straws. The aim was to evaluate the quality of boar spermatozoa cryopreserved in the presence of LDL. Motility of semen cryopreserved in LDL was analyzed and compared to semen cryopreserved with Tris-citric acid-glucose (TCG) and Tris-citric acid-fructose (TCF), two basic freezing extenders containing egg yolk. Similarly, acrosome and plasma membrane integrity were also evaluated and compared to semen cryopreserved with TCG and TCF. Analysis of sperm quality after freeze-thaw showed that the motility, acrosome and plasma membrane integrity were improved with LDL in the extender, as compared to the TCG and TCF. The highest post-thaw integrity of acrosome and plasma membrane and motility were obtained with 9% LDL (w/v). Consequently, the optimum LDL concentration in the extender was 9%. It is also suggested that the concentration of LDL addition is important for the effect on boar sperm protection during freezing and thawing. The percentage of motile spermatozoa was significantly higher after freezing in 9% LDL than in TCG and TCF 54.4% versus 30.4% and 30.1% (p<0.05), respectively. The integrity of acrosome and plasma membrane were also significantly higher at 70.3% and 50.5% respectively with semen frozen in 9% LDL extender compared to TCG at 37.8% and 30.3% and TCF at 36.4% and 29.9%, respectively (p<0.05),. In conclusion, we propose that extender containing LDL extracted from hen egg yolk could be used as a cryoprotective media with a better efficiency than TCG and TCF. LDL improved boar semen quality, allowing better spermatozoa motility, acrosome and plasma membrane integrity after the freeze-thaw process. Furthermore, we found out that the extender with 9% LDL concentration significantly enhanced motility, acrosome and plasma membrane integrity of boar sperm after freezing and thawing.

Analysis of Membrane Integrity and Removal Efficiency Considering Membrane Defect and Pore Size (막 파단 및 공극크기에 따른 막 완결성 및 제거효율 분석)

  • Hur, Hyun-chul;Rhee, Ok-jae;Lee, Kwang-jae;Kim, Kwang-ho;Choi, Young-june;Lee, Joo-hee;Hong, Seungkwan
    • Journal of Korean Society on Water Environment
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    • v.24 no.4
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    • pp.423-429
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    • 2008
  • Microfiltration (MF) and ultrafiltration (UF) processes for removal of particulate materials (i.e., turbidity, microorganisms and viruses) have been used to produce drinking water with higher quality. As membrane filtration technique has become widely applied for drinking water treatment, the importance of membrane integrity test (MIT) has also been increasingly emphasized. The results of pressure decay test (PDT) were presented in the paper to monitor membrane integrity. In this paper the PDT was carried out with deliberately-defected membrane fibers to evaluate the sensitivity of PDT on membrane fiber damage. Variation of pressure decay rate and removal rate were investigated to evaluate the impact of defection (defection ratio) and pore size of membrane. The membrane integrity could be successfully monitored by the PDT. The pressure decay rate varied from $0.002{\sim}0.189kg_f/cm^2hr$ with the initial pressure ranged from 0.2 to $1.0kg_f/cm^2hr$. Higher initial pressure which provided with higher pressure decay rate was preferred to evaluate the defection of membrane fiber. As for the particle removal rate, the Log Removal Rate (LRV) of kaolin solution decreased significantly from 3.78 to 2.31 when one fiber out of 3,200 fibers was cut. The membranes with different pore size were tested to evaluate virus removal efficiency. The virus removal rate of the MF membrane ($0.1{\mu}m$) was about 30% although the poliovirus was smaller than the pore size of the MF membrane, indicating that the removal rate was much lower than Korea Water Works Association (KWWA) certificate LRV of 1.5.

Effect of bicarbonate and progesterone on plasma membrane integrity, acrosome reaction and proportion of fatty acids in boar sperm

  • Park, Choon-Keun;Lee, Sang-Hee
    • Journal of Animal Reproduction and Biotechnology
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    • v.37 no.3
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    • pp.202-208
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    • 2022
  • This study investigated the influence of sodium bicarbonate (NaHCO3) and progesterone on acrosome reaction and proportion of polyunsaturated fatty acid (PUFA) composition boar sperm. The sperm were diluted with semen extender and incubated with NaHCO3 and progesterone at 38℃, 5% CO2 for 6 h. Plasma membrane integrity and acrosome reaction were analyzed using SYBR14/propidium iodide (PI) and FITC-PNA/PI doubling staining method, and proportion of PUFA was analyzed using gas chromatography. In results, Plasma membrane integrity was significantly decreased in 50 mM NaHCO3 group and acrosome reaction was significantly increased by over the 100 mM NaHCO3 group compared to control group (p < 0.05). In addition, progesterone significantly increased decreased plasma membrane integrity at 100 mM progesterone and acrosome reaction at over the 5.0 µM progesterone (p < 0.05), but there was no difference among the 5.0 to 100 µM groups. PUFAs were significantly decreased in 100 mM NaHCO3 and 50 µM progesterone treatments compared to control group. In summary NaHCO3 and progesterone induce acrosome reaction and reduce PUFA composition in boar sperm, therefore, the results maybe help to understand basically knowledge for the acrosome reaction and PUFA composition in boar sperm.