Computational molecular modeling and structural rationalization for the design of a drug-loaded PLLA/PVA biopolymeric membrane

International Nuclear Information System (INIS)

Sibeko, B; Pillay, V; Choonara, Y E; Khan, R A; Danckwerts, M P; Modi, G; Iyuke, S E; Naidoo, D

2009-01-01

The purpose of this study was to design, characterize and assess the influence of triethanolamine (TEA) on the physicomechanical properties and release of methotrexate (MTX) from a composite biopolymeric membrane. Conjugated poly(L-lactic acid) (PLLA) and poly(vinyl alcohol) (PVA) membranes were prepared by immersion precipitation with and without the addition of TEA. Drug entrapment efficiency (DEE) and release studies were performed in phosphate buffered saline (pH 7.4, 37 deg. C). Scanning electron microscopy elucidated the membrane surface morphology. Computational and structural molecular modeling rationalized the potential mechanisms of membrane formation and MTX release. Bi-axial force-distance (F-D) extensibility profiles were generated to determine the membrane toughness, elasticity and fracturability. Membranes were significantly toughened by the addition of TEA as a discrete rubbery phase within the co-polymer matrix. MTX-TEA-PLLA-PVA membranes were tougher (F = 89 N) and more extensible (D = 8.79 mm) compared to MTX-PLLA-PVA (F = 35 N, D = 3.7 mm) membranes as a greater force of extension and fracture distance were required (N = 10). DEE values were relatively high (>80%, N = 5) for both formulations. Photomicrographs revealed distinct crystalline layered morphologies with macro-pores. MTX was released by tri-phasic kinetics with a lower fractional release of MTX from MTX-TEA-PLLA-PVA membranes compared to MTX-PLLA-PVA. TEA provided a synergistic approach to improving the membrane physicomechanical properties and modulation of MTX release. The composite biopolymeric membrane may therefore be suitable for the novel delivery of MTX in the treatment of chronic primary central nervous system lymphoma.

Structure and Orientation of Bovine Lactoferrampin in the Mimetic Bacterial Membrane as Revealed by Solid-State NMR and Molecular Dynamics Simulation

Science.gov (United States)

Tsutsumi, Atsushi; Javkhlantugs, Namsrai; Kira, Atsushi; Umeyama, Masako; Kawamura, Izuru; Nishimura, Katsuyuki; Ueda, Kazuyoshi; Naito, Akira

2012-01-01

Bovine lactoferrampin (LFampinB) is a newly discovered antimicrobial peptide found in the N1-domain of bovine lactoferrin (268–284), and consists of 17 amino-acid residues. It is important to determine the orientation and structure of LFampinB in bacterial membranes to reveal the antimicrobial mechanism. We therefore performed 13C and 31P NMR, 13C-31P rotational echo double resonance (REDOR), potassium ion-selective electrode, and quartz-crystal microbalance measurements for LFampinB with mimetic bacterial membrane and molecular-dynamics simulation in acidic membrane. 31P NMR results indicated that LFampinB caused a defect in mimetic bacterial membranes. Ion-selective electrode measurements showed that ion leakage occurred for the mimetic bacterial membrane containing cardiolipin. Quartz-crystal microbalance measurements revealed that LFampinB had greater affinity to acidic phospholipids than that to neutral phospholipids. 13C DD-MAS and static NMR spectra showed that LFampinB formed an α-helix in the N-terminus region and tilted 45° to the bilayer normal. REDOR dephasing patterns between carbonyl carbon nucleus in LFampinB and phosphorus nuclei in lipid phosphate groups were measured by 13C-31P REDOR and the results revealed that LFampinB is located in the interfacial region of the membrane. Molecular-dynamics simulation showed the tilt angle to be 42° and the rotation angle to be 92.5° for Leu3, which are in excellent agreement with the experimental values. PMID:23083717

Axionic membranes

International Nuclear Information System (INIS)

Aurilia, A.; Spallucci, E.

1992-01-01

A metal ring removed from a soap-water solution encloses a film of soap which can be mathematically described as a minimal surface having the ring as its only boundary. This is known to everybody. In this letter we suggest a relativistic extension of the above fluidodynamic system where the soap film is replaced by a Kalb-Ramand gauge potential B μν (x) and the ring by a closed string. The interaction between the B μν field and the string current excites a new configuration of the system consisting of a relativistic membrane bounded by the string. We call such a classical solution of the equation of motion an axionic membrane. As a dynamical system, the axionic membrane admits a Hamilton-Jacobi formulation which is an extension of the HJ theory of electromagnetic strings. (orig.)

Proteomic analysis of Herbaspirillum seropedicae reveals ammonium-induced AmtB-dependent membrane sequestration of PII proteins.

Science.gov (United States)

Huergo, Luciano F; Noindorf, Lilian; Gimenes, Camila; Lemgruber, Renato S P; Cordellini, Daniela F; Falarz, Lucas J; Cruz, Leonardo M; Monteiro, Rose A; Pedrosa, Fábio O; Chubatsu, Leda S; Souza, Emanuel M; Steffens, Maria B R

2010-07-01

This study was aimed at describing the spectrum and dynamics of proteins associated with the membrane in the nitrogen-fixing bacterium Herbaspirillum seropedicae according to the availability of fixed nitrogen. Using two-dimensional electrophoresis we identified 79 protein spots representing 45 different proteins in the membrane fraction of H. seropedicae. Quantitative analysis of gel images of membrane extracts indicated two spots with increased levels when cells were grown under nitrogen limitation in comparison with nitrogen sufficiency; these spots were identified as the GlnK protein and as a conserved noncytoplasmic protein of unknown function which was encoded in an operon together with GlnK and AmtB. Comparison of gel images of membrane extracts from cells grown under nitrogen limitation or under the same regime but collected after an ammonium shock revealed two proteins, GlnB and GlnK, with increased levels after the shock. The P(II) proteins were not present in the membrane fraction of an amtB mutant. The results reported here suggest that changes in the cellular localization of P(II) might play a role in the control of nitrogen metabolism in H. seropedicae.

Economics of membrane occupancy and respiro-fermentation

Science.gov (United States)

Zhuang, Kai; Vemuri, Goutham N; Mahadevan, Radhakrishnan

2011-01-01

The simultaneous utilization of efficient respiration and inefficient fermentation even in the presence of abundant oxygen is a puzzling phenomenon commonly observed in bacteria, yeasts, and cancer cells. Despite extensive research, the biochemical basis for this phenomenon remains obscure. We hypothesize that the outcome of a competition for membrane space between glucose transporters and respiratory chain (which we refer to as economics of membrane occupancy) proteins influences respiration and fermentation. By incorporating a sole constraint based on this concept in the genome-scale metabolic model of Escherichia coli, we were able to simulate respiro-fermentation. Further analysis of the impact of this constraint revealed differential utilization of the cytochromes and faster glucose uptake under anaerobic conditions than under aerobic conditions. Based on these simulations, we propose that bacterial cells manage the composition of their cytoplasmic membrane to maintain optimal ATP production by switching between oxidative and substrate-level phosphorylation. These results suggest that the membrane occupancy constraint may be a fundamental governing constraint of cellular metabolism and physiology, and establishes a direct link between cell morphology and physiology. PMID:21694717

Olefins-selective asymmetric carbon molecular sieve hollow fiber membranes for hybrid membrane-distillation processes for olefin/paraffin separations

KAUST Repository

Xu, Liren

2012-12-01

In this paper, the development of asymmetric carbon molecular sieve (CMS) hollow fiber membranes and advanced processes for olefin/paraffin separations based on the CMS membranes are reported. Membrane-based olefin/paraffin separations have been pursued extensively over the past decades. CMS membranes are promising to exceed the performance upper bound of polymer materials and have demonstrated excellent stability for gas separations. Previously, a substructure collapse phenomenon was found in Matrimid ® precursor derived CMS fiber. To overcome the permeance loss due to the increased separation layer thickness, 6FDA-DAM and 6FDA/BPDA-DAM precursors were selected as potential new precursors for carbon membrane formation. Defect-free asymmetric 6FDA-DAM and 6FDA/BPDA-DAM hollow fibers were successfully fabricated from a dry-jet/wet-quench spinning process. Polymer rigidity, glass-rubber transition and asymmetric morphology were correlated. CMS hollow fiber membranes produced from 6FDA-polymer precursors showed significant improvement in permeance for ethylene/ethane and propylene/propane separations. Further studies revealed that the CMS membranes are olefins-selective, which means the membranes are able to effectively separate olefins (ethylene and propylene) from paraffins (ethane and propane). This unique feature of CMS materials enables advanced hybrid membrane-distillation process designs. By using the olefins-selective membranes, these new processes may provide advantages over previously proposed retrofitting concepts. Further applications of the membranes are explored for hydrocarbons processes. Significant energy savings and even reduced footprint may be achieved in olefins production units. © 2012 Elsevier B.V.

Extensive in silico analysis of Mimivirus coded Rab GTPase homolog suggests a possible role in virion membrane biogenesis

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Amrutraj eZade

2015-09-01

Full Text Available Rab GTPases are the key regulators of intracellular membrane trafficking in eukaryotes. Many viruses and intracellular bacterial pathogens have evolved to hijack the host Rab GTPase functions, mainly through activators and effector proteins, for their benefit. Acanthamoeba polyphaga mimivirus (APMV is one of the largest viruses and belongs to the monophyletic clade of nucleo-cytoplasmic large DNA viruses (NCLDV. The inner membrane lining is integral to the APMV virion structure. APMV assembly involves extensive host membrane modifications, like vesicle budding and fusion, leading to the formation of a membrane sheet that is incorporated into the virion. Intriguingly, APMV and all group I members of the Mimiviridae family code for a putative Rab GTPase protein. APMV is the first reported virus to code for a Rab GTPase (encoded by R214 gene. Our thorough in silico analysis of the subfamily specific (SF region of Mimiviridae Rab GTPase sequences suggests that they are related to Rab5, a member of the group II Rab GTPases, of lower eukaryotes. Because of their high divergence from the existing three isoforms, A, B and C of the Rab5-family, we suggest that Mimiviridae Rabs constitute a new isoform, Rab5D. Phylogenetic analysis indicated probable horizontal acquisition from a lower eukaryotic ancestor followed by selection and divergence. Furthermore, interaction network analysis suggests that vps34 (a Class III P13K homolog, coded by APMV L615, Atg-8 and dynamin (host proteins are recruited by APMV Rab GTPase during capsid assembly. Based on these observations, we hypothesize that APMV Rab plays a role in the acquisition of inner membrane during virion assembly.

Periplasmic quality control in biogenesis of outer membrane proteins.

Science.gov (United States)

Lyu, Zhi Xin; Zhao, Xin Sheng

2015-04-01

The β-barrel outer membrane proteins (OMPs) are integral membrane proteins that reside in the outer membrane of Gram-negative bacteria and perform a diverse range of biological functions. Synthesized in the cytoplasm, OMPs must be transported across the inner membrane and through the periplasmic space before they are assembled in the outer membrane. In Escherichia coli, Skp, SurA and DegP are the most prominent factors identified to guide OMPs across the periplasm and to play the role of quality control. Although extensive genetic and biochemical analyses have revealed many basic functions of these periplasmic proteins, the mechanism of their collaboration in assisting the folding and insertion of OMPs is much less understood. Recently, biophysical approaches have shed light on the identification of the intricate network. In the present review, we summarize recent advances in the characterization of these key factors, with a special emphasis on the multifunctional protein DegP. In addition, we present our proposed model on the periplasmic quality control in biogenesis of OMPs.

Two-step membrane binding by the bacterial SRP receptor enable efficient and accurate Co-translational protein targeting.

Science.gov (United States)

Hwang Fu, Yu-Hsien; Huang, William Y C; Shen, Kuang; Groves, Jay T; Miller, Thomas; Shan, Shu-Ou

2017-07-28

The signal recognition particle (SRP) delivers ~30% of the proteome to the eukaryotic endoplasmic reticulum, or the bacterial plasma membrane. The precise mechanism by which the bacterial SRP receptor, FtsY, interacts with and is regulated at the target membrane remain unclear. Here, quantitative analysis of FtsY-lipid interactions at single-molecule resolution revealed a two-step mechanism in which FtsY initially contacts membrane via a Dynamic mode, followed by an SRP-induced conformational transition to a Stable mode that activates FtsY for downstream steps. Importantly, mutational analyses revealed extensive auto-inhibitory mechanisms that prevent free FtsY from engaging membrane in the Stable mode; an engineered FtsY pre-organized into the Stable mode led to indiscriminate targeting in vitro and disrupted FtsY function in vivo. Our results show that the two-step lipid-binding mechanism uncouples the membrane association of FtsY from its conformational activation, thus optimizing the balance between the efficiency and fidelity of co-translational protein targeting.

Membrane order in the plasma membrane and endocytic recycling compartment.

Science.gov (United States)

Iaea, David B; Maxfield, Frederick R

2017-01-01

The cholesterol content of membranes plays an important role in organizing membranes for signal transduction and protein trafficking as well as in modulating the biophysical properties of membranes. While the properties of model or isolated membranes have been extensively studied, there has been little evaluation of internal membranes in living cells. Here, we use a Nile Red based probe, NR12S, and ratiometric live cell imaging, to analyze the membrane order of the plasma membrane and endocytic recycling compartment. We find that after a brief incubation to allow endocytosis, NR12S is distributed between the plasma membrane and the endocytic recycling compartment. The NR12S reports that the endocytic recycling compartment is more highly ordered than the plasma membrane. We also find that the plasma membrane and the endocytic recycling compartment are differentially affected by altering cellular cholesterol levels. The membrane order of the plasma membrane, but not the endocytic recycling compartment, is altered significantly when cellular cholesterol content is increased or decreased by 20%. These results demonstrate that changes in cellular cholesterol differentially alter membrane order within different organelles.

Nanoscale Electric Characteristics and Oriented Assembly of Halobacterium salinarum Membrane Revealed by Electric Force Microscopy

Directory of Open Access Journals (Sweden)

Denghua Li

2016-11-01

Full Text Available Purple membranes (PM of the bacteria Halobacterium salinarum are a unique natural membrane where bacteriorhodopsin (BR can convert photon energy and pump protons. Elucidating the electronic properties of biomembranes is critical for revealing biological mechanisms and developing new devices. We report here the electric properties of PMs studied by using multi-functional electric force microscopy (EFM at the nanoscale. The topography, surface potential, and dielectric capacity of PMs were imaged and quantitatively measured in parallel. Two orientations of PMs were identified by EFM because of its high resolution in differentiating electrical characteristics. The extracellular (EC sides were more negative than the cytoplasmic (CP side by 8 mV. The direction of potential difference may facilitate movement of protons across the membrane and thus play important roles in proton pumping. Unlike the side-dependent surface potentials observed in PM, the EFM capacitive response was independent of the side and was measured to be at a dC/dz value of ~5.25 nF/m. Furthermore, by modification of PM with de novo peptides based on peptide-protein interaction, directional oriented PM assembly on silicon substrate was obtained for technical devices. This work develops a new method for studying membrane nanoelectronics and exploring the bioelectric application at the nanoscale.

Quantitative membrane proteomics reveals a role for tetraspanin enriched microdomains during entry of human cytomegalovirus.

Directory of Open Access Journals (Sweden)

Kasinath Viswanathan

Full Text Available Human cytomegalovirus (HCMV depends on and modulates multiple host cell membrane proteins during each stage of the viral life cycle. To gain a global view of the impact of HCMV-infection on membrane proteins, we analyzed HCMV-induced changes in the abundance of membrane proteins in fibroblasts using stable isotope labeling with amino acids (SILAC, membrane fractionation and protein identification by two-dimensional liquid chromatography and tandem mass spectrometry. This systematic approach revealed that CD81, CD44, CD98, caveolin-1 and catenin delta-1 were down-regulated during infection whereas GRP-78 was up-regulated. Since CD81 downregulation was also observed during infection with UV-inactivated virus we hypothesized that this tetraspanin is part of the viral entry process. Interestingly, additional members of the tetraspanin family, CD9 and CD151, were also downregulated during HCMV-entry. Since tetraspanin-enriched microdomains (TEM cluster host cell membrane proteins including known CMV receptors such as integrins, we studied whether TEMs are required for viral entry. When TEMs were disrupted with the cholesterol chelator methyl-β-cylcodextrin, viral entry was inhibited and this inhibition correlated with reduced surface levels of CD81, CD9 and CD151, whereas integrin levels remained unchanged. Furthermore, simultaneous siRNA-mediated knockdown of multiple tetraspanins inhibited viral entry whereas individual knockdown had little effect suggesting essential, but redundant roles for individual tetraspanins during entry. Taken together, our data suggest that TEM act as platforms for receptors utilized by HCMV for entry into cells.

Super-resolution microscopy reveals the insulin-resistance-regulated reorganization of GLUT4 on plasma membranes.

Science.gov (United States)

Gao, Lan; Chen, Junling; Gao, Jing; Wang, Hongda; Xiong, Wenyong

2017-01-15

GLUT4 (also known as SLC2A4) is essential for glucose uptake in skeletal muscles and adipocytes, which play central roles in whole-body glucose metabolism. Here, using direct stochastic optical reconstruction microscopy (dSTORM) to investigate the characteristics of plasma-membrane-fused GLUT4 at the single-molecule level, we have demonstrated that insulin and insulin resistance regulate the spatial organization of GLUT4 in adipocytes. Stimulation with insulin shifted the balance of GLUT4 on the plasma membrane toward a more dispersed configuration. In contrast, insulin resistance induced a more clustered distribution of GLUT4 and increased the mean number of molecules per cluster. Furthermore, our data demonstrate that the F 5 QQI motif and lipid rafts mediate the maintenance of GLUT4 clusters on the plasma membrane. Mutation of F 5 QQI (F 5 QQA-GLUT4) induced a more clustered distribution of GLUT4; moreover, destruction of lipid rafts in adipocytes expressing F 5 QQA-GLUT4 dramatically decreased the percentage of large clusters and the mean number of molecules per cluster. In conclusion, our data clarify the effects of insulin stimulation or insulin resistance on GLUT4 reorganization on the plasma membrane and reveal new pathogenic mechanisms of insulin resistance. © 2017. Published by The Company of Biologists Ltd.

Membrane distillation for milk concentration

NARCIS (Netherlands)

Moejes, S.N.; Romero Guzman, Maria; Hanemaaijer, J.H.; Barrera, K.H.; Feenstra, L.; Boxtel, van A.J.B.

2015-01-01

Membrane distillation is an emerging technology to concentrate liquid products while producing high quality water as permeate. Application for desalination has been studied extensively the past years, but membrane distillation has also potential to produce concentrated food products like

Dynamic membrane interactions of antibacterial and antifungal biomolecules, and amyloid peptides, revealed by solid-state NMR spectroscopy.

Science.gov (United States)

Naito, Akira; Matsumori, Nobuaki; Ramamoorthy, Ayyalusamy

2018-02-01

A variety of biomolecules acting on the cell membrane folds into a biologically active structure in the membrane environment. It is, therefore, important to determine the structures and dynamics of such biomolecules in a membrane environment. While several biophysical techniques are used to obtain low-resolution information, solid-state NMR spectroscopy is one of the most powerful means for determining the structure and dynamics of membrane bound biomolecules such as antibacterial biomolecules and amyloidogenic proteins; unlike X-ray crystallography and solution NMR spectroscopy, applications of solid-state NMR spectroscopy are not limited by non-crystalline, non-soluble nature or molecular size of membrane-associated biomolecules. This review article focuses on the applications of solid-state NMR techniques to study a few selected antibacterial and amyloid peptides. Solid-state NMR studies revealing the membrane inserted bent α-helical structure associated with the hemolytic activity of bee venom melittin and the chemical shift oscillation analysis used to determine the transmembrane structure (with α-helix and 3 10 -helix in the N- and C-termini, respectively) of antibiotic peptide alamethicin are discussed in detail. Oligomerization of an amyloidogenic islet amyloid polypeptide (IAPP, or also known as amylin) resulting from its aggregation in a membrane environment, molecular interactions of the antifungal natural product amphotericin B with ergosterol in lipid bilayers, and the mechanism of lipid raft formation by sphingomyelin studied using solid state NMR methods are also discussed in this review article. This article is part of a Special Issue entitled "Biophysical Exploration of Dynamical Ordering of Biomolecular Systems" edited by Dr. Koichi Kato. Copyright © 2017 Elsevier B.V. All rights reserved.

Membrane curvature enables N-Ras lipid anchor sorting to liquid-ordered membrane phases

DEFF Research Database (Denmark)

Larsen, Jannik Bruun; Jensen, Martin Borch; Bhatia, Vikram Kjøller

2015-01-01

Trafficking and sorting of membrane-anchored Ras GTPases are regulated by partitioning between distinct membrane domains. Here, in vitro experiments and microscopic molecular theory reveal membrane curvature as a new modulator of N-Ras lipid anchor and palmitoyl chain partitioning. Membrane...

Secretagogue stimulation of neurosecretory cells elicits filopodial extensions uncovering new functional release sites.

Science.gov (United States)

Papadopulos, Andreas; Martin, Sally; Tomatis, Vanesa M; Gormal, Rachel S; Meunier, Frederic A

2013-12-04

Regulated exocytosis in neurosecretory cells relies on the timely fusion of secretory granules (SGs) with the plasma membrane. Secretagogue stimulation leads to an enlargement of the cell footprint (surface area in contact with the coverslip), an effect previously attributed to exocytic fusion of SGs with the plasma membrane. Using total internal reflection fluorescence microscopy, we reveal the formation of filopodia-like structures in bovine chromaffin and PC12 cells driving the footprint expansion, suggesting the involvement of cortical actin network remodeling in this process. Using exocytosis-incompetent PC12 cells, we demonstrate that footprint enlargement is largely independent of SG fusion, suggesting that vesicular exocytic fusion plays a relatively minor role in filopodial expansion. The footprint periphery, including filopodia, undergoes extensive F-actin remodeling, an effect abolished by the actomyosin inhibitors cytochalasin D and blebbistatin. Imaging of both Lifeact-GFP and the SG marker protein neuropeptide Y-mCherry reveals that SGs actively translocate along newly forming actin tracks before undergoing fusion. Together, these data demonstrate that neurosecretory cells regulate the number of SGs undergoing exocytosis during sustained stimulation by controlling vesicular mobilization and translocation to the plasma membrane through actin remodeling. Such remodeling facilitates the de novo formation of fusion sites.

Methylation and in vivo expression of the surface-exposed Leptospira interrogans outer-membrane protein OmpL32.

Science.gov (United States)

Eshghi, Azad; Pinne, Marija; Haake, David A; Zuerner, Richard L; Frank, Ami; Cameron, Caroline E

2012-03-01

Recent studies have revealed that bacterial protein methylation is a widespread post-translational modification that is required for virulence in selected pathogenic bacteria. In particular, altered methylation of outer-membrane proteins has been shown to modulate the effectiveness of the host immune response. In this study, 2D gel electrophoresis combined with MALDI-TOF MS identified a Leptospira interrogans serovar Copenhageni strain Fiocruz L1-130 protein, corresponding to ORF LIC11848, which undergoes extensive and differential methylation of glutamic acid residues. Immunofluorescence microscopy implicated LIC11848 as a surface-exposed outer-membrane protein, prompting the designation OmpL32. Indirect immunofluorescence microscopy of golden Syrian hamster liver and kidney sections revealed expression of OmpL32 during colonization of these organs. Identification of methylated surface-exposed outer-membrane proteins, such as OmpL32, provides a foundation for delineating the role of this post-translational modification in leptospiral virulence.

Membrane potentials of membranes with fixed ionic sites

NARCIS (Netherlands)

van den Berg, Albert; van der Wal, P.D.; van der Wal, P.D.; Skowronska-ptasinska, M.; Sudhölter, E.J.R.; Sudholter, Ernst; Bergveld, Piet; Reinhoudt, David

1990-01-01

A theoretical model has been developed to simulate the formation of a membrane potential as a function of physically accessible parameters. The description is an extension of the well-known Teorell-Meyer-Sievers (TMS) model, now including free and fixed ionic sites and free and fixed neutral

Integrative Analysis of Subcellular Quantitative Proteomics Studies Reveals Functional Cytoskeleton Membrane-Lipid Raft Interactions in Cancer.

Science.gov (United States)

Shah, Anup D; Inder, Kerry L; Shah, Alok K; Cristino, Alexandre S; McKie, Arthur B; Gabra, Hani; Davis, Melissa J; Hill, Michelle M

2016-10-07

Lipid rafts are dynamic membrane microdomains that orchestrate molecular interactions and are implicated in cancer development. To understand the functions of lipid rafts in cancer, we performed an integrated analysis of quantitative lipid raft proteomics data sets modeling progression in breast cancer, melanoma, and renal cell carcinoma. This analysis revealed that cancer development is associated with increased membrane raft-cytoskeleton interactions, with ∼40% of elevated lipid raft proteins being cytoskeletal components. Previous studies suggest a potential functional role for the raft-cytoskeleton in the action of the putative tumor suppressors PTRF/Cavin-1 and Merlin. To extend the observation, we examined lipid raft proteome modulation by an unrelated tumor suppressor opioid binding protein cell-adhesion molecule (OPCML) in ovarian cancer SKOV3 cells. In agreement with the other model systems, quantitative proteomics revealed that 39% of OPCML-depleted lipid raft proteins are cytoskeletal components, with microfilaments and intermediate filaments specifically down-regulated. Furthermore, protein-protein interaction network and simulation analysis showed significantly higher interactions among cancer raft proteins compared with general human raft proteins. Collectively, these results suggest increased cytoskeleton-mediated stabilization of lipid raft domains with greater molecular interactions as a common, functional, and reversible feature of cancer cells.

Functionalization of a Hydrophilic Commercial Membrane Using Inorganic-Organic Polymers Coatings for Membrane Distillation

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Lies Eykens

2017-06-01

Full Text Available Membrane distillation is a thermal separation technique using a microporous hydrophobic membrane. One of the concerns with respect to the industrialization of the technique is the development of novel membranes. In this paper, a commercially available hydrophilic polyethersulfone membrane with a suitable structure for membrane distillation was modified using available hydrophobic coatings using ORMOCER® technology to obtain a hydrophobic membrane that can be applied in membrane distillation. The surface modification was performed using a selection of different components, concentrations, and application methods. The resulting membranes can have two hydrophobic surfaces or a hydrophobic and hydrophilic surface depending on the application method. An extensive characterization procedure confirmed the suitability of the coating technique and the obtained membranes for membrane distillation. The surface contact angle of water could be increased from 27° up to 110°, and fluxes comparable to membranes commonly used for membrane distillation were achieved under similar process conditions. A 100 h test demonstrated the stability of the coating and the importance of using sufficiently stable base membranes.

Revealing Linear Aggregates of Light Harvesting Antenna Proteins in Photosynthetic Membranes

OpenAIRE

He, Yufan; Zeng, Xiaohua; Mukherjee, Saptarshi; Rajapaksha, Suneth; Kaplan, Samuel; Lu, H. Peter

2010-01-01

How light energy is harvested in a natural photosynthetic membrane through energy transfer is closely related to the stoichiometry and arrangement of light harvesting antenna proteins in the membrane. The specific photosynthetic architecture facilitates a rapid and efficient energy transfer among the light harvesting proteins (LH2 and LH1) and to the reaction center. Here we report the identification of linear aggregates of light harvesting proteins, LH2, in the photosynthetic membranes under...

Fabrication of electrospun nanofibrous membranes for membrane distillation application

KAUST Repository

Francis, Lijo

2013-02-01

Nanofibrous membranes of Matrimid have been successfully fabricated using an electrospinning technique under optimized conditions. Nanofibrous membranes are found to be highly hydrophobic with a high water contact angle of 130°. Field emission scanning electron microscopy and pore size distribution analysis revealed the big pore size structure of electrospun membranes to be greater than 2 μm and the pore size distribution is found to be narrow. Flat sheet Matrimid membranes were fabricated via casting followed by phase inversion. The morphology, pore size distribution, and water contact angle were measured and compared with the electrospun membranes. Both membranes fabricated by electrospinning and phase inversion techniques were tested in a direct contact membrane distillation process. Electrospun membranes showed high water vapor flux of 56 kg/m2-h, which is very high compared to the casted membrane as well as most of the fabricated and commercially available highly hydrophobic membranes. ©2013 Desalination Publications.

Design of block copolymer membranes using segregation strength trend lines

KAUST Repository

Sutisna, Burhannudin

2016-05-18

Block copolymer self-assembly and non-solvent induced phase separation are now being combined to fabricate membranes with narrow pore size distribution and high porosity. The method has the potential to be used with a broad range of tailor-made block copolymers to control functionality and selectivity for specific separations. However, the extension of this process to any new copolymer is challenging and time consuming, due to the complex interplay of influencing parameters, such as solvent composition, polymer molecular weights, casting solution concentration, and evaporation time. We propose here an effective method for designing new block copolymer membranes. The method consists of predetermining a trend line for the preparation of isoporous membranes, obtained by computing solvent properties, interactions and copolymer block sizes for a set of successful systems and using it as a guide to select the preparation conditions for new membranes. We applied the method to membranes based on poly(styrene-b-ethylene oxide) diblocks and extended it to newly synthesized poly(styrene-b-2-vinyl pyridine-b-ethylene oxide) (PS-b-P2VP-b-PEO) terpolymers. The trend line method can be generally applied to other new systems and is expected to dramatically shorten the path of isoporous membrane manufacture. The PS-b-P2VP-b-PEO membrane formation was investigated by in situ Grazing Incident Small Angle X-ray Scattering (GISAXS), which revealed a hexagonal micelle order with domain spacing clearly correlated to the membrane interpore distances.

Analyses in zebrafish embryos reveal that nanotoxicity profiles are dependent on surface-functionalization controlled penetrance of biological membranes.

Science.gov (United States)

Paatero, Ilkka; Casals, Eudald; Niemi, Rasmus; Özliseli, Ezgi; Rosenholm, Jessica M; Sahlgren, Cecilia

2017-08-21

Mesoporous silica nanoparticles (MSNs) are extensively explored as drug delivery systems, but in depth understanding of design-toxicity relationships is still scarce. We used zebrafish (Danio rerio) embryos to study toxicity profiles of differently surface functionalized MSNs. Embryos with the chorion membrane intact, or dechoroniated embryos, were incubated or microinjected with amino (NH 2 -MSNs), polyethyleneimine (PEI-MSNs), succinic acid (SUCC-MSNs) or polyethyleneglycol (PEG-MSNs) functionalized MSNs. Toxicity was assessed by viability and cardiovascular function. NH 2 -MSNs, SUCC-MSNs and PEG-MSNs were well tolerated, 50 µg/ml PEI-MSNs induced 100% lethality 48 hours post fertilization (hpf). Dechoroniated embryos were more sensitive and 10 µg/ml PEI-MSNs reduced viability to 5% at 96hpf. Sensitivity to PEG- and SUCC-, but not NH 2 -MSNs, was also enhanced. Typically cardiovascular toxicity was evident prior to lethality. Confocal microscopy revealed that PEI-MSNs penetrated into the embryos whereas PEG-, NH2- and SUCC-MSNs remained aggregated on the skin surface. Direct exposure of inner organs by microinjecting NH 2 -MSNs and PEI-MSNs demonstrated that the particles displayed similar toxicity indicating that functionalization affects the toxicity profile by influencing penetrance through biological barriers. The data emphasize the need for careful analyses of toxicity mechanisms in relevant models and constitute an important knowledge step towards the development of safer and sustainable nanotherapies.

Diffusion of Integral Membrane Proteins in Protein-Rich Membranes

DEFF Research Database (Denmark)

Javanainen, Matti; Martinez-Seara, Hector; Metzler, Ralf

2017-01-01

of being protein-poor, native cell membranes are extremely crowded with proteins. On the basis of extensive molecular simulations, we here demonstrate that protein crowding of the membrane at physiological levels leads to deviations from the SD relation and to the emergence of a stronger Stokes......-like dependence D ∝ 1/R. We propose that this 1/R law mainly arises due to geometrical factors: smaller proteins are able to avoid confinement effects much better than their larger counterparts. The results highlight that the lateral dynamics in the crowded setting found in native membranes is radically different......The lateral diffusion of embedded proteins along lipid membranes in protein-poor conditions has been successfully described in terms of the Saffman-Delbrück (SD) model, which predicts that the protein diffusion coefficient D is weakly dependent on its radius R as D ∝ ln(1/R). However, instead...

Arsenic Removal by Liquid Membranes

Directory of Open Access Journals (Sweden)

Tiziana Marino

2015-03-01

Full Text Available Water contamination with harmful arsenic compounds represents one of the most serious calamities of the last two centuries. Natural occurrence of the toxic metal has been revealed recently for 21 countries worldwide; the risk of arsenic intoxication is particularly high in Bangladesh and India but recently also Europe is facing similar problem. Liquid membranes (LMs look like a promising alternative to the existing removal processes, showing numerous advantages in terms of energy consumption, efficiency, selectivity, and operational costs. The development of different LM configurations has been a matter of investigation by several researching groups, especially for the removal of As(III and As(V from aqueous solutions. Most of these LM systems are based on the use of phosphine oxides as carriers, when the metal removal is from sulfuric acid media. Particularly promising for water treatment is the hollow fiber supported liquid membrane (HFSLM configuration, which offers high selectivity, easy transport of the targeted metal ions, large surface area, and non-stop flow process. The choice of organic extractant(s plays an essential role in the efficiency of the arsenic removal. Emulsion liquid membrane (ELM systems have not been extensively investigated so far, although encouraging results have started to appear in the literature. For such LM configuration, the most relevant step toward efficiency is the choice of the surfactant type and its concentration.

Amyloid and membrane complexity: The toxic interplay revealed by AFM.

Science.gov (United States)

Canale, Claudio; Oropesa-Nuñez, Reinier; Diaspro, Alberto; Dante, Silvia

2018-01-01

Lipid membranes play a fundamental role in the pathological development of protein misfolding diseases. Several pieces of evidence suggest that the lipid membrane could act as a catalytic surface for protein aggregation. Furthermore, a leading theory indicates the interaction between the cell membrane and misfolded oligomer species as the responsible for cytotoxicity, hence, for neurodegeneration in disorders such as Alzheimer's and Parkinson's disease. The definition of the mechanisms that drive the interaction between pathological protein aggregates and plasma membrane is fundamental for the development of effective therapies for a large class of diseases. Atomic force microscopy (AFM) has been employed to study how amyloid aggregates affect the cell physiological properties. Considerable efforts were spent to characterize the interaction with model systems, i.e., planar supported lipid bilayers, but some works also addressed the problem directly on living cells. Here, an overview of the main works involving the use of the AFM on both model system and living cells will be provided. Different kind of approaches will be presented, as well as the main results derived from the AFM analysis. Copyright © 2017 Elsevier Ltd. All rights reserved.

Residue-specific membrane location of peptides and proteins using specifically and extensively deuterated lipids and {sup 13}C-{sup 2}H rotational-echo double-resonance solid-state NMR

Energy Technology Data Exchange (ETDEWEB)

Xie Li; Ghosh, Ujjayini; Schmick, Scott D.; Weliky, David P., E-mail: weliky@chemistry.msu.edu [Michigan State University, Department of Chemistry (United States)

2013-01-15

Residue-specific location of peptides in the hydrophobic core of membranes was examined using {sup 13}C-{sup 2}H REDOR and samples in which the lipids were selectively deuterated. The transmembrane topology of the KALP peptide was validated with this approach with substantial dephasing observed for deuteration in the bilayer center and reduced or no dephasing for deuteration closer to the headgroups. Insertion of {beta} sheet HIV and helical and {beta} sheet influenza virus fusion peptides into the hydrophobic core of the membrane was validated in samples with extensively deuterated lipids.

Membrane bioreactor wastewater treatment plants reveal diverse yeast and protist communities of potential significance in biofouling.

Science.gov (United States)

Liébana, Raquel; Arregui, Lucía; Belda, Ignacio; Gamella, Luis; Santos, Antonio; Marquina, Domingo; Serrano, Susana

2015-01-01

The yeast community was studied in a municipal full-scale membrane bioreactor wastewater treatment plant (MBR-WWTP). The unexpectedly high diversity of yeasts indicated that the activated sludge formed a suitable environment for them to proliferate, with cellular concentrations of 2.2 ± 0.8 × 10(3) CFU ml(-1). Sixteen species of seven genera were present in the biological reactor, with Ascomycetes being the most prevalent group (93%). Most isolates were able to grow in a synthetic wastewater medium, adhere to polyethylene surfaces, and develop biofilms of variable complexity. The relationship between yeast populations and the protists in the MBR-WWTP was also studied, revealing that some protist species preyed on and ingested yeasts. These results suggest that yeast populations may play a role in the food web of a WWTP and, to some extent, contribute to membrane biofouling in MBR systems.

High Performance Regenerated Cellulose Membranes from Trimethylsilyl Cellulose

KAUST Repository

Ali, Ola

2013-05-01

Regenerated cellulose (RC) membranes are extensively used in medical and pharmaceutical separation processes due to their biocompatibility, low fouling tendency and solvent resistant properties. They typically possess ultrafiltration and microfiltration separation characteristics, but recently, there have been attempts to widen their pool of applications in nanofiltration processes. In this work, a novel method for preparing high performance composite RC membranes was developed. These membranes reveal molecular weight cut-offs (MWCO) of less than 250 daltons, which possibly put them ahead of all commercial RC membranes and in competition with high performance nanofiltration membranes. The membranes were prepared by acidic hydrolysis of dip-coated trimethylsilyl cellulose (TMSC) films. TMSC, with a degree of silylation (DS) of 2.8, was prepared from microcrystalline cellulose by reaction with hexamethyldisilazane under the homogeneous conditions of LiCl/DMAC solvent system. Effects of parameters, such as coating solution concentration and drying rates, were investigated. It was concluded that higher TMSC concentrations as well as higher solvent evaporation rates favor better MWCOs, mainly due to increase in the selective layer thickness. Successful cross-linking of prepared membranes with glyoxal solutions, in the presence of boric acid as a catalyst, resulted in MWCOs less than 250 daltons. The suitability of this crosslinking reaction for large scale productions was already proven in the manufacturing of durable-press fabrics. For us, the inexpensive raw materials as well as the low reaction times and temperatures were of interest. Moreover, the non-toxic nature of glyoxal is a key advantage in medical and pharmaceutical applications. The membranes prepared in this work are strong candidates for separation of small organic solutes from organic solvents streams in pharmaceutical industries. Their hydrophilicity, compared to typical nanofiltration membranes, offer

Crystal structure of an orthomyxovirus matrix protein reveals mechanisms for self-polymerization and membrane association.

Science.gov (United States)

Zhang, Wenting; Zheng, Wenjie; Toh, Yukimatsu; Betancourt-Solis, Miguel A; Tu, Jiagang; Fan, Yanlin; Vakharia, Vikram N; Liu, Jun; McNew, James A; Jin, Meilin; Tao, Yizhi J

2017-08-08

Many enveloped viruses encode a matrix protein. In the influenza A virus, the matrix protein M1 polymerizes into a rigid protein layer underneath the viral envelope to help enforce the shape and structural integrity of intact viruses. The influenza virus M1 is also known to mediate virus budding as well as the nuclear export of the viral nucleocapsids and their subsequent packaging into nascent viral particles. Despite extensive studies on the influenza A virus M1 (FLUA-M1), only crystal structures of its N-terminal domain are available. Here we report the crystal structure of the full-length M1 from another orthomyxovirus that infects fish, the infectious salmon anemia virus (ISAV). The structure of ISAV-M1 assumes the shape of an elbow, with its N domain closely resembling that of the FLUA-M1. The C domain, which is connected to the N domain through a flexible linker, is made of four α-helices packed as a tight bundle. In the crystal, ISAV-M1 monomers form infinite 2D arrays with a network of interactions involving both the N and C domains. Results from liposome flotation assays indicated that ISAV-M1 binds membrane via electrostatic interactions that are primarily mediated by a positively charged surface loop from the N domain. Cryoelectron tomography reconstruction of intact ISA virions identified a matrix protein layer adjacent to the inner leaflet of the viral membrane. The physical dimensions of the virion-associated matrix layer are consistent with the 2D ISAV-M1 crystal lattice, suggesting that the crystal lattice is a valid model for studying M1-M1, M1-membrane, and M1-RNP interactions in the virion.

Structural changes in plasma membranes prepared from irradiated Chinese hamster V79 cells as revealed by Raman spectroscopy

International Nuclear Information System (INIS)

Verma, S.P.; Sonwalkar, N.

1991-01-01

The effect of gamma irradiation on the integrity of plasma membranes isolated from Chinese hamster V79 cells was investigated by Raman spectroscopy. Plasma membranes of control V79 cells show transitions between -10 and 5 degree C (low-temperature transition), 10 and 22 degree C (middle-temperature transition), and 32 and 40 degree C (high-temperature transition). Irradiation (5 Gy) alters these transitions markedly. First, the low-temperature transition shifts to higher temperature (onset and completion temperatures 4 and 14 degree C). Second, the middle-temperature transition shifts up to the range of about 20-32 degree C, but the width remains unchanged. Third, the higher temperature transition broadens markedly and shifts to the range of about 15-40 degree C. Protein secondary structure as determined by least-squares analysis of the amide I bands shows 36% total helix, 55% total beta-strand, and 9% turn plus undefined for control plasma membrane proteins. Plasma membrane proteins of irradiated V79 cells show an increase in total helix (40 and 45% at 5 and 10 Gy, respectively) and a decrease in the total beta-strand (48 and 44% at 5 and 10 Gy, respectively) structures. The qualitative analysis of the Raman features of plasma membranes and model compounds in the 1600 cm-1 region, assigned to tyrosine groups, revealed that irradiation alters the microenvironment of these groups. We conclude that the radiation dose used in the survival range of Chinese hamster V79 cells can cause damage to plasma membrane proteins without detectable lipid peroxidation, and that the altered proteins react differently with lipids, yielding a shift in the thermal transition properties

REDOR NMR Reveals Multiple Conformers for a Protein Kinase C Ligand in a Membrane Environment

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Hao Yang

2018-01-01

Full Text Available Bryostatin 1 (henceforth bryostatin is in clinical trials for the treatment of Alzheimer’s disease and for HIV/AIDS eradication. It is also a preclinical lead for cancer immunotherapy and other therapeutic indications. Yet nothing is known about the conformation of bryostatin bound to its protein kinase C (PKC target in a membrane microenvironment. As a result, efforts to design more efficacious, better tolerated, or more synthetically accessible ligands have been limited to structures that do not include PKC or membrane effects known to influence PKC–ligand binding. This problem extends more generally to many membrane-associated proteins in the human proteome. Here, we use rotational-echo double-resonance (REDOR solid-state NMR to determine the conformations of PKC modulators bound to the PKCδ-C1b domain in the presence of phospholipid vesicles. The conformationally limited PKC modulator phorbol diacetate (PDAc is used as an initial test substrate. While unanticipated partitioning of PDAc between an immobilized protein-bound state and a mobile state in the phospholipid assembly was observed, a single conformation in the bound state was identified. In striking contrast, a bryostatin analogue (bryolog was found to exist exclusively in a protein-bound state, but adopts a distribution of conformations as defined by three independent distance measurements. The detection of multiple PKCδ-C1b-bound bryolog conformers in a functionally relevant phospholipid complex reveals the inherent dynamic nature of cellular systems that is not captured with single-conformation static structures. These results indicate that binding, selectivity, and function of PKC modulators, as well as the design of new modulators, are best addressed using a dynamic multistate model, an analysis potentially applicable to other membrane-associated proteins.

Alternative energy efficient membrane bioreactor using reciprocating submerged membrane.

Science.gov (United States)

Ho, J; Smith, S; Roh, H K

2014-01-01

A novel membrane bioreactor (MBR) pilot system, using membrane reciprocation instead of air scouring, was operated at constant high flux and daily fluctuating flux to demonstrate its application under peak and diurnal flow conditions. Low and stable transmembrane pressure was achieved at 40 l/m(2)/h (LMH) by use of repetitive membrane reciprocation. The results reveal that the inertial forces acting on the membrane fibers effectively propel foulants from the membrane surface. Reciprocation of the hollow fiber membrane is beneficial for the constant removal of solids that may build up on the membrane surface and inside the membrane bundle. The membrane reciprocation in the reciprocating MBR pilot consumed less energy than coarse air scouring used in conventional MBR systems. Specific energy consumption for the membrane reciprocation was 0.072 kWh/m(3) permeate produced at 40 LMH flux, which is 75% less than for a conventional air scouring system as reported in literature without consideration of energy consumption for biological aeration (0.29 kWh/m(3)). The daily fluctuating flux test confirmed that the membrane reciprocation is effective to handle fluctuating flux up to 50 LMH. The pilot-scale reciprocating MBR system successfully demonstrated that fouling can be controlled via 0.43 Hz membrane reciprocation with 44 mm or higher amplitude.

Comparative Study of One-Step Cross-Linked Electrospun Chitosan-Based Membranes

Directory of Open Access Journals (Sweden)

Yanet E. Aguirre-Chagala

2017-01-01

Full Text Available Chitosan membranes are widely applied for tissue engineering; however, a major drawback is their low resistance in aqueous phases and therefore the structure collapses impeding their long-term use. Although there is extensive research, because of chitosan’s importance as a biomaterial, studies involving chitosan-based membranes are still needed. Herein, a detailed investigation of diverse chemical routes to cross-link fibers in situ by electrospinning process is described. In case of using genipin as cross-linker, a close relationship with the content and the mean diameter values is reported, suggesting a crucial effect over the design of nanostructures. Also, the physical resistance is enhanced for the combination of two types of methods, such as chemical and physical methods. Cross-linked fibers upon exposure to long wave ultraviolet A (UVA light change their morphology, but not their chemical composition. When they are incubated in aqueous phase for 70 days, they show an extensive improvement of their macrostructural integrity which makes them attractive candidates for tissue engineering application. As a result, the thermal properties of these materials reveal less crystallinity and higher temperature of degradation.

