• Title/Summary/Keyword: membrane integrity

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Comparative Phenotypic Analysis of Anabaena sp. PCC 7120 Mutants of Porin-like Genes

  • Schatzle, Hannah;Brouwer, Eva-Maria;Liebhart, Elisa;Stevanovic, Mara;Schleiff, Enrico
    • Journal of Microbiology and Biotechnology
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    • v.31 no.5
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    • pp.645-658
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    • 2021
  • Porins are essential for the viability of Gram-negative bacteria. They ensure the uptake of nutrients, can be involved in the maintenance of outer membrane integrity and define the antibiotic or drug resistance of organisms. The function and structure of porins in proteobacteria is well described, while their function in photoautotrophic cyanobacteria has not been systematically explored. We compared the domain architecture of nine putative porins in the filamentous cyanobacterium Anabaena sp. PCC 7120 and analyzed the seven candidates with predicted OprB-domain. Single recombinant mutants of the seven genes were created and their growth capacity under different conditions was analyzed. Most of the putative porins seem to be involved in the transport of salt and copper, as respective mutants were resistant to elevated concentrations of these substances. In turn, only the mutant of alr2231 was less sensitive to elevated zinc concentrations, while mutants of alr0834, alr4741 and all4499 were resistant to high manganese concentrations. Notably the mutant of alr4550 shows a high sensitivity against harmful compounds, which is indicative for a function related to the maintenance of outer membrane integrity. Moreover, the mutant of all5191 exhibited a phenotype which suggests either a higher nitrate demand or an inefficient nitrogen fixation. The dependency of porin membrane insertion on Omp85 proteins was tested exemplarily for Alr4550, and an enhanced aggregation of Alr4550 was observed in two omp85 mutants. The comparative analysis of porin mutants suggests that the proteins in parts perform distinct functions related to envelope integrity and solute uptake.

Antioxidative Effect of Pyruvate, Taurine and Melatonin, and Relationships of the Evaluation Methods on Boar Sperm Viability for In Vitro Storage (돼지 정자의 체외조본에 있어 Pyruvate, Taurine 및 Melatonin의 항산화효과와 평가방법의 상관관계 분석)

  • Jang, H.Y.;Park, C.K.;Cheong, H.T.;Kim, J.T.;Lee, H.K.;Im, S.K.;Yang, B.K.
    • Journal of Animal Science and Technology
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    • v.49 no.6
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    • pp.729-736
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    • 2007
  • The objective of this study was to investigate the anti-oxidative effects of pyruvate, taurine and melatonin on sperm characteristics(motility, membrane integrity) and lipid peroxidation(LPO) for in vitro storage of boar semen. Semen was treated with various antioxidants such as pyruvate(1mM), taurine(50mM) and melatonin(100nM) with or without 100uM H2O2. Antioxidant treatments were significantly increased the sperm motility when compare to control group in all incubation periods(P≤0.05). Hypoosmotic swelling test(HOST), membrane integrity was similar to the result of motility. In lipid peroxidation measurement by TBA reactions of spermatozoal plasma membrane, malondialdehyde(MDA) level in control and antioxidant treatments were lower than those of antioxidant plus H2O2 or H2O2 treatment for 3 to 6 h incubation period. Relationships of evaluation methods for sperm viability were investigated by motility, membrane integrity and lipid peroxidation. Among evaluation methods, LPO vs motility and membrane integrity vs LPO were negatively correlated(-0.23~-0.92 and -0.68~-0.85), but membrane integrity vs motility was positively correlated (0.53~0.94) in all treatments. These experiments indicate that supplementation of antioxidant to the semen extender can increase the sperm motility and membrane integrity and decrease the lipid peroxidation of spermatozoal plasma membrane. The HOST might be utilized to evaluate the sperm quality instead of lipid peroxidation or motility.

Combined Effects of Metal Coagulants and Monochloramine on Polyamide RO Membrane Performance (금속성 응집제와 모노클로라민의 상호작용이 Polyamide계 RO막 성능에 미치는 영향)

  • Kim, Kyunghwa;Hong, Seungkwan;Park, Chanhyuk;Yoon, Seongro;Hong, Seongpyuo;Lee, Jonghwa
    • Journal of Korean Society of Water and Wastewater
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    • v.20 no.4
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    • pp.637-643
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    • 2006
  • The bench-scale chlorine exposure study was performed to investigate the effect of pretreatment by free chlorine and monochloramine ($NH_2Cl$) on the performance of RO membranes made of polyamide (PA). Feed monochloramination at 2mg/L did not cause significant productivity loss compared to free chlorine. However, metal coagulants reacted with monochloramine, the PA membrane suffered from a gradual loss of membrane integrity by chlorine oxidation, which was characterized as a decrease in salt rejection. Especially, RO membranes exposed to alum coagulants with monochloramine revealed the salt rejection lower than those exposed to iron coagulants. XPS membrane surface analysis demonstrated that the chlorine uptake on the membrane surface increased and carbon peaks were shifted significantly when exposed to alum coagulants with monochloramine.