Analysis of global gene expression in Brachypodium distachyon reveals extensive network plasticity in response to abiotic stress.

Directory of Open Access Journals (Sweden)

Henry D Priest

Full Text Available Brachypodium distachyon is a close relative of many important cereal crops. Abiotic stress tolerance has a significant impact on productivity of agriculturally important food and feedstock crops. Analysis of the transcriptome of Brachypodium after chilling, high-salinity, drought, and heat stresses revealed diverse differential expression of many transcripts. Weighted Gene Co-Expression Network Analysis revealed 22 distinct gene modules with specific profiles of expression under each stress. Promoter analysis implicated short DNA sequences directly upstream of module members in the regulation of 21 of 22 modules. Functional analysis of module members revealed enrichment in functional terms for 10 of 22 network modules. Analysis of condition-specific correlations between differentially expressed gene pairs revealed extensive plasticity in the expression relationships of gene pairs. Photosynthesis, cell cycle, and cell wall expression modules were down-regulated by all abiotic stresses. Modules which were up-regulated by each abiotic stress fell into diverse and unique gene ontology GO categories. This study provides genomics resources and improves our understanding of abiotic stress responses of Brachypodium.

Two conformational states of the membrane-associated Bacillus thuringiensis Cry4Ba δ-endotoxin complex revealed by electron crystallography: Implications for toxin-pore formation

International Nuclear Information System (INIS)

Ounjai, Puey; Unger, Vinzenz M.; Sigworth, Fred J.; Angsuthanasombat, Chanan

2007-01-01

The insecticidal nature of Cry δ-endotoxins produced by Bacillus thuringiensis is generally believed to be caused by their ability to form lytic pores in the midgut cell membrane of susceptible insect larvae. Here we have analyzed membrane-associated structures of the 65-kDa dipteran-active Cry4Ba toxin by electron crystallography. The membrane-associated toxin complex was crystallized in the presence of DMPC via detergent dialysis. Depending upon the charge of the adsorbed surface, 2D crystals of the oligomeric toxin complex have been captured in two distinct conformations. The projection maps of those crystals have been generated at 17 A resolution. Both complexes appeared to be trimeric; as in one crystal form, its projection structure revealed a symmetrical pinwheel-like shape with virtually no depression in the middle of the complex. The other form revealed a propeller-like conformation displaying an obvious hole in the center region, presumably representing the toxin-induced pore. These crystallographic data thus demonstrate for the first time that the 65-kDa activated Cry4Ba toxin in association with lipid membranes could exist in at least two different trimeric conformations, conceivably implying the closed and open states of the pore

Membranes and Fluorescence microscopy

DEFF Research Database (Denmark)

Bagatolli, Luis

2009-01-01

Fluorescence spectroscopy-based techniques using conventional fluorimeters have been extensively applied since the late 1960s to study different aspects of membrane-related phenomena, i.e., mainly relating to lipid-lipid and lipid-protein (peptide) interactions. Even though fluorescence...

Assessment of Membrane Fluidity Fluctuations during Cellular Development Reveals Time and Cell Type Specificity

KAUST Repository

Noutsi, Bakiza Kamal

2016-06-30

Cell membrane is made up of a complex structure of lipids and proteins that diffuse laterally giving rise to what we call membrane fluidity. During cellular development, such as differentiation cell membranes undergo dramatic fluidity changes induced by proteins such as ARC and Cofilin among others. In this study we used the generalized polarization (GP) property of fluorescent probe Laurdan using two-photon microscopy to determine membrane fluidity as a function of time and for various cell lines. A low GP value corresponds to a higher fluidity and a higher GP value is associated with a more rigid membrane. Four different cell lines were monitored such as hN2, NIH3T3, HEK293 and L6 cells. Membrane fluidity was measured at 12h, 72h and 92 h. Our results show significant changes in membrane fluidity among all cell types at different time points. GP values tend to increase significantly within 92 h in hN2 cells and 72 h in NIH3T3 cells and only at 92 h in HEK293 cells. L6 showed a marked decrease in membrane fluidity at 72 h and starts to increase at 92 h. As expected, NIH3T3 cells have more rigid membrane at earlier time points. On the other hand, neurons tend to have the highest membrane fluidity at early time points emphasizing its correlation with plasticity and the need for this malleability during differentiation. This study sheds light on the involvement of membrane fluidity during neuronal differentiation and development of other cell lines.

Assessment of Membrane Fluidity Fluctuations during Cellular Development Reveals Time and Cell Type Specificity

KAUST Repository

Noutsi, Bakiza Kamal; Gratton, Enrico; Chaieb, Saharoui

2016-01-01

Cell membrane is made up of a complex structure of lipids and proteins that diffuse laterally giving rise to what we call membrane fluidity. During cellular development, such as differentiation cell membranes undergo dramatic fluidity changes induced by proteins such as ARC and Cofilin among others. In this study we used the generalized polarization (GP) property of fluorescent probe Laurdan using two-photon microscopy to determine membrane fluidity as a function of time and for various cell lines. A low GP value corresponds to a higher fluidity and a higher GP value is associated with a more rigid membrane. Four different cell lines were monitored such as hN2, NIH3T3, HEK293 and L6 cells. Membrane fluidity was measured at 12h, 72h and 92 h. Our results show significant changes in membrane fluidity among all cell types at different time points. GP values tend to increase significantly within 92 h in hN2 cells and 72 h in NIH3T3 cells and only at 92 h in HEK293 cells. L6 showed a marked decrease in membrane fluidity at 72 h and starts to increase at 92 h. As expected, NIH3T3 cells have more rigid membrane at earlier time points. On the other hand, neurons tend to have the highest membrane fluidity at early time points emphasizing its correlation with plasticity and the need for this malleability during differentiation. This study sheds light on the involvement of membrane fluidity during neuronal differentiation and development of other cell lines.

Extensive structural variations between mitochondrial genomes of CMS and normal peppers (Capsicum annuum L.) revealed by complete nucleotide sequencing.

Science.gov (United States)

Jo, Yeong Deuk; Choi, Yoomi; Kim, Dong-Hwan; Kim, Byung-Dong; Kang, Byoung-Cheorl

2014-07-04

Cytoplasmic male sterility (CMS) is an inability to produce functional pollen that is caused by mutation of the mitochondrial genome. Comparative analyses of mitochondrial genomes of lines with and without CMS in several species have revealed structural differences between genomes, including extensive rearrangements caused by recombination. However, the mitochondrial genome structure and the DNA rearrangements that may be related to CMS have not been characterized in Capsicum spp. We obtained the complete mitochondrial genome sequences of the pepper CMS line FS4401 (507,452 bp) and the fertile line Jeju (511,530 bp). Comparative analysis between mitochondrial genomes of peppers and tobacco that are included in Solanaceae revealed extensive DNA rearrangements and poor conservation in non-coding DNA. In comparison between pepper lines, FS4401 and Jeju mitochondrial DNAs contained the same complement of protein coding genes except for one additional copy of an atp6 gene (ψatp6-2) in FS4401. In terms of genome structure, we found eighteen syntenic blocks in the two mitochondrial genomes, which have been rearranged in each genome. By contrast, sequences between syntenic blocks, which were specific to each line, accounted for 30,380 and 17,847 bp in FS4401 and Jeju, respectively. The previously-reported CMS candidate genes, orf507 and ψatp6-2, were located on the edges of the largest sequence segments that were specific to FS4401. In this region, large number of small sequence segments which were absent or found on different locations in Jeju mitochondrial genome were combined together. The incorporation of repeats and overlapping of connected sequence segments by a few nucleotides implied that extensive rearrangements by homologous recombination might be involved in evolution of this region. Further analysis using mtDNA pairs from other plant species revealed common features of DNA regions around CMS-associated genes. Although large portion of sequence context was

Studies on lactoferricin-derived Escherichia coli membrane-active peptides reveal differences in the mechanism of N-acylated versus nonacylated peptides.

Science.gov (United States)

Zweytick, Dagmar; Deutsch, Günter; Andrä, Jörg; Blondelle, Sylvie E; Vollmer, Ekkehard; Jerala, Roman; Lohner, Karl

2011-06-17

To improve the low antimicrobial activity of LF11, an 11-mer peptide derived from human lactoferricin, mutant sequences were designed based on the defined structure of LF11 in the lipidic environment. Thus, deletion of noncharged polar residues and strengthening of the hydrophobic N-terminal part upon adding a bulky hydrophobic amino acid or N-acylation resulted in enhanced antimicrobial activity against Escherichia coli, which correlated with the peptides' degree of perturbation of bacterial membrane mimics. Nonacylated and N-acylated peptides exhibited different effects at a molecular level. Nonacylated peptides induced segregation of peptide-enriched and peptide-poor lipid domains in negatively charged bilayers, although N-acylated peptides formed small heterogeneous domains resulting in a higher degree of packing defects. Additionally, only N-acylated peptides perturbed the lateral packing of neutral lipids and exhibited increased permeability of E. coli lipid vesicles. The latter did not correlate with the extent of improvement of the antimicrobial activity, which could be explained by the fact that elevated binding of N-acylated peptides to lipopolysaccharides of the outer membrane of gram-negative bacteria seems to counteract the elevated membrane permeabilization, reflected in the respective minimal inhibitory concentration for E. coli. The antimicrobial activity of the peptides correlated with an increase of membrane curvature stress and hence bilayer instability. Transmission electron microscopy revealed that only the N-acylated peptides induced tubular protrusions from the outer membrane, whereas all peptides caused detachment of the outer and inner membrane of E. coli bacteria. Viability tests demonstrated that these bacteria were dead before onset of visible cell lysis.

Studies on Lactoferricin-derived Escherichia coli Membrane-active Peptides Reveal Differences in the Mechanism of N-Acylated Versus Nonacylated Peptides*

Science.gov (United States)

Zweytick, Dagmar; Deutsch, Günter; Andrä, Jörg; Blondelle, Sylvie E.; Vollmer, Ekkehard; Jerala, Roman; Lohner, Karl

2011-01-01

To improve the low antimicrobial activity of LF11, an 11-mer peptide derived from human lactoferricin, mutant sequences were designed based on the defined structure of LF11 in the lipidic environment. Thus, deletion of noncharged polar residues and strengthening of the hydrophobic N-terminal part upon adding a bulky hydrophobic amino acid or N-acylation resulted in enhanced antimicrobial activity against Escherichia coli, which correlated with the peptides' degree of perturbation of bacterial membrane mimics. Nonacylated and N-acylated peptides exhibited different effects at a molecular level. Nonacylated peptides induced segregation of peptide-enriched and peptide-poor lipid domains in negatively charged bilayers, although N-acylated peptides formed small heterogeneous domains resulting in a higher degree of packing defects. Additionally, only N-acylated peptides perturbed the lateral packing of neutral lipids and exhibited increased permeability of E. coli lipid vesicles. The latter did not correlate with the extent of improvement of the antimicrobial activity, which could be explained by the fact that elevated binding of N-acylated peptides to lipopolysaccharides of the outer membrane of Gram-negative bacteria seems to counteract the elevated membrane permeabilization, reflected in the respective minimal inhibitory concentration for E. coli. The antimicrobial activity of the peptides correlated with an increase of membrane curvature stress and hence bilayer instability. Transmission electron microscopy revealed that only the N-acylated peptides induced tubular protrusions from the outer membrane, whereas all peptides caused detachment of the outer and inner membrane of E. coli bacteria. Viability tests demonstrated that these bacteria were dead before onset of visible cell lysis. PMID:21515687

Global Proteomic Analysis Reveals an Exclusive Role of Thylakoid Membranes in Bioenergetics of a Model Cyanobacterium

Energy Technology Data Exchange (ETDEWEB)

Liberton, Michelle; Saha, Rajib; Jacobs, Jon M.; Nguyen, Amelia Y.; Gritsenko, Marina A.; Smith, Richard D.; Koppenaal, David W.; Pakrasi, Himadri B.

2016-04-07

Cyanobacteria are photosynthetic microbes with highly differentiated membrane systems. These organisms contain an outer membrane, plasma membrane, and an internal system of thylakoid membranes where the photosynthetic and respiratory machinery are found. This existence of compartmentalization and differentiation of membrane systems poses a number of challenges for cyanobacterial cells in terms of organization and distribution of proteins to the correct membrane system. Proteomics studies have long sought to identify the components of the different membrane systems, and to date about 450 different proteins have been attributed to either the plasma membrane or thylakoid membrane. Given the complexity of these membranes, many more proteins remain to be identified in these membrane systems, and a comprehensive catalog of plasma membrane and thylakoid membrane proteins is needed. Here we describe the identification of 635 proteins in Synechocystis sp. PCC 6803 by quantitative iTRAQ isobaric labeling; of these, 459 proteins were localized to the plasma membrane and 176 were localized to the thylakoid membrane. Surprisingly, we found over 2.5 times the number of unique proteins identified in the plasma membrane compared to the thylakoid membrane. This suggests that the protein composition of the thylakoid membrane is more homogeneous than the plasma membrane, consistent with the role of the plasma membrane in diverse cellular processes including protein trafficking and nutrient import, compared to a more specialized role for the thylakoid membrane in cellular energetics. Overall, the protein composition of the Synechocystis 6803 plasma membrane and thylakoid membrane is quite similar to the E.coli plasma membrane and Arabidopsis thylakoid membrane, respectively. Synechocystis 6803 can therefore be described as a gram-negative bacterium that has an additional internal membrane system that fulfils the energetic requirements of the cell.

Global Proteomic Analysis Reveals an Exclusive Role of Thylakoid Membranes in Bioenergetics of a Model Cyanobacterium

Energy Technology Data Exchange (ETDEWEB)

Liberton, Michelle; Saha, Rajib; Jacobs, Jon M.; Nguyen, Amelia Y.; Gritsenko, Marina A.; Smith, Richard D.; Koppenaal, David W.; Pakrasi, Himadri B.

2016-04-07

Cyanobacteria are photosynthetic microbes with highlydifferentiated membrane systems. These organisms contain an outer membrane, plasma membrane, and an internal system of thylakoid membranes where the photosynthetic and respiratory machinery are found. This existence of compartmentalization and differentiation of membrane systems poses a number of challenges for cyanobacterial cells in terms of organization and distribution of proteins to the correct membrane system. Proteomics studies have long sought to identify the components of the different membrane systems in cyanobacteria, and to date about 450 different proteins have been attributed to either the plasma membrane or thylakoid membrane. Given the complexity of these membranes, many more proteins remain to be identified, and a comprehensive catalogue of plasma membrane and thylakoid membrane proteins is needed. Here we describe the identification of 635 differentially localized proteins in Synechocystis sp. PCC 6803 by quantitative iTRAQ isobaric labeling; of these, 459 proteins were localized to the plasma membrane and 176 were localized to the thylakoid membrane. Surprisingly, we found over 2.5 times the number of unique proteins identified in the plasma membrane compared with the thylakoid membrane. This suggests that the protein composition of the thylakoid membrane is more homogeneous than the plasma membrane, consistent with the role of the plasma membrane in diverse cellular processes including protein trafficking and nutrient import, compared with a more specialized role for the thylakoid membrane in cellular energetics. Thus, our data clearly define the two membrane systems with distinct functions. Overall, the protein compositions of the Synechocystis 6803 plasma membrane and thylakoid membrane are quite similar to that of the plasma membrane of Escherichia coli and thylakoid membrane of Arabidopsis chloroplasts, respectively. Synechocystis 6803 can therefore be described as a Gram

Fluorinated polyimides grafted with poly(ethylene glycol) side chains by the RAFT-mediated process and their membranes

International Nuclear Information System (INIS)

Chen Yiwang; Chen Lie; Nie Huarong; Kang, E.T.; Vora, R.H.

2005-01-01

Graft polymerization of poly(ethylene glycol) methyl ether methacrylate (PEGMA) from fluorinated polyimide (FPI) was carried out by the reversible addition-fragmentation chain transfer (RAFT)-mediated process. The peroxides generated by the ozone treatment on FPI facilitated the thermally-initiated graft copolymerization from FPI backbone. The 'living' character of the graft chain growing was ascertained in the subsequent chain extension of PEGMA. Nuclear magnetic resonance (NMR) and molecular weight measurements were used to characterize the chemical composition and structure of the copolymers. Microfiltration (MF) membranes were fabricated from the FPI-g-PEGMA comb copolymers by phase inversion in aqueous media. Surface composition analysis of the membranes scanned by X-ray photoelectron spectroscopy (XPS) revealed a substantial surface enrichment of the hydrophilic components. The pore size distribution of the resulting membranes was found to be much more uniform than that of the corresponding membranes cast from FPI-g-PEGMA prepared by the conventional radical polymerization process in the absence of the chain transfer agent. The morphology of the membranes was characterized by scanning electron microscopy (SEM)

Cell surface area and membrane folding in glioblastoma cell lines differing in PTEN and p53 status.

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Simon Memmel

Full Text Available Glioblastoma multiforme (GBM is characterized by rapid growth, invasion and resistance to chemo-/radiotherapy. The complex cell surface morphology with abundant membrane folds, microvilli, filopodia and other membrane extensions is believed to contribute to the highly invasive behavior and therapy resistance of GBM cells. The present study addresses the mechanisms leading to the excessive cell membrane area in five GBM lines differing in mutational status for PTEN and p53. In addition to scanning electron microscopy (SEM, the membrane area and folding were quantified by dielectric measurements of membrane capacitance using the single-cell electrorotation (ROT technique. The osmotic stability and volume regulation of GBM cells were analyzed by video microscopy. The expression of PTEN, p53, mTOR and several other marker proteins involved in cell growth and membrane synthesis were examined by Western blotting. The combined SEM, ROT and osmotic data provided independent lines of evidence for a large variability in membrane area and folding among tested GBM lines. Thus, DK-MG cells (wild type p53 and wild type PTEN exhibited the lowest degree of membrane folding, probed by the area-specific capacitance C m = 1.9 µF/cm(2. In contrast, cell lines carrying mutations in both p53 and PTEN (U373-MG and SNB19 showed the highest C m values of 3.7-4.0 µF/cm(2, which corroborate well with their heavily villated cell surface revealed by SEM. Since PTEN and p53 are well-known inhibitors of mTOR, the increased membrane area/folding in mutant GBM lines may be related to the enhanced protein and lipid synthesis due to a deregulation of the mTOR-dependent downstream signaling pathway. Given that membrane folds and extensions are implicated in tumor cell motility and metastasis, the dielectric approach presented here provides a rapid and simple tool for screening the biophysical cell properties in studies on targeting chemo- or radiotherapeutically the

Correlative STED and Atomic Force Microscopy on Live Astrocytes Reveals Plasticity of Cytoskeletal Structure and Membrane Physical Properties during Polarized Migration

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Nathalie Rouach

2017-04-01

Full Text Available The plasticity of the cytoskeleton architecture and membrane properties is important for the establishment of cell polarity, adhesion and migration. Here, we present a method which combines stimulated emission depletion (STED super-resolution imaging and atomic force microscopy (AFM to correlate cytoskeletal structural information with membrane physical properties in live astrocytes. Using STED compatible dyes for live cell imaging of the cytoskeleton, and simultaneously mapping the cell surface topology with AFM, we obtain unprecedented detail of highly organized networks of actin and microtubules in astrocytes. Combining mechanical data from AFM with optical imaging of actin and tubulin further reveals links between cytoskeleton organization and membrane properties. Using this methodology we illustrate that scratch-induced migration induces cytoskeleton remodeling. The latter is caused by a polarization of actin and microtubule elements within astroglial cell processes, which correlates strongly with changes in cell stiffness. The method opens new avenues for the dynamic probing of the membrane structural and functional plasticity of living brain cells. It is a powerful tool for providing new insights into mechanisms of cell structural remodeling during physiological or pathological processes, such as brain development or tumorigenesis.

Plant lipid environment and membrane enzymes: the case of the plasma membrane H+-ATPase.

Science.gov (United States)

Morales-Cedillo, Francisco; González-Solís, Ariadna; Gutiérrez-Angoa, Lizbeth; Cano-Ramírez, Dora Luz; Gavilanes-Ruiz, Marina

2015-04-01

Several lipid classes constitute the universal matrix of the biological membranes. With their amphipathic nature, lipids not only build the continuous barrier that confers identity to every cell and organelle, but they are also active actors that modulate the activity of the proteins immersed in the lipid bilayer. The plasma membrane H(+)-ATPase, an enzyme from plant cells, is an excellent example of a transmembrane protein whose activity is influenced by the hydrophilic compartments at both sides of the membrane and by the hydrophobic domains of the lipid bilayer. As a result, an extensive documentation of the effect of numerous amphiphiles in the enzyme activity can be found. Detergents, membrane glycerolipids, and sterols can produce activation or inhibition of the enzyme activity. In some cases, these effects are associated with the lipids of the membrane bulk, but in others, a direct interaction of the lipid with the protein is involved. This review gives an account of reports related to the action of the membrane lipids on the H(+)-ATPase activity.

Evolution and development of model membranes for physicochemical and functional studies of the membrane lateral heterogeneity.

Science.gov (United States)

Morigaki, Kenichi; Tanimoto, Yasushi

2018-03-14

One of the main questions in the membrane biology is the functional roles of membrane heterogeneity and molecular localization. Although segregation and local enrichment of protein/lipid components (rafts) have been extensively studied, the presence and functions of such membrane domains still remain elusive. Along with biochemical, cell observation, and simulation studies, model membranes are emerging as an important tool for understanding the biological membrane, providing quantitative information on the physicochemical properties of membrane proteins and lipids. Segregation of fluid lipid bilayer into liquid-ordered (Lo) and liquid-disordered (Ld) phases has been studied as a simplified model of raft in model membranes, including giant unilamellar vesicles (GUVs), giant plasma membrane vesicles (GPMVs), and supported lipid bilayers (SLB). Partition coefficients of membrane proteins between Lo and Ld phases were measured to gauze their affinities to lipid rafts (raftophilicity). One important development in model membrane is patterned SLB based on the microfabrication technology. Patterned Lo/Ld phases have been applied to study the partition and function of membrane-bound molecules. Quantitative information of individual molecular species attained by model membranes is critical for elucidating the molecular functions in the complex web of molecular interactions. The present review gives a short account of the model membranes developed for studying the lateral heterogeneity, especially focusing on patterned model membranes on solid substrates. Copyright © 2018 Elsevier B.V. All rights reserved.

Crystal Structure of the Herpesvirus Nuclear Egress Complex Provides Insights into Inner Nuclear Membrane Remodeling

Directory of Open Access Journals (Sweden)

Tzviya Zeev-Ben-Mordehai

2015-12-01

Full Text Available Although nucleo-cytoplasmic transport is typically mediated through nuclear pore complexes, herpesvirus capsids exit the nucleus via a unique vesicular pathway. Together, the conserved herpesvirus proteins pUL31 and pUL34 form the heterodimeric nuclear egress complex (NEC, which, in turn, mediates the formation of tight-fitting membrane vesicles around capsids at the inner nuclear membrane. Here, we present the crystal structure of the pseudorabies virus NEC. The structure revealed that a zinc finger motif in pUL31 and an extensive interaction network between the two proteins stabilize the complex. Comprehensive mutational analyses, characterized both in situ and in vitro, indicated that the interaction network is not redundant but rather complementary. Fitting of the NEC crystal structure into the recently determined cryoEM-derived hexagonal lattice, formed in situ by pUL31 and pUL34, provided details on the molecular basis of NEC coat formation and inner nuclear membrane remodeling.

Site-directed fluorescence labeling reveals a revised N-terminal membrane topology and functional periplasmic residues in the Escherichia coli cell division protein FtsK.

Science.gov (United States)

Berezuk, Alison M; Goodyear, Mara; Khursigara, Cezar M

2014-08-22

In Escherichia coli, FtsK is a large integral membrane protein that coordinates chromosome segregation and cell division. The N-terminal domain of FtsK (FtsKN) is essential for division, and the C terminus (FtsKC) is a well characterized DNA translocase. Although the function of FtsKN is unknown, it is suggested that FtsK acts as a checkpoint to ensure DNA is properly segregated before septation. This may occur through modulation of protein interactions between FtsKN and other division proteins in both the periplasm and cytoplasm; thus, a clear understanding of how FtsKN is positioned in the membrane is required to characterize these interactions. The membrane topology of FtsKN was initially determined using site-directed reporter fusions; however, questions regarding this topology persist. Here, we report a revised membrane topology generated by site-directed fluorescence labeling. The revised topology confirms the presence of four transmembrane segments and reveals a newly identified periplasmic loop between the third and fourth transmembrane domains. Within this loop, four residues were identified that, when mutated, resulted in the appearance of cellular voids. High resolution transmission electron microscopy of these voids showed asymmetric division of the cytoplasm in the absence of outer membrane invagination or visible cell wall ingrowth. This uncoupling reveals a novel role for FtsK in linking cell envelope septation events and yields further evidence for FtsK as a critical checkpoint of cell division. The revised topology of FtsKN also provides an important platform for future studies on essential interactions required for this process. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

Role for chlamydial inclusion membrane proteins in inclusion membrane structure and biogenesis.

Directory of Open Access Journals (Sweden)

Jeffrey Mital

Full Text Available The chlamydial inclusion membrane is extensively modified by the insertion of type III secreted effector proteins. These inclusion membrane proteins (Incs are exposed to the cytosol and share a common structural feature of a long, bi-lobed hydrophobic domain but little or no primary amino acid sequence similarity. Based upon secondary structural predictions, over 50 putative inclusion membrane proteins have been identified in Chlamydia trachomatis. Only a limited number of biological functions have been defined and these are not shared between chlamydial species. Here we have ectopically expressed several C. trachomatis Incs in HeLa cells and find that they induce the formation of morphologically distinct membranous vesicular compartments. Formation of these vesicles requires the bi-lobed hydrophobic domain as a minimum. No markers for various cellular organelles were observed in association with these vesicles. Lipid probes were incorporated by the Inc-induced vesicles although the lipids incorporated were dependent upon the specific Inc expressed. Co-expression of Inc pairs indicated that some colocalized in the same vesicle, others partially overlapped, and others did not associate at all. Overall, it appears that Incs may have an intrinsic ability to induce membrane formation and that individual Incs can induce membranous structures with unique properties.

In Situ and Real-Time SFG Measurements Revealing Organization and Transport of Cholesterol Analogue 6-Ketocholestanol in a Cell Membrane.

Science.gov (United States)

Ma, Sulan; Li, Hongchun; Tian, Kangzhen; Ye, Shuji; Luo, Yi

2014-02-06

Cholesterol organization and transport within a cell membrane are essential for human health and many cellular functions yet remain elusive so far. Using cholesterol analogue 6-ketocholestanol (6-KC) as a model, we have successfully exploited sum frequency generation vibrational spectroscopy (SFG-VS) to track the organization and transport of cholesterol in a membrane by combining achiral-sensitive ssp (ppp) and chiral-sensitive psp polarization measurements. It is found that 6-KC molecules are aligned at the outer leaflet of the DMPC lipid bilayer with a tilt angle of about 10°. 6-KC organizes itself by forming an α-β structure at low 6-KC concentration and most likely a β-β structure at high 6-KC concentration. Among all proposed models, our results favor the so-called umbrella model with formation of a 6-KC cluster. Moreover, we have found that the long anticipated flip-flop motion of 6-KC in the membrane takes time to occur, at least much longer than previously thought. All of these interesting findings indicate that it is critical to explore in situ, real-time, and label-free methodologies to obtain a precise molecular description of cholesterol's behavior in membranes. This study represents the first application of SFG to reveal the cholesterol-lipid interaction mechanism at the molecular level.

Improved surface property of PVDF membrane with amphiphilic zwitterionic copolymer as membrane additive

Energy Technology Data Exchange (ETDEWEB)

Li Jianhua, E-mail: jhli_2005@163.com [Institute of Biomedical and Pharmaceutical Technology and College of Chemistry and Chemical Engineering, Fuzhou University, Fuzhou 350001 (China); Li Mizi; Miao Jing; Wang Jiabin; Shao Xisheng [Institute of Biomedical and Pharmaceutical Technology and College of Chemistry and Chemical Engineering, Fuzhou University, Fuzhou 350001 (China); Zhang Qiqing, E-mail: zhangqiq@126.com [Institute of Biomedical and Pharmaceutical Technology and College of Chemistry and Chemical Engineering, Fuzhou University, Fuzhou 350001 (China) and Institute of Biomedical Engineering, Chinese Academy of Medical Science, Peking Union Medical College, Tianjin 300192 (China)

2012-06-15

An attempt to improve hydrophilicity and anti-fouling properties of PVDF membranes, a novel amphiphilic zwitterionic copolymer poly(vinylidene fluoride)-graft-poly(sulfobetaine methacrylate) (PVDF-g-PSBMA) was firstly synthesized by atom transfer radical polymerization (ATRP) and used as amphiphilic copolymer additive in the preparation of PVDF membranes. The PVDF-g-PSBMA/PVDF blend membranes were prepared by immersion precipitation process. Fourier transform infrared attenuated reflection spectroscopy (FTIR-ATR) and X-ray photoelectronic spectroscopy (XPS) measurements confirmed that PSBMA brushes from amphiphilic additives were preferentially segregated to membrane-coagulant interface during membrane formation. The morphology of membranes was characterized by scanning electron microscopy (SEM). Water contact angle measurements showed that the surface hydrophilicity of PVDF membranes was improved significantly with the increasing of amphiphilic copolymer PVDF-g-PSBMA in cast solution. Protein static adsorption experiment and dynamic fouling resistance experiment revealed that the surface enrichment of PSBMA brush endowed PVDF blend membrane great improvement of surface anti-fouling ability.

Improved surface property of PVDF membrane with amphiphilic zwitterionic copolymer as membrane additive

International Nuclear Information System (INIS)

Li Jianhua; Li Mizi; Miao Jing; Wang Jiabin; Shao Xisheng; Zhang Qiqing

2012-01-01

An attempt to improve hydrophilicity and anti-fouling properties of PVDF membranes, a novel amphiphilic zwitterionic copolymer poly(vinylidene fluoride)-graft-poly(sulfobetaine methacrylate) (PVDF-g-PSBMA) was firstly synthesized by atom transfer radical polymerization (ATRP) and used as amphiphilic copolymer additive in the preparation of PVDF membranes. The PVDF-g-PSBMA/PVDF blend membranes were prepared by immersion precipitation process. Fourier transform infrared attenuated reflection spectroscopy (FTIR-ATR) and X-ray photoelectronic spectroscopy (XPS) measurements confirmed that PSBMA brushes from amphiphilic additives were preferentially segregated to membrane-coagulant interface during membrane formation. The morphology of membranes was characterized by scanning electron microscopy (SEM). Water contact angle measurements showed that the surface hydrophilicity of PVDF membranes was improved significantly with the increasing of amphiphilic copolymer PVDF-g-PSBMA in cast solution. Protein static adsorption experiment and dynamic fouling resistance experiment revealed that the surface enrichment of PSBMA brush endowed PVDF blend membrane great improvement of surface anti-fouling ability.

Unique thylakoid membrane architecture of a unicellular N2-fixing cyanobacterium revealed by electron tomography

Energy Technology Data Exchange (ETDEWEB)

Liberton, Michelle; Austin II, Jotham R; Berg, R. Howard; Pakrasi, Himadri B

2011-04-01

Cyanobacteria, descendants of the endosymbiont that gave rise to modern-day chloroplasts, are vital contributors to global biological energy conversion processes. A thorough understanding of the physiology of cyanobacteria requires detailed knowledge of these organisms at the level of cellular architecture and organization. In these prokaryotes, the large membrane protein complexes of the photosynthetic and respiratory electron transport chains function in the intracellular thylakoid membranes. Like plants, the architecture of the thylakoid membranes in cyanobacteria has direct impact on cellular bioenergetics, protein transport, and molecular trafficking. However, whole-cell thylakoid organization in cyanobacteria is not well understood. Here we present, by using electron tomography, an in-depth analysis of the architecture of the thylakoid membranes in a unicellular cyanobacterium, Cyanothece sp. ATCC 51142. Based on the results of three-dimensional tomographic reconstructions of near-entire cells, we determined that the thylakoids in Cyanothece 51142 form a dense and complex network that extends throughout the entire cell. This thylakoid membrane network is formed from the branching and splitting of membranes and encloses a single lumenal space. The entire thylakoid network spirals as a peripheral ring of membranes around the cell, an organization that has not previously been described in a cyanobacterium. Within the thylakoid membrane network are areas of quasi-helical arrangement with similarities to the thylakoid membrane system in chloroplasts. This cyanobacterial thylakoid arrangement is an efficient means of packing a large volume of membranes in the cell while optimizing intracellular transport and trafficking.

Unique Thylakoid Membrane Architecture of a Unicellular N2-Fixing Cyanobacterium Revealed by Electron Tomography

Energy Technology Data Exchange (ETDEWEB)

Liberton, Michelle L.; Austin, Jotham R.; Berg, R. H.; Pakrasi, Himadri B.

2011-04-01

Cyanobacteria, descendants of the endosymbiont that gave rise to modern-day chloroplasts, are vital contributors to global biological energy conversion processes. A thorough understanding of the physiology of cyanobacteria requires detailed knowledge of these organisms at the level of cellular architecture and organization. In these prokaryotes, the large membrane protein complexes of the photosynthetic and respiratory electron transport chains function in the intracellular thylakoid membranes. Like plants, the architecture of the thylakoid membranes in cyanobacteria has direct impact on cellular bioenergetics, protein transport, and molecular trafficking. However, whole-cell thylakoid organization in cyanobacteria is not well understood. Here we present, by using electron tomography, an in-depth analysis of the architecture of the thylakoid membranes in a unicellular cyanobacterium, Cyanothece sp. ATCC 51142. Based on the results of three-dimensional tomographic reconstructions of near-entire cells, we determined that the thylakoids in Cyanothece 51142 form a dense and complex network that extends throughout the entire cell. This thylakoid membrane network is formed from the branching and splitting of membranes and encloses a single lumenal space. The entire thylakoid network spirals as a peripheral ring of membranes around the cell, an organization that has not previously been described in a cyanobacterium. Within the thylakoid membrane network are areas of quasi-helical arrangement with similarities to the thylakoid membrane system in chloroplasts. This cyanobacterial thylakoid arrangement is an efficient means of packing a large volume of membranes in the cell while optimizing intracellular transport and trafficking.

Reversible membrane reorganizations during photosynthesis in vivo: revealed by small-angle neutron scattering

DEFF Research Database (Denmark)

Nagy, Gergely; Posselt, Dorthe; Kovacs, Laszlo

2011-01-01

In the present study, we determined characteristic repeat distances of the photosynthetic membranes in living cyanobacterial and eukaryotic algal cells, and in intact thylakoid membranes isolated from higher plants with time-resolved small-angle neutron scattering. This non-invasive technique...

Photoresponsive nanostructured membranes

KAUST Repository

Madhavan, Poornima

2016-07-26

The perspective of adding stimuli-response to isoporous membranes stimulates the development of separation devices with pores, which would open or close under control of environment chemical composition, temperature or exposure to light. Changes in pH and temperature have been previously investigated. In this work, we demonstrate for the first time the preparation of photoresponsive isoporous membranes, applying self-assembly non-solvent induced phase separation to a new light responsive block copolymer. First, we optimized the membrane formation by using poly(styrene-b-anthracene methyl methacrylate-b-methylmethacrylate) (PS-b-PAnMMA-b-PMMA) copolymer, identifying the most suitable solvent, copolymer block length, and other parameters. The obtained final triblock copolymer membrane morphologies were characterized using atomic force and electron microscopy. The microscopic analysis reveals that the PS-b-PAnMMA-b-PMMA copolymer can form both lamellar and ordered hexagonal nanoporous structures on the membrane top layer in appropriate solvent compositions. The nanostructured membrane emits fluorescence due to the presence of the anthracene mid-block. On irradiation of light the PS-b-PAnMMA-b-PMMA copolymer membranes has an additional stimuli response. The anthracene group undergoes conformational changes by forming [4 + 4] cycloadducts and this alters the membrane\\'s water flux and solute retention. © 2016 The Royal Society of Chemistry.

Molecular Structure of Membrane Tethers

NARCIS (Netherlands)

Baoukina, Svetlana; Marrink, Siewert J.; Tieleman, D. Peter

2012-01-01

Membrane tethers are nanotubes formed by a lipid bilayer. They play important functional roles in cell biology and provide an experimental window on lipid properties. Tethers have been studied extensively in experiments and described by theoretical models, but their molecular structure remains

Membrane Compartmentalization Reducing the Mobility of Lipids and Proteins within a Model Plasma Membrane.

Science.gov (United States)

Koldsø, Heidi; Reddy, Tyler; Fowler, Philip W; Duncan, Anna L; Sansom, Mark S P

2016-09-01

The cytoskeleton underlying cell membranes may influence the dynamic organization of proteins and lipids within the bilayer by immobilizing certain transmembrane (TM) proteins and forming corrals within the membrane. Here, we present coarse-grained resolution simulations of a biologically realistic membrane model of asymmetrically organized lipids and TM proteins. We determine the effects of a model of cytoskeletal immobilization of selected membrane proteins using long time scale coarse-grained molecular dynamics simulations. By introducing compartments with varying degrees of restraints within the membrane models, we are able to reveal how compartmentalization caused by cytoskeletal immobilization leads to reduced and anomalous diffusional mobility of both proteins and lipids. This in turn results in a reduced rate of protein dimerization within the membrane and of hopping of membrane proteins between compartments. These simulations provide a molecular realization of hierarchical models often invoked to explain single-molecule imaging studies of membrane proteins.

The preterm cervix reveals a transcriptomic signature in the presence of premature prelabor rupture of membranes.

Science.gov (United States)

Makieva, Sofia; Dubicke, Aurelija; Rinaldi, Sara F; Fransson, Emma; Ekman-Ordeberg, Gunvor; Norman, Jane E

2017-06-01

Premature prelabor rupture of fetal membranes accounts for 30% of all premature births and is associated with detrimental long-term infant outcomes. Premature cervical remodeling, facilitated by matrix metalloproteinases, may trigger rupture at the zone of the fetal membranes overlying the cervix. The similarities and differences underlying cervical remodeling in premature prelabor rupture of fetal membranes and spontaneous preterm labor with intact membranes are unexplored. We aimed to perform the first transcriptomic assessment of the preterm human cervix to identify differences between premature prelabor rupture of fetal membranes and preterm labor with intact membranes and to compare the enzymatic activities of matrix metalloproteinases-2 and -9 between premature prelabor rupture of fetal membranes and preterm labor with intact membranes. Cervical biopsies were collected following preterm labor with intact membranes (n = 6) and premature prelabor rupture of fetal membranes (n = 5). Biopsies were also collected from reference groups at term labor (n = 12) or term not labor (n = 5). The Illumina HT-12 version 4.0 BeadChips microarray was utilized, and a novel network graph approach determined the specificity of changes between premature prelabor rupture of fetal membranes and preterm labor with intact membranes. Quantitative reverse transcription-polymerase chain reaction and Western blotting confirmed the microarray findings. Immunofluorescence was used for localization studies and gelatin zymography to assess matrix metalloproteinase activity. PML-RARA-regulated adapter molecule 1, FYVE-RhoGEF and PH domain-containing protein 3 and carcinoembryonic antigen-ralated cell adhesion molecule 3 were significantly higher, whereas N-myc downstream regulated gene 2 was lower in the premature prelabor rupture of fetal membranes cervix when compared with the cervix in preterm labor with intact membranes, term labor, and term not labor. PRAM1 and CEACAM3 were localized

Isogeometric Shape Optimization of Vibrating Membranes

DEFF Research Database (Denmark)

Nguyen, Dang Manh; Evgrafov, Anton; Gersborg, Allan Roulund

2011-01-01

We consider a model problem of isogeometric shape optimization of vibrating membranes whose shapes are allowed to vary freely. The main obstacle we face is the need for robust and inexpensive extension of a B-spline parametrization from the boundary of a domain onto its interior, a task which has...... perform a number of numerical experiments with our isogeometric shape optimization algorithm and present smooth, optimized membrane shapes. Our conclusion is that isogeometric analysis fits well with shape optimization....

Photoresponsive nanostructured membranes

KAUST Repository

Madhavan, Poornima; Sutisna, Burhannudin; Sougrat, Rachid; Nunes, Suzana Pereira

2016-01-01

The perspective of adding stimuli-response to isoporous membranes stimulates the development of separation devices with pores, which would open or close under control of environment chemical composition, temperature or exposure to light. Changes in pH and temperature have been previously investigated. In this work, we demonstrate for the first time the preparation of photoresponsive isoporous membranes, applying self-assembly non-solvent induced phase separation to a new light responsive block copolymer. First, we optimized the membrane formation by using poly(styrene-b-anthracene methyl methacrylate-b-methylmethacrylate) (PS-b-PAnMMA-b-PMMA) copolymer, identifying the most suitable solvent, copolymer block length, and other parameters. The obtained final triblock copolymer membrane morphologies were characterized using atomic force and electron microscopy. The microscopic analysis reveals that the PS-b-PAnMMA-b-PMMA copolymer can form both lamellar and ordered hexagonal nanoporous structures on the membrane top layer in appropriate solvent compositions. The nanostructured membrane emits fluorescence due to the presence of the anthracene mid-block. On irradiation of light the PS-b-PAnMMA-b-PMMA copolymer membranes has an additional stimuli response. The anthracene group undergoes conformational changes by forming [4 + 4] cycloadducts and this alters the membrane's water flux and solute retention. © 2016 The Royal Society of Chemistry.

Cell membrane conformation at vertical nanowire array interface revealed by fluorescence imaging

International Nuclear Information System (INIS)

Berthing, Trine; Bonde, Sara; Rostgaard, Katrine R; Martinez, Karen L; Madsen, Morten Hannibal; Sørensen, Claus B; Nygård, Jesper

2012-01-01

The perspectives offered by vertical arrays of nanowires for biosensing applications in living cells depend on the access of individual nanowires to the cell interior. Recent results on electrical access and molecular delivery suggest that direct access is not always obtained. Here, we present a generic approach to directly visualize the membrane conformation of living cells interfaced with nanowire arrays, with single nanowire resolution. The method combines confocal z-stack imaging with an optimized cell membrane labelling strategy which was applied to HEK293 cells interfaced with 2–11 μm long and 3–7 μm spaced nanowires with various surface coatings (bare, aminosilane-coated or polyethyleneimine-coated indium arsenide). We demonstrate that, for all commonly used nanowire lengths, spacings and surface coatings, nanowires generally remain enclosed in a membrane compartment, and are thereby not in direct contact with the cell interior. (paper)

Early Detection of Biofouling on Water Purification Membranes by Ambient Ionization Mass Spectrometry Imaging.

Science.gov (United States)

Jakka Ravindran, Swathy; Kumar, Ramesh; Srimany, Amitava; Philip, Ligy; Pradeep, Thalappil

2018-01-02

By direct analysis of water purification membranes using ambient ionization mass spectrometry, an attempt has been made to understand the molecular signatures of bacterial fouling. Membrane based purification methods are used extensively in water treatment, and a major challenge for them is biofouling. The buildup of microbes and their extracellular polymeric matrix clog the purification membranes and reduce their efficiency. To understand the early stages of bacterial fouling on water purification membranes, we have used desorption electrospray ionization mass spectrometry (DESI MS), where ion formation occurs in ambient conditions and the ionization event is surface sensitive. Biosurfactants at the air-water interface generated by microorganisms as a result of quorum sensing, influence the water-membrane interface and are important for the bacterial attachment. We show that these biosurfactants produced by bacteria can be indicator molecular species signifying initiation of biofilms on membrane surfaces, demonstrated by specific DESI MS signatures. In Pseudomonas aeruginosa, one of the best studied models for biofilm formation, this process is mediated by rhamnolipids forewarning bacterial fouling. Species dependent variation of such molecules can be used for the precise identification of the microorganisms, as revealed by studies on P. aeroginosa (ATCC 25619). The production of biosurfactants is tightly regulated at the transcriptional level by the quorum-sensing (QS) response. Thus, secretion of these extracellular molecules across the membrane surface allows rapid screening of the biofilm community. We show that, the ambient ionization mass spectrometry can detect certain toxic heavy metals present in water, using surfactant-metal complexes as analytes. We believe that such studies conducted on membranes in various input water streams will help design suitable membrane processes specific to the input streams.

Controlled Bulk Properties of Composite Polymeric Solutions for Extensive Structural Order of Honeycomb Polysulfone Membranes.

Science.gov (United States)

Gugliuzza, Annarosa; Perrotta, Maria Luisa; Drioli, Enrico

2016-05-16

This work provides additional insights into the identification of operating conditions necessary to overcome a current limitation to the scale-up of the breath figure method, which is regarded as an outstanding manufacturing approach for structurally ordered porous films. The major restriction concerns, indeed, uncontrolled touching droplets at the boundary. Herein, the bulk of polymeric solutions are properly managed to generate honeycomb membranes with a long-range structurally ordered texture. Water uptake and dynamics are explored as chemical environments are changed with the intent to modify the hydrophilic/hydrophobic balance and local water floatation. In this context, a model surfactant such as the polyoxyethylene sorbitan monolaurate is used in combination with alcohols at different chain length extents and a traditional polymer such as the polyethersufone. Changes in the interfacial tension and kinematic viscosity taking place in the bulk of composite solutions are explored and examined in relation to competitive droplet nucleation and growth rate. As a result, extensive structurally ordered honeycomb textures are obtained with the rising content of the surfactant while a broad range of well-sized pores is targeted as a function of the hydrophilic-hydrophobic balance and viscosity of the composite polymeric mixture. The experimental findings confirm the consistency of the approach and are expected to give propulsion to the commercially production of breath figures films shortly.

Controlled Bulk Properties of Composite Polymeric Solutions for Extensive Structural Order of Honeycomb Polysulfone Membranes

Directory of Open Access Journals (Sweden)

Annarosa Gugliuzza

2016-05-01

Full Text Available This work provides additional insights into the identification of operating conditions necessary to overcome a current limitation to the scale-up of the breath figure method, which is regarded as an outstanding manufacturing approach for structurally ordered porous films. The major restriction concerns, indeed, uncontrolled touching droplets at the boundary. Herein, the bulk of polymeric solutions are properly managed to generate honeycomb membranes with a long-range structurally ordered texture. Water uptake and dynamics are explored as chemical environments are changed with the intent to modify the hydrophilic/hydrophobic balance and local water floatation. In this context, a model surfactant such as the polyoxyethylene sorbitan monolaurate is used in combination with alcohols at different chain length extents and a traditional polymer such as the polyethersufone. Changes in the interfacial tension and kinematic viscosity taking place in the bulk of composite solutions are explored and examined in relation to competitive droplet nucleation and growth rate. As a result, extensive structurally ordered honeycomb textures are obtained with the rising content of the surfactant while a broad range of well-sized pores is targeted as a function of the hydrophilic-hydrophobic balance and viscosity of the composite polymeric mixture. The experimental findings confirm the consistency of the approach and are expected to give propulsion to the commercially production of breath figures films shortly.

Recent achievements in facilitated transport membranes for separation processes

Directory of Open Access Journals (Sweden)

H. C. Ferraz

2007-03-01

Full Text Available Membrane separation processes have been extensively used for some important industrial separations, substituting traditional methods. However, some applications require the development of new membranes. In this work, we discuss recent progress achieved in this field, focusing on gas and liquid separation using facilitated transport membranes. The advantages of using a carrier species either in a liquid membrane or fixed in a polymer matrix to enhance both the flux and the selectivity of the transport are summarized. The most probable transport mechanisms in these membranes are presented and the improvements needed to spread this technology are also discussed. As examples, we discuss our very successful experiences in air fractioning, olefin/paraffin separation and sugar recovery using liquid and fixed carrier membranes.

Tracking Glideosome-associated protein 50 reveals the development and organization of the inner membrane complex of Plasmodium falciparum.

Science.gov (United States)

Yeoman, Jeffrey A; Hanssen, Eric; Maier, Alexander G; Klonis, Nectarios; Maco, Bohumil; Baum, Jake; Turnbull, Lynne; Whitchurch, Cynthia B; Dixon, Matthew W A; Tilley, Leann

2011-04-01

The most deadly of the human malaria parasites, Plasmodium falciparum, has different stages specialized for invasion of hepatocytes, erythrocytes, and the mosquito gut wall. In each case, host cell invasion is powered by an actin-myosin motor complex that is linked to an inner membrane complex (IMC) via a membrane anchor called the glideosome-associated protein 50 (PfGAP50). We generated P. falciparum transfectants expressing green fluorescent protein (GFP) chimeras of PfGAP50 (PfGAP50-GFP). Using immunoprecipitation and fluorescence photobleaching, we show that C-terminally tagged PfGAP50-GFP can form a complex with endogenous copies of the linker protein PfGAP45 and the myosin A tail domain-interacting protein (MTIP). Full-length PfGAP50-GFP is located in the endoplasmic reticulum in early-stage parasites and then redistributes to apical caps during the formation of daughter merozoites. In the final stage of schizogony, the PfGAP50-GFP profile extends further around the merozoite surface. Three-dimensional (3D) structured illumination microscopy reveals the early-stage IMC as a doubly punctured flat ellipsoid that separates to form claw-shaped apposed structures. A GFP fusion of PfGAP50 lacking the C-terminal membrane anchor is misdirected to the parasitophorous vacuole. Replacement of the acid phosphatase homology domain of PfGAP50 with GFP appears to allow correct trafficking of the chimera but confers a growth disadvantage.

Structure and interactions in biomaterials based on membrane-biopolymer self-assembly

Science.gov (United States)

Koltover, Ilya

Physical and chemical properties of artificial pure lipid membranes have been extensively studied during the last two decades and are relatively well understood. However, most real membrane systems of biological and biotechnological importance incorporate macromolecules either embedded into the membranes or absorbed onto their surfaces. We have investigated three classes of self-assembled membrane-biopolymer biomaterials: (i) Structure, interactions and stability of the two-dimensional crystals of the integral membrane protein bacteriorhodopsin (bR). We have conducted a synchrotron x-ray diffraction study of oriented bR multilayers. The important findings were as follows: (1) the protein 2D lattice exhibited diffraction patterns characteristic of a 2D solid with power-law decay of in-plane positional correlations, which allowed to measure the elastic constants of protein crystal; (2) The crystal melting temperature was a function of the multilayer hydration, reflecting the effect of inter-membrane repulsion on the stability of protein lattice; (3) Preparation of nearly perfect (mosaicity gene therapy applications. We have established that DNA complexes with cationic lipid (DOTAP) and a neutral lipid (DOPC) have a compact multilayer liquid crystalline structure ( L ca ) with DNA intercalated between the lipid bilayers in a periodic 2D smectic phase. Furthermore, a different 2D columnar phase of complexes was found in mixtures with a transfectionen-hancing lipid DOPE. This structure ( HcII ) derived from synchrotron x-ray diffraction consists of DNA coated by cationic lipid monolayers and arranged on a two-dimensional hexagonal lattice. Optical microscopy revealed that the L ca complexes bind stably to anionic vesicles (models of cellular membranes), whereas the more transfectant HcII complexes are unstable, rapidly fusing and releasing DNA upon adhering to anionic vesicles.

Comparative Analysis of Membrane Vesicles from Three Piscirickettsia salmonis Isolates Reveals Differences in Vesicle Characteristics.

Directory of Open Access Journals (Sweden)

Julia I Tandberg

Full Text Available Membrane vesicles (MVs are spherical particles naturally released from the membrane of Gram-negative bacteria. Bacterial MV production is associated with a range of phenotypes including biofilm formation, horizontal gene transfer, toxin delivery, modulation of host immune responses and virulence. This study reports comparative profiling of MVs from bacterial strains isolated from three widely disperse geographical areas. Mass spectrometry identified 119, 159 and 142 proteins in MVs from three different strains of Piscirickettsia salmonis isolated from salmonids in Chile (LF-89, Norway (NVI 5692 and Canada (NVI 5892, respectively. MV comparison revealed several strain-specific differences related to higher virulence capability for LF-89 MVs, both in vivo and in vitro, and stronger similarities between the NVI 5692 and NVI 5892 MV proteome. The MVs were similar in size and appearance as analyzed by electron microscopy and dynamic light scattering. The MVs from all three strains were internalized by both commercial and primary immune cell cultures, which suggest a potential role of the MVs in the bacterium's utilization of leukocytes. When MVs were injected into an adult zebrafish infection model, an upregulation of several pro-inflammatory genes were observed in spleen and kidney, indicating a modulating effect on the immune system. The present study is the first comparative analysis of P. salmonis derived MVs, highlighting strain-specific vesicle characteristics. The results further illustrate that the MV proteome from one bacterial strain is not representative of all bacterial strains within one species.

High Performance Regenerated Cellulose Membranes from Trimethylsilyl Cellulose

KAUST Repository

Ali, Ola

2013-01-01

Regenerated cellulose (RC) membranes are extensively used in medical and pharmaceutical separation processes due to their biocompatibility, low fouling tendency and solvent resistant properties. They typically possess ultrafiltration

Membrane skeletal proteins and their integral membrane protein anchors are targets for tyrosine and threonine kinases in Euglena.

Science.gov (United States)

Fazio, M J; Da Silva, A C; Rosiere, T K; Bouck, G B

1995-01-01

Proteins of the membrane skeleton of Euglena gracilis were extensively phosphorylated in vivo and in vitro after incubation with [32P]-orthophosphate or gamma-[32P] ATP. Endogenous protein threonine/serine activity phosphorylated the major membrane skeletal proteins (articulins) and the putative integral membrane protein (IP39) anchor for articulins. The latter was also the major target for endogenous protein tyrosine kinase activity. A cytoplasmic domain of IP39 was specifically phosphorylated, and removal of this domain with papain eliminated the radiolabeled phosphoamino acids and eliminated or radically shifted the PI of the multiple isoforms of IP39. In gel kinase assays IP39 autophosphorylated and a 25 kDa protein which does not autophosphorylate was identified as a threonine/serine (casein) kinase. Plasma membranes from the membrane skeletal protein complex contained threonine/serine (casein) kinase activity, and cross-linking experiments suggested that IP39 was the likely source for this membrane activity. pH optima, cation requirements and heparin sensitivity of the detergent solubilized membrane activity were determined. Together these results suggest that protein kinases may be important modulators of protein assembly and function of the membrane skeleton of these protistan cells.

Functional dynamics of cell surface membrane proteins.

Science.gov (United States)

Nishida, Noritaka; Osawa, Masanori; Takeuchi, Koh; Imai, Shunsuke; Stampoulis, Pavlos; Kofuku, Yutaka; Ueda, Takumi; Shimada, Ichio

2014-04-01

Cell surface receptors are integral membrane proteins that receive external stimuli, and transmit signals across plasma membranes. In the conventional view of receptor activation, ligand binding to the extracellular side of the receptor induces conformational changes, which convert the structure of the receptor into an active conformation. However, recent NMR studies of cell surface membrane proteins have revealed that their structures are more dynamic than previously envisioned, and they fluctuate between multiple conformations in an equilibrium on various timescales. In addition, NMR analyses, along with biochemical and cell biological experiments indicated that such dynamical properties are critical for the proper functions of the receptors. In this review, we will describe several NMR studies that revealed direct linkage between the structural dynamics and the functions of the cell surface membrane proteins, such as G-protein coupled receptors (GPCRs), ion channels, membrane transporters, and cell adhesion molecules. Copyright © 2013 Elsevier Inc. All rights reserved.

Integration of computational modeling with membrane transport studies reveals new insights into amino acid exchange transport mechanisms

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Widdows, Kate L.; Panitchob, Nuttanont; Crocker, Ian P.; Please, Colin P.; Hanson, Mark A.; Sibley, Colin P.; Johnstone, Edward D.; Sengers, Bram G.; Lewis, Rohan M.; Glazier, Jocelyn D.

2015-01-01

Uptake of system L amino acid substrates into isolated placental plasma membrane vesicles in the absence of opposing side amino acid (zero-trans uptake) is incompatible with the concept of obligatory exchange, where influx of amino acid is coupled to efflux. We therefore hypothesized that system L amino acid exchange transporters are not fully obligatory and/or that amino acids are initially present inside the vesicles. To address this, we combined computational modeling with vesicle transport assays and transporter localization studies to investigate the mechanisms mediating [14C]l-serine (a system L substrate) transport into human placental microvillous plasma membrane (MVM) vesicles. The carrier model provided a quantitative framework to test the 2 hypotheses that l-serine transport occurs by either obligate exchange or nonobligate exchange coupled with facilitated transport (mixed transport model). The computational model could only account for experimental [14C]l-serine uptake data when the transporter was not exclusively in exchange mode, best described by the mixed transport model. MVM vesicle isolates contained endogenous amino acids allowing for potential contribution to zero-trans uptake. Both L-type amino acid transporter (LAT)1 and LAT2 subtypes of system L were distributed to MVM, with l-serine transport attributed to LAT2. These findings suggest that exchange transporters do not function exclusively as obligate exchangers.—Widdows, K. L., Panitchob, N., Crocker, I. P., Please, C. P., Hanson, M. A., Sibley, C. P., Johnstone, E. D., Sengers, B. G., Lewis, R. M., Glazier, J. D. Integration of computational modeling with membrane transport studies reveals new insights into amino acid exchange transport mechanisms. PMID:25761365

Revealing the Raft Domain Organization in the Plasma Membrane by Single-Molecule Imaging of Fluorescent Ganglioside Analogs.

Science.gov (United States)

Suzuki, Kenichi G N; Ando, Hiromune; Komura, Naoko; Konishi, Miku; Imamura, Akihiro; Ishida, Hideharu; Kiso, Makoto; Fujiwara, Takahiro K; Kusumi, Akihiro

2018-01-01

Gangliosides have been implicated in a variety of physiological processes, particularly in the formation and function of raft domains in the plasma membrane. However, the scarcity of suitable fluorescent ganglioside analogs had long prevented us from determining exactly how gangliosides perform their functions in the live-cell plasma membrane. With the development of new fluorescent ganglioside analogs, as described by Komura et al. (2017), this barrier has been broken. We can now address the dynamic behaviors of gangliosides in the live-cell plasma membrane, using fluorescence microscopy, particularly by single-fluorescent molecule imaging and tracking. Single-molecule tracking of fluorescent GM1 and GM3 revealed that these molecules are transiently and dynamically recruited to monomers (monomer-associated rafts) and homodimer rafts of the raftophilic GPI-anchored protein CD59 in quiescent cells, with exponential residency times of 12 and 40ms, respectively, in a manner dependent on raft-lipid interactions. Upon CD59 stimulation, which induces CD59-cluster signaling rafts, the fluorescent GM1 and GM3 analogs were recruited to the signaling rafts, with a lifetime of 48ms. These results represent the first direct evidence that GPI-anchored receptors and gangliosides interact in a cholesterol-dependent manner. Furthermore, they show that gangliosides continually move in and out of rafts that contain CD59 in an extremely dynamic manner, with much higher frequency than expected previously. Such studies would not have been possible without fluorescent ganglioside probes, which exhibit native-like behavior and single-molecule tracking. In this chapter, we review the methods for single-molecule tracking of fluorescent ganglioside analogs and the results obtained by applying these methods. © 2018 Elsevier Inc. All rights reserved.

Protein transport across and into cell membranes in bacteria and archaea

NARCIS (Netherlands)

Yuan, Jijun; Zweers, Jessica C.; van Dijl, Jan Maarten; Dalbey, Ross E.

In the three domains of life, the Sec, YidC/Oxa1, and Tat translocases play important roles in protein translocation across membranes and membrane protein insertion. While extensive studies have been performed on the endoplasmic reticular and Escherichia coli systems, far fewer studies have been

Potassium accumulation by the glial membrane pump as revealed by membrane potential recording from isolated rabbit retinal Müller cells.

Science.gov (United States)

Reichenbach, A; Nilius, B; Eberhardt, W

1986-01-30

Müller (glial) cells were isolated from rabbit retinae by papaine and mechanical dissociation. In a special perfusion chamber, the cells were penetrated with a recording electrode. When high-K+ solutions were applied into the environment of the cells by means of a second micropipette, the cell membrane depolarized strongly. During prolonged application of high-K+ solutions, however, there occurred a marked repolarization, and after cessation of high-K+ application, a strong hyperpolarization was observed. Both effects disappeared under the influence of ouabain, suggesting the accumulation of intracellular K+ by an active membrane pump. The data were used for calculation of the membrane's Na+:K+ permeability ratio, the intracellular K+ concentration, the pump rate and the mean pump site density. The calculated values are in good agreement with published data from mammalian astrocytes and are compared with those from amphibian Müller cells.

Proteomic amino-termini profiling reveals targeting information for protein import into complex plastids.

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Pitter F Huesgen

Full Text Available In organisms with complex plastids acquired by secondary endosymbiosis from a photosynthetic eukaryote, the majority of plastid proteins are nuclear-encoded, translated on cytoplasmic ribosomes, and guided across four membranes by a bipartite targeting sequence. In-depth understanding of this vital import process has been impeded by a lack of information about the transit peptide part of this sequence, which mediates transport across the inner three membranes. We determined the mature N-termini of hundreds of proteins from the model diatom Thalassiosira pseudonana, revealing extensive N-terminal modification by acetylation and proteolytic processing in both cytosol and plastid. We identified 63 mature N-termini of nucleus-encoded plastid proteins, deduced their complete transit peptide sequences, determined a consensus motif for their cleavage by the stromal processing peptidase, and found evidence for subsequent processing by a plastid methionine aminopeptidase. The cleavage motif differs from that of higher plants, but is shared with other eukaryotes with complex plastids.

Physics of smectic membranes

Science.gov (United States)

Pieranski, P.; Beliard, L.; Tournellec, J.-Ph.; Leoncini, X.; Furtlehner, C.; Dumoulin, H.; Riou, E.; Jouvin, B.; Fénerol, J.-P.; Palaric, Ph.; Heuving, J.; Cartier, B.; Kraus, I.

1993-03-01

Due to their layered structure, smectic liquid crystals can form membranes, similar to soap bubbles, that can be spanned on frames. Such smectic membranes have been used extensively as samples in many structural X-ray studies of smectic liquid crystals. In this context they have been considered as very convenient and highly perfect samples but little attention has been paid to the reasons for their existence and to the process of their formation. Our aim here is to address a first list of questions, which are the most urgent to answer. We will also describe experiments and models that have been conceived especially in order to understand the physics of these fascinating systems.

Development of novel nano-composite membranes as introduction systems for mass spectrometers: Contrasting nano-composite membranes and conventional inlet systems

Science.gov (United States)

Miranda, Luis Diego

This dissertation presents the development of novel nano-composite membranes as introduction systems for mass spectrometers. These nano-composite membranes incorporate anodic aluminum oxide (AAO) membranes as templates that can be used by themselves or modified by a variety of chemical deposition processes. Two types of nano-composite membranes are presented. The first nano-composite membrane has carbon deposited within the pores of an AAO membrane. The second nano-composite membrane is made by coating an AAO membrane with a thin polymer film. The following chapters describe the transmission properties these nano-composite membranes and compare them to conventional mass spectrometry introduction systems. The nano- composite membranes were finally coupled to the inlet system of an underwater mass spectrometer revealing their utility in field deployments.

[Formation of purple membranes during salt bacteria cultivation].

Science.gov (United States)

Chekulaeva, L N; Korolev, Iu N; Telegin, N L; Rikhireva, G T

1975-01-01

Experiments have been carried out on cultivation of halophile with probe selection in the interval of 1--2 hours to record the spectra of repeated disturbed completed inner reflection. Periodicity in the changes of spectral characteristics of the culture with the interval of 20--24 hours is revealed. A clearly expressed dichroism of the amid II band of the membrane complex is found, the absence of this dichroism in the protein isolated from the membrane complex is stated. It is suggested that dichroism revealed is a specific feature of the presence of purpuric membranes in the cells. Spontaneous plane orientation of protein macromolecules in purpuric membranes is established. The level of dichroism of amid II band is shown to depend on fermentation conditions of salt bacteria.

Membrane-bound 2,3-diphosphoglycerate phosphatase of human erythrocytes.

Science.gov (United States)

Schröter, W; Neuvians, M

1970-12-01

Gradual osmotic hemolysis of human erythrocytes reduces the cell content of whole protein, hemoglobin, 2,3-diphosphoglycerate and triosephosphate isomerase extensively, but not that of membrane protein and 2,3-diphosphoglycerate phosphatase. After the refilling of the ghosts with 2,3-diphosphoglycerate and reconstitution of the membrane, the 2,3-diphosphoglycerate phosphatase activity equals that of intact red cells. The membrane-bound 2,3-diphosphoglycerate phosphatase can be activated by sodium hyposulfite. The enzyme system of ghosts seems to differ from that of intact red cells with regard to the optima of pH and temperature. It remains to be elucidated if the membrane binding of the 2,3-diphosphoglycerate phosphatase is related to the transfer of inorganic phosphate across the red cell membrane.

Flapping tail membrane in bats produces potentially important thrust during horizontal takeoffs and very slow flight.

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Rick A Adams

Full Text Available Historically, studies concerning bat flight have focused primarily on the wings. By analyzing high-speed video taken on 48 individuals of five species of vespertilionid bats, we show that the capacity to flap the tail-membrane (uropatagium in order to generate thrust and lift during takeoffs and minimal-speed flight (<1 m (s-1 was largely underestimated. Indeed, bats flapped the tail-membrane by extensive dorso-ventral fanning motions covering as much as 135 degrees of arc consistent with thrust generation by air displacement. The degree of dorsal extension of the tail-membrane, and thus the potential amount of thrust generated during platform launches, was significantly correlated with body mass (P = 0.02. Adduction of the hind limbs during upstrokes collapsed the tail-membrane thereby reducing its surface area and minimizing negative lift forces. Abduction of the hind limbs during the downstroke fully expanded the tail-membrane as it was swept ventrally. The flapping kinematics of the tail-membrane is thus consistent with expectations for an airfoil. Timing offsets between the wings and tail-membrane during downstrokes was as much as 50%, suggesting that the tail-membrane was providing thrust and perhaps lift when the wings were retracting through the upstoke phase of the wing-beat cycle. The extent to which the tail-membrane was used during takeoffs differed significantly among four vespertilionid species (P = 0.01 and aligned with predictions derived from bat ecomorphology. The extensive fanning motion of the tail membrane by vespertilionid bats has not been reported for other flying vertebrates.

SYNOVIAL MEMBRANE CHANGES IN PATIENTS WITH RHEUMATOID ARTHRITIS REVEALED DURING ARTHROSCOPY

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E. B. Komarova

2017-01-01

Full Text Available Exudative or proliferative processes accompanied by joint destruction may predominate in the synovial membrane (SM in different stages of rheumatoid arthritis (RA. The features of SM changes should be considered in the early diagnosis of RA and in the development of combination treatment for this disease.Objective: to study arthroscopic SM changes in patients with different RA durations and different blood anti-cyclic citrullinated peptide (anti-CCP antibody levels.Subjects and methods. 37 patients with RA underwent arthroscopy with an arthroscope of a 2.4-mm diameter and 30° angle (Karl Storz GmbH, Germany. RA duration was <2 years in 17 patients and ranged from 2 to 8 years in 20; the blood level of anti-CCP antibodies did not exceed 60 IU/ml in 15 patients and it was higher in 22. SM changes were assessed by a semiquantitative method.Results and discussion. Arthroscopy revealed a preponderance of SM hyperemia with an increased vascular pattern (p<0.01 and fibrin deposits (p<0.05 in patients with a RA duration of < 2 years and that of villous hyperplasia with the formation of club-shaped villi (p<0.05 in those with a RA duration of >2 years. The increase in anti-CCP by >60 U/ml was associated with a predominance of inflammatory hyperplasia (p < 0.01, SM hyperemia with a pronounced vascular pattern (p<0.05, as well as with the presence of pannus (pp<0.01.

Molecular view of the structural reorganization of water in DPPC multilamellar membranes induced by L-cysteine methyl ester

Science.gov (United States)

Arias, Juan Marcelo; Tuttolomondo, María Eugenia; Díaz, Sonia Beatriz; Altabef, Aida Ben

2018-03-01

In order to study the interaction between L-cysteine methyl ester (CM) and multilamellar vesicles (MLV's) of DPPC, an extensive study was made by various techniques such as Infrared and Raman spectroscopy and Differential Scanning Calorimetry (DSC). Our results revealed by the different techniques used that CM interacts with the DPPC in the region of the polar head, specifying with the phosphate groups, replacing water molecules of hydration by modifying the hydration of the polar head. By Infrared spectroscopy and DSC we observed an increase in the main transition temperature (Tm) and a gradual loss of the pre-transition (Tp) with the increase of the molar ratio CM:DPPC. Of the analyzed, we can conclude that the interaction of CM with DPPC alters the degree of hydration of the membrane altering properties of the same as the transition temperature. Moreover, the results of the thiol site behavior in CM interacting in the CM/DPPC complex will be reveal the possibility of unknown functional roles of the lipidic components of the membrane.

Nanoclustering as a dominant feature of plasma membrane organization

NARCIS (Netherlands)

Garcia-Parajo, M.F.; Cambi, A.; Torreno-Pina, J.A.; Thompson, N.; Jacobson, K.

2014-01-01

Early studies have revealed that some mammalian plasma membrane proteins exist in small nanoclusters. The advent of super-resolution microscopy has corroborated and extended this picture, and led to the suggestion that many, if not most, membrane proteins are clustered at the plasma membrane at

Atomic force microscopy reveals multiple patterns of antenna organization in purple bacteria: implications for energy transduction mechanisms and membrane modeling.

Science.gov (United States)

Sturgis, James N; Niederman, Robert A

2008-01-01

Recent topographs of the intracytoplasmic membrane (ICM) of purple bacteria obtained by atomic force microscopy (AFM) have provided the first surface views of the native architecture of a multicomponent biological membrane at submolecular resolution, representing an important landmark in structural biology. A variety of species-dependent, closely packed arrangements of light-harvesting (LH) complexes was revealed: the most highly organized was found in Rhodobacter sphaeroides in which the peripheral LH2 antenna was seen either in large clusters or in fixed rows interspersed among ordered arrays of dimeric LH1-reaction center (RC) core complexes. A more random organization was observed in other species containing both the LH1 and LH2 complexes, as typified by Rhododspirillum photometricum with randomly packed monomeric LH1-RC core complexes intermingled with large, paracrystalline domains of LH2 antenna. Surprisingly, no structures that could be identified as the ATP synthase or cytochrome bc (1) complexes were observed, which may reflect their localization at ICM vesicle poles or in curved membrane areas, out of view from the flat regions imaged by AFM. This possible arrangement of energy transducing complexes has required a reassessment of energy tranduction mechanisms which place the cytochrome bc (1) complex in close association with the RC. Instead, more plausible proposals must account for the movement of quinone redox species over considerable membrane distances on appropriate time scales. AFM, together with atomic resolution structures are also providing the basis for molecular modeling of the ICM that is leading to an improved picture of the supramolecular organization of photosynthetic complexes, as well as the forces that drive their segregation into distinct domains.

Chain scission and anti fungal effect of electron beam on cellulose membrane

International Nuclear Information System (INIS)

Wanichapichart, Pikul; Taweepreeda, Wirach; Nawae, Safitree; Choomgan, Pastraporn; Yasenchak, Dan

2012-01-01

Two types of bacterial cellulose (BC) membranes were produced under a modified H and S medium using sucrose as a carbon source, with (CCB) and without (SHB) coconut juice supplement. Both membranes showed similar crystallinity of 69.24 and 71.55%. After being irradiated with E-beams under oxygen limited and ambient condition, the results from water contact angle showed that only the irradiated membrane CCB was increased from 30 to 40 degrees, and irradiation under oxygen ambient condition provided the greatest value. Comparing with the control membranes, smaller water flux was the cases after electron beam irradiation which indicated a reduction of membrane pore area. However, the results from molecular weight cut off (MWCO) revealed that chain scission was greater for membrane SHB and its cut off was increased from 28,000 Da to more than 35,000 Da. FTIR analysis revealed some changes in membrane functional groups, corresponding with the above results. These changes initiated new property of cellulose membranes, an anti-fungal food wrap. - Highlights: ► Electron beam irradiation increased membrane hydrophobicity and molecular weight cut off. ► The irradiation caused chain scissoring and anti fungal property of cellulose membrane. ► FT-IR studies revealed changes in functional groups causing a decrease in membrane moisture. ► Anti fungal test of cellulose membrane showed the same shelf life as polyethylene sheet.

Fluxes of water through aquaporin 9 weaken membrane-cytoskeleton anchorage and promote formation of membrane protrusions.

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Thommie Karlsson

Full Text Available All modes of cell migration require rapid rearrangements of cell shape, allowing the cell to navigate within narrow spaces in an extracellular matrix. Thus, a highly flexible membrane and a dynamic cytoskeleton are crucial for rapid cell migration. Cytoskeleton dynamics and tension also play instrumental roles in the formation of different specialized cell membrane protrusions, viz. lamellipodia, filopodia, and membrane blebs. The flux of water through membrane-anchored water channels, known as aquaporins (AQPs has recently been implicated in the regulation of cell motility, and here we provide novel evidence for the role of AQP9 in the development of various forms of membrane protrusion. Using multiple imaging techniques and cellular models we show that: (i AQP9 induced and accumulated in filopodia, (ii AQP9-associated filopodial extensions preceded actin polymerization, which was in turn crucial for their stability and dynamics, and (iii minute, local reductions in osmolarity immediately initiated small dynamic bleb-like protrusions, the size of which correlated with the reduction in osmotic pressure. Based on this, we present a model for AQP9-induced membrane protrusion, where the interplay of water fluxes through AQP9 and actin dynamics regulate the cellular protrusive and motile activity of cells.

Anti-glomerular basement membrane disease superimposed on membranous nephropathy: a case report and review of the literature

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Nivera Noel

2010-08-01

Full Text Available Abstract Introduction Anti-glomerular basement membrane disease is a rare autoimmune disorder characterized by pulmonary hemorrhage, crescentic glomerulonephritis and the presence of circulating anti-glomerular basement membrane antibodies. The simultaneous occurrence of both anti-glomerular basement membrane disease and membranous nephropathy is rare. Case presentation A 59-year-old Hispanic man presented with acute onset of nausea and vomiting and was found to have renal insufficiency. Work-up included a kidney biopsy, which revealed anti-glomerular basement membrane disease with underlying membranous nephropathy. He was treated with emergent hemodialysis, intravenous corticosteroids, plasmapheresis, and cyclophosphamide without improvement in his renal function. Conclusion Simultaneous anti-glomerular basement membrane disease and membranous nephropathy is very rare. There have been 16 previous case reports in the English language literature that have been associated with a high mortality and morbidity, and a very high rate of renal failure resulting in hemodialysis. Co-existence of membranous nephropathy and anti-glomerular basement membrane disease may be immune-mediated, although the exact mechanism is not clear.

Evaluation of thin film ceria membranes for syngas membrane reactors—Preparation, characterization and testing

DEFF Research Database (Denmark)

Kaiser, Andreas; Foghmoes, Søren Preben Vagn; Chatzichristodoulou, Christodoulos

2011-01-01

Gadolinium doped ceria (Ce0.1Gd0.9O1.95−δ, CGO10) was investigated as oxygen separation membrane material for application in syngas production. Planar, thin film CGO10 membranes were fabricated by tape casting and lamination on porous NiO-YSZ supports and subsequent co-sintering. High oxygen fluxes......-stoichiometry profile in the 30μm thin CGO membrane under operation reveal that due to oxygen permeation in the membrane the largest non-stoichiometry at the permeate (fuel) side is more than a factor of 6 times smaller at 850°C than that expected for CGO10 at equilibrium. The related relative expansion of the thin...... film CGO membrane should therefore lie below the expansion limit of 0.1% expected to be critical for mechanical stability and thereby allows for operation at high temperatures and low oxygen partial pressures....

'Boomerang'-like insertion of a fusogenic peptide in a lipid membrane revealed by solid-state 19F NMR.

Science.gov (United States)

Afonin, Sergii; Dürr, Ulrich H N; Glaser, Ralf W; Ulrich, Anne S

2004-02-01

Solid state (19)F NMR revealed the conformation and alignment of the fusogenic peptide sequence B18 from the sea urchin fertilization protein bindin embedded in flat phospholipid bilayers. Single (19)F labels were introduced into nine distinct positions along the wild-type sequence by substituting each hydrophobic amino acid, one by one, with L-4-fluorophenylglycine. Their anisotropic chemical shifts were measured in uniaxially oriented membrane samples and used as orientational constraints to model the peptide structure in the membrane-bound state. Previous (1)H NMR studies of B18 in 30% TFE and in detergent micelles had shown that the peptide structure consists of two alpha-helical segments that are connected by a flexible hinge. This helix-break-helix motif was confirmed here by the solid-state (19)F NMR data, while no other secondary structure (beta-sheet, 3(10)-helix) was compatible with the set of orientational constraints. For both alpha-helical segments we found that the helical conformation extends all the way to the respective N- and C-termini of the peptide. Analysis of the corresponding tilt and azimuthal rotation angles showed that the N-terminal helix of B18 is immersed obliquely into the bilayer (at a tilt angle tau approximately 54 degrees), whereas the C-terminus is peripherally aligned (tau approximately 91 degrees). The azimuthal orientation of the two segments is consistent with the amphiphilic distribution of side-chains. The observed 'boomerang'-like mode of insertion into the membrane may thus explain how peptide binding leads to lipid dehydration and acyl chain perturbation as a prerequisite for bilayer fusion to occur. Copyright 2004 John Wiley & Sons, Ltd.

Separator Membrane from Crosslinked Poly(Vinyl Alcohol and Poly(Methyl Vinyl Ether-alt-Maleic Anhydride

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Charu Vashisth Rohatgi

2015-03-01

Full Text Available In this work, we report separator membranes from crosslinking of two polymers, such as poly vinyl alcohol (PVA with an ionic polymer poly(methyl vinyl ether-alt-maleic anhydride (PMVE-MA. Such interpolymer-networked systems were extensively used for biomedical and desalination applications but they were not examined for their potential use as membranes or separators for batteries. Therefore, the chemical interactions between these two polymers and the influence of such crosslinking on physicochemical properties of the membrane are systematically investigated through rheology and by critical gel point study. The hydrogen bonding and the chemical interaction between PMVE-MA and PVA resulted in highly cross-linked membranes. Effect of the molecular weight of PVA on the membrane properties was also examined. The developed membranes were extensively characterized by studying their physicochemical properties (water uptake, swelling ratio, and conductivity, thermal and electrochemical properties using differential scanning calorimetry (DSC, dynamic mechanical analysis (DMA, thermo-gravimetric analysis (TGA and electrochemical impedance spectroscopy (EIS. The DSC study shows the presence of a single Tg in the membranes indicating compatibility of the two polymers in flexible and transparent films. The membranes show good stability and ion conductivity suitable for separator applications.

Separator Membrane from Crosslinked Poly(Vinyl Alcohol) and Poly(Methyl Vinyl Ether-alt-Maleic Anhydride)

Science.gov (United States)

Rohatgi, Charu Vashisth; Dutta, Naba K.; Choudhury, Namita Roy

2015-01-01

In this work, we report separator membranes from crosslinking of two polymers, such as poly vinyl alcohol (PVA) with an ionic polymer poly(methyl vinyl ether-alt-maleic anhydride) (PMVE-MA). Such interpolymer-networked systems were extensively used for biomedical and desalination applications but they were not examined for their potential use as membranes or separators for batteries. Therefore, the chemical interactions between these two polymers and the influence of such crosslinking on physicochemical properties of the membrane are systematically investigated through rheology and by critical gel point study. The hydrogen bonding and the chemical interaction between PMVE-MA and PVA resulted in highly cross-linked membranes. Effect of the molecular weight of PVA on the membrane properties was also examined. The developed membranes were extensively characterized by studying their physicochemical properties (water uptake, swelling ratio, and conductivity), thermal and electrochemical properties using differential scanning calorimetry (DSC), dynamic mechanical analysis (DMA), thermo-gravimetric analysis (TGA) and electrochemical impedance spectroscopy (EIS). The DSC study shows the presence of a single Tg in the membranes indicating compatibility of the two polymers in flexible and transparent films. The membranes show good stability and ion conductivity suitable for separator applications. PMID:28347019

Antifouling membranes for sustainable water purification: strategies and mechanisms.

Science.gov (United States)

Zhang, Runnan; Liu, Yanan; He, Mingrui; Su, Yanlei; Zhao, Xueting; Elimelech, Menachem; Jiang, Zhongyi

2016-10-24

One of the greatest challenges to the sustainability of modern society is an inadequate supply of clean water. Due to its energy-saving and cost-effective features, membrane technology has become an indispensable platform technology for water purification, including seawater and brackish water desalination as well as municipal or industrial wastewater treatment. However, membrane fouling, which arises from the nonspecific interaction between membrane surface and foulants, significantly impedes the efficient application of membrane technology. Preparing antifouling membranes is a fundamental strategy to deal with pervasive fouling problems from a variety of foulants. In recent years, major advancements have been made in membrane preparation techniques and in elucidating the antifouling mechanisms of membrane processes, including ultrafiltration, nanofiltration, reverse osmosis and forward osmosis. This review will first introduce the major foulants and the principal mechanisms of membrane fouling, and then highlight the development, current status and future prospects of antifouling membranes, including antifouling strategies, preparation techniques and practical applications. In particular, the strategies and mechanisms for antifouling membranes, including passive fouling resistance and fouling release, active off-surface and on-surface strategies, will be proposed and discussed extensively.

A membrane-bound vertebrate globin.

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Miriam Blank

Full Text Available The family of vertebrate globins includes hemoglobin, myoglobin, and other O(2-binding proteins of yet unclear functions. Among these, globin X is restricted to fish and amphibians. Zebrafish (Danio rerio globin X is expressed at low levels in neurons of the central nervous system and appears to be associated with the sensory system. The protein harbors a unique N-terminal extension with putative N-myristoylation and S-palmitoylation sites, suggesting membrane-association. Intracellular localization and transport of globin X was studied in 3T3 cells employing green fluorescence protein fusion constructs. Both myristoylation and palmitoylation sites are required for correct targeting and membrane localization of globin X. To the best of our knowledge, this is the first time that a vertebrate globin has been identified as component of the cell membrane. Globin X has a hexacoordinate binding scheme and displays cooperative O(2 binding with a variable affinity (P(50∼1.3-12.5 torr, depending on buffer conditions. A respiratory function of globin X is unlikely, but analogous to some prokaryotic membrane-globins it may either protect the lipids in cell membrane from oxidation or may act as a redox-sensing or signaling protein.

Site directed spin labeling studies of Escherichia coli dihydroorotate dehydrogenase N-terminal extension

Energy Technology Data Exchange (ETDEWEB)

Couto, Sheila G. [Instituto de Fisica de Sao Carlos, Universidade de Sao Paulo, Av. Trabalhador Sao-carlense 400, C.P. 369, 13560-970, Sao Carlos, SP (Brazil); Grupo de Biofisica e Fisica Aplicada a Medicina, Instituto de Fisica, Universidade Federal de Goias, Campus Samambaia, C.P. 131, 74001-970, Goiania, GO (Brazil); Cristina Nonato, M. [Laboratorio de Cristalografia de Proteinas, Faculdade de Ciencias Farmaceuticas de Ribeirao Preto, Universidade de Sao Paulo, Av. do Cafe S/N, 14040-903, Ribeirao Preto, SP (Brazil); Costa-Filho, Antonio J., E-mail: ajcosta@ffclrp.usp.br [Instituto de Fisica de Sao Carlos, Universidade de Sao Paulo, Av. Trabalhador Sao-carlense 400, C.P. 369, 13560-970, Sao Carlos, SP (Brazil); Departamento de Fisica, Faculdade de Filosofia, Ciencias e Letras de Ribeirao Preto, Av. Bandeirantes 3900, 14040-901, Ribeirao Preto, SP (Brazil)

2011-10-28

Highlights: Black-Right-Pointing-Pointer EcDHODH is a membrane-associated enzyme and a promising target for drug design. Black-Right-Pointing-Pointer Enzyme's N-terminal extension is responsible for membrane association. Black-Right-Pointing-Pointer N-terminal works as a molecular lid regulating access to the protein interior. -- Abstract: Dihydroorotate dehydrogenases (DHODHs) are enzymes that catalyze the fourth step of the de novo synthesis of pyrimidine nucleotides. In this reaction, DHODH converts dihydroorotate to orotate, using a flavine mononucleotide as a cofactor. Since the synthesis of nucleotides has different pathways in mammals as compared to parasites, DHODH has gained much attention as a promising target for drug design. Escherichia coli DHODH (EcDHODH) is a family 2 DHODH that interacts with cell membranes in order to promote catalysis. The membrane association is supposedly made via an extension found in the enzyme's N-terminal. In the present work, we used site directed spin labeling (SDSL) to specifically place a magnetic probe at positions 2, 5, 19, and 21 within the N-terminal and thus monitor, by using Electron Spin Resonance (ESR), dynamics and structural changes in this region in the presence of a membrane model system. Overall, our ESR spectra show that the N-terminal indeed binds to membranes and that it experiences a somewhat high flexibility that could be related to the role of this region as a molecular lid controlling the entrance of the enzyme's active site and thus allowing the enzyme to give access to quinones that are dispersed in the membrane and that are necessary for the catalysis.

Activation of Rab GTPase Sec4 by its GEF Sec2 is required for prospore membrane formation during sporulation in yeast Saccharomyces cerevisiae.

Science.gov (United States)

Suda, Yasuyuki; Tachikawa, Hiroyuki; Inoue, Ichiro; Kurita, Tomokazu; Saito, Chieko; Kurokawa, Kazuo; Nakano, Akihiko; Irie, Kenji

2018-02-01

Sec2 activates Sec4 Rab GTPase as a guanine nucleotide exchange factor for the recruitment of downstream effectors to facilitate tethering and fusion of post-Golgi vesicles at the plasma membrane. During the meiosis and sporulation of budding yeast, post-Golgi vesicles are transported to and fused at the spindle pole body (SPB) to form a de novo membrane, called the prospore membrane. Previous studies have revealed the role of the SPB outer surface called the meiotic outer plaque (MOP) in docking and fusion of post-Golgi vesicles. However, the upstream molecular machinery for post-Golgi vesicular fusion that facilitates prospore membrane formation remains enigmatic. Here, we demonstrate that the GTP exchange factor for Sec4, Sec2, participates in the formation of the prospore membrane. A conditional mutant in which the SEC2 expression is shut off during sporulation showed sporulation defects. Inactivation of Sec2 caused Sec4 targeting defects along the prospore membranes, thereby causing insufficient targeting of downstream effectors and cargo proteins to the prospore membrane. These results suggest that the activation of Sec4 by Sec2 is required for the efficient supply of post-Golgi vesicles to the prospore membrane and thus for prospore membrane formation/extension and subsequent deposition of spore wall materials. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Molecular Dynamics Simulations Reveal the Conformational Flexibility of Lipid II and Its Loose Association with the Defensin Plectasin in the Staphylococcus aureus Membrane

DEFF Research Database (Denmark)

Witzke, Sarah; Petersen, Michael; Carpenter, Timothy S.

2016-01-01

dynamics simulation study of the conformational dynamics of Lipid II within a detailed model of the Staphylococcus aureus cell membrane. We show that Lipid II is able to adopt a range of conformations, even within the packed lipidic environment of the membrane. Our simulations also reveal dimerization...... the biosynthesis of the cell wall. Given the urgent need for development of novel antibiotics to counter the growing threat of bacterial infection resistance, it is imperative that a thorough molecular-level characterization of the molecules targeted by antibiotics be achieved. To this end, we present a molecular...... of Lipid II mediated by cations. In the presence of the defensin peptide plectasin, the conformational lability of Lipid II allows it to form loose complexes with the protein, via a number of different binding modes....

Improved antifouling potential of polyether sulfone polymeric membrane containing silver nanoparticles: self-cleaning membranes.

Science.gov (United States)

Rana, Sidra; Nazar, Umair; Ali, Jafar; Ali, Qurat Ul Ain; Ahmad, Nasir M; Sarwar, Fiza; Waseem, Hassan; Jamil, Syed Umair Ullah

2018-06-01

A new strategy to enhance the antifouling potential of polyether sulfone (PES) membrane is presented. Chemically synthesized silver nanoparticles (AgNPs) were used to prepare a mixed-matrix PES membrane by the phase inversion technique. Primarily, AgNPs synthesis was confirmed by surface plasmon resonance at 410-430 nm using UV-Visible spectroscopy. X-ray diffraction analysis revealed that AgNPs were crystalline with a diameter of 21 ± 2 nm. Furthermore, PES membranes were characterized by energy dispersive X-ray spectroscopy to confirm the incorporation of AgNPs in membranes. Hydrophilicity of the membranes was enhanced, whereas roughness, mechanical strength and biofouling were relatively reduced after embedding the AgNPs. Antibacterial potential of AgNPs was evaluated for E. coli in the disc diffusion and colony-forming unit (CFU) count method. All of the membranes were assessed for antifouling activity by filtering a control dilution (10 6  CFU/ml) of E. coli and by counting CFU. Anti-biofouling activity of the membrane was observed with different concentrations of AgNPs. Maximum reduction (66%) was observed in membrane containing 1.5% of AgNPs. The addition of antibiotic ceftriaxone enhanced the antibacterial effect of AgNPs in PES membranes. Our practicable antifouling strategy may be applied to other polymeric membranes which may pave the new way to achieve sustainable and self-cleaning membrane reactors on large scale.

Lipid engineering reveals regulatory roles for membrane fluidity in yeast flocculation and oxygen-limited growth

DEFF Research Database (Denmark)

Degreif, Daniel; de Rond, Tristan; Bertl, Adam

2017-01-01

Cells modulate lipid metabolism in order to maintain membrane homeostasis. Here we use a metabolic engineering approach to manipulate the stoichiometry of fatty acid unsaturation, a regulator of cell membrane fluidity, in Saccharomyces cerevisiae. Unexpectedly, reduced lipid unsaturation triggere...

Membrane Protein Mobility and Orientation Preserved in Supported Bilayers Created Directly from Cell Plasma Membrane Blebs.