Accelerating repaired basement membrane after bevacizumab treatment on alkali-burned mouse cornea

  • Lee, Koon-Ja;Lee, Ji-Young;Lee, Sung Ho;Choi, Tae Hoon
    • BMB Reports
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    • v.46 no.4
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    • pp.195-200
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    • 2013
  • To understand the corneal regeneration induced by bevacizumab, we investigated the structure changes of stroma and basement membrane regeneration. A Stick soaked in 0.5 N NaOH onto the mouse cornea and 2.5 mg/ml of bevacizumab was delivered into an alkali-burned cornea (2 ${\mu}l$) by subconjunctival injections at 1 hour and 4 days after injury. At 7 days after injury, basement membrane regeneration was observed by transmission electron microscope. Uneven and thin epithelial basement membrane, light density of hemidesmosomes, and edematous collagen fibril bundles are shown in the alkali-burned cornea. Injured epithelial basement membrane and hemidesmosomes and edematous collagen fibril bundles resulting from alkali-burned mouse cornea was repaired by bevacizumab treatment. This study demonstrates that bevacizumab can play an important role in wound healing in the cornea by accelerating the reestablishment of basement membrane integrity that leads to barriers for scar formation.

Development of a robust bench-scale testing unit for low-pressure membranes used in water treatment

  • Huang, Haiou;Schwab, Kellogg;Jacangelo, Joseph G.
    • Membrane and Water Treatment
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    • v.2 no.2
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    • pp.121-136
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    • 2011
  • A bench-scale test has recently been proposed as a predictive tool to minimize the scope of pilot-scale testing or to optimize the operation of full-scale membrane filtration systems. Consequently, a bench-scale testing unit was developed for this purpose and systematically evaluated in this study. This unit was capable of accommodating commercially available, low pressure, hollow fiber (LPHF) membranes with various configurations for testing under conditions comparable to real-world applications. Reproducibility of this unit in assessing membrane fouling and microbial removal efficiency of LPHF membranes was tested and statistically comparable results were obtained. This unit serves as a useful apparatus for academic researchers and utilities to evaluate the performance of LPHF membranes used for water treatment.

p-Anisaldehyde Exerts Its Antifungal Activity Against Penicillium digitatum and Penicillium italicum by Disrupting the Cell Wall Integrity and Membrane Permeability

  • Che, Jinxin;Chen, Xiumei;Ouyang, Qiuli;Tao, Nengguo
    • Journal of Microbiology and Biotechnology
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    • v.30 no.6
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    • pp.878-884
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    • 2020
  • Penicillium digitatum and P. italicum are the two important postharvest pathogens in citrus, causing about 90% of the total loss of citrus fruit during storage and transportation. Natural fungicides such as essential oils have been widely used instead of chemical fungicides for preventing and controlling postharvest diseases. In this research, p-anisaldehyde exhibited a strong inhibitory effect on P. digitatum and P. italicum, with the minimum inhibitory concentration and minimum fungicidal concentration values of both being 2.00 μl/ml. Additionally, p-anisaldehyde visibly inhibited both the green mold and blue mold development of citrus fruits inoculated with P. digitatum and P. italicum. The mycelia morphologies of these pathogens were greatly altered, and the membrane permeability and cell wall integrity of mycelia were severely disrupted under p-anisaldehyde treatment. These results suggest that the antifungal activity of p-anisaldehyde against P. digitatum and P. italicum can be attributed to the disruption of the cell wall integrity.

The Effect of Cold Shock on Function and Morphology of Dog Epididymal Spermatozoa (개에서 cold shock이 정소상체유래의 정자의 기능과 형태에 미치는 영향)

  • Yu Il-jeoung
    • Journal of Veterinary Clinics
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    • v.21 no.4
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    • pp.329-335
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    • 2004
  • Dog spermatozoa were recovered from the caudae epididymides of 23 domestic dogs which were 11 pure breed and 12 mix-breed dogs ranging in age from 0.6 to 3 years. The experimental designs were as follows: 1) the effect of chilling to 0. 10 or 37$^{\circ}C$. 2) the kinetics of chilling injury at 0 or 4$^{\circ}C$, and 3) the effect of sugars at $0^{\circ}C$. Viable spermatozoa were recovered by percoll gradient separation and adjusted to 5${\times}$10$^{7}$ spermatozoa/ml. In experiment 1, spermatozoa were diluted with 0.33 M glucose supplemented with 3% BSA (G-BSA) at 1:2 dilution. Spermatozoa were loaded into straws and exposed at 0, 10 or 37$^{\circ}C$ for 30 min. In experiment 2, spermatozoa were prepared as the experiment 1 and exposed for 0.5, 5, 15, or 30 min at 0 or 4$^{\circ}C$. In experiment 3, spermatozoa were diluted with different sugars (0.33 M galactose, glucose, fructose, mannitol, lactose, sucrose, raffinose) and cooled to $0^{\circ}C$ for 30 min. Sperm membrane integrity, motility and acrosome integrity were assayed after rewarming at 37$^{\circ}C$ for 5 min. Sperm motility and membrane integrity abruptly decreased with decreasing temperature but acrosome integrity gradually decreased (P<0.05). Sperm motility was more sensitive to cold shock than membrane integrity and acrosome integrity. Spermatozoa cooled to $0^{\circ}C$ were more damaged than those at 4$^{\circ}C$. Sperm motility was not different among exposed times at both. 0 and 4$^{\circ}C$. However, membrane integrity of spermatozoa exposed for 30 min at both 0 and 4$^{\circ}C$ was significantly lower (P<0.05). Spermatozoa diluted in 0.33 M fructose or galactose showed lower motility and higher morphological abnormality with coiled tail at $0^{\circ}C$. These sperm characteristics were strongly related. These results indicate that dog epididymal spermatozoa are relatively sensitive to rapid cooling and higher morphological abnormality at $0^{\circ}C$ was shown in spermatozoa diluted in fructose and galactose.