Science.gov (United States)

Richards, Mark J; Hsia, Chih-Yun; Singh, Rohit R; Haider, Huma; Kumpf, Julia; Kawate, Toshimitsu; Daniel, Susan

2016-03-29

Membrane protein interactions with lipids are crucial for their native biological behavior, yet traditional characterization methods are often carried out on purified protein in the absence of lipids. We present a simple method to transfer membrane proteins expressed in mammalian cells to an assay-friendly, cushioned, supported lipid bilayer platform using cell blebs as an intermediate. Cell blebs, expressing either GPI-linked yellow fluorescent proteins or neon-green fused transmembrane P2X2 receptors, were induced to rupture on glass surfaces using PEGylated lipid vesicles, which resulted in planar supported membranes with over 50% mobility for multipass transmembrane proteins and over 90% for GPI-linked proteins. Fluorescent proteins were tracked, and their diffusion in supported bilayers characterized, using single molecule tracking and moment scaling spectrum (MSS) analysis. Diffusion was characterized for individual proteins as either free or confined, revealing details of the local lipid membrane heterogeneity surrounding the protein. A particularly useful result of our bilayer formation process is the protein orientation in the supported planar bilayer. For both the GPI-linked and transmembrane proteins used here, an enzymatic assay revealed that protein orientation in the planar bilayer results in the extracellular domains facing toward the bulk, and that the dominant mode of bleb rupture is via the "parachute" mechanism. Mobility, orientation, and preservation of the native lipid environment of the proteins using cell blebs offers advantages over proteoliposome reconstitution or disrupted cell membrane preparations, which necessarily result in significant scrambling of protein orientation and typically immobilized membrane proteins in SLBs. The bleb-based bilayer platform presented here is an important step toward integrating membrane proteomic studies on chip, especially for future studies aimed at understanding fundamental effects of lipid interactions

Small RNAs controlling outer membrane porins

DEFF Research Database (Denmark)

Valentin-Hansen, Poul; Johansen, Jesper; Rasmussen, Anders A

2007-01-01

are key regulators of environmental stress. Recent work has revealed an intimate interplay between small RNA regulation of outer membrane proteins and the stress-induced sigmaE-signalling system, which has an essential role in the maintenance of the integrity of the outer membrane.......Gene regulation by small non-coding RNAs has been recognized as an important post-transcriptional regulatory mechanism for several years. In Gram-negative bacteria such as Escherichia coli and Salmonella, these RNAs control stress response and translation of outer membrane proteins and therefore...

Phosphoproteome analysis of functional mitochondria isolated from resting human muscle reveals extensive phosphorylation of inner membrane protein complexes and enzymes

DEFF Research Database (Denmark)

Zhao, Xiaolu; Leon, Ileana R; Bak, Steffen

2011-01-01

. In skeletal muscle, mitochondrial dysfunction is linked to insulin resistance in humans with obesity and type 2 diabetes. We performed a phosphoproteomic study of functional mitochondria isolated from human muscle biopsies with the aim to obtain a comprehensive overview of mitochondrial phosphoproteins...... in insulin resistance. We also assigned phosphorylation sites in mitochondrial proteins involved in amino acid degradation, importers and transporters, calcium homeostasis, and apoptosis. Bioinformatics analysis of kinase motifs revealed that many of these mitochondrial phosphoproteins are substrates....... Future comparative phosphoproteome analysis of mitochondria from healthy and diseased individuals will provide insights into the role of abnormal phosphorylation in pathologies, such as type 2 diabetes....

Toward the Structure of Dynamic Membrane-Anchored Actin Networks

Science.gov (United States)

Weber, Igor

2007-01-01

In the cortex of a motile cell, membrane-anchored actin filaments assemble into structures of varying shape and function. Filopodia are distinguished by a core of bundled actin filaments within finger-like extensions of the membrane. In a recent paper by Medalia et al1 cryo-electron tomography has been used to reconstruct, from filopodia of Dictyostelium cells, the 3-dimensional organization of actin filaments in connection with the plasma membrane. A special arrangement of short filaments converging toward the filopod's tip has been called a “terminal cone”. In this region force is applied for protrusion of the membrane. Here we discuss actin organization in the filopodia of Dictyostelium in the light of current views on forces that are generated by polymerizing actin filaments, and on the resistance of membranes against deformation that counteracts these forces. PMID:19262130

NASAL MUCOUS MEMBRANE MICROFLORA IN PATIENTS WITH POLYPOUS RHINOSINUSITIS

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O. A. Kolenchukova

2016-01-01

Full Text Available Nasal mucous membrane microbiocenosis Research amongst patients with polypous rhinosinusitis is conducted. Patients with polypous rhinosinusitis (PRS, n = 58 aged from 18 till 64 years and group of control (n = 156. For an microflora assessment of nasal mucous membrane during an exacerbation of a disease carried out crops of microorga nisms on nutrient differential and diagnostic agars. When studying the microflora received from nasal mucous membrane 407 cultures of microorganisms at PRS were revealed. In control group of 174 microorganisms cultures are revealed. Among isolates were established 6 genera of 9 species of bacteria at PRS against 6 genera and 8 species in group of control. Microflora quantitative structure research of nasal mucous membrane at PRS of rather control group considerable prevalence of the microorganisms belonging to the sorts Staphylococcus and Streptococcus, and also to Enterobacteriaceae was revealed. It is also revealed increase S. pneumoniae. When determining specific accessory of the microorganisms relating to the Staphylococcus genera in PRS group concerning control the increase in total number of the strains of S. aureus relating to coagulase-positive and S. haemolyticus, S. epidermidis relating to coagulase-negative staphylococcus was established. A big variety the coagulase-negative of Staphylococcus is also revealed: S. epidermidis, S. haemolyticus, S. hominis, S. cohnii, S. capitis, S. hyicius and S. xylosus. In control group of such types as S. capitis and S. hyicius it isn’t revealed. Thus at a polypous rhinosinusitis nasal mucous membrane the expressed dysbacteriosis takes place. The analysis of frequency of occurrence of the microorganisms belonging to different childbirth showed that in PRS group bacteria of the sort Streptococcus and this Nesseria, and also S. pneumoniae, S. pyogenes were most often allocated concerning control. The frequency prevalence analysis of the gene ra Staphylococcus related

Does Chlorination of Seawater Reverse Osmosis Membranes Control Biofouling?

KAUST Repository

Khan, Muhammad Tariq; Hong, Pei-Ying; Nada, Nabil; Croue, Jean Philippe

2015-01-01

Biofouling is the major problem of reverse osmosis (RO) membranes used for desalting seawater (SW). The use of chlorine is a conventional and common practice to control/prevent biofouling. Unlike polyamide RO membranes, cellulose triacetate (CTA) RO membranes display a high chlorine tolerance. Due to this characteristic, CTA membranes are used in most of the RO plants located in the Middle East region where the elevated seawater temperature and water quality promote the risk of membrane biofouling. However, there is no detailed study on the investigation/characterization of CTA-RO membrane fouling. In this investigation, the fouling profile of a full–scale SWRO desalination plant operating with not only continuous chlorination of raw seawater but also intermittent chlorination of CTA-RO membranes was studied. Detailed water quality and membrane fouling analyses were conducted. Profiles of microbiological, inorganic, and organic constituents of analysed fouling layers were extensively discussed. Our results clearly identified biofilm development on these membranes. The incapability of chlorination on preventing biofilm formation on SWRO membranes could be assigned to its failure in effectively reaching throughout the different regions of the permeators. This failure could have occurred due to three main factors: plugging of membrane fibers, chlorine consumption by organics accumulated on the front side fibers, or chlorine adaptation of certain bacterial populations.

Does Chlorination of Seawater Reverse Osmosis Membranes Control Biofouling?

KAUST Repository

Khan, Muhammad Tariq

2015-04-01

Biofouling is the major problem of reverse osmosis (RO) membranes used for desalting seawater (SW). The use of chlorine is a conventional and common practice to control/prevent biofouling. Unlike polyamide RO membranes, cellulose triacetate (CTA) RO membranes display a high chlorine tolerance. Due to this characteristic, CTA membranes are used in most of the RO plants located in the Middle East region where the elevated seawater temperature and water quality promote the risk of membrane biofouling. However, there is no detailed study on the investigation/characterization of CTA-RO membrane fouling. In this investigation, the fouling profile of a full–scale SWRO desalination plant operating with not only continuous chlorination of raw seawater but also intermittent chlorination of CTA-RO membranes was studied. Detailed water quality and membrane fouling analyses were conducted. Profiles of microbiological, inorganic, and organic constituents of analysed fouling layers were extensively discussed. Our results clearly identified biofilm development on these membranes. The incapability of chlorination on preventing biofilm formation on SWRO membranes could be assigned to its failure in effectively reaching throughout the different regions of the permeators. This failure could have occurred due to three main factors: plugging of membrane fibers, chlorine consumption by organics accumulated on the front side fibers, or chlorine adaptation of certain bacterial populations.

Activation of PVDF membranes through facile hydroxylation of the polymeric dope

KAUST Repository

Al-Gharabli, Samer; Mavukkandy, Musthafa O.; Kujawa, Joanna; Nunes, Suzana Pereira; Arafat, Hassan A.

2017-01-01

A method comprising a two-step alkali/acid treatment of poly (vinylidene fluoride) (PVDF) polymer is developed for the fabrication of flat-sheet PVDF membranes functionalized with labile hydroxyl groups. This method involves the application of a short-duration modification in alkali medium (5% KOH). Extensive characterizations were performed on the prepared membranes. Modification of the polymer altered the crystallinity of the PVDF from a mixture of both α and β phases to a predominant β phase. Lower work of adhesion of the modified membrane indicated the formation of a more hydrophobic and wetting-resistant membrane surface. Centrifugation of the polymer dope after the modification had a pronounced impact on the properties of the resultant membranes. This protocol could be utilized in fine-tuning the properties of PVDF membranes for various target-specific applications such as membrane distillation. This method can also be used in functionalizing PVDF membranes further by exploiting the labile –OH group present on the membrane surface.

Activation of PVDF membranes through facile hydroxylation of the polymeric dope

KAUST Repository

Al-Gharabli, Samer

2017-10-30

A method comprising a two-step alkali/acid treatment of poly (vinylidene fluoride) (PVDF) polymer is developed for the fabrication of flat-sheet PVDF membranes functionalized with labile hydroxyl groups. This method involves the application of a short-duration modification in alkali medium (5% KOH). Extensive characterizations were performed on the prepared membranes. Modification of the polymer altered the crystallinity of the PVDF from a mixture of both α and β phases to a predominant β phase. Lower work of adhesion of the modified membrane indicated the formation of a more hydrophobic and wetting-resistant membrane surface. Centrifugation of the polymer dope after the modification had a pronounced impact on the properties of the resultant membranes. This protocol could be utilized in fine-tuning the properties of PVDF membranes for various target-specific applications such as membrane distillation. This method can also be used in functionalizing PVDF membranes further by exploiting the labile –OH group present on the membrane surface.

Membrane Sculpting by F-BAR Domains Studied by Molecular Dynamics Simulations

Science.gov (United States)

Yu, Hang; Schulten, Klaus

2013-01-01

Interplay between cellular membranes and their peripheral proteins drives many processes in eukaryotic cells. Proteins of the Bin/Amphiphysin/Rvs (BAR) domain family, in particular, play a role in cellular morphogenesis, for example curving planar membranes into tubular membranes. However, it is still unclear how F-BAR domain proteins act on membranes. Electron microscopy revealed that, in vitro, F-BAR proteins form regular lattices on cylindrically deformed membrane surfaces. Using all-atom and coarse-grained (CG) molecular dynamics simulations, we show that such lattices, indeed, induce tubes of observed radii. A 250 ns all-atom simulation reveals that F-BAR domain curves membranes via the so-called scaffolding mechanism. Plasticity of the F-BAR domain permits conformational change in response to membrane interaction, via partial unwinding of the domains 3-helix bundle structure. A CG simulation covering more than 350 µs provides a dynamic picture of membrane tubulation by lattices of F-BAR domains. A series of CG simulations identified the optimal lattice type for membrane sculpting, which matches closely the lattices seen through cryo-electron microscopy. PMID:23382665

Utilization pattern of extension tools and methods by Agricultural Extension Agents

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M Surudhi

2018-05-01

Full Text Available A study was conducted in Krishnagiri district of Tamil Nadu state to understand the utilization pattern of extension tools and methods by the agricultural extension agents. As ICT revolution is slowly conquering the rural sector, it becomes imperative that the agricultural extension agents transform themselves to the changing times and develop competencies in utilizing these ICTs.  The study explored the usage of various extension tools and methods by the change agents and the constraints faced in utilizing them. The findings revealed that the extension functionaries frequently used the individual contact methods viz., telephone, office calls and farm and home visits in the process of transfer of technology. Least efforts were shown in sending SMS based communication. Meetings were the common and frequently adopted group contact method. Demonstrations, farmer field school, farmer’s interest groups, field trips and farmer training programmes were moderately adopted. Posters, leaflets and pre-season campaigns were the widely adopted mass contact methods. They possess least skill in utilizing farm magazines, presenting television and radio programmes, which are among the most popular and most efficient mass contact methods. The extension functionaries need to be trained adequately on the wider use of electronic communication methods like e mails, and SMS in the local language. Efforts should be taken up to sensitize the importance and train the extension agents in the usage of different group and mass contact methods.

Complex Dynamic Development of Poliovirus Membranous Replication Complexes

Science.gov (United States)

Nair, Vinod; Hansen, Bryan T.; Hoyt, Forrest H.; Fischer, Elizabeth R.; Ehrenfeld, Ellie

2012-01-01

Replication of all positive-strand RNA viruses is intimately associated with membranes. Here we utilize electron tomography and other methods to investigate the remodeling of membranes in poliovirus-infected cells. We found that the viral replication structures previously described as “vesicles” are in fact convoluted, branching chambers with complex and dynamic morphology. They are likely to originate from cis-Golgi membranes and are represented during the early stages of infection by single-walled connecting and branching tubular compartments. These early viral organelles gradually transform into double-membrane structures by extension of membranous walls and/or collapsing of the luminal cavity of the single-membrane structures. As the double-membrane regions develop, they enclose cytoplasmic material. At this stage, a continuous membranous structure may have double- and single-walled membrane morphology at adjacent cross-sections. In the late stages of the replication cycle, the structures are represented mostly by double-membrane vesicles. Viral replication proteins, double-stranded RNA species, and actively replicating RNA are associated with both double- and single-membrane structures. However, the exponential phase of viral RNA synthesis occurs when single-membrane formations are predominant in the cell. It has been shown previously that replication complexes of some other positive-strand RNA viruses form on membrane invaginations, which result from negative membrane curvature. Our data show that the remodeling of cellular membranes in poliovirus-infected cells produces structures with positive curvature of membranes. Thus, it is likely that there is a fundamental divergence in the requirements for the supporting cellular membrane-shaping machinery among different groups of positive-strand RNA viruses. PMID:22072780

Computational Approaches for Revealing the Structure of Membrane Transporters: Case Study on Bilitranslocase

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Katja Venko

Full Text Available The structural and functional details of transmembrane proteins are vastly underexplored, mostly due to experimental difficulties regarding their solubility and stability. Currently, the majority of transmembrane protein structures are still unknown and this present a huge experimental and computational challenge. Nowadays, thanks to X-ray crystallography or NMR spectroscopy over 3000 structures of membrane proteins have been solved, among them only a few hundred unique ones. Due to the vast biological and pharmaceutical interest in the elucidation of the structure and the functional mechanisms of transmembrane proteins, several computational methods have been developed to overcome the experimental gap. If combined with experimental data the computational information enables rapid, low cost and successful predictions of the molecular structure of unsolved proteins. The reliability of the predictions depends on the availability and accuracy of experimental data associated with structural information. In this review, the following methods are proposed for in silico structure elucidation: sequence-dependent predictions of transmembrane regions, predictions of transmembrane helix–helix interactions, helix arrangements in membrane models, and testing their stability with molecular dynamics simulations. We also demonstrate the usage of the computational methods listed above by proposing a model for the molecular structure of the transmembrane protein bilitranslocase. Bilitranslocase is bilirubin membrane transporter, which shares similar tissue distribution and functional properties with some of the members of the Organic Anion Transporter family and is the only member classified in the Bilirubin Transporter Family. Regarding its unique properties, bilitranslocase is a potentially interesting drug target. Keywords: Membrane proteins, Bilitranslocase, 3D protein structure, Transmembrane region predictors, Helix–helix interactions

Binding of Neurotransmitters to Lipid Membranes

DEFF Research Database (Denmark)

Peters, Günther H.J.; Werge, Mikkel; Elf-Lind, Maria Northved

2014-01-01

/ acetylated g-aminobutyrate (GABAneu) with a dipalmitoylphosphatidylcholine (DPPC) bilayer. This study was motivated by recent research results that suggested that neural transmission may also be affected by nonspecific interactions of NTs with the lipid matrix of the synaptic membrane. Our results revealed...... backbone of the phospholipids. It is surprising that hydrophilic solutes can deeply penetrate into the membrane pointing to the fact that membrane affinity is governed by specific interactions. Our MD simulations identified the salt-bridge between the primary amine of NTs and the lipid phosphate group...

Cell membrane softening in human breast and cervical cancer cells

Science.gov (United States)

Händel, Chris; Schmidt, B. U. Sebastian; Schiller, Jürgen; Dietrich, Undine; Möhn, Till; Kießling, Tobias R.; Pawlizak, Steve; Fritsch, Anatol W.; Horn, Lars-Christian; Briest, Susanne; Höckel, Michael; Zink, Mareike; Käs, Josef A.

2015-08-01

Biomechanical properties are key to many cellular functions such as cell division and cell motility and thus are crucial in the development and understanding of several diseases, for instance cancer. The mechanics of the cellular cytoskeleton have been extensively characterized in cells and artificial systems. The rigidity of the plasma membrane, with the exception of red blood cells, is unknown and membrane rigidity measurements only exist for vesicles composed of a few synthetic lipids. In this study, thermal fluctuations of giant plasma membrane vesicles (GPMVs) directly derived from the plasma membranes of primary breast and cervical cells, as well as breast cell lines, are analyzed. Cell blebs or GPMVs were studied via thermal membrane fluctuations and mass spectrometry. It will be shown that cancer cell membranes are significantly softer than their non-malignant counterparts. This can be attributed to a loss of fluid raft forming lipids in malignant cells. These results indicate that the reduction of membrane rigidity promotes aggressive blebbing motion in invasive cancer cells.

Application of MD Simulations to Predict Membrane Properties of MOFs

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Elda Adatoz

2015-01-01

Full Text Available Metal organic frameworks (MOFs are a new group of nanomaterials that have been widely examined for various chemical applications. Gas separation using MOF membranes has become an increasingly important research field in the last years. Several experimental studies have shown that thin-film MOF membranes can outperform well known polymer and zeolite membranes due to their higher gas permeances and selectivities. Given the very large number of available MOF materials, it is impractical to fabricate and test the performance of every single MOF membrane using purely experimental techniques. In this study, we used molecular simulations, Monte Carlo and Molecular Dynamics, to estimate both single-gas and mixture permeances of MOF membranes. Predictions of molecular simulations were compared with the experimental gas permeance data of MOF membranes in order to validate the accuracy of our computational approach. Results show that computational methodology that we described in this work can be used to accurately estimate membrane properties of MOFs prior to extensive experimental efforts.

The plant membrane surrounding powdery mildew haustoria shares properties with the endoplasmic reticulum membrane

DEFF Research Database (Denmark)

Kwaaitaal, Mark Adrianus Cornelis J; Nielsen, Mads Eggert; Böhlenius, Henrik

2017-01-01

Many filamentous plant pathogens place specialized feeding structures, called haustoria, inside living host cells. As haustoria grow, they are believed to manipulate plant cells to generate a specialized, still enigmatic extrahaustorial membrane (EHM) around them. Here, we focused on revealing...... properties of the EHM. With the help of membranespecific dyes and transient expression of membrane-associated proteins fused to fluorescent tags, we studied the nature of the EHM generated by barley leaf epidermal cells around powdery mildew haustoria. Observations suggesting that endoplasmic reticulum (ER...... that it is not a continuum of the ER. Furthermore, GDP-locked Sar1 and a nucleotide-free RabD2a, which block ER to Golgi exit, did not hamper haustorium formation. These results indicated that the EHM shares features with the plant ER membrane, but that the EHM membrane is not dependent on conventional secretion...

Sensing voltage across lipid membranes

Science.gov (United States)

Swartz, Kenton J.

2009-01-01

The detection of electrical potentials across lipid bilayers by specialized membrane proteins is required for many fundamental cellular processes such as the generation and propagation of nerve impulses. These membrane proteins possess modular voltage-sensing domains, a notable example being the S1-S4 domains of voltage-activated ion channels. Ground-breaking structural studies on these domains explain how voltage sensors are designed and reveal important interactions with the surrounding lipid membrane. Although further structures are needed to fully understand the conformational changes that occur during voltage sensing, the available data help to frame several key concepts that are fundamental to the mechanism of voltage sensing. PMID:19092925

Quantitative Proteomics Reveals Membrane Protein-Mediated Hypersaline Sensitivity and Adaptation in Halophilic Nocardiopsis xinjiangensis.

Science.gov (United States)

Zhang, Yao; Li, Yanchang; Zhang, Yongguang; Wang, Zhiqiang; Zhao, Mingzhi; Su, Na; Zhang, Tao; Chen, Lingsheng; Wei, Wei; Luo, Jing; Zhou, Yanxia; Xu, Yongru; Xu, Ping; Li, Wenjun; Tao, Yong

2016-01-04

The genus Nocardiopsis is one of the most dominant Actinobacteria that survives in hypersaline environments. However, the adaptation mechanisms for halophilism are still unclear. Here, we performed isobaric tags for relative and absolute quantification based quantitative proteomics to investigate the functions of the membrane proteome after salt stress. A total of 683 membrane proteins were identified and quantified, of which 126 membrane proteins displayed salt-induced changes in abundance. Intriguingly, bioinformatics analyses indicated that these differential proteins showed two expression patterns, which were further validated by phenotypic changes and functional differences. The majority of ABC transporters, secondary active transporters, cell motility proteins, and signal transduction kinases were up-regulated with increasing salt concentration, whereas cell differentiation, small molecular transporter (ions and amino acids), and secondary metabolism proteins were significantly up-regulated at optimum salinity, but down-regulated or unchanged at higher salinity. The small molecule transporters and cell differentiation-related proteins acted as sensing proteins that played a more important biological role at optimum salinity. However, the ABC transporters for compatible solutes, Na(+)-dependent transporters, and cell motility proteins acted as adaptive proteins that actively counteracted higher salinity stress. Overall, regulation of membrane proteins may provide a major protection strategy against hyperosmotic stress.

Mitigation of membrane biofouling by a quorum quenching bacterium for membrane bioreactors.

Science.gov (United States)

Ham, So-Young; Kim, Han-Shin; Cha, Eunji; Park, Jeong-Hoon; Park, Hee-Deung

2018-06-01

In this study, a quorum-quenching (QQ) bacterium named HEMM-1 was isolated at a membrane bioreactor (MBR) plant. HEMM-1 has diplococcal morphology and 99% sequence identity to Enterococcus species. The HEMM-1 cell-free supernatant (CFS) showed higher QQ activities than the CFS of other QQ bacteria, mostly by degrading N-acyl homoserine lactones (AHLs) with short acyl chains. Instrumental analyses revealed that HEMM-1 CFS degraded AHLs via lactonase activity. Under static, flow, and shear conditions, the HEMM-1 CFS was effective in reducing bacterial and activated-sludge biofilms formed on membrane surfaces. In conclusion, the HEMM-1 isolate is a QQ bacterium applicable to the control of biofouling in MBRs via inhibition of biofilm formation on membrane surfaces. Copyright © 2018 Elsevier Ltd. All rights reserved.

BSM-MBR: a benchmark simulation model to compare control and operational strategies for membrane bioreactors.

Science.gov (United States)

Maere, Thomas; Verrecht, Bart; Moerenhout, Stefanie; Judd, Simon; Nopens, Ingmar

2011-03-01

A benchmark simulation model for membrane bioreactors (BSM-MBR) was developed to evaluate operational and control strategies in terms of effluent quality and operational costs. The configuration of the existing BSM1 for conventional wastewater treatment plants was adapted using reactor volumes, pumped sludge flows and membrane filtration for the water-sludge separation. The BSM1 performance criteria were extended for an MBR taking into account additional pumping requirements for permeate production and aeration requirements for membrane fouling prevention. To incorporate the effects of elevated sludge concentrations on aeration efficiency and costs a dedicated aeration model was adopted. Steady-state and dynamic simulations revealed BSM-MBR, as expected, to out-perform BSM1 for effluent quality, mainly due to complete retention of solids and improved ammonium removal from extensive aeration combined with higher biomass levels. However, this was at the expense of significantly higher operational costs. A comparison with three large-scale MBRs showed BSM-MBR energy costs to be realistic. The membrane aeration costs for the open loop simulations were rather high, attributed to non-optimization of BSM-MBR. As proof of concept two closed loop simulations were run to demonstrate the usefulness of BSM-MBR for identifying control strategies to lower operational costs without compromising effluent quality. Copyright © 2011 Elsevier Ltd. All rights reserved.

Comparative proteome analysis reveals pathogen specific outer membrane proteins of Leptospira.

Science.gov (United States)

Dhandapani, Gunasekaran; Sikha, Thoduvayil; Rana, Aarti; Brahma, Rahul; Akhter, Yusuf; Gopalakrishnan Madanan, Madathiparambil

2018-04-10

Proteomes of pathogenic Leptospira interrogans and L. borgpetersenii and the saprophytic L. biflexa were filtered through computational tools to identify Outer Membrane Proteins (OMPs) that satisfy the required biophysical parameters for their presence on the outer membrane. A total of 133, 130, and 144 OMPs were identified in L. interrogans, L. borgpetersenii, and L. biflexa, respectively, which forms approximately 4% of proteomes. A holistic analysis of transporting and pathogenic characteristics of OMPs together with Clusters of Orthologous Groups (COGs) among the OMPs and their distribution across 3 species was made and put forward a set of 21 candidate OMPs specific to pathogenic leptospires. It is also found that proteins homologous to the candidate OMPs were also present in other pathogenic species of leptospires. Six OMPs from L. interrogans and 2 from L. borgpetersenii observed to have similar COGs while those were not found in any intermediate or saprophytic forms. These OMPs appears to have role in infection and pathogenesis and useful for anti-leptospiral strategies. © 2018 Wiley Periodicals, Inc.

Metallic Membranes for High Temperature Hydrogen Separation

DEFF Research Database (Denmark)

Ma, Y.H.; Catalano, Jacopo; Guazzone, Federico

2013-01-01

membrane fabrication methods have matured over the last decades, and the deposition of very thin films (1–5 µm) of Pd over porous ceramics or modified porous metal supports is quite common. The H2 permeances and the selectivities achieved at 400–500 °C were in the order of 50–100 Nm3/m/h/bar0.5 and greater......Composite palladium membranes have extensively been studied in laboratories and, more recently, in small pilot industrial applications for the high temperature separation of hydrogen from reactant mixtures such as water-gas shift (WGS) reaction or methane steam reforming (MSR). Composite Pd...... than 1000, respectively. This chapter describes in detail composite Pd-based membrane preparation methods, which consist of the grading of the support and the deposition of the dense metal layer, their performances, and their applications in catalytic membrane reactors (CMRs) at high temperatures (400...

Amphotericin B channels in phospholipid membrane-coated nanoporous silicon surfaces: implications for photovoltaic driving of ions across membranes.

Science.gov (United States)

Yilma, Solomon; Liu, Nangou; Samoylov, Alexander; Lo, Ting; Brinker, C Jeffrey; Vodyanoy, Vitaly

2007-03-15

The antimycotic agent amphotericin B (AmB) functions by forming complexes with sterols to form ion channels that cause membrane leakage. When AmB and cholesterol mixed at 2:1 ratio were incorporated into phospholipid bilayer membranes formed on the tip of patch pipettes, ion channel current fluctuations with characteristic open and closed states were observed. These channels were also functional in phospholipid membranes formed on nanoporous silicon surfaces. Electrophysiological studies of AmB-cholesterol mixtures that were incorporated into phospholipid membranes formed on the surface of nanoporous (6.5 nm pore diameter) silicon plates revealed large conductance ion channels ( approximately 300 pS) with distinct open and closed states. Currents through the AmB-cholesterol channels on nanoporous silicon surfaces can be driven by voltage applied via conventional electrical circuits or by photovoltaic electrical potential entirely generated when the nanoporous silicon surface is illuminated with a narrow laser beam. Electrical recordings made during laser illumination of AmB-cholesterol containing membrane-coated nanoporous silicon surfaces revealed very large conductance ion channels with distinct open and closed states. Our findings indicate that nanoporous silicon surfaces can serve as mediums for ion-channel-based biosensors. The photovoltaic properties of nanoporous silicon surfaces show great promise for making such biosensors addressable via optical technologies.

Insertion of Neurotransmitters into a Lipid Bilayer Membrane and Its Implication on Membrane Stability: A Molecular Dynamics Study.

Science.gov (United States)

Shen, Chun; Xue, Minmin; Qiu, Hu; Guo, Wanlin

2017-03-17

The signaling molecules in neurons, called neurotransmitters, play an essential role in the transportation of neural signals, during which the neurotransmitters interact with not only specific receptors, but also cytomembranes, such as synaptic vesicle membranes and postsynaptic membranes. Through extensive molecular dynamics simulations, the atomic-scale insertion dynamics of typical neurotransmitters, including methionine enkephalin (ME), leucine enkephalin (LE), dopamine (DA), acetylcholine (ACh), and aspartic acid (ASP), into lipid bilayers is investigated. The results show that the first three neurotransmitters (ME, LE, and DA) are able to diffuse freely into both 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE) membranes, and are guided by the aromatic residues Tyr and Phe. Only a limited number of these neurotransmitters are allowed to penetrate into the membrane, which suggests an intrinsic mechanism by which the membrane is protected from being destroyed by excessive inserted neurotransmitters. After spontaneous insertion, the neurotransmitters disturb the surrounding phospholipids in the membrane, as indicated by the altered distribution of components in lipid leaflets and the disordered lipid tails. In contrast, the last two neurotransmitters (ACh and ASP) cannot enter the membrane, but instead always diffuse freely in solution. These findings provide an understanding at the atomic level of how neurotransmitters interact with the surrounding cytomembrane, as well as their impact on membrane behavior. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

Development of ZIF-8 membranes: opportunities and challenges for commercial applications

KAUST Repository

Lai, Zhiping

2018-03-24

Metal organic framework (MOF) membranes have attracted significant attentions in recent years because of their potentials in gas and liquid separations and other applications such as catalysis and chemical sensors, etc. More than half of the MOF membrane publications up to date are related to ZIF-8 system, because of its easy synthesis, relatively high stability, and excellent gas separation performance, which allows many novel ideas to be easily implemented. Extensive studies have shown that ZIF-8 membranes hold great potentials in gas separations, but may face great challenges in liquid separations mainly because of their poor stability. This is also a common observation for other MOF membranes. As such, in this article we use ZIF-8 membrane as a prototype and focus on its development in gas separations for the discussions of the most concerned issues related to membrane commercialization including membrane synthesis, separation performance, stability, process reproducibility, and finally on the opportunities and challenges that MOF membranes may face in industrial applications.

Development of ZIF-8 membranes: opportunities and challenges for commercial applications

KAUST Repository

Lai, Zhiping

2018-01-01

Metal organic framework (MOF) membranes have attracted significant attentions in recent years because of their potentials in gas and liquid separations and other applications such as catalysis and chemical sensors, etc. More than half of the MOF membrane publications up to date are related to ZIF-8 system, because of its easy synthesis, relatively high stability, and excellent gas separation performance, which allows many novel ideas to be easily implemented. Extensive studies have shown that ZIF-8 membranes hold great potentials in gas separations, but may face great challenges in liquid separations mainly because of their poor stability. This is also a common observation for other MOF membranes. As such, in this article we use ZIF-8 membrane as a prototype and focus on its development in gas separations for the discussions of the most concerned issues related to membrane commercialization including membrane synthesis, separation performance, stability, process reproducibility, and finally on the opportunities and challenges that MOF membranes may face in industrial applications.

Synthesis and properties of a novel sulfonated poly(arylene ether ketone sulfone) membrane with a high β-value for direct methanol fuel cell applications

International Nuclear Information System (INIS)

Xu, Jingmei; Ma, Li; Han, Hailan; Ni, Hongzhe; Wang, Zhe; Zhang, Huixuan

2014-01-01

Highlights: • Introduction of carboxyl groups into copolymers resulted in extensive hydrogen bond. • The C-SPAEKS membranes had obviously hydrophilic/hydrophobic phase separation. • The membranes showed low methanol permeability and high β values. • The membranes exhibited good thermal property and desirable mechanical performance. - Abstract: Sulfonated poly(arylene ether ketone sulfone) membranes containing carboxylic acid groups (C-SPAEKS) with different degrees of sulfonation were synthesized by the nucleophilic aromatic substitution reactions of 4-carboxylphenyl hydroquinone (4C-PH), bisphenol A, 3,3′-disulfonated 4,4′-dichlorodiphenyl sulfone, and 4,4′-difluorobenzophenone. The Fourier transform infrared and 1 H NMR analyses of C-SPAEKS revealed the presence of carboxylic acid groups in the C-SPAEKS membranes. The membranes exhibited a low swelling degree and methanol crossover level. The effects of different degrees of sulfonation on the water uptake, proton conductivity, and methanol permeability coefficient of the membranes were studied. The maximum proton conductivity of C-SPAEKS-80 membrane at room temperature was 0.069 S cm −1 , which was higher than that of Nafion ® 117 membrane. The methanol permeability coefficient of C-SPAEKS-80 membrane was 9.15 × 10 −7 cm 2 s −1 at 20 °C, much lower than that of Nafion 117 membrane (22.9 × 10 −7 cm 2 s −1 ). Furthermore, the carboxyl group-containing membranes exhibited a high β-value, further confirming that this series of membranes possess excellent comprehensive performance and can be applied in direct methanol fuel cells

Performance evaluation and mass transfer study of CO2 absorption in flat sheet membrane contactor using novel porous polysulfone membrane

International Nuclear Information System (INIS)

Nabian, Nima; Ghoreyshi, Ali Asghar; Rahimpour, Ahmad; Shakeri, Mohsen

2015-01-01

The performance of gas-liquid membrane contactor for CO 2 capture was investigated using a novel polysulfone (PSF) flat membrane prepared via non-solvent phase inversion method. Polyvinyl pyrrolidone (PVP) was used as an additive in the dope solution of PSF membranes. Morphological studies by scanning electron microscopy (SEM) analysis revealed that PSF membrane with PVP has a finger-like structure, but the PSF membrane without PVP has a sponge-like structure. Also, characterization results through atomic force microscopy (AFM) and contact angle measurement demonstrated that the porosity, surface roughness and hydrophobicity of the PSF membrane increased with addition of PVP to the dope solution. Mass transfer resistance analysis, based on CO 2 absorption flux, displayed that addition of PVP to the dope solution of PSF membrane decreased membrane mass transfer resistance, and significantly improved CO 2 absorption flux up to 2.7 and 1.8 times of absorption fluxes of PSF membrane without PVP and commercial PVDF, respectively.

Perineural extension of facial melanoma

Energy Technology Data Exchange (ETDEWEB)

Kalina, Peter [Mayo Clinic, Department of Radiology, Rochester, Minnesota (United States); Bevilacqua, Paula

2005-05-01

A 64-year-old man presented with a pigmented cutaneous lesion on the right side of his face along with right facial numbness. Histological examination revealed malignant melanoma. Magnetic resonance imaging (MRI) revealed perineural extension along the entire course of the maxillary division of the right trigeminal nerve. This is a rare but important manifestation of the spread of head and neck malignancy. (orig.)

A Pilot-Scale System for Carbon Molecular Sieve Hollow Fiber Membrane Manufacturing

KAUST Repository

Karvan, O.

2012-12-21

Carbon molecular sieve (CMS) membranes offer advantages over traditional polymeric membrane materials, but scale-up of manufacturing systems has not received much attention. In the recent decade, there has been a dramatic increase in fundamental research on these materials with a variety of applications being studied. The results from a pilot-scale CMS production system are presented. This system was designed based on extensive laboratory research, and hollow fiber membranes produced in this system show similar performance compared to membranes produced using a smaller bench-scale system. After optimizing the system design, a 93% recovery of the precursor fibers for use in membrane module preparation were obtained. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Constitutively active ezrin increases membrane tension, slows migration, and impedes endothelial transmigration of lymphocytes in vivo in mice.

Science.gov (United States)

Liu, Yin; Belkina, Natalya V; Park, Chung; Nambiar, Raj; Loughhead, Scott M; Patino-Lopez, Genaro; Ben-Aissa, Khadija; Hao, Jian-Jiang; Kruhlak, Michael J; Qi, Hai; von Andrian, Ulrich H; Kehrl, John H; Tyska, Matthew J; Shaw, Stephen

2012-01-12

ERM (ezrin, radixin moesin) proteins in lymphocytes link cortical actin to plasma membrane, which is regulated in part by ERM protein phosphorylation. To assess whether phosphorylation of ERM proteins regulates lymphocyte migration and membrane tension, we generated transgenic mice whose T-lymphocytes express low levels of ezrin phosphomimetic protein (T567E). In these mice, T-cell number in lymph nodes was reduced by 27%. Lymphocyte migration rate in vitro and in vivo in lymph nodes decreased by 18% to 47%. Lymphocyte membrane tension increased by 71%. Investigations of other possible underlying mechanisms revealed impaired chemokine-induced shape change/lamellipod extension and increased integrin-mediated adhesion. Notably, lymphocyte homing to lymph nodes was decreased by 30%. Unlike most described homing defects, there was not impaired rolling or sticking to lymph node vascular endothelium but rather decreased migration across that endothelium. Moreover, decreased numbers of transgenic T cells in efferent lymph suggested defective egress. These studies confirm the critical role of ERM dephosphorylation in regulating lymphocyte migration and transmigration. Of particular note, they identify phospho-ERM as the first described regulator of lymphocyte membrane tension, whose increase probably contributes to the multiple defects observed in the ezrin T567E transgenic mice.

Architectural switches in plant thylakoid membranes.

Science.gov (United States)

Kirchhoff, Helmut

2013-10-01

Recent progress in elucidating the structure of higher plants photosynthetic membranes provides a wealth of information. It allows generation of architectural models that reveal well-organized and complex arrangements not only on whole membrane level, but also on the supramolecular level. These arrangements are not static but highly responsive to the environment. Knowledge about the interdependency between dynamic structural features of the photosynthetic machinery and the functionality of energy conversion is central to understanding the plasticity of photosynthesis in an ever-changing environment. This review summarizes the architectural switches that are realized in thylakoid membranes of green plants.

Retention measurements of nanofiltration membranes with electrolyte solutions

NARCIS (Netherlands)

Peeters, J.M.M.; Peeters, J.M.M.; Boom, J.P.; Boom, J.P.; Mulder, M.H.V.; Strathmann, H.

1998-01-01

Retention measurements with single salt solutions of CaCl2, NaCl and Na2SO4 revealed that the rejection mechanism of commercial polymeric nanofiltration membranes investigated in this study may be divided into two categories: 1. Membranes for which Donnan exclusion seems to play an important role.

Minocycline-induced hyperpigmentation of tympanic membrane, sclera, teeth, and pinna.

Science.gov (United States)

Reese, Stephen; Grundfast, Kenneth

2015-11-01

A 40-year-old woman was referred by her primary care physician for evaluation after a routine physical exam revealed bilateral brownish pigmentation of the tympanic membrane. Head and neck examination in the otolaryngology clinic revealed bluish hue of both sclera, teeth, and portions of her pinnae. A hearing test revealed bilateral mild sensorineural hearing loss. The patient had a history of taking minocycline for 14 years, and the hyperpigmentation that she had is known to be a rare complication of prolonged minocycline use. However, to our knowledge, this is the first case showing photographic evidence of minocycline-induced tympanic membrane hyperpigmentation. Minocycline-induced hyperpigmentation should be considered when a patient presents with brown or blue discoloration of the tympanic membrane. © 2015 The American Laryngological, Rhinological and Otological Society, Inc.

Membrane bioreactors' potential for ethanol and biogas production: a review.

Science.gov (United States)

Ylitervo, Päivi; Akinbomia, Julius; Taherzadeha, Mohammad J

2013-01-01

Companies developing and producing membranes for different separation purposes, as well as the market for these, have markedly increased in numbers over the last decade. Membrane and separation technology might well contribute to making fuel ethanol and biogas production from lignocellulosic materials more economically viable and productive. Combining biological processes with membrane separation techniques in a membrane bioreactor (MBR) increases cell concentrations extensively in the bioreactor. Such a combination furthermore reduces product inhibition during the biological process, increases product concentration and productivity, and simplifies the separation of product and/or cells. Various MBRs have been studied over the years, where the membrane is either submerged inside the liquid to be filtered, or placed in an external loop outside the bioreactor. All configurations have advantages and drawbacks, as reviewed in this paper. The current review presents an account of the membrane separation technologies, and the research performed on MBRs, focusing on ethanol and biogas production. The advantages and potentials of the technology are elucidated.

Extensive intestinal spirochaetosis in pigs challenged with Brachyspira pilosicoli

DEFF Research Database (Denmark)

Jensen, Tim Kåre; Boye, Mette; Møller, Kristian

2004-01-01

of the study). Mild mucosal reddening and flecks of pus characterized the gross lesions, while diffuse, catarrhal colitis was revealed microscopically in all animals. Intestinal spirochaetosis with moderate to densely packed end-attached B. pilosicoli organisms was revealed extensively on the mucosal surface...... of the large intestines by light microscopy and fluorescent in situ hybridization. This study is the first to report extensive intestinal spirochaetosis in pigs challenged with B. pilosicoli....

Amniotic membrane for burn trauma

International Nuclear Information System (INIS)

Jamaluddin Zainol; Hasim Mohammad

1999-01-01

Amniotic membranes are derived from human placentae at birth. They have two layers mainly the amniotic and the chorionic surfaces which are separated by a thin layer of connective tissues. The two layers are separated during procurement, the placenta and the chorionic side are discarded and the amnion membranes are then further processed. Amnion membranes are normally procured from placentae which are normally free of infections, i.e; the mothers are antenatally screened for sexually transmitted diseases or AlDs related diseases. Intrapartum the mother should not be having chorioamnionitis or jaundice. Sometimes the amniotic membranes are acquired from fresh elective caeserian sections. After processing, the amniotic membranes are packed in two layers of polypropylene and radiated with cobalt 60 at a dose of about 25 kGy. The amniotic membranes are clinically used to cover burn surfaces especially effective for superficial or partial thickness burns. The thin membranes adhered well to the trauma areas and peeled off automatically by the second week. No change of dressing were necessary during these times because of the close adherence, there were less chance of external contamination or infections of these wounds. Due to their flexibility they are very useful to cover difference contours of the human body for example the face, body, elbows or knees. However our experience revealed that amniotic membranes are not useful for third degree bums because the membranes dissolves by the enzymes present in the wounds

Organization of Subunits in the Membrane Domain of the Bovine F-ATPase Revealed by Covalent Cross-linking.

Science.gov (United States)

Lee, Jennifer; Ding, ShuJing; Walpole, Thomas B; Holding, Andrew N; Montgomery, Martin G; Fearnley, Ian M; Walker, John E

2015-05-22

The F-ATPase in bovine mitochondria is a membrane-bound complex of about 30 subunits of 18 different kinds. Currently, ∼85% of its structure is known. The enzyme has a membrane extrinsic catalytic domain, and a membrane intrinsic domain where the turning of the enzyme's rotor is generated from the transmembrane proton-motive force. The domains are linked by central and peripheral stalks. The central stalk and a hydrophobic ring of c-subunits in the membrane domain constitute the enzyme's rotor. The external surface of the catalytic domain and membrane subunit a are linked by the peripheral stalk, holding them static relative to the rotor. The membrane domain contains six additional subunits named ATP8, e, f, g, DAPIT (diabetes-associated protein in insulin-sensitive tissues), and 6.8PL (6.8-kDa proteolipid), each with a single predicted transmembrane α-helix, but their orientation and topography are unknown. Mutations in ATP8 uncouple the enzyme and interfere with its assembly, but its roles and the roles of the other five subunits are largely unknown. We have reacted accessible amino groups in the enzyme with bifunctional cross-linking agents and identified the linked residues. Cross-links involving the supernumerary subunits, where the structures are not known, show that the C terminus of ATP8 extends ∼70 Å from the membrane into the peripheral stalk and that the N termini of the other supernumerary subunits are on the same side of the membrane, probably in the mitochondrial matrix. These experiments contribute significantly toward building up a complete structural picture of the F-ATPase. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

Organization of Subunits in the Membrane Domain of the Bovine F-ATPase Revealed by Covalent Cross-linking*

Science.gov (United States)

Lee, Jennifer; Ding, ShuJing; Walpole, Thomas B.; Holding, Andrew N.; Montgomery, Martin G.; Fearnley, Ian M.; Walker, John E.

2015-01-01

The F-ATPase in bovine mitochondria is a membrane-bound complex of about 30 subunits of 18 different kinds. Currently, ∼85% of its structure is known. The enzyme has a membrane extrinsic catalytic domain, and a membrane intrinsic domain where the turning of the enzyme's rotor is generated from the transmembrane proton-motive force. The domains are linked by central and peripheral stalks. The central stalk and a hydrophobic ring of c-subunits in the membrane domain constitute the enzyme's rotor. The external surface of the catalytic domain and membrane subunit a are linked by the peripheral stalk, holding them static relative to the rotor. The membrane domain contains six additional subunits named ATP8, e, f, g, DAPIT (diabetes-associated protein in insulin-sensitive tissues), and 6.8PL (6.8-kDa proteolipid), each with a single predicted transmembrane α-helix, but their orientation and topography are unknown. Mutations in ATP8 uncouple the enzyme and interfere with its assembly, but its roles and the roles of the other five subunits are largely unknown. We have reacted accessible amino groups in the enzyme with bifunctional cross-linking agents and identified the linked residues. Cross-links involving the supernumerary subunits, where the structures are not known, show that the C terminus of ATP8 extends ∼70 Å from the membrane into the peripheral stalk and that the N termini of the other supernumerary subunits are on the same side of the membrane, probably in the mitochondrial matrix. These experiments contribute significantly toward building up a complete structural picture of the F-ATPase. PMID:25851905

Impact of the antimicrobial peptide Novicidin on membrane structure and integrity

DEFF Research Database (Denmark)

Nielsen, Søren B; Otzen, Daniel Erik

2010-01-01

We have studied the impact of an 18-residue cationic antimicrobial peptide Novicidin (Nc) on the structure and integrity of partially anionic lipid membranes using oriented circular dichroism (OCD), quartz crystal microbalance with dissipation (QCM-D), dual polarization interferometry (DPI......), calcein dye leakage and fluorescence spectroscopy. OCD consistently showed that Nc is bound in an alpha-helical, surface bound state over a range of peptide to lipid (P/L) ratios up to approximately 1:15. Realignment of Nc at higher P/L ratios correlates to loss of membrane integrity as shown by Laurdan...... concentration, probably through formation of transient pores or transient disruption of the membrane integrity, followed by more extensive membrane disintegration at higher P/L ratios....

Ballistic impact response of lipid membranes.

Science.gov (United States)

Zhang, Yao; Meng, Zhaoxu; Qin, Xin; Keten, Sinan

2018-03-08

Therapeutic agent loaded micro and nanoscale particles as high-velocity projectiles can penetrate cells and tissues, thereby serving as gene and drug delivery vehicles for direct and rapid internalization. Despite recent progress in developing micro/nanoscale ballistic tools, the underlying biophysics of how fast projectiles deform and penetrate cell membranes is still poorly understood. To understand the rate and size-dependent penetration processes, we present coarse-grained molecular dynamics simulations of the ballistic impact of spherical projectiles on lipid membranes. Our simulations reveal that upon impact, the projectile can pursue one of three distinct pathways. At low velocities below the critical penetration velocity, projectiles rebound off the surface. At intermediate velocities, penetration occurs after the projectile deforms the membrane into a tubular thread. At very high velocities, rapid penetration occurs through localized membrane deformation without tubulation. Membrane tension, projectile velocity and size govern which phenomenon occurs, owing to their positive correlation with the reaction force generated between the projectile and the membrane during impact. Two critical membrane tension values dictate the boundaries among the three pathways for a given system, due to the rate dependence of the stress generated in the membrane. Our findings provide broad physical insights into the ballistic impact response of soft viscous membranes and guide design strategies for drug delivery through lipid membranes using micro/nanoscale ballistic tools.

Differential Effect of Plant Lipids on Membrane Organization

Science.gov (United States)

Grosjean, Kevin; Mongrand, Sébastien; Beney, Laurent; Simon-Plas, Françoise; Gerbeau-Pissot, Patricia

2015-01-01

The high diversity of the plant lipid mixture raises the question of their respective involvement in the definition of membrane organization. This is particularly the case for plant plasma membrane, which is enriched in specific lipids, such as free and conjugated forms of phytosterols and typical phytosphingolipids, such as glycosylinositolphosphoceramides. This question was here addressed extensively by characterizing the order level of membrane from vesicles prepared using various plant lipid mixtures and labeled with an environment-sensitive probe. Fluorescence spectroscopy experiments showed that among major phytosterols, campesterol exhibits a stronger ability than β-sitosterol and stigmasterol to order model membranes. Multispectral confocal microscopy, allowing spatial analysis of membrane organization, demonstrated accordingly the strong ability of campesterol to promote ordered domain formation and to organize their spatial distribution at the membrane surface. Conjugated sterol forms, alone and in synergy with free sterols, exhibit a striking ability to order membrane. Plant sphingolipids, particularly glycosylinositolphosphoceramides, enhanced the sterol-induced ordering effect, emphasizing the formation and increasing the size of sterol-dependent ordered domains. Altogether, our results support a differential involvement of free and conjugated phytosterols in the formation of ordered domains and suggest that the diversity of plant lipids, allowing various local combinations of lipid species, could be a major contributor to membrane organization in particular through the formation of sphingolipid-sterol interacting domains. PMID:25575593

The organisation of Ebola virus reveals a capacity for extensive, modular polyploidy.

Directory of Open Access Journals (Sweden)

Daniel R Beniac

Full Text Available BACKGROUND: Filoviruses, including Ebola virus, are unusual in being filamentous animal viruses. Structural data on the arrangement, stoichiometry and organisation of the component molecules of filoviruses has until now been lacking, partially due to the need to work under level 4 biological containment. The present study provides unique insights into the structure of this deadly pathogen. METHODOLOGY AND PRINCIPAL FINDINGS: We have investigated the structure of Ebola virus using a combination of cryo-electron microscopy, cryo-electron tomography, sub-tomogram averaging, and single particle image processing. Here we report the three-dimensional structure and architecture of Ebola virus and establish that multiple copies of the RNA genome can be packaged to produce polyploid virus particles, through an extreme degree of length polymorphism. We show that the helical Ebola virus inner nucleocapsid containing RNA and nucleoprotein is stabilized by an outer layer of VP24-VP35 bridges. Elucidation of the structure of the membrane-associated glycoprotein in its native state indicates that the putative receptor-binding site is occluded within the molecule, while a major neutralizing epitope is exposed on its surface proximal to the viral envelope. The matrix protein VP40 forms a regular lattice within the envelope, although its contacts with the nucleocapsid are irregular. CONCLUSIONS: The results of this study demonstrate a modular organization in Ebola virus that accommodates a well-ordered, symmetrical nucleocapsid within a flexible, tubular membrane envelope.

Mixed hydrocarbon/fluoropolymer membrane/ionomer MEAs for durability studies

Energy Technology Data Exchange (ETDEWEB)

Li, Bo [Los Alamos National Laboratory; Kim, Yu Seung [Los Alamos National Laboratory; Mukundan, Rangachary [Los Alamos National Laboratory; Borup, Rodney L [Los Alamos National Laboratory; Wilson, Mahlon S [Los Alamos National Laboratory; Welch, Cynthia [Los Alamos National Laboratory; Fenton, James [FLORIDA SOLAR ENERGY CENTER

2010-01-01

The durability of polymer electrolyte membrane (PEM) fuel cells is a major barrier to the commercialization of these systems for stationary and transportation power applications. Commercial viability depends on improving the durability of the fuel cell components to increase the system reliability. The aim of this work is to separate ionomer degradation from membrane degradation via mixed membrane/ionomer MEA experiments. The challenges of mixed MEA fabrication due to the incompatibility of the membrane and the electrode are addressed. OCV accelerated testing experiment (AST) were performed. Development of in situ diagnostics and unique experiments to characterize the performance and properties of the ionomer in the electrode as a function of time is reported. These measurements, along with extensive ex situ and post-mortem characterization, can delineate the degradation mechanisms in order to develop more durable fuel cells and fuel cell components.

Physiologically-Relevant Modes of Membrane Interactions by the Human Antimicrobial Peptide, LL-37, Revealed by SFG Experiments

Science.gov (United States)

Ding, Bei; Soblosky, Lauren; Nguyen, Khoi; Geng, Junqing; Yu, Xinglong; Ramamoorthy, Ayyalusamy; Chen, Zhan

2013-05-01

Antimicrobial peptides (AMPs) could become the next generation antibiotic compounds which can overcome bacterial resistance by disrupting cell membranes and it is essential to determine the factors underlying its mechanism of action. Although high-resolution NMR and other biological studies have provided valuable insights, it has been a major challenge to follow the AMP-membrane interactions at physiologically-relevant low peptide concentrations. In this study, we demonstrate a novel approach to overcome this major limitation by performing Sum Frequency Generation (SFG) vibrational spectroscopic experiments on lipid bilayers containing an AMP, LL-37. Our results demonstrate the power of SFG to study non-linear helical peptides and also infer that lipid-peptide interaction and the peptide orientation depend on the lipid membrane composition. The observed SFG signal changes capture the aggregating process of LL-37 on membrane. In addition, our SFG results on cholesterol-containing lipid bilayers indicate the inhibition effect of cholesterol on peptide-induced membrane permeation process.

Foulant analysis of hollow fine fiber (HFF) membranes in Red Sea SWRO plants using membrane punch autopsy (MPA)

KAUST Repository

Green, Troy N.

2017-06-12

Membrane punch autopsy (MPA) is a procedure for quantitative foulant analysis of hollow fine fiber (HFF) permeators. In the past, quantitative autopsies of membranes were restricted to spiral wound. This procedure was developed at SWCC laboratories and tested on permeators of two commercial Red Sea reverse osmosis plants. For membrane autopsies, stainless steel hollow bore picks were penetrated to membrane cores and fibers extracted for foulant analysis. Quantitative analysis of extracted materials contained inorganic and organic foulants including bacteria. Fourier transform infrared spectroscopy analysis confirmed the presence of organic fouling functional groups and scanning electron microscopy with energy dispersive X-ray spectroscopy in the presence of diatoms and silica most likely not from particulate sand. API analysis revealed the presence of Shewanella and two Vibrio microbial species confirmed by 16S rDNA sequence library. It was observed that fouling content of HFF cellulose triacetate (CTA) membranes were more than 800 times than polyamide spiral wound membranes.

Electrospun polyacrylonitrile nanofibrous membranes with varied fiber diameters and different membrane porosities as lithium-ion battery separators

International Nuclear Information System (INIS)

Ma, Xiaojing; Kolla, Praveen; Yang, Ruidong; Wang, Zhao; Zhao, Yong; Smirnova, Alevtina L.; Fong, Hao

2017-01-01

Highlights: • Nine types of electrospun polyacrylonitrile nanofibrous membranes were prepared. • These membranes had varied fiber diameters and different membrane porosities. • The membranes were explored as innovative Li-ion battery (LIB) separators. • The hot-pressed membrane with thin fibers had superior performance as LIB separator. - Abstract: In this study, nine types of polyacrylonitrile (PAN) nanofibrous membranes with varied fiber diameters and different membrane porosities are prepared by electrospinning followed by hot-pressing. Subsequently, these membranes are explored as Li-ion battery (LIB) separators. The impacts of fiber diameter and membrane porosity on electrolyte uptake, Li"+ ion transport through the membrane, electrochemical oxidation potential, and membrane performance as LIB separator (during charge/discharge cycling and rate capability tests of a cathodic half-cell) have been investigated. When compared to commercial Celgard PP separator, hot-pressed electrospun PAN nanofibrous membranes exhibit larger electrolyte uptake, higher thermal stability, wider electrochemical potential window, higher Li"+ ion permeability, and better electrochemical performance in LiMn_2O_4/separator/Li half-cell. The results also indicate that the PAN-based membrane/separator with small fiber diameters of 200–300 nm and hot-pressed under high pressure of 20 MPa surpasses all other membranes/separators and demonstrates the best performance, leading to the highest discharge capacity (89.5 mA h g"−"1 at C/2 rate) and cycle life (with capacity retention ratio being 97.7%) of the half-cell. In summary, this study has revealed that the hot-pressed electrospun PAN nanofibrous membranes (particularly those consisting of thin nanofibers) are promising as high-performance LIB separators.

Nanoscale Membrane Domain Formation Driven by Cholesterol

DEFF Research Database (Denmark)

Javanainen, Matti; Martinez-Seara, Hector; Vattulainen, Ilpo

2017-01-01

Biological membranes generate specific functions through compartmentalized regions such as cholesterol-enriched membrane nanodomains that host selected proteins. Despite the biological significance of nanodomains, details on their structure remain elusive. They cannot be observed via microscopic...... dipalmitoylphosphatidylcholine and cholesterol - the "minimal standard" for nanodomain formation. The simulations reveal how cholesterol drives the formation of fluid cholesterol-rich nanodomains hosting hexagonally packed cholesterol-poor lipid nanoclusters, both of which show registration between the membrane leaflets....... The complex nanodomain substructure forms when cholesterol positions itself in the domain boundary region. Here cholesterol can also readily flip-flop across the membrane. Most importantly, replacing cholesterol with a sterol characterized by a less asymmetric ring region impairs the emergence of nanodomains...

Biofouling of reverse osmosis membranes: effects of cleaning on biofilm microbial communities, membrane performance, and adherence of extracellular polymeric substances.

Science.gov (United States)

Al Ashhab, Ashraf; Sweity, Amer; Bayramoglu, Bihter; Herzberg, Moshe; Gillor, Osnat

2017-05-01

Laboratory-scale reverse osmosis (RO) flat-sheet systems were used with two parallel flow cells, one treated with cleaning agents and a control (ie undisturbed). The cleaning efforts increased the affinity of extracellular polymeric substances (EPS) to the RO membrane and altered the biofilm surface structure. Analysis of the membrane biofilm community composition revealed the dominance of Proteobacteria. However, within the phylum Proteobacteria, γ-Proteobacteria dominated the cleaned membrane biofilm, while β-Proteobacteria dominated the control biofilm. The composition of the fungal phyla was also altered by cleaning, with enhancement of Ascomycota and suppression of Basidiomycota. The results suggest that repeated cleaning cycles select for microbial groups that strongly attach to the RO membrane surface by producing rigid and adhesive EPS that hampers membrane performance.

Membrane biofilm communities in full-scale membrane bioreactors are not randomly assembled and consist of a core microbiome

KAUST Repository

Matar, Gerald Kamil

2017-06-21

Finding efficient biofouling control strategies requires a better understanding of the microbial ecology of membrane biofilm communities in membrane bioreactors (MBRs). Studies that characterized the membrane biofilm communities in lab-and pilot-scale MBRs are numerous, yet similar studies in full-scale MBRs are limited. Also, most of these studies have characterized the mature biofilm communities with very few studies addressing early biofilm communities. In this study, five full-scale MBRs located in Seattle (Washington, U.S.A.) were selected to address two questions concerning membrane biofilm communities (early and mature): (i) Is the assembly of biofilm communities (early and mature) the result of random immigration of species from the source community (i.e. activated sludge)? and (ii) Is there a core membrane biofilm community in full-scale MBRs? Membrane biofilm (early and mature) and activated sludge (AS) samples were collected from the five MBRs, and 16S rRNA gene sequencing was applied to investigate the bacterial communities of AS and membrane biofilms (early and mature). Alpha and beta diversity measures revealed clear differences in the bacterial community structure between the AS and biofilm (early and mature) samples in the five full-scale MBRs. These differences were mainly due to the presence of large number of unique but rare operational taxonomic units (∼13% of total reads in each MBR) in each sample. In contrast, a high percentage (∼87% of total reads in each MBR) of sequence reads was shared between AS and biofilm samples in each MBR, and these shared sequence reads mainly belong to the dominant taxa in these samples. Despite the large fraction of shared sequence reads between AS and biofilm samples, simulated biofilm communities from random sampling of the respective AS community revealed that biofilm communities differed significantly from the random assemblages (P < 0.001 for each MBR), indicating that the biofilm communities (early

Plasma membrane wounding and repair in pulmonary diseases.

Science.gov (United States)

Cong, Xiaofei; Hubmayr, Rolf D; Li, Changgong; Zhao, Xiaoli

2017-03-01

Various pathophysiological conditions such as surfactant dysfunction, mechanical ventilation, inflammation, pathogen products, environmental exposures, and gastric acid aspiration stress lung cells, and the compromise of plasma membranes occurs as a result. The mechanisms necessary for cells to repair plasma membrane defects have been extensively investigated in the last two decades, and some of these key repair mechanisms are also shown to occur following lung cell injury. Because it was theorized that lung wounding and repair are involved in the pathogenesis of acute respiratory distress syndrome (ARDS) and idiopathic pulmonary fibrosis (IPF), in this review, we summarized the experimental evidence of lung cell injury in these two devastating syndromes and discuss relevant genetic, physical, and biological injury mechanisms, as well as mechanisms used by lung cells for cell survival and membrane repair. Finally, we discuss relevant signaling pathways that may be activated by chronic or repeated lung cell injury as an extension of our cell injury and repair focus in this review. We hope that a holistic view of injurious stimuli relevant for ARDS and IPF could lead to updated experimental models. In addition, parallel discussion of membrane repair mechanisms in lung cells and injury-activated signaling pathways would encourage research to bridge gaps in current knowledge. Indeed, deep understanding of lung cell wounding and repair, and discovery of relevant repair moieties for lung cells, should inspire the development of new therapies that are likely preventive and broadly effective for targeting injurious pulmonary diseases. Copyright © 2017 the American Physiological Society.

Performance of diatomite/iron oxide modified nonwoven membrane used in membrane bioreactor process for wastewater reclamation.

Science.gov (United States)

He, Yueling; Zhang, Wenqi; Rao, Pinhua; Jin, Peng

2014-01-01

This study describes an approach for surface modification of a nonwoven membrane by diatomite/iron oxide to examine its filterability. Analysis results showed that nonwoven hydrophilicity is enhanced. Static contact angle decreases dramatically from 122.66° to 39.33°. Scanning electron micrograph images show that diatomite/iron oxide is attached on nonwoven fiber. X-ray diffraction analysis further proves that the compound is mostly magnetite. Fourier transformed infrared spectra results reveal that two new absorption peaks might be attributed to Si-O and Fe-O, respectively. Modified and original membranes were used in double nonwoven membrane bioreactors (MBRs) for synthetic wastewater treatment. High critical flux, long filtration time, slow trans-membrane pressure rise and stable sludge volume index confirmed the advantages of modified nonwoven. Comparing with original nonwoven, similar effluent qualities are achieved, meeting the requirements for wastewater reclamation.

Perceived Factors Affecting Performance Of Extension Workers In ...

African Journals Online (AJOL)

The study focused on perceived factors affecting performance of extension workers in Imo State, Nigeria. Data for the study was collected from 83 Extension agents from the Imo State Agricultural Development Programme (ADP). Results of the study revealed that the organizational factors that affect performance are ...

The Impact Of Information And Communication In Extension ...

African Journals Online (AJOL)

The article examines the impact of information and communication in extension services to rural farmers in Niger- Delta. Questionnaire, interview and personal observation methods were employed to elicit information on the impact information and communication in extension services to rural farmers. The study reveals the ...

KSHV cell attachment sites revealed by ultra sensitive tyramide signal amplification (TSA) localize to membrane microdomains that are up-regulated on mitotic cells.

Science.gov (United States)

Garrigues, H Jacques; Rubinchikova, Yelena E; Rose, Timothy M

2014-03-01

Cell surface structures initiating attachment of Kaposi's sarcoma-associated herpesvirus (KSHV) were characterized using purified hapten-labeled virions visualized by confocal microscopy with a sensitive fluorescent enhancement using tyramide signal amplification (TSA). KSHV attachment sites were present in specific cellular domains, including actin-based filopodia, lamellipodia, ruffled membranes, microvilli and intercellular junctions. Isolated microdomains were identified on the dorsal surface, which were heterogeneous in size with a variable distribution that depended on cellular confluence and cell cycle stage. KSHV binding domains ranged from scarce on interphase cells to dense and continuous on mitotic cells, and quantitation of bound virus revealed a significant increase on mitotic compared to interphase cells. KSHV also bound to a supranuclear domain that was distinct from microdomains in confluent and interphase cells. These results suggest that rearrangement of the cellular membrane during mitosis induces changes in cell surface receptors implicated in the initial attachment stage of KSHV entry. Copyright © 2014 Elsevier Inc. All rights reserved.

Membrane raft association is a determinant of plasma membrane localization.

Science.gov (United States)

Diaz-Rohrer, Blanca B; Levental, Kandice R; Simons, Kai; Levental, Ilya

2014-06-10

The lipid raft hypothesis proposes lateral domains driven by preferential interactions between sterols, sphingolipids, and specific proteins as a central mechanism for the regulation of membrane structure and function; however, experimental limitations in defining raft composition and properties have prevented unequivocal demonstration of their functional relevance. Here, we establish a quantitative, functional relationship between raft association and subcellular protein sorting. By systematic mutation of the transmembrane and juxtamembrane domains of a model transmembrane protein, linker for activation of T-cells (LAT), we generated a panel of variants possessing a range of raft affinities. These mutations revealed palmitoylation, transmembrane domain length, and transmembrane sequence to be critical determinants of membrane raft association. Moreover, plasma membrane (PM) localization was strictly dependent on raft partitioning across the entire panel of unrelated mutants, suggesting that raft association is necessary and sufficient for PM sorting of LAT. Abrogation of raft partitioning led to mistargeting to late endosomes/lysosomes because of a failure to recycle from early endosomes. These findings identify structural determinants of raft association and validate lipid-driven domain formation as a mechanism for endosomal protein sorting.

Membrane Fusion Involved in Neurotransmission: Glimpse from Electron Microscope and Molecular Simulation

Directory of Open Access Journals (Sweden)

Zhiwei Yang

2017-06-01

Full Text Available Membrane fusion is one of the most fundamental physiological processes in eukaryotes for triggering the fusion of lipid and content, as well as the neurotransmission. However, the architecture features of neurotransmitter release machinery and interdependent mechanism of synaptic membrane fusion have not been extensively studied. This review article expounds the neuronal membrane fusion processes, discusses the fundamental steps in all fusion reactions (membrane aggregation, membrane association, lipid rearrangement and lipid and content mixing and the probable mechanism coupling to the delivery of neurotransmitters. Subsequently, this work summarizes the research on the fusion process in synaptic transmission, using electron microscopy (EM and molecular simulation approaches. Finally, we propose the future outlook for more exciting applications of membrane fusion involved in synaptic transmission, with the aid of stochastic optical reconstruction microscopy (STORM, cryo-EM (cryo-EM, and molecular simulations.

Membrane Fusion Involved in Neurotransmission: Glimpse from Electron Microscope and Molecular Simulation

Science.gov (United States)

Yang, Zhiwei; Gou, Lu; Chen, Shuyu; Li, Na; Zhang, Shengli; Zhang, Lei

2017-01-01

Membrane fusion is one of the most fundamental physiological processes in eukaryotes for triggering the fusion of lipid and content, as well as the neurotransmission. However, the architecture features of neurotransmitter release machinery and interdependent mechanism of synaptic membrane fusion have not been extensively studied. This review article expounds the neuronal membrane fusion processes, discusses the fundamental steps in all fusion reactions (membrane aggregation, membrane association, lipid rearrangement and lipid and content mixing) and the probable mechanism coupling to the delivery of neurotransmitters. Subsequently, this work summarizes the research on the fusion process in synaptic transmission, using electron microscopy (EM) and molecular simulation approaches. Finally, we propose the future outlook for more exciting applications of membrane fusion involved in synaptic transmission, with the aid of stochastic optical reconstruction microscopy (STORM), cryo-EM (cryo-EM), and molecular simulations. PMID:28638320

A Peptidomimetic Antibiotic Targets Outer Membrane Proteins and Disrupts Selectively the Outer Membrane in Escherichia coli.

Science.gov (United States)

Urfer, Matthias; Bogdanovic, Jasmina; Lo Monte, Fabio; Moehle, Kerstin; Zerbe, Katja; Omasits, Ulrich; Ahrens, Christian H; Pessi, Gabriella; Eberl, Leo; Robinson, John A

2016-01-22

Increasing antibacterial resistance presents a major challenge in antibiotic discovery. One attractive target in Gram-negative bacteria is the unique asymmetric outer membrane (OM), which acts as a permeability barrier that protects the cell from external stresses, such as the presence of antibiotics. We describe a novel β-hairpin macrocyclic peptide JB-95 with potent antimicrobial activity against Escherichia coli. This peptide exhibits no cellular lytic activity, but electron microscopy and fluorescence studies reveal an ability to selectively disrupt the OM but not the inner membrane of E. coli. The selective targeting of the OM probably occurs through interactions of JB-95 with selected β-barrel OM proteins, including BamA and LptD as shown by photolabeling experiments. Membrane proteomic studies reveal rapid depletion of many β-barrel OM proteins from JB-95-treated E. coli, consistent with induction of a membrane stress response and/or direct inhibition of the Bam folding machine. The results suggest that lethal disruption of the OM by JB-95 occurs through a novel mechanism of action at key interaction sites within clusters of β-barrel proteins in the OM. These findings open new avenues for developing antibiotics that specifically target β-barrel proteins and the integrity of the Gram-negative OM. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

Biocompatibility of epoxidized styrene-butadiene-styrene block copolymer membrane

International Nuclear Information System (INIS)

Yang, Jen Ming; Tsai, Shih Chang

2010-01-01

Styrene-butadiene-styrene block copolymer (SBS) membrane was prepared by solution casting method and then was epoxidized with peroxyformic acid generated in situ to yield the epoxidized styrene-butadiene-styrene block copolymer membrane (ESBS). The structure and properties of ESBS were characterized with infrared spectroscopy, Universal Testing Machine, differential scanning calorimetry (DSC), and thermogravimetry analysis (TGA). The performances of contact angle, water content, protein adsorption, and water vapor transmission rate on ESBS membrane were determined. After epoxidation, the hydrophilicity of the membrane increased. The water vapor transmission rate of ESBS membrane is similar to human skin. The biocompatibility of ESBS membrane was evaluated with the cell culture of fibroblasts on the membrane. It revealed that the cells not only remained viable but also proliferated on the surface of the various ESBS membranes and the population doubling time for fibroblast culture decreased.

Nafion–clay nanocomposite membranes: Morphology and properties

KAUST Repository

Herrera Alonso, Rafael; Estevez, Luis; Lian, Huiqin; Kelarakis, Antonios; Giannelis, Emmanuel P.

2009-01-01

A series of Nafion-clay nanocomposite membranes were synthesized and characterized. To minimize any adverse effects on ionic conductivity the clay nanoparticles were H+ exchanged prior to mixing with Nafion. Well-dispersed, mechanically robust, free-standing nanocomposite membranes were prepared by casting from a water suspension at 180 °C under pressure. SAXS profiles reveal a preferential orientation of Nafion aggregates parallel to the membrane surface, or normal plane. This preferred orientation is induced by the platy nature of the clay nanoparticles, which tend to align parallel to the surface of the membrane. The nanocomposite membranes show dramatically reduced methanol permeability, while maintaining high levels of proton conductivity. The hybrid films are much stiffer and can withstand much higher temperatures compared to pure Nafion. The superior thermomechanical, electrochemical and barrier properties of the nanocomposite membranes are of significant interest for direct methanol fuel cell applications. © 2009 Elsevier Ltd. All rights reserved.

Nafion–clay nanocomposite membranes: Morphology and properties

KAUST Repository

Herrera Alonso, Rafael

2009-05-01

A series of Nafion-clay nanocomposite membranes were synthesized and characterized. To minimize any adverse effects on ionic conductivity the clay nanoparticles were H+ exchanged prior to mixing with Nafion. Well-dispersed, mechanically robust, free-standing nanocomposite membranes were prepared by casting from a water suspension at 180 °C under pressure. SAXS profiles reveal a preferential orientation of Nafion aggregates parallel to the membrane surface, or normal plane. This preferred orientation is induced by the platy nature of the clay nanoparticles, which tend to align parallel to the surface of the membrane. The nanocomposite membranes show dramatically reduced methanol permeability, while maintaining high levels of proton conductivity. The hybrid films are much stiffer and can withstand much higher temperatures compared to pure Nafion. The superior thermomechanical, electrochemical and barrier properties of the nanocomposite membranes are of significant interest for direct methanol fuel cell applications. © 2009 Elsevier Ltd. All rights reserved.

Subaortic membrane mimicking hypertrophic cardiomyopathy.

Science.gov (United States)

Anderson, Mark Joseph; Arruda-Olson, Adelaide; Gersh, Bernard; Geske, Jeffrey

2015-11-04

A 34-year-old man was referred for progressive angina and exertional dyspnoea refractory to medical therapy, with a presumptive diagnosis of hypertrophic cardiomyopathy (HCM). Transthoracic echocardiography (TTE) revealed asymmetric septal hypertrophy without systolic anterior motion of the mitral valve leaflet and with no dynamic left ventricular outflow tract (LVOT) obstruction. However, the LVOT velocity was elevated at rest as well as with provocation, without the characteristic late peaking obstruction seen in HCM. Focused TTE to evaluate for suspected fixed obstruction demonstrated a subaortic membrane 2.2 cm below the aortic valve. Coronary CT angiography confirmed the presence of the subaortic membrane and was negative for concomitant coronary artery disease. Surgical resection of the subaortic membrane and septal myectomy resulted in significant symptomatic relief and lower LVOT velocities on postoperative TTE. This case reminds the clinician to carefully evaluate for alternative causes of LVOT obstruction, especially subaortic membrane, as a cause of symptoms mimicking HCM. 2015 BMJ Publishing Group Ltd.

A Membrane Model from Implicit Elasticity Theory

Science.gov (United States)

Freed, A. D.; Liao, J.; Einstein, D. R.

2014-01-01

A Fungean solid is derived for membranous materials as a body defined by isotropic response functions whose mathematical structure is that of a Hookean solid where the elastic constants are replaced by functions of state derived from an implicit, thermodynamic, internal-energy function. The theory utilizes Biot’s (1939) definitions for stress and strain that, in 1-dimension, are the stress/strain measures adopted by Fung (1967) when he postulated what is now known as Fung’s law. Our Fungean membrane model is parameterized against a biaxial data set acquired from a porcine pleural membrane subjected to three, sequential, proportional, planar extensions. These data support an isotropic/deviatoric split in the stress and strain-rate hypothesized by our theory. These data also demonstrate that the material response is highly non-linear but, otherwise, mechanically isotropic. These data are described reasonably well by our otherwise simple, four-parameter, material model. PMID:24282079

Changes in physicochemical and transport properties of a reverse osmosis membrane exposed to chloraminated seawater

KAUST Repository

Valentino, Lauren; Renkens, Tennie; Maugin, Thomas; Crouè , Jean-Philippe Philippe; Mariñ as, Benito J.

2015-01-01

This study contributed to improving our understanding of how disinfectants, applied to control biofouling of reverse osmosis (RO) membranes, result in membrane performance degradation. We investigated changes in physicochemical properties and permeation performance of a RO membrane with fully aromatic polyamide (PA) active layer. Membrane samples were exposed to varying concentrations of monochloramine, bromide, and iodide in both synthetic and natural seawater. Elemental analysis of the membrane active layer by Rutherford backscattering spectrometry (RBS) revealed the incorporation of bromine and iodine into the polyamide. The kinetics of polyamide bromination were first order with respect to the concentration of the secondary oxidizing agent Br2 for the conditions investigated. Halogenated membranes were characterized after treatment with a reducing agent and heavy ion probes to reveal the occurrence of irreversible ring halogenation and an increase in carboxylic groups, the latter produced as a result of amide bond cleavage. Finally, permeation experiments revealed increases in both water permeability and salt passage as a result of oxidative damage.

Changes in physicochemical and transport properties of a reverse osmosis membrane exposed to chloraminated seawater

KAUST Repository

Valentino, Lauren

2015-02-17

This study contributed to improving our understanding of how disinfectants, applied to control biofouling of reverse osmosis (RO) membranes, result in membrane performance degradation. We investigated changes in physicochemical properties and permeation performance of a RO membrane with fully aromatic polyamide (PA) active layer. Membrane samples were exposed to varying concentrations of monochloramine, bromide, and iodide in both synthetic and natural seawater. Elemental analysis of the membrane active layer by Rutherford backscattering spectrometry (RBS) revealed the incorporation of bromine and iodine into the polyamide. The kinetics of polyamide bromination were first order with respect to the concentration of the secondary oxidizing agent Br2 for the conditions investigated. Halogenated membranes were characterized after treatment with a reducing agent and heavy ion probes to reveal the occurrence of irreversible ring halogenation and an increase in carboxylic groups, the latter produced as a result of amide bond cleavage. Finally, permeation experiments revealed increases in both water permeability and salt passage as a result of oxidative damage.

Image Restoration and Analysis of Influenza Virions Binding to Membrane Receptors Reveal Adhesion-Strengthening Kinetics.

Directory of Open Access Journals (Sweden)

Donald W Lee

Full Text Available With the development of single-particle tracking (SPT microscopy and host membrane mimics called supported lipid bilayers (SLBs, stochastic virus-membrane binding interactions can be studied in depth while maintaining control over host receptor type and concentration. However, several experimental design challenges and quantitative image analysis limitations prevent the widespread use of this approach. One main challenge of SPT studies is the low signal-to-noise ratio of SPT videos, which is sometimes inevitable due to small particle sizes, low quantum yield of fluorescent dyes, and photobleaching. These situations could render current particle tracking software to yield biased binding kinetic data caused by intermittent tracking error. Hence, we developed an effective image restoration algorithm for SPT applications called STAWASP that reveals particles with a signal-to-noise ratio of 2.2 while preserving particle features. We tested our improvements to the SPT binding assay experiment and imaging procedures by monitoring X31 influenza virus binding to α2,3 sialic acid glycolipids. Our interests lie in how slight changes to the peripheral oligosaccharide structures can affect the binding rate and residence times of viruses. We were able to detect viruses binding weakly to a glycolipid called GM3, which was undetected via assays such as surface plasmon resonance. The binding rate was around 28 folds higher when the virus bound to a different glycolipid called GD1a, which has a sialic acid group extending further away from the bilayer surface than GM3. The improved imaging allowed us to obtain binding residence time distributions that reflect an adhesion-strengthening mechanism via multivalent bonds. We empirically fitted these distributions using a time-dependent unbinding rate parameter, koff, which diverges from standard treatment of koff as a constant. We further explain how to convert these models to fit ensemble-averaged binding data

Spontaneous Absorption of Extensive Subinternal Limiting Membrane Hemorrhage in Shaken Baby Syndrome

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Tatiana Tarules Azzi

2014-01-01

Full Text Available The Shaken Baby Syndrome (SBS is characterized by subdural hematomas (SH, retinal hemorrhages (RH, and multiple fractures of long bones without external evidence of head trauma. Subinternal limiting membrane (ILM hemorrhage, also known as macular schisis, is a characteristic finding of this entity. There is no guideline on the right time to indicate surgical treatment. This report describes an abused child with massive sub-ILM hemorrhage, which showed spontaneous absorption after less than two months of follow-up. Due to the possible spontaneous resolution, we suggest an initial conservative treatment in cases of sub-ILM hemorrhage related to SBS.

Membrane contactors for CO2 capture processes - critical review

Science.gov (United States)

Nogalska, Adrianna; Trojanowska, Anna; Garcia-Valls, Ricard

2017-07-01

The use of membrane contactor in industrial processes is wide, and lately it started to be used in CO2 capture process mainly for gas purification or to reduce the emission. Use of the membrane contactor provides high contact surface area so the size of the absorber unit significantly decreases, which is an important factor for commercialization. The research has been caried out regarding the use of novel materials for the membrane production and absorbent solution improvements. The present review reveals the progress in membrane contactor systems for CO2 capture processes concerning solution for ceramic membrane wetting, comparison study of different polymers used for fabrication and methods of enzyme immobilization for biocomposite membrane. Also information about variety of absorbent solutions is described.

Integral membrane protein structure determination using pseudocontact shifts

Energy Technology Data Exchange (ETDEWEB)

Crick, Duncan J.; Wang, Jue X. [University of Cambridge, Department of Biochemistry (United Kingdom); Graham, Bim; Swarbrick, James D. [Monash University, Monash Institute of Pharmaceutical Sciences (Australia); Mott, Helen R.; Nietlispach, Daniel, E-mail: dn206@cam.ac.uk [University of Cambridge, Department of Biochemistry (United Kingdom)

2015-04-15

Obtaining enough experimental restraints can be a limiting factor in the NMR structure determination of larger proteins. This is particularly the case for large assemblies such as membrane proteins that have been solubilized in a membrane-mimicking environment. Whilst in such cases extensive deuteration strategies are regularly utilised with the aim to improve the spectral quality, these schemes often limit the number of NOEs obtainable, making complementary strategies highly beneficial for successful structure elucidation. Recently, lanthanide-induced pseudocontact shifts (PCSs) have been established as a structural tool for globular proteins. Here, we demonstrate that a PCS-based approach can be successfully applied for the structure determination of integral membrane proteins. Using the 7TM α-helical microbial receptor pSRII, we show that PCS-derived restraints from lanthanide binding tags attached to four different positions of the protein facilitate the backbone structure determination when combined with a limited set of NOEs. In contrast, the same set of NOEs fails to determine the correct 3D fold. The latter situation is frequently encountered in polytopical α-helical membrane proteins and a PCS approach is thus suitable even for this particularly challenging class of membrane proteins. The ease of measuring PCSs makes this an attractive route for structure determination of large membrane proteins in general.

YSZ-Reinforced Alumina Multi-Channel Capillary Membranes for Micro-Filtration

Directory of Open Access Journals (Sweden)

Bo Wang

2015-12-01

Full Text Available The combined phase-inversion and sintering method not only produces ceramic hollow fibre membranes with much lower fabrication costs than conventional methods, but these membranes can also be designed to have greatly reduced transport resistances for filtration processes. The bottleneck of this technique is the weak mechanical property of the fibres, due to the small dimensions and the brittle nature of the ceramic materials. In this study, yttrium stabilised zirconia (YSZ reinforced alumina seven-channel capillary microfiltration membranes were prepared with a pore size of ~230 nm and their mechanical property and permeation characteristics were studied. It is found that the addition of YSZ can effectively enhance the mechanical property of the membrane and also increase pure water permeation flux. The Al2O3-YSZ seven-channel capillary membranes could reach a fracture load of 23.4 N and a bending extension of 0.54 mm when being tested with a 6 cm span, to meet the requirements for most industrial microfiltration applications.

YSZ-Reinforced Alumina Multi-Channel Capillary Membranes for Micro-Filtration.

Science.gov (United States)

Wang, Bo; Lee, Melanie; Li, Kang

2015-12-30

The combined phase-inversion and sintering method not only produces ceramic hollow fibre membranes with much lower fabrication costs than conventional methods, but these membranes can also be designed to have greatly reduced transport resistances for filtration processes. The bottleneck of this technique is the weak mechanical property of the fibres, due to the small dimensions and the brittle nature of the ceramic materials. In this study, yttrium stabilised zirconia (YSZ) reinforced alumina seven-channel capillary microfiltration membranes were prepared with a pore size of ~230 nm and their mechanical property and permeation characteristics were studied. It is found that the addition of YSZ can effectively enhance the mechanical property of the membrane and also increase pure water permeation flux. The Al₂O₃-YSZ seven-channel capillary membranes could reach a fracture load of 23.4 N and a bending extension of 0.54 mm when being tested with a 6 cm span, to meet the requirements for most industrial microfiltration applications.

Ceramic membrane defouling (cleaning) by air Nano Bubbles.

Science.gov (United States)

Ghadimkhani, Aliasghar; Zhang, Wen; Marhaba, Taha

2016-03-01

Ceramic membranes are among the most promising technologies for membrane applications, owing to their excellent resistance to mechanical, chemical, and thermal stresses. However, membrane fouling is still an issue that hampers the applications at large scales. Air Nano Bubbles (NBs), due to high mass transfer efficiency, could potentially prevent fouling of ceramic membrane filtration processes. In this study, bench and pilot scale ceramic membrane filtration was performed with air NBs to resist fouling. To simulate fouling, humic acid, as an organic foulant, was applied to the membrane flat sheet surface. Complete membrane clogging was achieved in less than 6 h. Membrane defouling (cleaning) was performed by directly feeding of air NBs to the membrane cells. The surface of the ceramic membrane was superbly cleaned by air NBs, as revealed by atomic force microscope (AFM) images before and after the treatment. The permeate flux recovered to its initial level (e.g., 26.7 × 10(-9) m(3)/m(2)/s at applied pressure of 275.8 kPa), which indicated that NBs successfully unclogged the pores of the membrane. The integrated ceramic membrane and air NBs system holds potential as an innovative sustainable technology. Copyright © 2015 Elsevier Ltd. All rights reserved.

Superdiffusive motion of membrane-targeting C2 domains

Science.gov (United States)

Campagnola, Grace; Nepal, Kanti; Schroder, Bryce W.; Peersen, Olve B.; Krapf, Diego

2015-12-01

Membrane-targeting domains play crucial roles in the recruitment of signalling molecules to the plasma membrane. For most peripheral proteins, the protein-to-membrane interaction is transient. After proteins dissociate from the membrane they have been observed to rebind following brief excursions in the bulk solution. Such membrane hops can have broad implications for the efficiency of reactions on membranes. We study the diffusion of membrane-targeting C2 domains using single-molecule tracking in supported lipid bilayers. The ensemble-averaged mean square displacement (MSD) exhibits superdiffusive behaviour. However, traditional time-averaged MSD analysis of individual trajectories remains linear and does not reveal superdiffusion. Our observations are explained in terms of bulk excursions that introduce jumps with a heavy-tail distribution. These hopping events allow proteins to explore large areas in a short time. The experimental results are shown to be consistent with analytical models of bulk-mediated diffusion and numerical simulations.

Argon and nitrogen beams influencing membrane permeate fluxes and microbial growth

International Nuclear Information System (INIS)

Wanichapichart, P.; Taweepreeda, W.; Choomgan, P.; Yu, L.D.

2010-01-01

Porous cellulose and dense chitosan membranes were bombarded with argon and nitrogen-ion beams using two energy levels, 30 and 120 keV, of the same fluency of 5x10 14 ions/cm 2 for a comparison study. The results revealed that both beam types reduced the hydraulic permeability of the membranes. Using a NaCl solution of 4000 ppm concentration as feed, the ability to reject salt of dense chitosan membrane was reduced only if it was pretreated with 120 keV nitrogen-ion beams. A Fourier Transform Infrared Spectroscopy study showed that molecular weight of chitosan was possibly decreased after the bombardment with 120 keV beams. The analysis of the cellulose membranes revealed that a dense structure was created without affecting the OH functional groups. This study found that only chitosan membranes possessed an anti-fungi property if being implanted with positive charges of nitrogen or argon ions of 120 keV.

Proteomics of plasma membranes from poplar trees reveals tissue distribution of transporters, receptors, and proteins in cell wall formation.

Science.gov (United States)

Nilsson, Robert; Bernfur, Katja; Gustavsson, Niklas; Bygdell, Joakim; Wingsle, Gunnar; Larsson, Christer

2010-02-01

By exploiting the abundant tissues available from Populus trees, 3-4 m high, we have been able to isolate plasma membranes of high purity from leaves, xylem, and cambium/phloem at a time (4 weeks after bud break) when photosynthesis in the leaves and wood formation in the xylem should have reached a steady state. More than 40% of the 956 proteins identified were found in the plasma membranes of all three tissues and may be classified as "housekeeping" proteins, a typical example being P-type H(+)-ATPases. Among the 213 proteins predicted to be integral membrane proteins, transporters constitute the largest class (41%) followed by receptors (14%) and proteins involved in cell wall and carbohydrate metabolism (8%) and membrane trafficking (8%). ATP-binding cassette transporters (all members of subfamilies B, C, and G) and receptor-like kinases (four subfamilies) were two of the largest protein families found, and the members of these two families showed pronounced tissue distribution. Leaf plasma membranes were characterized by a very high proportion of transporters, constituting almost half of the integral proteins. Proteins involved in cell wall synthesis (such as cellulose and sucrose synthases) and membrane trafficking were most abundant in xylem plasma membranes in agreement with the role of the xylem in wood formation. Twenty-five integral proteins and 83 soluble proteins were exclusively found in xylem plasma membranes, which identifies new candidates associated with cell wall synthesis and wood formation. Among the proteins uniquely found in xylem plasma membranes were most of the enzymes involved in lignin biosynthesis, which suggests that they may exist as a complex linked to the plasma membrane.

Herpesvirus glycoproteins undergo multiple antigenic changes before membrane fusion.

Directory of Open Access Journals (Sweden)

Daniel L Glauser

Full Text Available Herpesvirus entry is a complicated process involving multiple virion glycoproteins and culminating in membrane fusion. Glycoprotein conformation changes are likely to play key roles. Studies of recombinant glycoproteins have revealed some structural features of the virion fusion machinery. However, how the virion glycoproteins change during infection remains unclear. Here using conformation-specific monoclonal antibodies we show in situ that each component of the Murid Herpesvirus-4 (MuHV-4 entry machinery--gB, gH/gL and gp150--changes in antigenicity before tegument protein release begins. Further changes then occurred upon actual membrane fusion. Thus virions revealed their final fusogenic form only in late endosomes. The substantial antigenic differences between this form and that of extracellular virions suggested that antibodies have only a limited opportunity to block virion membrane fusion.

Oxidative degradation of polybenzimidazole membranes as electrolytes for high temperature proton exchange membrane fuel cells

DEFF Research Database (Denmark)

Liao, J.H.; Li, Qingfeng; Rudbeck, H.C.

2011-01-01

the oxidative degradation of the polymer membrane was studied under the Fenton test conditions by the weight loss, intrinsic viscosity, size exclusion chromatography, scanning electron microscopy and Fourier transform infrared spectroscopy. During the Fenton test, significant weight losses depending...... on the initial molecular weight of the polymer were observed. At the same time, viscosity and SEC measurements revealed a steady decrease in molecular weight. The degradation of acid doped PBI membranes under Fenton test conditions is proposed to start by the attack of hydroxyl radicals at the carbon atom......Polybenzimidazole membranes imbibed with acid are emerging as a suitable electrolyte material for high-temperature polymer electrolyte fuel cells. The oxidative stability of polybenzimidazole has been identified as an important issue for the long-term durability of such cells. In this paper...