Effect of Alpha-Linolenic Acid with Bovine Serum Albumin or Methyl-Beta-Cyclodextrin on Membrane Integrity and Oxidative Stress of Frozen-Thawed Boar Sperm

  • Lee, Won-Hee;Kim, Wook-Hwan;Cheong, Hee-Tae;Yang, Boo-Keun;Park, Choon-Keun
    • Development and Reproduction
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    • v.23 no.1
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    • pp.11-19
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    • 2019
  • The study was conducted to investigate the effects of alpha-linolenic acid (ALA) combined with bovine serum albumin (BSA) or methyl-beta-cyclodextrin (MBCD) on plasma and acrosomal membrane damages, mitochondrial activity, morphological abnormality, motility, and oxidative stress in frozen-thawed boar sperm. In previous our study, 3 ng/mL ALA had been shown protective effect during freezing process of boar sperm. Therefore, we used 3 ng/mL ALA in present study and ALA was combined with same molar ratio of BSA or MBCD (ALA+BSA and ALA+MBCD, respectively). To confirm the effect of two carrier proteins, same volume of BSA and MBCD without ALA were added during cryopreservation. Membrane damage, mitochondrial activity, reactive oxygen species (ROS) and lipid peroxidation (LPO) levels were measured using flow cytometry, and movement of sperm tail as motility parameter and morphological abnormality were observed under light microscope. In results, all of sperm parameters were enhanced by ALA combined with BSA or MBCD compared to control groups (p<0.05). Mitochondrial activity, morphological abnormality, ROS and LPO levels in ALA+BSA or MBCD groups were no significant difference compared with ALA, BSA and MBCD treatment groups. On the other hand, plasma and acrosomal membrane intact, and sperm motility in ALA+MBCD group were higher than single treatment groups (p<0.05), whereas ALA+BSA did not differ. Our findings indicate that carrier proteins such as BSA and MBCD could improve the effect of ALA during cryopreservation of boar sperm, and treatment of ALA with carrier proteins enhance membrane integrity, mitochondrial activity through reduction of ROS-induced LPO.

Selectivity and structural integrity of a nanofiltration membrane for treatment of liquid waste containing uranium

  • Oliveira, Elizabeth E.M.;Barbosa, Celina C.R.;Afonso, Julio C.
    • Membrane and Water Treatment
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    • v.3 no.4
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    • pp.231-242
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    • 2012
  • The performance of a nanofiltration membrane for treatment of a low-level radioactive liquid waste was investigated through static and dynamic tests. The liquid waste ("carbonated water") was obtained during conversion of $UF_6$ to $UO_2$. In the static tests membrane samples were immersed in the waste for 24, 48 or 72 h. The transport properties of the samples (hydraulic permeability, permeate flow, selectivity) were evaluated before and after immersion in the waste. In the dynamic tests the waste was permeated in a permeation flow front system under 0.5 MPa, to determine the selectivity of NF membranes to uranium. The surface layer of the membrane was characterized by zeta potential, field emission microscopy, atomic force spectroscopy and infrared spectroscopy. The static test showed that the pore size distribution of the selective layer was altered, but the membrane surface charge was not significantly changed. 99% of uranium was rejected after the dynamic tests.

Effect of Feed Monochloramination on Performance of RO Membranes (NH2Cl 사용으로 인한 RO 막의 성능 향상)

  • Hong, Seungkwan;Reiss, Robert;Taylor, James S.;Beverly, Sharon
    • Journal of Korean Society of Water and Wastewater
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    • v.19 no.3
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    • pp.312-317
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    • 2005
  • The 15 month pilot study was performed to investigate the effect of pretreatment by monochloramine ($NH_2Cl$) on the performance of RO membranes made of cellulose acetate (CA) and polyamide (PA). Both RO membranes experienced severe biological fouling without any pretreatment during the treatment of highly organic surface water in Florida, USA. Feed monochloramination at 5 mg/L significantly minimized productivity loss by effective control of biofouling. The CA membrane did not show any structural damages by monochloramine, while the PA membrane suffered from a gradual loss of membrane integrity by chlorine oxidation, which was characterized as an increase in productivity and a decrease in selectivity. The degradation of PA membrane increased with increasing monochloramine dose.