Transcriptome and membrane fatty acid analyses reveal different strategies for responding to permeating and non-permeating solutes in the bacterium Sphingomonas wittichii

Directory of Open Access Journals (Sweden)

Johnson David R

2011-11-01

opposite effect and decreased after perturbation with PEG8000. Conclusions A combination of growth assays, transcriptome profiling, and membrane fatty acid analyses revealed that permeating and non-permeating solutes trigger different adaptive responses in strain RW1, suggesting these solutes affect cells in fundamentally different ways. Future work is now needed that connects these responses with the responses observed in more realistic scenarios of soil desiccation.

The future of hemodialysis membranes.

Science.gov (United States)

Humes, H D; Fissell, W H; Tiranathanagul, K

2006-04-01

Hemodialytic treatment of patients with either acute or chronic renal failure has had a dramatic impact on the mortality rates of these patients. Unfortunately, this membrane-based therapy is still incomplete renal replacement, as the mortality and morbidity of these patients remain unacceptably high. Much progress must be made to improve the biocompatibility of hemodialysis membranes as well as their hydraulic and permselective properties to remove small solutes and 'middle molecules' in compact cartridges. The next directions of development will leverage materials and mechanical engineering technology, including microfluidics and nanofabrication, to further improve the clearance functions of the kidney to replicate glomerular permselectivity while retaining high rates of hydraulic permeability. The extension of membrane technology to biohybrid devices utilizing progenitor/stem cells will be another substantive advance for renal replacement therapy. The ability to not only replace solute and water clearance but also active reabsorptive transport and metabolic activity will add additional benefit to the therapy of patients suffering from renal failure. This area of translational research is rich in creative opportunities to improve the unmet medical needs of patients with either chronic or acute renal failure.

Flavivirus infection from mosquitoes in vitro reveals cell entry at the plasma membrane

International Nuclear Information System (INIS)

Vancini, Ricardo; Kramer, Laura D.; Ribeiro, Mariana; Hernandez, Raquel; Brown, Dennis

2013-01-01

Dengue and West Nile viruses are enveloped RNA viruses that belong to genus Flavivirus (family Flaviviridae) and are considered important mosquito-borne viral pathogenic agents worldwide. A potential target for intervention strategies is the virus cell entry mechanism. Previous studies of flavivirus entry have focused on the effects of biochemical and molecular inhibitors on viral entry leading to controversial conclusions suggesting that the process is dependent upon endocytosis and low pH mediated membrane fusion. In this study we analyzed the early events in the infection process by means of electron microscopy and immuno-gold labeling of viral particles during cell entry, and used as a new approach for infecting cells with viruses obtained directly from mosquitoes. The results show that Dengue and West Nile viruses may infect cells by a mechanism that involves direct penetration of the host cell plasma membrane as proposed for alphaviruses.

Flavivirus infection from mosquitoes in vitro reveals cell entry at the plasma membrane

Energy Technology Data Exchange (ETDEWEB)

Vancini, Ricardo [Department of Molecular and Structural Biochemistry, North Carolina State University, Raleigh, NC (United States); Kramer, Laura D. [Wadsworth Center, New York State Department of Health, and School of Public Health, State University of New York at Albany, Albany, NY (United States); Ribeiro, Mariana; Hernandez, Raquel [Department of Molecular and Structural Biochemistry, North Carolina State University, Raleigh, NC (United States); Brown, Dennis, E-mail: dennis_brown@ncsu.edu [Department of Molecular and Structural Biochemistry, North Carolina State University, Raleigh, NC (United States)

2013-01-20

Dengue and West Nile viruses are enveloped RNA viruses that belong to genus Flavivirus (family Flaviviridae) and are considered important mosquito-borne viral pathogenic agents worldwide. A potential target for intervention strategies is the virus cell entry mechanism. Previous studies of flavivirus entry have focused on the effects of biochemical and molecular inhibitors on viral entry leading to controversial conclusions suggesting that the process is dependent upon endocytosis and low pH mediated membrane fusion. In this study we analyzed the early events in the infection process by means of electron microscopy and immuno-gold labeling of viral particles during cell entry, and used as a new approach for infecting cells with viruses obtained directly from mosquitoes. The results show that Dengue and West Nile viruses may infect cells by a mechanism that involves direct penetration of the host cell plasma membrane as proposed for alphaviruses.

Meningococcal outer membrane vesicle composition-dependent activation of the innate immune response

NARCIS (Netherlands)

Zariri, Afshin; Beskers, Joep; van de Waterbeemd, Bas; Hamstra, Hendrik Jan; Bindels, Tim H E; van Riet, Elly; van Putten, Jos P M; van der Ley, Peter

2016-01-01

Meningococcal outer membrane vesicles (OMVs) have been extensively investigated and successfully implemented as vaccines. They contain pathogen associated molecular patterns including lipopolysaccharide (LPS), capable of triggering innate immunity. However, Neisseria meningitidis contains an

The actin homologue MreB organizes the bacterial cell membrane.

Science.gov (United States)

Strahl, Henrik; Bürmann, Frank; Hamoen, Leendert W

2014-03-07

The eukaryotic cortical actin cytoskeleton creates specific lipid domains, including lipid rafts, which determine the distribution of many membrane proteins. Here we show that the bacterial actin homologue MreB displays a comparable activity. MreB forms membrane-associated filaments that coordinate bacterial cell wall synthesis. We noticed that the MreB cytoskeleton influences fluorescent staining of the cytoplasmic membrane. Detailed analyses combining an array of mutants, using specific lipid staining techniques and spectroscopic methods, revealed that MreB filaments create specific membrane regions with increased fluidity (RIFs). Interference with these fluid lipid domains (RIFs) perturbs overall lipid homeostasis and affects membrane protein localization. The influence of MreB on membrane organization and fluidity may explain why the active movement of MreB stimulates membrane protein diffusion. These novel MreB activities add additional complexity to bacterial cell membrane organization and have implications for many membrane-associated processes.

Hydrophilic Phage-Mimicking Membrane Active Antimicrobials Reveal Nanostructure-Dependent Activity and Selectivity.

Science.gov (United States)

Jiang, Yunjiang; Zheng, Wan; Kuang, Liangju; Ma, Hairong; Liang, Hongjun

2017-09-08

The prevalent wisdom on developing membrane active antimicrobials (MAAs) is to seek a delicate, yet unquantified, cationic-hydrophobic balance. Inspired by phages that use nanostructured protein devices to invade bacteria efficiently and selectively, we study here the antibiotic role of nanostructures by designing spherical and rod-like polymer molecular brushes (PMBs) that mimic the two basic structural motifs of bacteriophages. Three model PMBs with different well-defined geometries consisting of multiple, identical copies of densely packed poly(4-vinyl-N-methylpyridine iodide) branches are synthesized by controlled/"living" polymerization, reminiscent of the viral structural motifs comprised of multiple copies of protein subunits. We show that, while the individual linear-chain polymer branch that makes up the PMBs is hydrophilic and a weak antimicrobial, amphiphilicity is not a required antibiotic trait once nanostructures come into play. The nanostructured PMBs induce an unusual topological transition of bacterial but not mammalian membranes to form pores. The sizes and shapes of the nanostructures further help define the antibiotic activity and selectivity of the PMBs against different families of bacteria. This study highlights the importance of nanostructures in the design of MAAs with high activity, low toxicity, and target specificity.

Organization and Dynamics of Receptor Proteins in a Plasma Membrane.

Science.gov (United States)

Koldsø, Heidi; Sansom, Mark S P

2015-11-25

The interactions of membrane proteins are influenced by their lipid environment, with key lipid species able to regulate membrane protein function. Advances in high-resolution microscopy can reveal the organization and dynamics of proteins and lipids within living cells at resolutions membranes of in vivo-like complexity. We explore the dynamics of proteins and lipids in crowded and complex plasma membrane models, thereby closing the gap in length and complexity between computations and experiments. Our simulations provide insights into the mutual interplay between lipids and proteins in determining mesoscale (20-100 nm) fluctuations of the bilayer, and in enabling oligomerization and clustering of membrane proteins.

Fabrication of high flux and antifouling mixed matrix fumarate-alumoxane/PAN membranes via electrospinning for application in membrane bioreactors

Science.gov (United States)

Moradi, Golshan; Zinadini, Sirus; Rajabi, Laleh; Dadari, Soheil

2018-01-01

The nanofibrous Polyacrylonitrile (PAN) membranes embedded with fumarate-alumoxane (Fum-A) nanoparticles were prepared via electrospinning technique as high flux and antifouling membranes for membrane bioreactor (MBR) applications. The effect of Fum-A nanoparticles on membrane morphology, surface hydrophilicity, pure water flux, effluent turbidity and the antifouling property was investigated. Fum-A is a carboxylate-alumoxane nanoparticle covered by extra hydroxyl and carboxylate groups on its surface. By embedding Fum-A nanoparticles into the spinning solution, the surface hydrophilicity and pure water flux of the resulted membranes were improved. The smooth surface of fibers at the low amount of nanoparticles and the agglomeration of nanoparticles at their high concentration were shown in SEM images of the membranes surface. The energy dispersive spectroscopy (EDS) and Fourier transform infrared spectroscopy (FTIR) analysis of the prepared Fum-A/PAN membrane confirmed the presence of carboxylate and hydroxyl functional groups of Fum-A nanoparticles on the surface of the Fum-A nanoparticles containing membrane. The results obtained from the filtration of activated sludge suspension revealed that by addition of a low amount of Fum-A nanoparticles, the irreversible fouling was significantly decreased due to the higher hydrophilicity. The Fum-A/PAN membranes showed superior permeate flux and antifouling properties compared to bare electrospun PAN membrane. Finally, 2 wt.% Fum-A/PAN membrane exhibited the highest FRR of 96% and the lowest irreversible fouling of 4% with excellent durability of antifouling property during twenty repeated activated sludge filtrations.

The Human Pathogen Streptococcus pyogenes Releases Lipoproteins as Lipoprotein-rich Membrane Vesicles.

Science.gov (United States)

Biagini, Massimiliano; Garibaldi, Manuela; Aprea, Susanna; Pezzicoli, Alfredo; Doro, Francesco; Becherelli, Marco; Taddei, Anna Rita; Tani, Chiara; Tavarini, Simona; Mora, Marirosa; Teti, Giuseppe; D'Oro, Ugo; Nuti, Sandra; Soriani, Marco; Margarit, Immaculada; Rappuoli, Rino; Grandi, Guido; Norais, Nathalie

2015-08-01

Bacterial lipoproteins are attractive vaccine candidates because they represent a major class of cell surface-exposed proteins in many bacteria and are considered as potential pathogen-associated molecular patterns sensed by Toll-like receptors with built-in adjuvanticity. Although Gram-negative lipoproteins have been extensively characterized, little is known about Gram-positive lipoproteins. We isolated from Streptococcus pyogenes a large amount of lipoproteins organized in vesicles. These vesicles were obtained by weakening the bacterial cell wall with a sublethal concentration of penicillin. Lipid and proteomic analysis of the vesicles revealed that they were enriched in phosphatidylglycerol and almost exclusively composed of lipoproteins. In association with lipoproteins, a few hypothetical proteins, penicillin-binding proteins, and several members of the ExPortal, a membrane microdomain responsible for the maturation of secreted proteins, were identified. The typical lipidic moiety was apparently not necessary for lipoprotein insertion in the vesicle bilayer because they were also recovered from the isogenic diacylglyceryl transferase deletion mutant. The vesicles were not able to activate specific Toll-like receptor 2, indicating that lipoproteins organized in these vesicular structures do not act as pathogen-associated molecular patterns. In light of these findings, we propose to name these new structures Lipoprotein-rich Membrane Vesicles. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

A Two-Dimensional Lamellar Membrane: MXene Nanosheet Stacks.

Science.gov (United States)

Ding, Li; Wei, Yanying; Wang, Yanjie; Chen, Hongbin; Caro, Jürgen; Wang, Haihui

2017-02-06

Two-dimensional (2D) materials are promising candidates for advanced water purification membranes. A new kind of lamellar membrane is based on a stack of 2D MXene nanosheets. Starting from compact Ti 3 AlC 2 , delaminated nanosheets of the composition Ti 3 C 2 T x with the functional groups T (O, OH, and/or F) can be produced by etching and ultrasonication and stapled on a porous support by vacuum filtration. The MXene membrane supported on anodic aluminum oxide (AAO) substrate shows excellent water permeance (more than 1000 L m -2  h -1  bar -1 ) and favorable rejection rate (over 90 %) for molecules with sizes larger than 2.5 nm. The water permeance through the MXene membrane is much higher than that of the most membranes with similar rejections. Long-time operation also reveals the outstanding stability of the MXene membrane for water purification. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

Modes of Interaction of Pleckstrin Homology Domains with Membranes: Toward a Computational Biochemistry of Membrane Recognition.

Science.gov (United States)

Naughton, Fiona B; Kalli, Antreas C; Sansom, Mark S P

2018-02-02

Pleckstrin homology (PH) domains mediate protein-membrane interactions by binding to phosphatidylinositol phosphate (PIP) molecules. The structural and energetic basis of selective PH-PIP interactions is central to understanding many cellular processes, yet the molecular complexities of the PH-PIP interactions are largely unknown. Molecular dynamics simulations using a coarse-grained model enables estimation of free-energy landscapes for the interactions of 12 different PH domains with membranes containing PIP 2 or PIP 3 , allowing us to obtain a detailed molecular energetic understanding of the complexities of the interactions of the PH domains with PIP molecules in membranes. Distinct binding modes, corresponding to different distributions of cationic residues on the PH domain, were observed, involving PIP interactions at either the "canonical" (C) and/or "alternate" (A) sites. PH domains can be grouped by the relative strength of their C- and A-site interactions, revealing that a higher affinity correlates with increased C-site interactions. These simulations demonstrate that simultaneous binding of multiple PIP molecules by PH domains contributes to high-affinity membrane interactions, informing our understanding of membrane recognition by PH domains in vivo. Copyright © 2017. Published by Elsevier Ltd.

Fabrication of asymmetric alumina membranes

International Nuclear Information System (INIS)

Firouzghalb, H.; Falamaki, C.

2010-01-01

The effect of SrO addition on the thermal stabilization of transition aluminas with the aim of producing membrane layers (supported and unsupported) has been investigated. Al 2 O 3 -x wt.% SrO composite powders (x = 1, 3, 5, 8) were synthesized by co-precipitation of the hydroxides from solutions of AlCl 3 and Sr(NO 3 ) 2 salts using NH 4 OH as a precipitating agent. Optimum SrO dopant concentration regarding the transition aluminas stabilization effect was determined to be 5 wt.% based on XRD analysis. STA analysis showed a 30 deg. C shift versus higher temperatures in the transformation of final transitional alumina (θ-Al 2 O 3 ) to stable alpha phase due to addition of 5 wt.% SrO. The mechanism of transition aluminas thermal stabilization as a result of SrO addition is thoroughly discussed. Unsupported alumina membranes were prepared by drying boehmite sols at 600, 800, 1000 and 1100 deg. C. The effect of calcination temperature on surface area, pore size distribution of unsupported membranes containing 5 wt.% SrO has been investigated. The microstructure of unsupported and supported membranes revealed quite different. Smaller grains in the supported layers were attributed to the interaction between support and membrane.

Thermoelectric integrated membrane evaporation water recovery technology

Science.gov (United States)

Roebelen, G. J., Jr.; Winkler, H. E.; Dehner, G. F.

1982-01-01

The recently developed Thermoelectric Integrated Membrane Evaporation Subsystem (TIMES) offers a highly competitive approach to water recovery from waste fluids for future on-orbit stations such as the Space Operations Center. Low power, compactness and gravity insensitive operation are featured in this vacuum distillation subsystem that combines a hollow fiber membrane evaporator with a thermoelectric heat pump. The hollow fiber elements provide positive liquid/gas phase control with no moving parts other than pumps and an accumulator, thus solving problems inherent in other reclamation subsystem designs. In an extensive test program, over 850 hours of operation were accumulated during which time high quality product water was recovered from both urine and wash water at an average steady state production rate of 2.2 pounds per hour.

Effect of gamma irradiation on membranes of normal and pathological erythrocytes (beta-thalassemia)

International Nuclear Information System (INIS)

Sportelli, L.; Bonincontro, A.; Cametti, C.; Consiglio Nazionale delle Ricerche, Rome

1987-01-01

The influence of ionizing radiation on the membrane of human normal erythrocytes has extensively been studied and a variety of effects including changes in the cation fluxes or in non-electrolytes permeability, in membrane fluidity, in peroxidation of unsaturated lipids as well as chemical composition or structural modifications has been observed. However, only few studies deal with the effects of ionizing radiation on pathological red blood cells. In this work, we have investigated by means of electron spin resonance (ESR) spectroscopy the effects of 60 Co γ-radiation on the normal and homozygous β-thalassemic human erythrocyte membranes. (orig.)

Crystallization and preliminary X-ray analysis of membrane-bound pyrophosphatases.

Science.gov (United States)

Kellosalo, Juho; Kajander, Tommi; Honkanen, Riina; Goldman, Adrian

2013-02-01

Membrane-bound pyrophosphatases (M-PPases) are enzymes that enhance the survival of plants, protozoans and prokaryotes in energy constraining stress conditions. These proteins use pyrophosphate, a waste product of cellular metabolism, as an energy source for sodium or proton pumping. To study the structure and function of these enzymes we have crystallized two membrane-bound pyrophosphatases recombinantly produced in Saccharomyces cerevisae: the sodium pumping enzyme of Thermotoga maritima (TmPPase) and the proton pumping enzyme of Pyrobaculum aerophilum (PaPPase). Extensive crystal optimization has allowed us to grow crystals of TmPPase that diffract to a resolution of 2.6 Å. The decisive step in this optimization was in-column detergent exchange during the two-step purification procedure. Dodecyl maltoside was used for high temperature solubilization of TmPPase and then exchanged to a series of different detergents. After extensive screening, the new detergent, octyl glucose neopentyl glycol, was found to be the optimal for TmPPase but not PaPPase.

Discussion on cleaning and maintenance of YA system reverse osmosis membrane

International Nuclear Information System (INIS)

Zhu Yidong

2012-01-01

According to the overproof of pollution data of YA system reverse osmosis membrane in extension project, the daily maintenance company is using chemical cleaning on reverse osmosis unit to eliminate the pollution blindly, the fixed prescription, fixed dosage and high frequency of the chemical cleaning. The writer analyzed the cause of the membrane pollution and commended several chemical cleaning methods by the long-period study of the system, and also some suggestion, according to the status of operational site, for the daily maintenance. (author)

High temperature polymer electrolyte membrane fuel cells: Approaches, status, and perspectives

DEFF Research Database (Denmark)

This book is a comprehensive review of high-temperature polymer electrolyte membrane fuel cells (PEMFCs). PEMFCs are the preferred fuel cells for a variety of applications such as automobiles, cogeneration of heat and power units, emergency power and portable electronics. The first 5 chapters...... of and motivated extensive research activity in the field. The last 11 chapters summarize the state-of-the-art of technological development of high temperature-PEMFCs based on acid doped PBI membranes including catalysts, electrodes, MEAs, bipolar plates, modelling, stacking, diagnostics and applications....

Membrane tension regulates clathrin-coated pit dynamics

Science.gov (United States)

Liu, Allen

2014-03-01

Intracellular organization depends on close communication between the extracellular environment and a network of cytoskeleton filaments. The interactions between cytoskeletal filaments and the plasma membrane lead to changes in membrane tension that in turns help regulate biological processes. Endocytosis is thought to be stimulated by low membrane tension and the removal of membrane increases membrane tension. While it is appreciated that the opposing effects of exocytosis and endocytosis have on keeping plasma membrane tension to a set point, it is not clear how membrane tension affects the dynamics of clathrin-coated pits (CCPs), the individual functional units of clathrin-mediated endocytosis. Furthermore, although it was recently shown that actin dynamics counteracts membrane tension during CCP formation, it is not clear what roles plasma membrane tension plays during CCP initiation. Based on the notion that plasma membrane tension is increased when the membrane area increases during cell spreading, we designed micro-patterned surfaces of different sizes to control the cell spreading sizes. Total internal reflection fluorescence microscopy of living cells and high content image analysis were used to quantify the dynamics of CCPs. We found that there is an increased proportion of CCPs with short (<20s) lifetime for cells on larger patterns. Interestingly, cells on larger patterns have higher CCP initiation density, an effect unexpected based on the conventional view of decreasing endocytosis with increasing membrane tension. Furthermore, by analyzing the intensity profiles of CCPs that were longer-lived, we found CCP intensity decreases with increasing cell size, indicating that the CCPs are smaller with increasing membrane tension. Finally, disruption of actin dynamics significantly increased the number of short-lived CCPs, but also decreased CCP initiation rate. Together, our study reveals new mechanistic insights into how plasma membrane tension regulates

Polymethylmethacrylate/Polyacrylonitrile Membranes via Centrifugal Spinning as Separator in Li-Ion Batteries

Directory of Open Access Journals (Sweden)

Meltem Yanilmaz

2015-04-01

Full Text Available Electrospun nanofiber membranes have been extensively studied as separators in Li-ion batteries due to their large porosity, unique pore structure, and high electrolyte uptake. However, the electrospinning process has some serious drawbacks, such as low spinning rate and high production cost. The centrifugal spinning technique can be used as a fast, cost-effective and safe technique to fabricate high-performance fiber-based separators. In this work, polymethylmethacrylate (PMMA/polyacrylonitrile (PAN membranes with different blend ratios were produced via centrifugal spinning and characterized by using different electrochemical techniques for use as separators in Li-ion batteries. Compared with commercial microporous polyolefin membrane, centrifugally-spun PMMA/PAN membranes had larger ionic conductivity, higher electrochemical oxidation limit, and lower interfacial resistance with lithium. Centrifugally-spun PMMA/PAN membrane separators were assembled into Li/LiFePO4 cells and these cells delivered high capacities and exhibited good cycling performance at room temperature. In addition, cells using centrifugally-spun PMMA/PAN membrane separators showed superior C-rate performance compared to those using microporous polypropylene (PP membranes. It is, therefore, demonstrated that centrifugally-spun PMMA/PAN membranes are promising separator candidate for high-performance Li-ion batteries.

FRET imaging in living maize cells reveals that plasma membrane aquaporins interact to regulate their subcellular localization

NARCIS (Netherlands)

Zelazny, E.; Borst, J.W.; Muylaert, M.; Batoko, H.; Hemminga, M.A.; Chaumont, F.

2007-01-01

Zea mays plasma membrane intrinsic proteins (ZmPIPs) fall into two groups, ZmPIP1s and ZmPIP2s, that exhibit different water channel activities when expressed in Xenopus oocytes. ZmPIP1s are inactive, whereas ZmPIP2s induce a marked increase in the membrane osmotic water permeability coefficient,

Microfluidic devices for investigation of biomimetic membranes for sensor and separation applications

DEFF Research Database (Denmark)

Pszon-Bartosz, Kamila Justyna

to microfluidic designs involving protein delivery to biomimetic membranes developed for sensor and separation applications. Finally, an OMP functionality modulation with β-cyclodextrin (β-CD) was shown and revealed the protein potential application as a sensor. Moreover, the β-CD blocker may be used to prevent...... for industrial applications. Among them are the inherent fragility of lipid membranes, the challenge of up-scaling the effective membrane area and the quantification of the protein delivery to the lipid membrane which may determined the biomimetic membrane application. This PhD thesis addresses the above...

Plants and fungi in the era of heterogeneous plasma membranes.

Science.gov (United States)

Opekarová, M; Malinsky, J; Tanner, W

2010-09-01

Examples from yeast and plant cells are described that show that their plasma membrane is laterally compartmented. Distinct lateral domains encompassing both specific lipids and integral proteins coexist within the plane of the plasma membrane. The compartments are either spatially stable and include distinct sets of proteins, or they are transiently formed to accomplish diverse functions. They are not related to lipid rafts or their clusters, as defined for mammalian cells. This review summarises only well-documented compartments of plasma membranes from plants and fungi, which have been recognised using microscopic approaches. In several cases, physiological functions of the membrane compartmentation are revealed.

Comparative proteomic analysis reveals a dynamic pollen plasma membrane protein map and the membrane landscape of receptor-like kinases and transporters important for pollen tube growth and interaction with pistils in rice.

Science.gov (United States)

Yang, Ning; Wang, Tai

2017-01-05

The coordination of pollen tube (PT) growth, guidance and timely growth arrest and rupture mediated by PT-pistil interaction is crucial for the PT to transport sperm cells into ovules for double fertilization. The plasma membrane (PM) represents an important interface for cell-cell interaction, and PM proteins of PTs are pioneers for mediating PT integrity and interaction with pistils. Thus, understanding the mechanisms underlying these events is important for proteomics. Using the efficient aqueous polymer two-phase system and alkali buffer treatment, we prepared high-purity PM from mature and germinated pollen of rice. We used iTRAQ quantitative proteomic methods and identified 1,121 PM-related proteins (PMrPs) (matched to 899 loci); 192 showed differential expression in the two pollen cell types, 119 increased and 73 decreased in abundance during germination. The PMrP and differentially expressed PMrP sets all showed a functional skew toward signal transduction, transporters, wall remodeling/metabolism and membrane trafficking. Their genomic loci had strong chromosome bias. We found 37 receptor-like kinases (RLKs) from 8 kinase subfamilies and 209 transporters involved in flux of diversified ions and metabolites. In combination with the rice pollen transcriptome data, we revealed that in general, the protein expression of these PMrPs disagreed with their mRNA expression, with inconsistent mRNA expression for 74% of differentially expressed PMrPs. This study identified genome-wide pollen PMrPs, and provided insights into the membrane profile of receptor-like kinases and transporters important for pollen tube growth and interaction with pistils. These pollen PMrPs and their mRNAs showed discordant expression. This work provides resource and knowledge to further dissect mechanisms by which pollen or the PT controls PMrP abundance and monitors interactions and ion and metabolite exchanges with female cells in rice.

Extensive Turnover of Compatible Solutes in Cyanobacteria Revealed by Deuterium Oxide (D_2O) Stable Isotope Probing

International Nuclear Information System (INIS)

Baran, Richard; Lau, Rebecca; Bowen, Benjamin P.; Diamond, Spencer; Jose, Nick

2017-01-01

In diverse environments on a global scale cyanobacteria are important primary producers of organic matter. Moreover, while mechanisms of CO_2 fixation are well understood, the distribution of the flow of fixed organic carbon within individual cells and complex microbial communities is less well characterized. To obtain a general overview of metabolism, we describe the use of deuterium oxide (D_2O) to measure deuterium incorporation into the intracellular metabolites of two physiologically diverse cyanobacteria: a terrestrial filamentous strain (Microcoleus vaginatus PCC 9802) and a euryhaline unicellular strain (Synechococcus sp. PCC 7002). D_2O was added to the growth medium during different phases of the diel cycle. Incorporation of deuterium into metabolites at nonlabile positions, an indicator of metabolite turnover, was assessed using liquid chromatography mass spectrometry. Expectedly, large differences in turnover among metabolites were observed. Some metabolites, such as fatty acids, did not show significant turnover over 12–24 h time periods but did turn over during longer time periods. Unexpectedly, metabolites commonly regarded to act as compatible solutes, including glutamate, glucosylglycerol, and a dihexose, showed extensive turnover compared to most other metabolites already after 12 h, but only during the light phase in the cycle. We observed extensive turnover and found it surprising considering the conventional view on compatible solutes as biosynthetic end points given the relatively slow growth and constant osmotic conditions. Our suggests the possibility of a metabolic sink for some compatible solutes (e.g., into glycogen) that allows for rapid modulation of intracellular osmolarity. To investigate this, uniformly "1"3C-labeled Synechococcus sp. PCC 7002 were exposed to "1"2C glucosylglycerol. Following metabolite extraction, amylase treatment of methanol-insoluble polymers revealed "1"2C labeling of glycogen. Overall, our work shows that D_2

Towards structural and functional analysis of the plant plasma membrane proton pump

DEFF Research Database (Denmark)

Justesen, Bo Højen

The plasma membrane H+-ATPase is a proton pump essential for several physiological important processes in plants. Through the extrusion of protons from the cell, the PM H+-ATPase establishes and maintains a proton gradient used by proton coupled transporters and secondary active transport...... of nutrients and metabolites across the plasma membrane. Additional processes involving the PM H+-ATPase includes plant growth, development, and response to biotic and abiotic stresses. Extensive efforts have been made in attempts to elucidate the detailed physiological role and biochemical characteristics...... of plasma membrane H+-ATPases. Studies on the plasma membrane H+-ATPases have involved both in vivo and in vitro approaches, with the latter employing either solubilisation by detergent micelles, or reconstitution into lipid vesicles. Despite resulting in a large body of information on structure, function...

Elasto-plasticity in wrinkled polymerized lipid membranes

KAUST Repository

Chaieb, Sahraoui

2014-01-15

Biomembranes shown to behave like elastic sheets, can also suffer plastic deformations. Neutron scattering experiments on partially polymerised wrinkled membranes revealed that when a critical degree of polymerisation is crossed, the wrinkled membranes do not resume their spherical shapes. Instead they remain wrinkled and rigid while their non-polymerised counterparts resume their spherical floppy shapes. The yield stress of these membranes, measured for the first time via the fractal dimension, is intimately related to the degree of polymerisation probably through a 2D disorder that quenches the lateral diffusion of the lipid molecules. This work might shed light on the physical reason behind the irreversible deformation of echinocytes, acanthocytes and malaria infected red blood cells.

Elasto-plasticity in wrinkled polymerized lipid membranes

KAUST Repository

Chaieb, Saharoui

2014-01-01

Biomembranes shown to behave like elastic sheets, can also suffer plastic deformations. Neutron scattering experiments on partially polymerised wrinkled membranes revealed that when a critical degree of polymerisation is crossed, the wrinkled membranes do not resume their spherical shapes. Instead they remain wrinkled and rigid while their non-polymerised counterparts resume their spherical floppy shapes. The yield stress of these membranes, measured for the first time via the fractal dimension, is intimately related to the degree of polymerisation probably through a 2D disorder that quenches the lateral diffusion of the lipid molecules. This work might shed light on the physical reason behind the irreversible deformation of echinocytes, acanthocytes and malaria infected red blood cells.

2,2'-Bis(monoacylglycero) PO4 (BMP), but Not 3,1'-BMP, increases membrane curvature stress to enhance α-tocopherol transfer protein binding to membranes.

Science.gov (United States)

Baptist, Matilda; Panagabko, Candace; Nickels, Jonathan D; Katsaras, John; Atkinson, Jeffrey

2015-03-01

Previous work revealed that α-tocopherol transfer protein (α-TTP) co-localizes with bis(monoacylglycero)phosphate (BMP) in late endosomes. BMP is a lipid unique to late endosomes and is believed to induce membrane curvature and support the multivesicular nature of this organelle. We examined the effect of BMP on α-TTP binding to membranes using dual polarization interferometry and vesicle-binding assay. α-TTP binding to membranes is increased by the curvature-inducing lipid BMP. α-TTP binds to membranes with greater affinity when they contain the 2,2'-BMP versus 3,1'-BMP isomers.

N-3 fatty acids and membrane microdomains: from model membranes to lymphocyte function.

Science.gov (United States)

Shaikh, Saame Raza; Teague, Heather

2012-12-01

This article summarizes the author's research on fish oil derived n-3 fatty acids, plasma membrane organization and B cell function. We first cover basic model membrane studies that investigated how docosahexaenoic acid (DHA) targeted the organization of sphingolipid-cholesterol enriched lipid microdomains. A key finding here was that DHA had a relatively poor affinity for cholesterol. This work led to a model that predicted DHA acyl chains in cells would manipulate lipid-protein microdomain organization and thereby function. We then review how the predictions of the model were tested with B cells in vitro followed by experiments using mice fed fish oil. These studies reveal a highly complex picture on how n-3 fatty acids target lipid-protein organization and B cell function. Key findings are as follows: (1) n-3 fatty acids target not just the plasma membrane but also endomembrane organization; (2) DHA, but not eicosapentaenoic acid (EPA), disrupts microdomain spatial distribution (i.e. clustering), (3) DHA alters protein lateral organization and (4) changes in membrane organization are accompanied by functional effects on both innate and adaptive B cell function. Altogether, the research over the past 10 years has led to an evolution of the original model on how DHA reorganizes membrane microdomains. The work raises the intriguing possibility of testing the model at the human level to target health and disease. Copyright © 2012 Elsevier Ltd. All rights reserved.

Lipid engineering reveals regulatory roles for membrane fluidity in yeast flocculation and oxygen-limited growth

Energy Technology Data Exchange (ETDEWEB)

Degreif, Daniel [Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Technical Univ. of Darmstadt (Germany); de Rond, Tristan [Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Univ. of California, Berkeley, CA (United States); Bertl, Adam [Technical Univ. of Darmstadt (Germany); Keasling, Jay D. [Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Univ. of California, Berkeley, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Technical Univ. of Denmark, Lyngby (Denmark); Budin, Itay [Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Univ. of California, Berkeley, CA (United States)

2017-03-18

Cells modulate lipid metabolism in order to maintain membrane homeostasis. In this paper, we use a metabolic engineering approach to manipulate the stoichiometry of fatty acid unsaturation, a regulator of cell membrane fluidity, in Saccharomyces cerevisiae. Unexpectedly, reduced lipid unsaturation triggered cell-cell adhesion (flocculation), a phenomenon characteristic of industrial yeast but uncommon in laboratory strains. We find that ER lipid saturation sensors induce expression of FLO1 – encoding a cell wall polysaccharide binding protein – independently of its canonical regulator. In wild-type cells, Flo1p-dependent flocculation occurs under oxygen-limited growth, which reduces unsaturated lipid synthesis and thus serves as the environmental trigger for flocculation. Transcriptional analysis shows that FLO1 is one of the most highly induced genes in response to changes in lipid unsaturation, and that the set of membrane fluidity-sensitive genes is globally activated as part of the cell's long-term response to hypoxia during fermentation. Finally, our results show how the lipid homeostasis machinery of budding yeast is adapted to carry out a broad response to an environmental stimulus important in biotechnology.

Composite plasma polymerized sulfonated polystyrene membrane for PEMFC

Energy Technology Data Exchange (ETDEWEB)

Nath, Bhabesh Kumar; Khan, Aziz; Chutia, Joyanti, E-mail: jchutiaiasst@gmail.com

2015-10-15

Highlights: • Methyl methane sulfonate (MMS) is used as the sulfonating agent. • The proton conductivity of the membrane is found to be 0.141 S cm{sup −1}. • Power density of fuel cell with styrene/MMS membrane is 0.5 W cm{sup −2}. • The membrane exhibits thermal stability up to 140 °C. - Abstract: This work presents the introduction of an organic compound methyl methane sulfonate (MMS) for the first time in fabrication of polystyrene based proton exchange membrane (PEM) by plasma polymerization process. The membrane is fabricated by co-polymerizing styrene and MMS in capacitively coupled continuous RF plasma. The chemical composition of the plasma polymerized polymer membrane is investigated using Fourier Transform Infrared Spectroscopy which reveals the formation of composite structure of styrene and MMS. The surface morphology studied using AFM and SEM depicts the effect of higher partial pressure of MMS on surface topography of the membrane. The proton transport property of the membrane studied using electrochemical impedance spectroscopy shows the achievement of maximum proton conductivity of 0.141 S cm{sup −1} which is comparable to Nafion 117 membrane. Fuel cell performance test of the synthesized membrane shows a maximum power density of 500 mW cm{sup −2} and current density of 0.62 A cm{sup −2} at 0.6 V.

Research into topology optimization and the FDM method for a space cracked membrane

Science.gov (United States)

Hu, Qingxi; Li, Wanyuan; Zhang, Haiguang; Liu, Dali; Peng, Fujun; Duan, Yongchao

2017-07-01

The problem that the space membranes are easily torn open is the main focus in this paper, and a bionic strengthening-ribs structure is proposed for a space membrane based on interdisciplinary strengths, such as topology optimization, composite materials, and rapid prototyping. The optimization method and modeling method of membranes with bionic strengthening-ribs was studied. The PEEK and SCF/PEEK composite material which are applied to the space environment are chosen, and FDM technology is used. Through topology optimization, bionic strengthening-ribs with good tensile and tear capacities were obtained. Cracked membranes, cracked membranes with PEEK strengthening-ribs and SCF/PEEK strengthening-ribs were tested and test data were obtained. An extension situation and tension fracture were compared for three cases. The experimental results showed that membranes with the bionic strengthening-ribs structure have better mechanical properties, and the strength of the membranes with PEEK and SCF/PEEK strengthening-ribs were raised, respectively, up to 266.9% and 185.9%. The strengthening-ribs structure greatly improves the capacity to halt membrane crack-growth, which has an important significance to avoid membrane tear, and to ensure the spacecraft orbital lifetime.

Surface interactions and fouling properties of Micrococcus luteus with microfiltration membranes.

Science.gov (United States)

Feng, Lei; Li, Xiufen; Song, Ping; Du, Guocheng; Chen, Jian

2011-11-01

This study was conducted to investigate microbial adhesion of Micrococcus luteus to polypropylene (PP) and polyvinylidene fluoride (PVDF) membranes in relation to the variation of the interfacial energies in the membrane-bacteria systems, for revealing effects of short-range surface interactions on filtration behavior. Both the membranes and M. luteus showed typical strong electron donors and hydrophilic properties. The AB component was dominant in the interfacial energies of the two membrane-bacteria systems. M. luteus presented larger negative U(mlb)(XDLVO) to the PP membrane than to the PVDF membrane. The adhesion experiments also proved that M. luteus had higher adhesion percentage to the PP membrane. This study demonstrated that the adhesion potentials of M. luteus to the PP and PVDF membranes might be explained in terms of bacterium, membrane, and intervening medium surface properties, which are mainly determined by the interfacial energies in the systems according to the XDLVO theory.

Reduced evolutionary rates in HIV-1 reveal extensive latency periods among replicating lineages.

Science.gov (United States)

Immonen, Taina T; Leitner, Thomas

2014-10-16

HIV-1 can persist for the duration of a patient's life due in part to its ability to hide from the immune system, and from antiretroviral drugs, in long-lived latent reservoirs. Latent forms of HIV-1 may also be disproportionally involved in transmission. Thus, it is important to detect and quantify latency in the HIV-1 life cycle. We developed a novel molecular clock-based phylogenetic tool to investigate the prevalence of HIV-1 lineages that have experienced latency. The method removes alternative sources that may affect evolutionary rates, such as hypermutation, recombination, and selection, to reveal the contribution of generation-time effects caused by latency. Our method was able to recover latent lineages with high specificity and sensitivity, and low false discovery rates, even on relatively short branches on simulated phylogenies. Applying the tool to HIV-1 sequences from 26 patients, we show that the majority of phylogenetic lineages have been affected by generation-time effects in every patient type, whether untreated, elite controller, or under effective or failing treatment. Furthermore, we discovered extensive effects of latency in sequence data (gag, pol, and env) from reservoirs as well as in the replicating plasma population. To better understand our phylogenetic findings, we developed a dynamic model of virus-host interactions to investigate the proportion of lineages in the actively replicating population that have ever been latent. Assuming neutral evolution, our dynamic modeling showed that under most parameter conditions, it is possible for a few activated latent viruses to propagate so that in time, most HIV-1 lineages will have been latent at some time in their past. These results suggest that cycling in and out of latency plays a major role in the evolution of HIV-1. Thus, no aspect of HIV-1 evolution can be fully understood without considering latency - including treatment, drug resistance, immune evasion, transmission, and pathogenesis.

Mechanics of membrane-cytoskeleton attachment in Paramecium

Science.gov (United States)

Campillo, C.; Jerber, J.; Fisch, C.; Simoes-Betbeder, M.; Dupuis-Williams, P.; Nassoy, P.; Sykes, C.

2012-12-01

In this paper we assess the role of the protein MKS1 (Meckel syndrome type 1) in the cortical membrane mechanics of the ciliated protist Paramecium. This protein is known to be crucial in the process of cilium formation, and we investigate its putative role in membrane-cytoskeleton attachment. Therefore, we compare cells where the gene coding for MKS1 is silenced to wild-type cells. We found that scanning electron microscopy observation of the cell surface reveals a cup-like structure in wild-type cells that is lost in silenced cells. Since this structure is based on the underlying cytoskeleton, one hypothesis to explain this observation is a disruption of membrane attachment to the cytoskeleton in the absence of MKS1 that should affect plasma membrane mechanics. We test this by probing the mechanics of wild-type and silenced cells by micropipette aspiration. Strikingly, we observe that, at the same aspiration pressure, the membrane of silenced cells is easily aspirated by the micropipette whereas that of wild-type cells enters only at a moderate velocity, an effect that suggests a detachment of the membrane from the underlying cytoskeleton in silenced cells. We quantify this detachment by measuring the deformation of the cell cortex and the rate of cell membrane entry in the micropipette. This study offers a new perspective for the characterization of membrane-cytoskeleton attachment in protists and paves the way for a better understanding of the role of membrane-cortex attachment in cilium formation.

Fouling distribution in forward osmosis membrane process.

Science.gov (United States)

Lee, Junseok; Kim, Bongchul; Hong, Seungkwan

2014-06-01

Fouling behavior along the length of membrane module was systematically investigated by performing simple modeling and lab-scale experiments of forward osmosis (FO) membrane process. The flux distribution model developed in this study showed a good agreement with experimental results, validating the robustness of the model. This model demonstrated, as expected, that the permeate flux decreased along the membrane channel due to decreasing osmotic pressure differential across the FO membrane. A series of fouling experiments were conducted under the draw and feed solutions at various recoveries simulated by the model. The simulated fouling experiments revealed that higher organic (alginate) fouling and thus more flux decline were observed at the last section of a membrane channel, as foulants in feed solution became more concentrated. Furthermore, the water flux in FO process declined more severely as the recovery increased due to more foulants transported to membrane surface with elevated solute concentrations at higher recovery, which created favorable solution environments for organic adsorption. The fouling reversibility also decreased at the last section of the membrane channel, suggesting that fouling distribution on FO membrane along the module should be carefully examined to improve overall cleaning efficiency. Lastly, it was found that such fouling distribution observed with co-current flow operation became less pronounced in counter-current flow operation of FO membrane process. Copyright © 2014 The Research Centre for Eco-Environmental Sciences, Chinese Academy of Sciences. Published by Elsevier B.V. All rights reserved.

A Shotgun Proteomic Approach Reveals That Fe Deficiency Causes Marked Changes in the Protein Profiles of Plasma Membrane and Detergent-Resistant Microdomain Preparations from Beta vulgaris Roots.

Science.gov (United States)

Gutierrez-Carbonell, Elain; Takahashi, Daisuke; Lüthje, Sabine; González-Reyes, José Antonio; Mongrand, Sébastien; Contreras-Moreira, Bruno; Abadía, Anunciación; Uemura, Matsuo; Abadía, Javier; López-Millán, Ana Flor

2016-08-05

In the present study we have used label-free shotgun proteomic analysis to examine the effects of Fe deficiency on the protein profiles of highly pure sugar beet root plasma membrane (PM) preparations and detergent-resistant membranes (DRMs), the latter as an approach to study microdomains. Altogether, 545 proteins were detected, with 52 and 68 of them changing significantly with Fe deficiency in PM and DRM, respectively. Functional categorization of these proteins showed that signaling and general and vesicle-related transport accounted for approximately 50% of the differences in both PM and DRM, indicating that from a qualitative point of view changes induced by Fe deficiency are similar in both preparations. Results indicate that Fe deficiency has an impact in phosphorylation processes at the PM level and highlight the involvement of signaling proteins, especially those from the 14-3-3 family. Lipid profiling revealed Fe-deficiency-induced decreases in phosphatidic acid derivatives, which may impair vesicle formation, in agreement with the decreases measured in proteins related to intracellular trafficking and secretion. The modifications induced by Fe deficiency in the relative enrichment of proteins in DRMs revealed the existence of a group of cytoplasmic proteins that appears to be more attached to the PM in conditions of Fe deficiency.

Quantitative Proteomics Reveals Distinct Differences in the Protein Content of Outer Membrane Vesicle Vaccines

NARCIS (Netherlands)

Waterbeemd, van de B.; Mommen, G.P.M.; Pennings, J.L.A.; Eppink, M.H.M.; Wijffels, R.H.; Pol, van der L.A.; Jong, de A.P.J.M.

2013-01-01

At present, only vaccines containing outer membrane vesicles (OMV) have successfully stopped Neisseria meningitidis serogroup B epidemics. These vaccines however require detergent-extraction to remove endotoxin, which changes immunogenicity and causes production difficulties. To investigate this in

Integrating complex functions: coordination of nuclear pore complex assembly and membrane expansion of the nuclear envelope requires a family of integral membrane proteins.

Science.gov (United States)

Schneiter, Roger; Cole, Charles N

2010-01-01

The nuclear envelope harbors numerous large proteinaceous channels, the nuclear pore complexes (NPCs), through which macromolecular exchange between the cytosol and the nucleoplasm occurs. This double-membrane nuclear envelope is continuous with the endoplasmic reticulum and thus functionally connected to such diverse processes as vesicular transport, protein maturation and lipid synthesis. Recent results obtained from studies in Saccharomyces cerevisiae indicate that assembly of the nuclear pore complex is functionally dependent upon maintenance of lipid homeostasis of the ER membrane. Previous work from one of our laboratories has revealed that an integral membrane protein Apq12 is important for the assembly of functional nuclear pores. Cells lacking APQ12 are viable but cannot grow at low temperatures, have aberrant NPCs and a defect in mRNA export. Remarkably, these defects in NPC assembly can be overcome by supplementing cells with a membrane fluidizing agent, benzyl alcohol, suggesting that Apq12 impacts the flexibility of the nuclear membrane, possibly by adjusting its lipid composition when cells are shifted to a reduced temperature. Our new study now expands these findings and reveals that an essential membrane protein, Brr6, shares at least partially overlapping functions with Apq12 and is also required for assembly of functional NPCs. A third nuclear envelope membrane protein, Brl1, is related to Brr6, and is also required for NPC assembly. Because maintenance of membrane homeostasis is essential for cellular survival, the fact that these three proteins are conserved in fungi that undergo closed mitoses, but are not found in metazoans or plants, may indicate that their functions are performed by proteins unrelated at the primary sequence level to Brr6, Brl1 and Apq12 in cells that disassemble their nuclear envelopes during mitosis.

A balance between membrane elasticity and polymerization energy sets the shape of spherical clathrin coats

Science.gov (United States)

Saleem, Mohammed; Morlot, Sandrine; Hohendahl, Annika; Manzi, John; Lenz, Martin; Roux, Aurélien

2015-02-01

In endocytosis, scaffolding is one of the mechanisms to create membrane curvature by moulding the membrane into the spherical shape of the clathrin cage. However, the impact of membrane elastic parameters on the assembly and shape of clathrin lattices has never been experimentally evaluated. Here, we show that membrane tension opposes clathrin polymerization. We reconstitute clathrin budding in vitro with giant unilamellar vesicles (GUVs), purified adaptors and clathrin. By changing the osmotic conditions, we find that clathrin coats cause extensive budding of GUVs under low membrane tension while polymerizing into shallow pits under moderate tension. High tension fully inhibits polymerization. Theoretically, we predict the tension values for which transitions between different clathrin coat shapes occur. We measure the changes in membrane tension during clathrin polymerization, and use our theoretical framework to estimate the polymerization energy from these data. Our results show that membrane tension controls clathrin-mediated budding by varying the membrane budding energy.

Refractive-index-based screening of membrane-protein-mediated transfer across biological membranes.

Science.gov (United States)

Brändén, Magnus; Tabaei, Seyed R; Fischer, Gerhard; Neutze, Richard; Höök, Fredrik

2010-07-07

Numerous membrane-transport proteins are major drug targets, and therefore a key ingredient in pharmaceutical development is the availability of reliable, efficient tools for membrane transport characterization and inhibition. Here, we present the use of evanescent-wave sensing for screening of membrane-protein-mediated transport across lipid bilayer membranes. This method is based on a direct recording of the temporal variations in the refractive index that occur upon a transfer-dependent change in the solute concentration inside liposomes associated to a surface plasmon resonance (SPR) active sensor surface. The applicability of the method is demonstrated by a functional study of the aquaglyceroporin PfAQP from the malaria parasite Plasmodium falciparum. Assays of the temperature dependence of facilitated diffusion of sugar alcohols on a single set of PfAQP-reconstituted liposomes reveal that the activation energies for facilitated diffusion of xylitol and sorbitol are the same as that previously measured for glycerol transport in the aquaglyceroporin of Escherichia coli (5 kcal/mole). These findings indicate that the aquaglyceroporin selectivity filter does not discriminate sugar alcohols based on their length, and that the extra energy cost of dehydration of larger sugar alcohols, upon entering the pore, is compensated for by additional hydrogen-bond interactions within the aquaglyceroporin pore. Copyright 2010 Biophysical Society. Published by Elsevier Inc. All rights reserved.

Comparative Genomics Analyses Reveal Extensive Chromosome Colinearity and Novel Quantitative Trait Loci in Eucalyptus

Science.gov (United States)

Weng, Qijie; Li, Mei; Yu, Xiaoli; Guo, Yong; Wang, Yu; Zhang, Xiaohong; Gan, Siming

2015-01-01

Dense genetic maps, along with quantitative trait loci (QTLs) detected on such maps, are powerful tools for genomics and molecular breeding studies. In the important woody genus Eucalyptus, the recent release of E. grandis genome sequence allows for sequence-based genomic comparison and searching for positional candidate genes within QTL regions. Here, dense genetic maps were constructed for E. urophylla and E. tereticornis using genomic simple sequence repeats (SSR), expressed sequence tag (EST) derived SSR, EST-derived cleaved amplified polymorphic sequence (EST-CAPS), and diversity arrays technology (DArT) markers. The E. urophylla and E. tereticornis maps comprised 700 and 585 markers across 11 linkage groups, totaling at 1,208.2 and 1,241.4 cM in length, respectively. Extensive synteny and colinearity were observed as compared to three earlier DArT-based eucalypt maps (two maps with E. grandis × E. urophylla and one map of E. globulus) and with the E. grandis genome sequence. Fifty-three QTLs for growth (10–56 months of age) and wood density (56 months) were identified in 22 discrete regions on both maps, in which only one colocalizaiton was found between growth and wood density. Novel QTLs were revealed as compared with those previously detected on DArT-based maps for similar ages in Eucalyptus. Eleven to 585 positional candidate genes were obained for a 56-month-old QTL through aligning QTL confidence interval with the E. grandis genome. These results will assist in comparative genomics studies, targeted gene characterization, and marker-assisted selection in Eucalyptus and the related taxa. PMID:26695430

Comparative Genomics Analyses Reveal Extensive Chromosome Colinearity and Novel Quantitative Trait Loci in Eucalyptus.

Directory of Open Access Journals (Sweden)

Fagen Li

Full Text Available Dense genetic maps, along with quantitative trait loci (QTLs detected on such maps, are powerful tools for genomics and molecular breeding studies. In the important woody genus Eucalyptus, the recent release of E. grandis genome sequence allows for sequence-based genomic comparison and searching for positional candidate genes within QTL regions. Here, dense genetic maps were constructed for E. urophylla and E. tereticornis using genomic simple sequence repeats (SSR, expressed sequence tag (EST derived SSR, EST-derived cleaved amplified polymorphic sequence (EST-CAPS, and diversity arrays technology (DArT markers. The E. urophylla and E. tereticornis maps comprised 700 and 585 markers across 11 linkage groups, totaling at 1,208.2 and 1,241.4 cM in length, respectively. Extensive synteny and colinearity were observed as compared to three earlier DArT-based eucalypt maps (two maps with E. grandis × E. urophylla and one map of E. globulus and with the E. grandis genome sequence. Fifty-three QTLs for growth (10-56 months of age and wood density (56 months were identified in 22 discrete regions on both maps, in which only one colocalizaiton was found between growth and wood density. Novel QTLs were revealed as compared with those previously detected on DArT-based maps for similar ages in Eucalyptus. Eleven to 585 positional candidate genes were obained for a 56-month-old QTL through aligning QTL confidence interval with the E. grandis genome. These results will assist in comparative genomics studies, targeted gene characterization, and marker-assisted selection in Eucalyptus and the related taxa.

A Pilot-Scale System for Carbon Molecular Sieve Hollow Fiber Membrane Manufacturing

KAUST Repository

Karvan, O.; Johnson, J. R.; Williams, P. J.; Koros, W. J.

2012-01-01

research on these materials with a variety of applications being studied. The results from a pilot-scale CMS production system are presented. This system was designed based on extensive laboratory research, and hollow fiber membranes produced in this system

Membrane Curvature Induced by Aggregates of LH2s and Monomeric LH1s

OpenAIRE

Chandler, Danielle E.; Gumbart, James; Stack, John D.; Chipot, Christophe; Schulten, Klaus

2009-01-01

The photosynthetic apparatus of purple bacteria is contained within organelles called chromatophores, which form as extensions of the cytoplasmic membrane. The shape of these chromatophores can be spherical (as in Rhodobacter sphaeroides), lamellar (as in Rhodopseudomonas acidophila and Phaeospirillum molischianum), or tubular (as in certain Rb. sphaeroides mutants). Chromatophore shape is thought to be influenced by the integral membrane proteins Light Harvesting Complexes I and II (LH1 and ...

A 39-kD plasma membrane protein (IP39) is an anchor for the unusual membrane skeleton of Euglena gracilis

International Nuclear Information System (INIS)

Rosiere, T.K.; Marrs, J.A.; Bouck, G.B.

1990-01-01

The major integral plasma membrane protein (IP39) of Euglena gracilis was radiolabeled, peptide mapped, and dissected with proteases to identify cytoplasmic domains that bind and anchor proteins of the cell surface. When plasma membranes were radioiodinated and extracted with octyl glucoside, 98% of the extracted label was found in IP39 or the 68- and 110-kD oligomers of IP39. The octyl glucoside extracts were incubated with unlabeled cell surface proteins immobilized on nitrocellulose (overlays). Radiolabel from the membrane extract bound one (80 kD) of the two (80 and 86 kD) major membrane skeletal protein bands. Resolubilization of the bound label yielded a radiolabeled polypeptide identical in Mr to IP39. Intact plasma membranes were also digested with papain before or after radioiodination, thereby producing a cytoplasmically truncated IP39. The octyl glucoside extract of truncated IP39 no longer bound to the 80-kD membrane skeletal protein in the nitrocellulose overlays. EM of intact or trypsin digested plasma membranes incubated with membrane skeletal proteins under stringent conditions similar to those used in the nitrocellulose overlays revealed a partially reformed membrane skeletal layer. Little evidence of a membrane skeletal layer was found, however, when plasma membranes were predigested with papain before reassociation. A candidate 80-kD binding domain of IP39 has been tentatively identified as a peptide fragment that was present after trypsin digestion of plasma membranes, but was absent after papain digestion in two-dimensional peptide maps of IP39. Together, these data suggest that the unique peripheral membrane skeleton of Euglena binds to the plasma membrane through noncovalent interactions between the major 80-kD membrane skeletal protein and a small, papain sensitive cytoplasmic domain of IP39

Electrochemical acidification of Kraft black liquor by electrodialysis with bipolar membrane: Ion exchange membrane fouling identification and mechanisms.

Science.gov (United States)

Haddad, Maryam; Mikhaylin, Sergey; Bazinet, Laurent; Savadogo, Oumarou; Paris, Jean

2017-02-15

Integrated forest biorefinery offers promising pathways to sustainably diversify the revenue of pulp and paper industry. In this context, lignin can be extracted from a residual stream of Kraft pulping process, called black liquor, and subsequently converted into a wide spectrum of bio-based products. Electrochemical acidification of Kraft black liquor by electrodialysis with bipolar membrane results in lignin extraction and caustic soda production. Even though the implementation of this method requires less chemicals than the chemical acidification process, fouling of the ion exchange membranes and especially bipolar membrane impairs its productivity. Membrane thickness and ash content measurements along with scanning electron microscopy (SEM), elemental analysis (EDX) and X-ray photoelectron spectrometry (XPS) analysis were performed to identify the nature and mechanisms of the membrane fouling. The results revealed that the fouling layer mostly consisted of organic components and particularly lignin. Based on our proposed fouling mechanisms, throughout the electrodialysis process the pH of the black liquor gradually decreased and as a result more proton ions were available to trigger protonation reaction of lignin phenolic groups and decrease the lignin solubility. Due to the abundance of the proton ions on the surface of the cation exchange layers of the bipolar membrane, destabilized lignin macro-molecules started to self-aggregate and formed lignin clusters on its surface. Over the time, these lignin clusters covered the entire surface of the bipolar membrane and the spaces between the membranes and, eventually, attached to the surface of the cation exchange membrane. Copyright © 2016 Elsevier Inc. All rights reserved.

Extracellular membrane vesicles and immune regulation in the brain

Directory of Open Access Journals (Sweden)

Stefano ePluchino

2012-05-01

Full Text Available The brain is characterized by a complex and integrated network of interacting cells in which cell-to-cell communication is critical for proper development and function. Initially considered as an immune privileged site, the brain is now regarded as an immune specialized system. Accumulating evidence reveals the presence of immune components in the brain, as well as extensive bidirectional communication that takes place between the nervous and the immune system both under homeostatic and pathological conditions. In recent years the secretion of extracellular membrane vesicles (EMVs has been described as a new and evolutionary well-conserved mechanism of cell-to-cell communication, with EMVs influencing the microenvironment through the traffic of bioactive molecules that include proteins and nucleic acids, such as DNA, protein coding and non coding RNAs. Increasing evidence suggests that EMVs are a promising candidate to study cross-boundary cell-to-cell communication pathways. Herein we review the role of EMVs secreted by neural cells in modulating the immune response(s within the brain under physiological and pathological circumstances.

Carbon nanotube embedded PVDF membranes: Effect of solvent composition on the structural morphology for membrane distillation

Science.gov (United States)

Mapunda, Edgar C.; Mamba, Bhekie B.; Msagati, Titus A. M.

2017-08-01

Rapid population increase, growth in industrial and agricultural sectors and global climate change have added significant pressure on conventional freshwater resources. Tapping freshwater from non-conventional water sources such as desalination and wastewater recycling is considered as sustainable alternative to the fundamental challenges of water scarcity. However, affordable and sustainable technologies need to be applied for the communities to benefit from the treatment of non-conventional water source. Membrane distillation is a potential desalination technology which can be used sustainably for this purpose. In this work multi-walled carbon nanotube embedded polyvinylidene fluoride membranes for application in membrane distillation desalination were prepared via non-solvent induced phase separation method. The casting solution was prepared using mixed solvents (N, N-dimethylacetamide and triethyl phosphate) at varying ratios to study the effect of solvent composition on membrane morphological structures. Membrane morphological features were studied using a number of techniques including scanning electron microscope, atomic force microscope, SAXSpace tensile strength analysis, membrane thickness, porosity and contact angle measurements. It was revealed that membrane hydrophobicity, thickness, tensile strength and surface roughness were increasing as the composition of N, N-dimethylacetamide in the solvent was increasing with maximum values obtained between 40 and 60% N, N-dimethylacetamide. Internal morphological structures were changing from cellular structures to short finger-like and sponge-like pores and finally to large macro void type of pores when the amount of N, N-dimethylacetamide in the solvent was changed from low to high respectively. Multi-walled carbon nanotube embedded polyvinylidene fluoride membranes of desired morphological structures and physical properties can be synthesized by regulating the composition of solvents used to prepare the

Pore formation mechanism of porous poly(DL-lactic acid) matrix membrane

Energy Technology Data Exchange (ETDEWEB)

Phaechamud, Thawatchai, E-mail: tphaechamud011@yahoo.com; Chitrattha, Sasiprapa, E-mail: sasi_toey@hotmail.com

2016-04-01

Porous PLA structure has been widely used in cell transplantation, drug carrier and wound dressing. The porous structure can be controlled depending on the choice of the polymer, solvent, nonsolvent and preparation parameters. In this study, the porous PLA matrix membranes were prepared by adding PEG 400 in PLA solution using dichloromethane (DCM) as solvent prior to casting. The influence of other liquids as co-solvent on pore formation and the structural change during membrane formation were evaluated. The co-solvents affected both porous topography and mechanical properties of PLA membrane. The porous matrix were produced when the non-solvent of PLA was used as co-solvent. Cryo-SEM micrographs revealed that PEG 400 still remained in the PLA porous matrix membrane. From the tracking of the structural change during film formation, the PLA–PEG solution changed into porous structure by liquid liquid phase separation and solidification processes, respectively. Thermogravimetric analysis revealed that PLA–PEG in DCM solution exhibited the two-step of weight loss, the first step occurred from DCM evaporation and the second step occurred from the degradation of PLA–PEG matrix. The liquid–liquid phase separation and solidification started when the amount of DCM was higher than PEG 400 for 2.67 folds and DCM amount was equal to that of PEG 400, respectively. These results could clarify the pore formation mechanism of porous PLA membrane and will be useful for the further investigation and application. - Highlights: • Pore formation mechanism of PLA matrix membrane inducing by PEG 400 addition was investigated. • Cryo-scanning electron microscopy revealed the embedded PEG 400 in matrix membrane. • Tracking of structural change during membrane formation with stereomicroscope and thermogravimetric analysis could explain the pore formation mechanism. • Liquid-liquid phase separation of PLA-PEG 400 solution started when the amount of dichloromethane remained 2

Pore formation mechanism of porous poly(DL-lactic acid) matrix membrane

International Nuclear Information System (INIS)

Phaechamud, Thawatchai; Chitrattha, Sasiprapa

2016-01-01

Porous PLA structure has been widely used in cell transplantation, drug carrier and wound dressing. The porous structure can be controlled depending on the choice of the polymer, solvent, nonsolvent and preparation parameters. In this study, the porous PLA matrix membranes were prepared by adding PEG 400 in PLA solution using dichloromethane (DCM) as solvent prior to casting. The influence of other liquids as co-solvent on pore formation and the structural change during membrane formation were evaluated. The co-solvents affected both porous topography and mechanical properties of PLA membrane. The porous matrix were produced when the non-solvent of PLA was used as co-solvent. Cryo-SEM micrographs revealed that PEG 400 still remained in the PLA porous matrix membrane. From the tracking of the structural change during film formation, the PLA–PEG solution changed into porous structure by liquid liquid phase separation and solidification processes, respectively. Thermogravimetric analysis revealed that PLA–PEG in DCM solution exhibited the two-step of weight loss, the first step occurred from DCM evaporation and the second step occurred from the degradation of PLA–PEG matrix. The liquid–liquid phase separation and solidification started when the amount of DCM was higher than PEG 400 for 2.67 folds and DCM amount was equal to that of PEG 400, respectively. These results could clarify the pore formation mechanism of porous PLA membrane and will be useful for the further investigation and application. - Highlights: • Pore formation mechanism of PLA matrix membrane inducing by PEG 400 addition was investigated. • Cryo-scanning electron microscopy revealed the embedded PEG 400 in matrix membrane. • Tracking of structural change during membrane formation with stereomicroscope and thermogravimetric analysis could explain the pore formation mechanism. • Liquid-liquid phase separation of PLA-PEG 400 solution started when the amount of dichloromethane remained 2

Signature of a Nonharmonic Potential as Revealed from a Consistent Shape and Fluctuation Analysis of an Adherent Membrane

Directory of Open Access Journals (Sweden)

Daniel Schmidt

2014-05-01

Full Text Available The interaction of fluid membranes with a scaffold, which can be a planar surface or a more complex structure, is intrinsic to a number of systems from artificial supported bilayers and vesicles to cellular membranes. In principle, these interactions can be either discrete and protein mediated, or continuous. In the latter case, they emerge from ubiquitous intrinsic surface interaction potentials as well as nature-designed steric contributions of the fluctuating membrane or from the polymers of the glycocalyx. Despite the fact that these nonspecific potentials are omnipresent, their description has been a major challenge from experimental and theoretical points of view. Here, we show that a full understanding of the implications of the continuous interactions can be achieved only by expanding the standard superposition models commonly used to treat these types of systems, beyond the usual harmonic level of description. Supported by this expanded theoretical framework, we present three independent, yet mutually consistent, experimental approaches to measure the interaction potential strength and the membrane tension. Upon explicitly taking into account the nature of shot noise as well as the nature of finite experimental resolution, excellent agreement with the augmented theory is obtained, which finally provides a coherent view of the behavior of the membrane in the vicinity of a scaffold.

Membrane binding properties of EBV gp110 C-terminal domain; evidences for structural transition in the membrane environment

International Nuclear Information System (INIS)

Park, Sung Jean; Seo, Min-Duk; Lee, Suk Kyeong; Lee, Bong Jin

2008-01-01

Gp110 of Epstein-Barr virus (EBV) mainly localizes on nuclear/ER membranes and plays a role in the assembly of EBV nucleocapsid. The C-terminal tail domain (gp110 CTD) is essential for the function of gp110 and the nuclear/ER membranes localization of gp110 is ruled by its C-terminal unique nuclear localization signal (NLS), consecutive four arginines. In the present study, the structural properties of gp110 CTD in membrane mimics were investigated using CD, size-exclusion chromatography, and NMR, to elucidate the effect of membrane environment on the structural transition and to compare the structural feature of the protein in the solution state with that of the membrane-bound form. CD and NMR analysis showed that gp110 CTD in a buffer solution appears to adopt a stable folding intermediate which lacks compactness, and a highly helical structure is formed only in membrane environments. The helical content of gp110 CTD was significantly affected by the negative charge as well as the size of membrane mimics. Based on the elution profiles of the size-exclusion chromatography, we found that gp110 CTD intrinsically forms a trimer, revealing that a trimerization region may exist in the C-terminal domain of gp110 like the ectodomain of gp110. The mutation of NLS (RRRR) to RTTR does not affect the overall structure of gp110 CTD in membrane mimics, while the helical propensity in a buffer solution was slightly different between the wild-type and the mutant proteins. This result suggests that not only the helicity induced in membrane environment but also the local structure around NLS may be related to trafficking to the nuclear membrane. More detailed structural difference between the wild-type and the mutant in membrane environment was examined using synthetic two peptides including the wild-type NLS and the mutant NLS

Developmental mechanisms of the tympanic membrane in mammals and non-mammalian amniotes.

Science.gov (United States)

Takechi, Masaki; Kitazawa, Taro; Hirasawa, Tatsuya; Hirai, Tamami; Iseki, Sachiko; Kurihara, Hiroki; Kuratani, Shigeru

2016-01-01

The tympanic membrane is a thin layer that originates from the ectoderm, endoderm, and mesenchyme. Molecular-genetic investigations have revealed that interaction between epithelial and mesenchymal cells in the pharyngeal arches is essential for development of the tympanic membrane. We have recently reported that developmental mechanisms underlying the tympanic membrane seem to be different between mouse and chicken, suggesting that the tympanic membrane evolved independently in mammals and non-mammalian amniotes. In this review, we summarize previous studies of tympanic membrane formation in the mouse. We also discuss its formation in amniotes from an evolutionary point of view. © 2015 Japanese Teratology Society.

Proteomic analysis of GPI-anchored membrane proteins

DEFF Research Database (Denmark)

Jung, Hye Ryung; Jensen, Ole Nørregaard

2006-01-01

Glycosyl-phosphatidyl-inositol-anchored proteins (GPI-APs) represent a subset of post-translationally modified proteins that are tethered to the outer leaflet of the plasma membrane via a C-terminal GPI anchor. GPI-APs are found in a variety of eukaryote species, from pathogenic microorganisms...... to humans. GPI-APs confer important cellular functions as receptors, enzymes and scaffolding molecules. Specific enzymes and detergent extraction methods combined with separation technologies and mass spectrometry permit proteomic analysis of GPI-APs from plasma membrane preparations to reveal cell...

Nitric oxide treatment for the control of reverse osmosis membrane biofouling.

Science.gov (United States)

Barnes, Robert J; Low, Jiun Hui; Bandi, Ratnaharika R; Tay, Martin; Chua, Felicia; Aung, Theingi; Fane, Anthony G; Kjelleberg, Staffan; Rice, Scott A

2015-04-01

Biofouling remains a key challenge for membrane-based water treatment systems. This study investigated the dispersal potential of the nitric oxide (NO) donor compound, PROLI NONOate, on single- and mixed-species biofilms formed by bacteria isolated from industrial membrane bioreactor and reverse osmosis (RO) membranes. The potential of PROLI NONOate to control RO membrane biofouling was also examined. Confocal microscopy revealed that PROLI NONOate exposure induced biofilm dispersal in all but two of the bacteria tested and successfully dispersed mixed-species biofilms. The addition of 40 μM PROLI NONOate at 24-h intervals to a laboratory-scale RO system led to a 92% reduction in the rate of biofouling (pressure rise over a given period) by a bacterial community cultured from an industrial RO membrane. Confocal microscopy and extracellular polymeric substances (EPS) extraction revealed that PROLI NONOate treatment led to a 48% reduction in polysaccharides, a 66% reduction in proteins, and a 29% reduction in microbial cells compared to the untreated control. A reduction in biofilm surface coverage (59% compared to 98%, treated compared to control) and average thickness (20 μm compared to 26 μm, treated compared to control) was also observed. The addition of PROLI NONOate led to a 22% increase in the time required for the RO module to reach its maximum transmembrane pressure (TMP), further indicating that NO treatment delayed fouling. Pyrosequencing analysis revealed that the NO treatment did not significantly alter the microbial community composition of the membrane biofilm. These results present strong evidence for the application of PROLI NONOate for prevention of RO biofouling. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

Quantitative Proteome Analysis Reveals Increased Content of Basement Membrane Proteins in Arteries from Patients with Type 2 Diabetes and Lower Levels among Metformin Users

DEFF Research Database (Denmark)

Rørdam Preil, Simone; Kristensen, Lars P; Beck, Hans C

2015-01-01

hypothesized that metformin intake influences the protein composition. METHODS AND RESULTS: -We analyzed non-atherosclerotic repair arteries gathered at coronary by-pass operations from 30 patients with type 2 diabetes, as well as from 30 age- and gender-matched non-diabetic individuals. Quantitative proteome......BACKGROUND: -The increased risk of cardiovascular diseases (CVD) in type 2 diabetes has been extensively documented, but the origins of the association remain largely unknown. We sought to determine changes in protein expressions in arterial tissue from patients with type 2 diabetes and moreover...... analysis was done by iTRAQ-labelling and LC-MS/MS analysis on individual arterial samples. The amounts of the basement membrane (BM) components, alpha-1- and alpha-2- type IV collagen, gamma-1- and beta-2-laminin were significantly increased in patients with diabetes. Moreover, the expressions of basement...

Challenges of extension workers in reaching rural women farmers in ...

African Journals Online (AJOL)

The study examined the challenges of extension workers in reaching rural women farmers in Enugu State Nigeria. Questionnaire was used to collect data from a sample size of 52 extension workers. Data were analyzed using percentage, mean statistic, chart and factor analysis. Results revealed that training and visit ...

Surface modification of poly(vinylidene fluoride) hollow fibre membranes for biogas purification in a gas-liquid membrane contactor system.

Science.gov (United States)

Jin, Pengrui; Huang, Chuan; Li, Jiaxiang; Shen, Yadong; Wang, Liao

2017-11-01

The wetting of hollow fibre membranes decreases the performance of the liquid-gas membrane contactor for CO 2 capture in biogas upgrading. To solve this problem, in this work, a poly(vinylidene fluoride) (PVDF) hollow fibre membrane for a liquid-gas membrane contactor was coated with a superhydrophobic layer composed of a combination of hydrophobic SiO 2 nanoparticles and polydimethylsiloxane (PDMS) by the method of spray deposition. A rough layer of SiO 2 deposited on the PVDF membrane resulted in an enhanced surface hydrophobicity. The surface structure of the pristine PVDF significantly affected the homogeneity of the generated SiO 2 layer. A uniform surface coating on the PVDF upper layer resulted from the presence of micrometre and nanometre-sized roughness on the surface of the PVDF membrane, which was achieved with a SiO 2 concentration of 4.44 mg ml -1 (0.2 g/45 ml) in the coating solution. As a result, the water contact angle of the modified surface was recorded as 155 ± 3°, which is higher than that of the pristine surface. The high contact angle is advantageous for reducing the wetting of the membrane. Additional mass transfer resistance was introduced by the superhydrophobic layer. In addition, continuous CO 2 absorption tests were carried out in original and modified PVDF hollow fibre membrane contactors, using monoethanolamine (MEA) solution as the absorbent. A long-term stability test revealed that the modified PVDF hollow fibre membrane contactor was able to outperform the original membrane contactor and demonstrated outstanding long-term stability, suggesting that spray deposition is a promising approach to obtain superhydrophobic PVDF membranes for liquid-gas membrane absorption.

Correlation between extension-block K-wire insertion angle and postoperative extension loss in mallet finger fracture.

Science.gov (United States)

Lee, S K; Kim, Y H; Moon, K H; Choy, W S

2018-02-01

Extension-block pinning represents a simple and reliable surgical technique. Although this procedure is commonly performed successfully, some patients develop postoperative extension loss. To date, the relationship between extension-block Kirschner wire (K-wire) insertion angle and postoperative extension loss in mallet finger fracture remains unclear. We aimed to clarify this relationship and further evaluate how various operative and non-operative factors affect postoperative extension loss after extension-block pinning for mallet finger fracture. A retrospective study was conducted to investigate a relationship between extension block K-wire insertion angle and postoperative extension loss. The inclusion criteria were: (1) a dorsal intra-articular fracture fragment involving 30% of the base of the distal phalanx with or without volar subluxation of the distal phalanx; and (2) block K-wire insertion angle and fixation angle of the distal interphalangeal (DIP) joint were assessed using lateral radiograph at immediate postoperative time. Postoperative extension loss was assessed by using lateral radiograph at latest follow-up. Extension-block K-wire insertion angle was defined as the acute angle between extension block K-wire and longitudinal axis of middle phalangeal head. DIP joint fixation angle was defined as the acute angle between the distal phalanx and middle phalanx longitudinal axes. Seventy-five patients were included. The correlation analysis revealed that extension-block K-wire insertion angle had a negative correlation with postoperative extension loss, whereas fracture size and time to operation had a positive correlation (correlation coefficient for extension block K-wire angle: -0.66, facture size: +0.67, time to operation: +0.60). When stratifying patients in terms of negative and positive fixation angle of the DIP joint, the independent t-test showed that mean postoperative extension loss is -3.67° and +4.54° (DIP joint fixation angles of block

Endoplasmic Reticulum-Plasma Membrane Contact Sites.

Science.gov (United States)

Saheki, Yasunori; De Camilli, Pietro

2017-06-20

The endoplasmic reticulum (ER) has a broad localization throughout the cell and forms direct physical contacts with all other classes of membranous organelles, including the plasma membrane (PM). A number of protein tethers that mediate these contacts have been identified, and study of these protein tethers has revealed a multiplicity of roles in cell physiology, including regulation of intracellular Ca 2+ dynamics and signaling as well as control of lipid traffic and homeostasis. In this review, we discuss the cross talk between the ER and the PM mediated by direct contacts. We review factors that tether the two membranes, their properties, and their dynamics in response to the functional state of the cell. We focus in particular on the role of ER-PM contacts in nonvesicular lipid transport between the two bilayers mediated by lipid transfer proteins.

Membranes, methods of making membranes, and methods of separating gases using membranes

Science.gov (United States)

Ho, W. S. Winston

2012-10-02

Membranes, methods of making membranes, and methods of separating gases using membranes are provided. The membranes can include at least one hydrophilic polymer, at least one cross-linking agent, at least one base, and at least one amino compound. The methods of separating gases using membranes can include contacting a gas stream containing at least one of CO.sub.2, H.sub.2S, and HCl with one side of a nonporous and at least one of CO.sub.2, H.sub.2S, and HCl selectively permeable membrane such that at least one of CO.sub.2, H.sub.2S, and HCl is selectively transported through the membrane.

Prevention of intra-abdominal adhesion by bi-layer electrospun membrane.

Science.gov (United States)

Jiang, Shichao; Wang, Wei; Yan, Hede; Fan, Cunyi

2013-06-04

The aim of this study was to compare the anti-adhesion efficacy of a bi-layer electrospun fibrous membrane consisting of hyaluronic acid-loaded poly(ε-caprolactone) (PCL) fibrous membrane as the inner layer and PCL fibrous membrane as the outer layer with a single-layer PCL electrospun fibrous membrane in a rat cecum abrasion model. The rat model utilized a cecal abrasion and abdominal wall insult surgical protocol. The bi-layer and PCL membranes were applied between the cecum and the abdominal wall, respectively. Control animals did not receive any treatment. After postoperative day 14, a visual semiquantitative grading scale was used to grade the extent of adhesion. Histological analysis was performed to reveal the features of adhesion tissues. Bi-layer membrane treated animals showed significantly lower adhesion scores than control animals (p compared with the PCL membrane. Histological analysis of the bi-layer membrane treated rat rarely demonstrated tissue adhesion while that of the PCL membrane treated rat and control rat showed loose and dense adhesion tissues, respectively. Bi-layer membrane can efficiently prevent adhesion formation in abdominal cavity and showed a significantly decreased adhesion tissue formation compared with the control.

Meninges: from protective membrane to stem cell niche

OpenAIRE

Decimo, Ilaria; Fumagalli, Guido; Berton, Valeria; Krampera, Mauro; Bifari, Francesco

2012-01-01

Meninges are a three tissue membrane primarily known as coverings of the brain. More in depth studies on meningeal function and ultrastructure have recently changed the view of meninges as a merely protective membrane. Accurate evaluation of the anatomical distribution in the CNS reveals that meninges largely penetrate inside the neural tissue. Meninges enter the CNS by projecting between structures, in the stroma of choroid plexus and form the perivascular space (Virchow-Robin) of every pare...

Solutocapillary Convection Effects on Polymeric Membrane Morphology

Science.gov (United States)

Krantz, William B.; Todd, Paul W.; Kinagurthu, Sanjay

1996-01-01

Macro voids are undesirable large pores in membranes used for purification. They form when membranes are cast as thin films on a smooth surface by evaporating solvent (acetone) from a polymer solution. There are two un-tested hypotheses explaining the growth of macro voids. One states that diffusion of the non-solvent (water) is solely responsible, while the other states that solutocapillary convection is the primary cause of macro void growth. Solutocapillary convection is flow-caused by a concentration induced surface-tension gradient. Macrovoid growth in the former hypothesis is gravity independent, while in the latter it is opposed by gravity. To distinguish between these two hypotheses, experiments were designed to cast membranes in zero-gravity. A semi-automated apparatus was designed and built for casting membranes during the 20 secs of zero-g time available in parabolic aircraft flight such as NASA's KC-135. The phase changes were monitored optically, and membrane morphology was evaluated by scanning electron microscopy (SEM). These studies appear to be the first quantitative studies of membrane casting in micro-gravity which incorporate real-time data acquisition. Morphological studies of membranes cast at 0, 1, and 1.8 g revealed the presence of numerous, sparse and no macrovoids respectively. These results are consistent with the predictions of the solutocapillary hypothesis of macrovoid growth.

Monitoring changes in membrane polarity, membrane integrity, and intracellular ion concentrations in Streptococcus pneumoniae using fluorescent dyes.

Science.gov (United States)

Clementi, Emily A; Marks, Laura R; Roche-Håkansson, Hazeline; Håkansson, Anders P

2014-02-17

Membrane depolarization and ion fluxes are events that have been studied extensively in biological systems due to their ability to profoundly impact cellular functions, including energetics and signal transductions. While both fluorescent and electrophysiological methods, including electrode usage and patch-clamping, have been well developed for measuring these events in eukaryotic cells, methodology for measuring similar events in microorganisms have proven more challenging to develop given their small size in combination with the more complex outer surface of bacteria shielding the membrane. During our studies of death-initiation in Streptococcus pneumoniae (pneumococcus), we wanted to elucidate the role of membrane events, including changes in polarity, integrity, and intracellular ion concentrations. Searching the literature, we found that very few studies exist. Other investigators had monitored radioisotope uptake or equilibrium to measure ion fluxes and membrane potential and a limited number of studies, mostly in Gram-negative organisms, had seen some success using carbocyanine or oxonol fluorescent dyes to measure membrane potential, or loading bacteria with cell-permeant acetoxymethyl (AM) ester versions of ion-sensitive fluorescent indicator dyes. We therefore established and optimized protocols for measuring membrane potential, rupture, and ion-transport in the Gram-positive organism S. pneumoniae. We developed protocols using the bis-oxonol dye DiBAC4(3) and the cell-impermeant dye propidium iodide to measure membrane depolarization and rupture, respectively, as well as methods to optimally load the pneumococci with the AM esters of the ratiometric dyes Fura-2, PBFI, and BCECF to detect changes in intracellular concentrations of Ca(2+), K(+), and H(+), respectively, using a fluorescence-detection plate reader. These protocols are the first of their kind for the pneumococcus and the majority of these dyes have not been used in any other bacterial

Translocation mechanism of C{sub 60} and C{sub 60} derivations across a cell membrane

Energy Technology Data Exchange (ETDEWEB)

Liang, Lijun, E-mail: michael.lijunl@gmail.com [Hangzhou Dianzi University, College of Life Information Science and Instrument Engineering (China); Kang, Zhengzhong [Zhejiang University, Department of Chemistry (China); Shen, Jia-Wei, E-mail: shen.jiawei@hotmail.com [Hangzhou Normal University, School of Medicine (China)

2016-11-15

Carbon-based nanoparticles (NPs) such as fullerenes and nanotubes have been extensively studied for drug delivery in recent years. The permeation process of fullerene and its derivative molecules through membrane is essential to the utilization of fullerene-based drug delivery system, but the mechanism and the dynamics of permeation through cell membrane are still unclear. In this study, coarse-grained molecular dynamics simulations were performed to investigate the permeation process of functionalized fullerene molecules (ca. 0.72 nm) through the membrane. Our results show that single functionalized fullerene molecule in such nanoscale could permeate the lipid membrane in micro-second time scale. Pristine C{sub 60} molecules prefer to aggregate into several small clusters while C{sub 60}OH{sub 15} molecules could aggregate into one big cluster to permeate through the lipid membrane. After permeation of C{sub 60} or its derivatives into membrane, all C{sub 60} and C{sub 60}OH{sub 15} molecules disaggregated and monodispersed in the lipid membrane.

Chitosan-Coated Collagen Membranes Promote Chondrocyte Adhesion, Growth, and Interleukin-6 Secretion

Directory of Open Access Journals (Sweden)

Nabila Mighri

2015-11-01

Full Text Available Designing scaffolds made from natural polymers may be highly attractive for tissue engineering strategies. We sought to produce and characterize chitosan-coated collagen membranes and to assess their efficacy in promoting chondrocyte adhesion, growth, and cytokine secretion. Porous collagen membranes were placed in chitosan solutions then crosslinked with glutaraldehyde vapor. Fourier transform infrared (FTIR analyses showed elevated absorption at 1655 cm-1 of the carbon–nitrogen (N=C bonds formed by the reaction between the (NH2 of the chitosan and the (C=O of the glutaraldehyde. A significant peak in the amide II region revealed a significant deacetylation of the chitosan. Scanning electron microscopy (SEM images of the chitosan-coated membranes exhibited surface variations, with pore size ranging from 20 to 50 µm. X-ray photoelectron spectroscopy (XPS revealed a decreased C–C groups and an increased C–N/C–O groups due to the reaction between the carbon from the collagen and the NH2 from the chitosan. Increased rigidity of these membranes was also observed when comparing the chitosan-coated and uncoated membranes at dried conditions. However, under wet conditions, the chitosan coated collagen membranes showed lower rigidity as compared to dried conditions. Of great interest, the glutaraldehyde-crosslinked chitosan-coated collagen membranes promoted chondrocyte adhesion, growth, and interleukin (IL-6 secretion. Overall results confirm the feasibility of using designed chitosan-coated collagen membranes in future applications, such as cartilage repair.

Platinum porphyrins as ionophores in polymeric membrane electrodes

DEFF Research Database (Denmark)

Lvova, Larisa; Verrelli, Giorgio; Nardis, Sara

2011-01-01

A comparative study of Pt(II)- and Pt(IV)-porphyrins as novel ionophores for anion-selective polymeric membrane electrodes is performed. Polymeric membranes of different compositions, prepared by varying plasticizers, cationic and anionic additives and Pt porphyrins, have been examined...... within the electrode membranes, while those based on Pt(IV)TPPCl2 operate via a mixed mode carrier mechanism, evidencing also a partial reduction of the starting ionophore to Pt(II)TPP. Spectrophotometric measurements of thin polymeric films indicate that no spontaneous formation of hydroxide ion bridged...... porphyrin dimers occurs in the membrane plasticized both with high or low dielectric constant plasticizer, due to a low oxophilicity of central Pt. The computational study of various anion–Pt(IV)TPPCl2 complex formation by means of semi-empirical and density functional theory (DFT) methods revealed a good...

Application of dynamic membranes in anaerobic membranes in anaerobic membrane bioreactor systems

NARCIS (Netherlands)

Erşahin, M.E.

2015-01-01

Anaerobic membrane bioreactors (AnMBRs) physically ensure biomass retention by the application of a membrane filtration process. With growing application experiences from aerobic membrane bioreactors (MBRs), the combination of membrane and anaerobic processes has received much attention and become

Light-induced modification of plant plasma membrane ion transport.

Science.gov (United States)

Marten, I; Deeken, R; Hedrich, R; Roelfsema, M R G

2010-09-01

Light is not only the driving force for electron and ion transport in the thylakoid membrane, but also regulates ion transport in various other membranes of plant cells. Light-dependent changes in ion transport at the plasma membrane and associated membrane potential changes have been studied intensively over the last century. These studies, with various species and cell types, revealed that apart from regulation by chloroplasts, plasma membrane transport can be controlled by phytochromes, phototropins or channel rhodopsins. In this review, we compare light-dependent plasma membrane responses of unicellular algae (Eremosphaera and Chlamydomonas), with those of a multicellular alga (Chara), liverworts (Conocephalum), mosses (Physcomitrella) and several angiosperm cell types. Light-dependent plasma membrane responses of Eremosphaera and Chara are characterised by the dominant role of K(+) channels during membrane potential changes. In most other species, the Ca(2+)-dependent activation of plasma membrane anion channels represents a general light-triggered event. Cell type-specific responses are likely to have evolved by modification of this general response or through the development of additional light-dependent signalling pathways. Future research to elucidate these light-activated signalling chains is likely to benefit from the recent identification of S-type anion channel genes and proteins capable of regulating these channels.

Robust performance of a membrane bioreactor for removing antibiotic resistance genes exposed to antibiotics: Role of membrane foulants.

Science.gov (United States)

Zhu, Yijing; Wang, Yayi; Zhou, Shuai; Jiang, Xuxin; Ma, Xiao; Liu, Chao

2018-03-01

Antibiotic resistance genes (ARGs) are an emerging concern in wastewater treatment plants (WWTPs), as dissemination of ARGs can pose a serious risk to human health. Few studies, however, have quantified ARGs in membrane bioreactors (MBRs), although MBRs have been widely used for both municipal and industrial wastewater treatment. To reveal the capacity of MBRs for removal of ARGs and the response of membrane fouling after antibiotic exposure, five typical ARG subtypes (sulI, sulII, tetC, tetX and ereA) and int1 were quantified affiliated by systematic membrane foulants analysis in a laboratory-scale anoxic/aerobic membrane bioreactor (A/O-MBR). Sulfamethoxazole and tetracycline hydrochloride additions increased ARG abundances by 0.5-1.4 orders of magnitude in the activated sludge, while the ARG removal performance of the membrane module remained stable (or even increased with ARG absolute abundance in several cases), with the abundance of removed ARGs ranging from 0.6 to 5.6 orders of magnitude. Specifically, the distribution of ARGs in membrane foulants accounted for 13%-25% of the total absolute abundance of all tested MBR samples. Indeed, substantial fouling occurred after the antibiotic additions, with the mean concentrations of soluble microbial product (SMP) and extracellular polymeric substance (EPS) increasing by 340% and 220%, respectively, in a membrane fouling cycle; moreover, the contents of EPS and SMP in the membrane foulants were significantly correlated with the ARG absolute abundance of membrane foulants (p removal of ARGs in MBR systems, and highlight the contribution of membrane fouling to ARG removals in terms of the potential of MBR as an effective strategy to reduce ARG levels in WWTP effluent. Copyright © 2017 Elsevier Ltd. All rights reserved.

Modeling electrically active viscoelastic membranes.

Directory of Open Access Journals (Sweden)

Sitikantha Roy

Full Text Available The membrane protein prestin is native to the cochlear outer hair cell that is crucial to the ear's amplification and frequency selectivity throughout the whole acoustic frequency range. The outer hair cell exhibits interrelated dimensional changes, force generation, and electric charge transfer. Cells transfected with prestin acquire unique active properties similar to those in the native cell that have also been useful in understanding the process. Here we propose a model describing the major electromechanical features of such active membranes. The model derived from thermodynamic principles is in the form of integral relationships between the history of voltage and membrane resultants as independent variables and the charge density and strains as dependent variables. The proposed model is applied to the analysis of an active force produced by the outer hair cell in response to a harmonic electric field. Our analysis reveals the mechanism of the outer hair cell active (isometric force having an almost constant amplitude and phase up to 80 kHz. We found that the frequency-invariance of the force is a result of interplay between the electrical filtering associated with prestin and power law viscoelasticity of the surrounding membrane. Paradoxically, the membrane viscoelasticity boosts the force balancing the electrical filtering effect. We also consider various modes of electromechanical coupling in membrane with prestin associated with mechanical perturbations in the cell. We consider pressure or strains applied step-wise or at a constant rate and compute the time course of the resulting electric charge. The results obtained here are important for the analysis of electromechanical properties of membranes, cells, and biological materials as well as for a better understanding of the mechanism of hearing and the role of the protein prestin in this mechanism.

Uncovering homo-and hetero-interactions on the cell membrane using single particle tracking approaches

International Nuclear Information System (INIS)

Torreno-Pina, Juan A; Manzo, Carlo; Garcia-Parajo, Maria F

2016-01-01

The plasma membrane of eukaryotic cells is responsible for a myriad of functions that regulate cell physiology and plays a crucial role in a multitude of processes that include adhesion, migration, signaling recognition and cell–cell communication. This is accomplished by specific interactions between different membrane components such as lipids and proteins on the lipid bilayer but also through interactions with the underlying cortical actin cytoskeleton on the intracellular side and the glycocalyx matrix in close proximity to the extracellular side. Advanced biophysical techniques, including single particle tracking (SPT) have revealed that the lateral diffusion of molecular components on the plasma membrane represents a landmark manifestation of such interactions. Indeed, by studying changes in the diffusivity of individual membrane molecules, including sub-diffusion, confined diffusion and/or transient arrest of molecules in membrane compartments, it has been possible to gain insight on the nature of molecular interactions and to infer on its functional role for cell response. In this review, we will revise some exciting results where SPT has been crucial to reveal homo- and hetero-interactions on the cell membrane. (paper)

One-way membrane trafficking of SOS in receptor-triggered Ras activation.

Science.gov (United States)

Christensen, Sune M; Tu, Hsiung-Lin; Jun, Jesse E; Alvarez, Steven; Triplet, Meredith G; Iwig, Jeffrey S; Yadav, Kamlesh K; Bar-Sagi, Dafna; Roose, Jeroen P; Groves, Jay T

2016-09-01

SOS is a key activator of the small GTPase Ras. In cells, SOS-Ras signaling is thought to be initiated predominantly by membrane recruitment of SOS via the adaptor Grb2 and balanced by rapidly reversible Grb2-SOS binding kinetics. However, SOS has multiple protein and lipid interactions that provide linkage to the membrane. In reconstituted-membrane experiments, these Grb2-independent interactions were sufficient to retain human SOS on the membrane for many minutes, during which a single SOS molecule could processively activate thousands of Ras molecules. These observations raised questions concerning how receptors maintain control of SOS in cells and how membrane-recruited SOS is ultimately released. We addressed these questions in quantitative assays of reconstituted SOS-deficient chicken B-cell signaling systems combined with single-molecule measurements in supported membranes. These studies revealed an essentially one-way trafficking process in which membrane-recruited SOS remains trapped on the membrane and continuously activates Ras until being actively removed via endocytosis.

Synthesis and transfer of galactolipids in the chloroplast envelope membranes of Arabidopsis thaliana.

Science.gov (United States)

Kelly, Amélie A; Kalisch, Barbara; Hölzl, Georg; Schulze, Sandra; Thiele, Juliane; Melzer, Michael; Roston, Rebecca L; Benning, Christoph; Dörmann, Peter

2016-09-20

Galactolipids [monogalactosyldiacylglycerol (MGDG) and digalactosyldiacylglycerol (DGDG)] are the hallmark lipids of photosynthetic membranes. The galactolipid synthases MGD1 and DGD1 catalyze consecutive galactosyltransfer reactions but localize to the inner and outer chloroplast envelopes, respectively, necessitating intermembrane lipid transfer. Here we show that the N-terminal sequence of DGD1 (NDGD1) is required for galactolipid transfer between the envelopes. Different diglycosyllipid synthases (DGD1, DGD2, and Chloroflexus glucosyltransferase) were introduced into the dgd1-1 mutant of Arabidopsis in fusion with N-terminal extensions (NDGD1 and NDGD2) targeting to the outer envelope. Reconstruction of DGDG synthesis in the outer envelope membrane was observed only with diglycosyllipid synthase fusion proteins carrying NDGD1, indicating that NDGD1 enables galactolipid translocation between envelopes. NDGD1 binds to phosphatidic acid (PA) in membranes and mediates PA-dependent membrane fusion in vitro. These findings provide a mechanism for the sorting and selective channeling of lipid precursors between the galactolipid pools of the two envelope membranes.

Destabilization of the Outer and Inner Mitochondrial Membranes by Core and Linker Histones

Science.gov (United States)

Cascone, Annunziata; Bruelle, Celine; Lindholm, Dan; Bernardi, Paolo; Eriksson, Ove

2012-01-01

Background Extensive DNA damage leads to apoptosis. Histones play a central role in DNA damage sensing and may mediate signals of genotoxic damage to cytosolic effectors including mitochondria. Methodology/Principal Findings We have investigated the effects of histones on mitochondrial function and membrane integrity. We demonstrate that both linker histone H1 and core histones H2A, H2B, H3, and H4 bind strongly to isolated mitochondria. All histones caused a rapid and massive release of the pro-apoptotic intermembrane space proteins cytochrome c and Smac/Diablo, indicating that they permeabilize the outer mitochondrial membrane. In addition, linker histone H1, but not core histones, permeabilized the inner membrane with a collapse of the membrane potential, release of pyridine nucleotides, and mitochondrial fragmentation. Conclusions We conclude that histones destabilize the mitochondrial membranes, a mechanism that may convey genotoxic signals to mitochondria and promote apoptosis following DNA damage. PMID:22523586

High temperature proton exchange membranes based on polybenzimidazoles for fuel cells

DEFF Research Database (Denmark)

Li, Qingfeng; Jensen, Jens Oluf; Savinell, Robert F

2009-01-01

-term durability with a degradation rate of 5Vh−1 has been achieved under continuous operation with hydrogen and air at 150–160 ◦C. With load or thermal cycling, a performance loss of 300V per cycle or 40Vh−1 per operating hour was observed. Further improvement should be done by, e.g. optimizing the thermal...... in recent years motivated extensive research activities with great progress. This treatise is devoted to updating the development, covering polymer synthesis, membrane casting, physicochemical characterizations and fuel cell technologies. To optimize the membrane properties, high molecular weight polymers...

Low-pH-induced transformation of bilayer membrane into bicontinuous cubic phase in dioleoylphosphatidylserine/monoolein membranes.

Science.gov (United States)

Okamoto, Yoshihide; Masum, Shah Md; Miyazawa, Haruna; Yamazaki, Masahito

2008-04-01

Cubic biomembranes, nonbilayer membranes with connections in three-dimensional space that have a cubic symmetry, have been observed in various cells. Interconversion between the bilayer liquid-crystalline (L(alpha)) phase and cubic phases attracted much attention in terms of both biological and physicochemical aspects. Herein we report the pH effect on the phase and structure of dioleoylphosphatidylserine (DOPS)/monoolein (MO) membranes under a physiological ion concentration condition, which was revealed by small-angle X-ray scattering (SAXS) measurement. At neutral pH, DOPS/MO membranes containing high concentrations of DOPS were in the L(alpha) phase. First, the pH effect on the phase and structure of the multilamellar vesicles (MLVs) of the DOPS/MO membranes preformed at neutral pH was investigated by adding various low-pH buffers into the MLV suspension. For 20%-DOPS/80%-MO MLVs, at and below pH 2.9, a transition from the L(alpha) to cubic (Q(224)) phase occurred within 1 h. This phase transition was reversible; a subsequent increase in pH to a neutral one in the membrane suspension transformed the cubic phase into the original L(alpha) phase. Second, we found that a decrease in pH transformed large unilamellar vesicles of DOPS/MO membranes into the cubic phase under similar conditions. We have proposed the mechanism of the low-pH-induced phase transition and also made a quantitative analysis on the critical pH of the phase transition. This finding is the first demonstration that a change in pH can induce a reversible phase transition between the L(alpha) and cubic phases of lipid membranes within 1 h.

Real-life applications with membrane computing

CERN Document Server

Zhang, Gexiang; Gheorghe, Marian

2017-01-01

This book thoroughly investigates the underlying theoretical basis of membrane computing models, and reveals their latest applications. In addition, to date there have been no illustrative case studies or complex real-life applications that capitalize on the full potential of the sophisticated membrane systems computational apparatus; gaps that this book remedies. By studying various complex applications – including engineering optimization, power systems fault diagnosis, mobile robot controller design, and complex biological systems involving data modeling and process interactions – the book also extends the capabilities of membrane systems models with features such as formal verification techniques, evolutionary approaches, and fuzzy reasoning methods. As such, the book offers a comprehensive and up-to-date guide for all researchers, PhDs and undergraduate students in the fields of computer science, engineering and the bio-sciences who are interested in the applications of natural computing models.

Nanocomposite Electrospun Nanofiber Membranes for Environmental Remediation.

Science.gov (United States)

Homaeigohar, Shahin; Elbahri, Mady

2014-02-10

Rapid worldwide industrialization and population growth is going to lead to an extensive environmental pollution. Therefore, so many people are currently suffering from the water shortage induced by the respective pollution, as well as poor air quality and a huge fund is wasted in the world each year due to the relevant problems. Environmental remediation necessitates implementation of novel materials and technologies, which are cost and energy efficient. Nanomaterials, with their unique chemical and physical properties, are an optimum solution. Accordingly, there is a strong motivation in seeking nano-based approaches for alleviation of environmental problems in an energy efficient, thereby, inexpensive manner. Thanks to a high porosity and surface area presenting an extraordinary permeability (thereby an energy efficiency) and selectivity, respectively, nanofibrous membranes are a desirable candidate. Their functionality and applicability is even promoted when adopting a nanocomposite strategy. In this case, specific nanofillers, such as metal oxides, carbon nanotubes, precious metals, and smart biological agents, are incorporated either during electrospinning or in the post-processing. Moreover, to meet operational requirements, e.g., to enhance mechanical stability, decrease of pressure drop, etc. , nanofibrous membranes are backed by a microfibrous non-woven forming a hybrid membrane. The novel generation of nanocomposite/hybrid nanofibrous membranes can perform extraordinarily well in environmental remediation and control. This reality justifies authoring of this review paper.

Nanocomposite Electrospun Nanofiber Membranes for Environmental Remediation

Directory of Open Access Journals (Sweden)

Shahin Homaeigohar

2014-02-01

Full Text Available Rapid worldwide industrialization and population growth is going to lead to an extensive environmental pollution. Therefore, so many people are currently suffering from the water shortage induced by the respective pollution, as well as poor air quality and a huge fund is wasted in the world each year due to the relevant problems. Environmental remediation necessitates implementation of novel materials and technologies, which are cost and energy efficient. Nanomaterials, with their unique chemical and physical properties, are an optimum solution. Accordingly, there is a strong motivation in seeking nano-based approaches for alleviation of environmental problems in an energy efficient, thereby, inexpensive manner. Thanks to a high porosity and surface area presenting an extraordinary permeability (thereby an energy efficiency and selectivity, respectively, nanofibrous membranes are a desirable candidate. Their functionality and applicability is even promoted when adopting a nanocomposite strategy. In this case, specific nanofillers, such as metal oxides, carbon nanotubes, precious metals, and smart biological agents, are incorporated either during electrospinning or in the post-processing. Moreover, to meet operational requirements, e.g., to enhance mechanical stability, decrease of pressure drop, etc., nanofibrous membranes are backed by a microfibrous non-woven forming a hybrid membrane. The novel generation of nanocomposite/hybrid nanofibrous membranes can perform extraordinarily well in environmental remediation and control. This reality justifies authoring of this review paper.

Structure-Related Roles for the Conservation of the HIV-1 Fusion Peptide Sequence Revealed by Nuclear Magnetic Resonance.

Science.gov (United States)

Serrano, Soraya; Huarte, Nerea; Rujas, Edurne; Andreu, David; Nieva, José L; Jiménez, María Angeles

2017-10-17

Despite extensive characterization of the human immunodeficiency virus type 1 (HIV-1) hydrophobic fusion peptide (FP), the structure-function relationships underlying its extraordinary degree of conservation remain poorly understood. Specifically, the fact that the tandem repeat of the FLGFLG tripeptide is absolutely conserved suggests that high hydrophobicity may not suffice to unleash FP function. Here, we have compared the nuclear magnetic resonance (NMR) structures adopted in nonpolar media by two FP surrogates, wtFP-tag and scrFP-tag, which had equal hydrophobicity but contained wild-type and scrambled core sequences LFLGFLG and FGLLGFL, respectively. In addition, these peptides were tagged at their C-termini with an epitope sequence that folded independently, thereby allowing Western blot detection without interfering with FP structure. We observed similar α-helical FP conformations for both specimens dissolved in the low-polarity medium 25% (v/v) 1,1,1,3,3,3-hexafluoro-2-propanol (HFIP), but important differences in contact with micelles of the membrane mimetic dodecylphosphocholine (DPC). Thus, whereas wtFP-tag preserved a helix displaying a Gly-rich ridge, the scrambled sequence lost in great part the helical structure upon being solubilized in DPC. Western blot analyses further revealed the capacity of wtFP-tag to assemble trimers in membranes, whereas membrane oligomers were not observed in the case of the scrFP-tag sequence. We conclude that, beyond hydrophobicity, preserving sequence order is an important feature for defining the secondary structures and oligomeric states adopted by the HIV FP in membranes.

Revealing mechanisms of selective, concentration-dependent potentials of 4-hydroxy-2-nonenal to induce apoptosis in cancer cells through inactivation of membrane-associated catalase.

Science.gov (United States)

Bauer, Georg; Zarkovic, Neven

2015-04-01

Tumor cells generate extracellular superoxide anions and are protected against superoxide anion-mediated intercellular apoptosis-inducing signaling by the expression of membrane-associated catalase. 4-Hydroxy-2-nonenal (4-HNE), a versatile second messenger generated during lipid peroxidation, has been shown to induce apoptosis selectively in malignant cells. The findings described in this paper reveal the strong, concentration-dependent potential of 4-HNE to specifically inactivate extracellular catalase of tumor cells both indirectly and directly and to consequently trigger apoptosis in malignant cells through superoxide anion-mediated intercellular apoptosis-inducing signaling. Namely, 4-HNE caused apoptosis selectively in NOX1-expressing tumor cells through inactivation of their membrane-associated catalase, thus reactivating subsequent intercellular signaling through the NO/peroxynitrite and HOCl pathways, followed by the mitochondrial pathway of apoptosis. Concentrations of 4-HNE of 1.2 µM and higher directly inactivated membrane-associated catalase of tumor cells, whereas at lower concentrations, 4-HNE triggered a complex amplificatory pathway based on initial singlet oxygen formation through H2O2 and peroxynitrite interaction. Singlet-oxygen-dependent activation of the FAS receptor and caspase-8 increased superoxide anion generation by NOX1 and amplification of singlet oxygen generation, which allowed singlet-oxygen-dependent inactivation of catalase. 4-HNE and singlet oxygen cooperate in complex autoamplificatory loops during this process. The finding of these novel anticancer pathways may be useful for understanding the role of 4-HNE in the control of malignant cells and for the optimization of ROS-dependent therapeutic approaches including antioxidant treatments. Copyright © 2015 Elsevier Inc. All rights reserved.

Bi-directional exchange of membrane components occurs during co-culture of mesenchymal stem cells and nucleus pulposus cells.

Science.gov (United States)

Strassburg, Sandra; Hodson, Nigel W; Hill, Patrick I; Richardson, Stephen M; Hoyland, Judith A

2012-01-01

Mesenchymal stem cell (MSC)-based therapies have been proposed as novel treatments for intervertebral disc (IVD) degeneration. We have previously demonstrated that when MSCs are co-cultured with nucleus pulposus (NP) cells with direct cell-cell contact, they differentiate along the NP lineage and simultaneously stimulate the degenerate NP cell population to regain a normal (non-degenerate) phenotype, an effect which requires cell-cell communication. However, the mechanisms by which NP cells and MSCs interact in this system are currently unclear. Thus, in this study we investigated a range of potential mechanisms for exchange of cellular components or information that may direct these changes, including cell fusion, gap-junctional communication and exchange of membrane components by direct transfer or via microvesicle formation. Flow cytometry of fluorescently labeled MSCs and NP cells revealed evidence of some cell fusion and formation of gapjunctions, although at the three timepoints studied these phenomena were detectable only in a small proportion of cells. While these mechanisms may play a role in cell-cell communication, the data suggests they are not the predominant mechanism of interaction. However, flow cytometry of fluorescently dual-labeled cells showed that extensive bi-directional transfer of membrane components is operational during direct co-culture of MSCs and NP cells. Furthermore, there was also evidence for secretion and internalization of membrane-bound microvesicles by both cell types. Thus, this study highlights bi-directional intercellular transfer of membrane components as a possible mechanism of cellular communication between MSC and NP cells.

Discovery of novel membrane binding structures and functions

Science.gov (United States)

Kufareva, Irina; Lenoir, Marc; Dancea, Felician; Sridhar, Pooja; Raush, Eugene; Bissig, Christin; Gruenberg, Jean; Abagyan, Ruben; Overduin, Michael

2014-01-01

The function of a protein is determined by its intrinsic activity in the context of its subcellular distribution. Membranes localize proteins within cellular compartments and govern their specific activities. Discovering such membrane-protein interactions is important for understanding biological mechanisms, and could uncover novel sites for therapeutic intervention. Here we present a method for detecting membrane interactive proteins and their exposed residues that insert into lipid bilayers. Although the development process involved analysis of how C1b, C2, ENTH, FYVE, Gla, pleckstrin homology (PH) and PX domains bind membranes, the resulting Membrane Optimal Docking Area (MODA) method yields predictions for a given protein of known three dimensional structures without referring to canonical membrane-targeting modules. This approach was tested on the Arf1 GTPase, ATF2 acetyltransferase, von Willebrand factor A3 domain and Neisseria gonorrhoeae MsrB protein, and further refined with membrane interactive and non-interactive FAPP1 and PKD1 pleckstrin homology domains, respectively. Furthermore we demonstrate how this tool can be used to discover unprecedented membrane binding functions as illustrated by the Bro1 domain of Alix, which was revealed to recognize lysobisphosphatidic acid (LBPA). Validation of novel membrane-protein interactions relies on other techniques such as nuclear magnetic resonance spectroscopy (NMR) which was used here to map the sites of micelle interaction. Together this indicates that genome-wide identification of known and novel membrane interactive proteins and sites is now feasible, and provides a new tool for functional annotation of the proteome. PMID:25394204

Intramolecular dynamics within the N-Cap-SH3-SH2 regulatory unit of the c-Abl tyrosine kinase reveal targeting to the cellular membrane.

Science.gov (United States)

de Oliveira, Guilherme A P; Pereira, Elen G; Ferretti, Giulia D S; Valente, Ana Paula; Cordeiro, Yraima; Silva, Jerson L

2013-09-27

c-Abl is a key regulator of cell signaling and is under strict control via intramolecular interactions. In this study, we address changes in the intramolecular dynamics coupling within the c-Abl regulatory unit by presenting its N-terminal segment (N-Cap) with an alternative function in the cell as c-Abl becomes activated. Using small angle x-ray scattering, nuclear magnetic resonance, and confocal microscopy, we demonstrate that the N-Cap and the Src homology (SH) 3 domain acquire μs-ms motions upon N-Cap association with the SH2-L domain, revealing a stabilizing synergy between these segments. The N-Cap-myristoyl tether likely triggers the protein to anchor to the membrane because of these flip-flop dynamics, which occur in the μs-ms time range. This segment not only presents the myristate during c-Abl inhibition but may also trigger protein localization inside the cell in a functional and stability-dependent mechanism that is lost in Bcr-Abl(+) cells, which underlie chronic myeloid leukemia. This loss of intramolecular dynamics and binding to the cellular membrane is a potential therapeutic target.

Optimization Study of Small-Scale Solar Membrane Distillation Desalination Systems (s-SMDDS

Directory of Open Access Journals (Sweden)

Hsuan Chang

2014-11-01

Full Text Available Membrane distillation (MD, which can utilize low-grade thermal energy, has been extensively studied for desalination. By incorporating solar thermal energy, the solar membrane distillation desalination system (SMDDS is a potential technology for resolving energy and water resource problems. Small-scale SMDDS (s-SMDDS is an attractive and viable option for the production of fresh water for small communities in remote arid areas. The minimum cost design and operation of s-SMDDS are determined by a systematic method, which involves a pseudo-steady-state approach for equipment sizing and dynamic optimization using overall system mathematical models. Two s-SMDDS employing an air gap membrane distillation module with membrane areas of 11.5 m2 and 23 m2 are analyzed. The lowest water production costs are $5.92/m3 and $5.16/m3 for water production rates of 500 kg/day and 1000 kg/day, respectively. For these two optimal cases, the performance ratios are 0.85 and 0.91; the recovery ratios are 4.07% and 4.57%. The effect of membrane characteristics on the production cost is investigated. For the commercial membrane employed in this study, the increase of the membrane mass transfer coefficient up to two times is beneficial for cost reduction.

Smart membranes for monitoring membrane based desalination processes

KAUST Repository

Laleg-Kirati, Taous-Meriem

2017-10-12

Various examples are related to smart membranes for monitoring membrane based process such as, e.g., membrane distillation processes. In one example, a membrane, includes a porous surface and a plurality of sensors (e.g., temperature, flow and/or impedance sensors) mounted on the porous surface. In another example, a membrane distillation (MD) process includes the membrane. Processing circuitry can be configured to monitor outputs of the plurality of sensors. The monitored outputs can be used to determine membrane degradation, membrane fouling, or to provide an indication of membrane replacement or cleaning. The sensors can also provide temperatures or temperature differentials across the porous surface, which can be used to improve modeling or control the MD process.

Antimicrobial membrane surfaces via efficient polyethyleneimine immobilization and cationization

Science.gov (United States)

Qiu, Wen-Ze; Zhao, Zi-Shu; Du, Yong; Hu, Meng-Xin; Xu, Zhi-Kang

2017-12-01

Biofouling control is a major task in membrane separation processes for water treatment and biomedical applications. In this work, N-alkylated polyethylenimine (PEI) is facilely and efficiently introduced onto the membrane surfaces via the co-deposition of catechol (CCh) and PEI, followed by further grafting of PEIs (600 Da, 70 kDa and 750 kDa) and cationization with methyl iodide (CH3I). The physical and chemical properties of the constructed membrane surfaces are characterized with scanning electron microscopy, Fourier transform infrared spectroscopy, X-ray photoelectron spectroscopy, zeta potential and water contact angle measurements. Antibacterial assay reveals that the optimized membrane surfaces possess around 95% antibacterial efficiency against Gram-positive Staphylococcus aureus (S. aureus) with weak adhesion of bacteria cells after 24 h of bacterial contact. Additionally, the membrane surfaces also exhibit much enhanced antifouling property during the filtration of opposite charged bovine serum albumin (BSA). These results demonstrate a useful strategy for the surface modification of separation membranes by a kind of antimicrobial and antifouling coating.

Proton-Conducting Sulfonated and Phosphonated Polymers and Fuel Cell Membranes by Chemical Modification of Polysulfones

OpenAIRE

Lafitte, Benoit

2007-01-01

The proton exchange membrane fuel cell (PEMFC) is currently emerging as an efficient and environmentally friendly power source. The technology is very complex and relies ultimately on materials and components which need further development. One of the major hurdles for advancing the PEMFC technology is currently the demand for new durable low-cost polymeric membranes that will allow fuel cell operation at high temperatures without extensive humidification requirements. Thus, the design and pr...

Chimaerin suppresses Rac1 activation at the apical membrane to maintain the cyst structure.

Directory of Open Access Journals (Sweden)

Shunsuke Yagi

Full Text Available Epithelial organs are made of a well-polarized monolayer of epithelial cells, and their morphology is maintained strictly for their proper functions. Previously, we showed that Rac1 activation is suppressed at the apical membrane in the mature organoid, and that such spatially biased Rac1 activity is required for the polarity maintenance. Here we identify Chimaerin, a GTPase activating protein for Rac1, as a suppressor of Rac1 activity at the apical membrane. Depletion of Chimaerin causes over-activation of Rac1 at the apical membrane in the presence of hepatocyte growth factor (HGF, followed by luminal cell accumulation. Importantly, Chimaerin depletion did not inhibit extension formation at the basal membrane. These observations suggest that Chimaerin functions as the apical-specific Rac1 GAP to maintain epithelial morphology.

Recent advances on polymeric membranes for membrane reactors

KAUST Repository

Buonomenna, M. G.; Choi, Seung Hak

2012-01-01

. The successful use of membranes in membrane reactors is primary the result of two developments concerning: (i) membrane materials and (ii) membrane structures. The selection of a suited material and preparation technique depends on the application the membrane

An Interactome-Centered Protein Discovery Approach Reveals Novel Components Involved in Mitosome Function and Homeostasis in Giardia lamblia.

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Samuel Rout

2016-12-01

Full Text Available Protozoan parasites of the genus Giardia are highly prevalent globally, and infect a wide range of vertebrate hosts including humans, with proliferation and pathology restricted to the small intestine. This narrow ecological specialization entailed extensive structural and functional adaptations during host-parasite co-evolution. An example is the streamlined mitosomal proteome with iron-sulphur protein maturation as the only biochemical pathway clearly associated with this organelle. Here, we applied techniques in microscopy and protein biochemistry to investigate the mitosomal membrane proteome in association to mitosome homeostasis. Live cell imaging revealed a highly immobilized array of 30-40 physically distinct mitosome organelles in trophozoites. We provide direct evidence for the single giardial dynamin-related protein as a contributor to mitosomal morphogenesis and homeostasis. To overcome inherent limitations that have hitherto severely hampered the characterization of these unique organelles we applied a novel interaction-based proteome discovery strategy using forward and reverse protein co-immunoprecipitation. This allowed generation of organelle proteome data strictly in a protein-protein interaction context. We built an initial Tom40-centered outer membrane interactome by co-immunoprecipitation experiments, identifying small GTPases, factors with dual mitosome and endoplasmic reticulum (ER distribution, as well as novel matrix proteins. Through iterative expansion of this protein-protein interaction network, we were able to i significantly extend this interaction-based mitosomal proteome to include other membrane-associated proteins with possible roles in mitosome morphogenesis and connection to other subcellular compartments, and ii identify novel matrix proteins which may shed light on mitosome-associated metabolic functions other than Fe-S cluster biogenesis. Functional analysis also revealed conceptual conservation of protein

Bone density of defects treated with lyophilized amniotic membrane versus colagen membrane: a tomographic and histomorfogenic study in a rabbi´s femur.

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Liz Ríos

2014-09-01

Full Text Available The aim of this study was to compare the bone density of bone defects treated with lyophilizated amniotic membrane (LAM and collagen Membrane (CM, at 3 and 5 weeks. Two bone defects of 4mm in diameter and 6mm deep were created in left distal femoral diaphysis of New Zealand rabbits (n=12. The animals were randomly divided into 2 groups. One of the defects was covered with lyophilized amniotic membrane (Rosa Chambergo Tissue Bank/National Institute of Child Health-IPEN, Lima, Peru or collagen Membrane (Dentium Co, Seoul, Korea. The second was left uncovered (NC. The rabbits were killed after 3 and 5 weeks (3 rabbits/period. The results showed a high bone density and repair of the defect by new bone. The tomographic study revealed that the bone density of the defects treated with LAM at 3 weeks was equivalent to the density obtained with CM and higher density compared with NC (p0.05. The results show that lyophilizated amniotic membrane provides bone density equal or higher to the collagen membrane.

Recent advances on polymeric membranes for membrane reactors

KAUST Repository

Buonomenna, M. G.

2012-06-24

Membrane reactors are generally applied in high temperature reactions (>400 °C). In the field of fine chemical synthesis, however, much milder conditions are generally applicable and polymeric membranes were applied without their damage. The successful use of membranes in membrane reactors is primary the result of two developments concerning: (i) membrane materials and (ii) membrane structures. The selection of a suited material and preparation technique depends on the application the membrane is to be used in. In this chapter a review of up to date literature about polymers and configuration catalyst/ membranes used in some recent polymeric membrane reactors is given. The new emerging concept of polymeric microcapsules as catalytic microreactors has been proposed. © 2012 Bentham Science Publishers. All rights reserved.

Formation of carrageenan-CaCO{sub 3} bioactive membranes

Energy Technology Data Exchange (ETDEWEB)

Nogueira, Lucas F.B.; Maniglia, Bianca C.; Pereira, Lourivaldo S.; Tapia-Blácido, Delia R.; Ramos, Ana P., E-mail: anapr@ffclrp.usp.br

2016-01-01

The high biocompatibility and resorbability of polymeric membranes have encouraged their use to manufacture medical devices. Here, we report on the preparation of membranes consisting of carrageenan, a naturally occurring sulfated polysaccharide that forms helical structures in the presence of calcium ions. We incorporated CaCO{sub 3} particles into the membranes to enhance their bioactivity and mechanical properties. Infrared spectroscopy and X-ray diffraction data confirmed CaCO{sub 3} incorporation into the polymeric matrix. We tested the bioactivity of the samples by immersing them in a solution that mimics the ionic composition and pH of the human body fluid. The hybrid membranes generated hydroxyapatite, as attested by X-ray diffraction data. Scanning electron and atomic force microscopies aided investigation of membrane topography before and after CaCO{sub 3} deposition. The wettability and surface free energy, evaluated by contact angle measures, increased in the presence of CaCO{sub 3} particles. These parameters are important for membrane implantation in the body. Moreover, membrane stiffness was up to 110% higher in the presence of the inorganic particles, as revealed by Young's modulus. - Highlights: • Hybrid kappa and iota carrageenan-CaCO{sub 3} membranes were formed. • The hybrid membrane's origin hydroxyapatite after exposure to simulated body fluid • The carrageenan's specificity to bind Ca{sup 2+} ions tailors the surface properties.

Sonication-induced Ostwald ripening of ZIF-8 nanoparticles and formation of ZIF-8/polymer composite membranes

KAUST Repository

Thompson, Joshua A.

2012-08-01

The effect of typical membrane processing conditions on the structure, interfacial morphology, and gas separation performance of MOF/polymer nanocomposite membranes is investigated. In particular, the ZIF-8/Matrimid® nanocomposite membrane system is examined, and it is shown that ultrasonication - a commonly employed particle dispersion method - induces significant changes in the shape, size distribution, and structure of ZIF-8 particles suspended in an organic solvent during membrane processing. Dynamic light scattering and electron microscopy reveal that ZIF-8 nanoparticles undergo substantial Ostwald ripening when subjected to high intensity ultrasonication as often required in the formation of MOF/polymer nanocomposite membranes. Other characterization techniques reveal that the ripened particles exhibit lower pore volumes and lower surface areas compared to the as-made material. ZIF-8/Matrimid® composite membranes fabricated using two sonication methods show significant differences in microstructure. Permeation measurements show significant enhancement in permeability of CO 2 and increased CO 2/CH 4 selectivity in membranes fabricated with high-intensity sonication. In contrast, composite membranes prepared with low-intensity sonication are found to be defective. A careful evaluation of MOF membrane processing conditions, as well as knowledge of the properties of the MOF material after these membrane processing steps, are necessary to develop reliable processing-structure-property relations for MOF-containing membranes. © 2012 Elsevier Inc. All rights reserved.

Cellular pH measurements in Emiliania huxleyi reveal pronounced membrane proton permeability.

Science.gov (United States)

Suffrian, K; Schulz, K G; Gutowska, M A; Riebesell, U; Bleich, M

2011-05-01

• To understand the influence of changing surface ocean pH and carbonate chemistry on the coccolithophore Emiliania huxleyi, it is necessary to characterize mechanisms involved in pH homeostasis and ion transport. • Here, we measured effects of changes in seawater carbonate chemistry on the fluorescence emission ratio of BCECF (2',7'-bis-(2-carboxyethyl)-5-(and-6)-carboxyfluorescein) as a measure of intracellular pH (pH(i)). Out of equilibrium solutions were used to differentiate between membrane permeation pathways for H(+), CO(2) and HCO(3)(-). • Changes in fluorescence ratio were calibrated in single cells, resulting in a ratio change of 0.78 per pH(i) unit. pH(i) acutely followed the pH of seawater (pH(e)) in a linear fashion between pH(e) values of 6.5 and 9 with a slope of 0.44 per pH(e) unit. pH(i) was nearly insensitive to changes in seawater CO(2) at constant pH(e) and HCO(3)(-). An increase in extracellular HCO(3)(-) resulted in a slight intracellular acidification. In the presence of DIDS (4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid), a broad-spectrum inhibitor of anion exchangers, E. huxleyi acidified irreversibly. DIDS slightly reduced the effect of pH(e) on pH(i). • The data for the first time show the occurrence of a proton permeation pathway in E. huxleyi plasma membrane. pH(i) homeostasis involves a DIDS-sensitive mechanism. © 2011 The Authors. New Phytologist © 2011 New Phytologist Trust.

Relating membrane potential to impedance spectroscopy

Directory of Open Access Journals (Sweden)

Eugen Gheorghiu

2011-12-01

Full Text Available Non-invasive, label-free assessment of membrane potential of living cells is still a challenging task. The theory linking membrane potential to the low frequency α dispersion exhibited by suspensions of spherical shelled particles (presenting a net charge distribution on the inner side of the shell has been pioneered in our previous studies with emphasis on the permittivity spectra. Whereas α dispersion is related to a rather large variation exhibited by the permittivity spectrum, we report that the related decrement presented by the impedance magnitude spectrum is either extremely small, or occurs (for large cells at very small frequencies (~mHz explaining the lack of experimental bioimpedance data on the matter. We stress that appropriate choice of the parameters (as revealed by the microscopic model may enable access to membrane potential as well as to other relevant parameters when investigating living cells and charged lipid vesicles. We analyse the effect on the low frequency of the permittivity and impedance spectra of: I. Parameters pertaining to cell membrane i.e. (i membrane potential (through the amount of the net charge on the inner side of the membrane, (ii size of the cells/vesicles, (iii conductivity of the membrane; II. Parameters of the extra cellular medium (viscosity and conductivity. The applicability of the study has far reaching implications for basic (life sciences (providing non-invasive access to the dynamics of relevant cell parameters as well as for biosensing applications, e.g. assessment of cytotoxicity of a wide range of stimuli. doi:10.5617/jeb.214 J Electr Bioimp, vol. 2, pp. 93-97, 2011

Influence of post-casting treatments on sulphonated polyetheretherketone composite membranes

Energy Technology Data Exchange (ETDEWEB)

Carbone, Alessandra; Gatto, Irene; Passalacqua, Enza [CNR-ITAE, Institute for Advanced Energy Technologies ' ' N. Giordano' ' Via Salita S. Lucia sopra Contesse, 5 - Messina (Italy); Ohira, Akihiro; Wu, Libin [FC-CUBIC (Polymer Electrolyte Fuel Cell Cutting-Edge Research Center) AIST Tokyo Waterfront, 2-41-6, Aomi, Koto-ku, Tokyo 135-0064 (Japan)

2010-09-15

Since the post-casting treatments influence the water entrapped in polymeric matrix and consequently its proton conductivity, an evaluation of annealing at 200 C and acid treatments was conducted on previously developed composite s-PEEK (1.55 mequiv. g{sup -1}) membranes, containing a commercial aminopropyl-functionalised silica. DSC, WAXS, SEM-EDX and laser microscope measurements carried out on membranes swollen at different temperatures highlighted different membrane properties depending on post-casting treatments. It was found that composite membranes have different structural and morphological characteristics than pristine polymer membranes. The silica distribution was modified when different treatments are used. The state of water changed when silica was inserted into the membranes. Actually, contrary to the pristine membranes the presence of freezable water was revealed at temperature lower than 80 C. The proton conductivity was also affected by the presence and the amount of water trapped into the membranes and was particularly influenced by the post-casting treatments. The silica introduction reduced the swelling effect and improved the robustness of the membranes even if a higher water content in the freezable state was observed. Acid treatment leads to significant improvement in membrane properties, but the present work shows that annealing before acid treatment can affect the membrane morphology more strongly than other treatments resulting in a much better fuel cell performance. (author)

The influence of saponins on cell membrane cholesterol.

Science.gov (United States)

Böttger, Stefan; Melzig, Matthias F

2013-11-15

We studied the influence of structurally different saponins on the cholesterol content of cellular membranes. Therefore a cell culture model using ECV-304 urinary bladder carcinoma cells was developed. To measure the cholesterol content we used radiolabeled (3)H-cholesterol which is chemically and physiologically identical to natural cholesterol. The cells were pre-incubated with (3)H-cholesterol and after a medium change, they were treated with saponins to assess a saponin-induced cholesterol liberation from the cell membrane. In another experiment the cells were pre-incubated with saponins and after a medium change, they were treated with (3)H-cholesterol to assess a saponin-induced inhibition of cholesterol uptake into the cell membrane. Furthermore, the membrane toxicity of all applied saponins was analyzed using extracellular LDH quantification and the general cytotoxicity was analyzed using a colorimetric MTT-assay and DNA quantification. Our results revealed a correlation between membrane toxicity and general cytotoxicity. We also compared the results from the experiments on the saponin-induced cholesterol liberation as well as the saponin-induced inhibition of cholesterol uptake with the membrane toxicity. A significant reduction in the cell membrane cholesterol content was noted for those saponins who showed membrane toxicity (IC50 saponins either liberated (3)H-cholesterol from intact cell membranes or blocked the integration of supplemented (3)H-cholesterol into the cell membrane. Saponins with little influence on the cell membrane (IC50 >100 μM) insignificantly altered the cell membrane cholesterol content. The results suggested that the general cytotoxicity of saponins is mainly dependent on their membrane toxicity and that the membrane toxicity might be caused by the loss of cholesterol from the cell membrane. We also analyzed the influence of a significantly membrane toxic saponin on the cholesterol content of intracellular membranes such as those

Metagenomes reveal microbial structures, functional potentials, and biofouling-related genes in a membrane bioreactor.

Science.gov (United States)

Ma, Jinxing; Wang, Zhiwei; Li, Huan; Park, Hee-Deung; Wu, Zhichao

2016-06-01

Metagenomic sequencing was used to investigate the microbial structures, functional potentials, and biofouling-related genes in a membrane bioreactor (MBR). The results showed that the microbial community in the MBR was highly diverse. Notably, function analysis of the dominant genera indicated that common genes from different phylotypes were identified for important functional potentials with the observation of variation of abundances of genes in a certain taxon (e.g., Dechloromonas). Despite maintaining similar metabolic functional potentials with a parallel full-scale conventional activated sludge (CAS) system due to treating the identical wastewater, the MBR had more abundant nitrification-related bacteria and coding genes of ammonia monooxygenase, which could well explain its excellent ammonia removal in the low-temperature period. Furthermore, according to quantification of the genes involved in exopolysaccharide and extracellular polymeric substance (EPS) protein metabolism, the MBR did not show a much different potential in producing EPS compared to the CAS system, and bacteria from the membrane biofilm had lower abundances of genes associated with EPS biosynthesis and transport compared to the activated sludge in the MBR.

Calibration procedures for the quantitative determination of membrane potential in human cells using anionic dyes.

Science.gov (United States)

Klapperstück, Thomas; Glanz, Dagobert; Hanitsch, Stefan; Klapperstück, Manuela; Markwardt, Fritz; Wohlrab, Johannes

2013-07-01

Quantitative determinations of the cell membrane potential of lymphocytes (Wilson et al., J Cell Physiol 1985;125:72-81) and thymocytes (Krasznai et al., J Photochem Photobiol B 1995;28:93-99) using the anionic dye DiBAC4 (3) proved that dye depletion in the extracellular medium as a result of cellular uptake can be negligible over a wide range of cell densities. In contrast, most flow cytometric studies have not verified this condition but rather assumed it from the start. Consequently, the initially prepared extracellular dye concentration has usually been used for the calculation of the Nernst potential of the dye. In this study, however, external dye depletion could be observed in both large IGR-1 and small LCL-HO cells under experimental conditions, which have often been applied routinely in spectrofluorimetry and flow cytometry. The maximum cell density at which dye depletion could be virtually avoided was dependent on cell size and membrane potential and definitely needed to be taken into account to ensure reliable results. In addition, accepted calibration procedures based on the partition of sodium and potassium (Goldman-Hodgkin-Katz equation) or potassium alone (Nernst equation) were performed by flow cytometry on cell suspensions with an appropriately low cell density. The observed extensive lack of concordance between the correspondingly calculated membrane potential and the equilibrium potential of DiBAC4 (3) revealed that these methods require the additional measurement of cation parameters (membrane permeability and/or intracellular concentration). In contrast, due to the linear relation between fluorescence and low DiBAC4 (3) concentrations, the Nernst potential of the dye for totally depolarized cells can be reliably used for calibration with an essentially lower effort and expense. Copyright © 2013 International Society for Advancement of Cytometry.

Swift heavy ion induced modification in polycarbonate membrane for gas separation

International Nuclear Information System (INIS)

Rajesh Kumar; Prasad, Rajendra; Vijay, Y.K.; Das, D.

2003-01-01

Polymeric membranes are extensively used for commercial gas separation applications. Makrofol-KG (polycarbonate) is a glassy polymer. 40 μm thick sheet of Makrofol-KG was irradiated with 40 Ar (14.9 MeV/n) of fluence 10 3 ions/cm 2 and 20 μm thick sheet with 5.3 MeV α-particles of fluence 10 7 ions/cm 2 . The permeability of these polycarbonate membranes for H 2 and CO 2 was measured and also after etching in 6 N NaOH at 60 degC for different periods. Permeability is found to be increased with etching time. At a definite time, critical etching time, the permeability rapidly increases in PC. Positron annihilation lifetimes for unirradiated and irradiated membranes were measured with fast fast coincidence system to study the correlation of free volume hole concentration with gas separation properties. (author)

Rural Development And Agricultural Extension Administration In ...

African Journals Online (AJOL)

This paper reviewed the wide range of policies and approaches formulated and implemented to effect agricultural and rural development in Nigeria. The paper reveals that the common feature of all the strategies is the use of institutionalized agricultural extension service, devoted principally to augment smallholder ...

Proton-conducting ionic liquid-based proton exchange membrane fuel cell membranes: The key role of ionomer-ionic liquid interaction

Energy Technology Data Exchange (ETDEWEB)

Martinez, Mathieu; Cointeaux, Laure; Iojoiu, Cristina; Lepretre, Jean-Claude; Sanchez, Jean-Yves [LEPMI, UMR 5631, CNRS-INP-UJF, PHELMA-Campus, BP.75, 1130 rue de la Piscine, 38402 Saint-Martin-d' Heres Cedex (France); Molmeret, Yannick; El Kissi, Nadia [Laboratoire de Rheologie, UMR 5520 CNRS-INPG-UJF, ENSHMG, BP 53, 38041 Grenoble (France); Judeinstein, Patrick [Institut de Chimie Moleculaire et des Materiaux d' Orsay (UMR 8182), Batiment 410, Universite Paris-Sud 11, 91405 Orsay Cedex (France)

2010-09-15

The paper deals with the synthesis and characterisation of proton-conducting ionic liquids (PCILs) and their polymer electrolytes obtained by blending modified Nafion membranes with different concentrations of PCILs. The PCILs are obtained by the neutralization of triethylamine with different organic acids. The first part of the paper studies the influence of acidity and acid structure on PCIL thermal and electrochemical performance, while the second part examines membrane conductivity and reveals it to depend more on PCIL structure than on its intrinsic conductivity. At 130 C, conductivities exceeding 10 mS cm{sup -1} were obtained in fully anhydrous conditions. (author)

Assessing the nature of lipid raft membranes

DEFF Research Database (Denmark)

Niemelä, Perttu S; Ollila, Samuli; Hyvönen, Marja T

2007-01-01

of highly ordered lateral domains rich in sphingomyelin and cholesterol (CHOL). These domains, called functional lipid rafts, have been suggested to take part in a variety of dynamic cellular processes such as membrane trafficking, signal transduction, and regulation of the activity of membrane proteins......-scale simulations to elucidate the properties of ternary raft mixtures with CHOL, palmitoylsphingomyelin (PSM), and palmitoyloleoylphosphatidylcholine. We simulate two bilayers of 1,024 lipids for 100 ns in the liquid-ordered phase and one system of the same size in the liquid-disordered phase. The studies provide...... heterogeneity more difficult. The findings reveal aspects of the role of favored (specific) lipid-lipid interactions within rafts and clarify the prominent role of CHOL in altering the properties of the membrane locally in its neighborhood. Also, we show that the presence of PSM and CHOL in rafts leads...

Ligand and membrane-binding behavior of the phosphatidylinositol transfer proteins PITPα and PITPβ.

Science.gov (United States)

Baptist, Matilda; Panagabko, Candace; Cockcroft, Shamshad; Atkinson, Jeffrey

2016-12-01

Phosphatidylinositol transfer proteins (PITPs) are believed to be lipid transfer proteins because of their ability to transfer either phosphatidylinositol (PI) or phosphatidylcholine (PC) between membrane compartments, in vitro. However, the detailed mechanism of this transfer process is not fully established. To further understand the transfer mechanism of PITPs we examined the interaction of PITPs with membranes using dual polarization interferometry (DPI), which measures protein binding affinity on a flat immobilized lipid surface. In addition, a fluorescence resonance energy transfer (FRET)-based assay was also employed to monitor how quickly PITPs transfer their ligands to lipid vesicles. DPI analysis revealed that PITPβ had a higher affinity to membranes compared with PITPα. Furthermore, the FRET-based transfer assay revealed that PITPβ has a higher ligand transfer rate compared with PITPα. However, both PITPα and PITPβ demonstrated a preference for highly curved membrane surfaces during ligand transfer. In other words, ligand transfer rate was higher when the accepting vesicles were highly curved.

Positron annihilation lifetime study of Nafion/titanium dioxide nano-composite membranes

Science.gov (United States)

Lei, M.; Wang, Y. J.; Liang, C.; Huang, K.; Ye, C. X.; Wang, W. J.; Jin, S. F.; Zhang, R.; Fan, D. Y.; Yang, H. J.; Wang, Y. G.

2014-01-01

Positron annihilation lifetime (PAL) technique is applied for investigation of size and number density of free volumes in Nafion/TiO2-nanoparticles composite membrane. The proton transporting ability is correlated with the properties of free volume inside the membrane. It is revealed that composite membrane with 5 wt% of TiO2 nano-fillers exhibits good electrochemical performance under reduced humidity and it can be saturated with water at relative humidity of 50%, under which ionic clusters and proton transporting channels are formed, indicating that composite membranes with 5 wt% of TiO2 nano-fillers are effective electrolyte for fuel cells operated at reduced humidification levels. The results suggest that PAL can be a powerful tool for elucidating the relationship between microstructure and ion transport in polymer electrolyte membranes.

Phosphoric acid doped polysulfone membranes with aminopyridine pendant groups and imidazole cross-links

DEFF Research Database (Denmark)

Hink, Steffen; Elsøe, Katrine; Cleemann, Lars Nilausen

2015-01-01

Udel polysulfone based membranes with 4-aminopyridine pendant groups and cross-linking imidazole units are synthesized in a simple two step reaction. The ratio of 4-aminopyridine and imidazole is varied and the materials are extensively characterized. The average phosphoric acid uptake (in 85 wt%...

Demonstration of biased membrane static figure mapping by optical beam subpixel centroid shift

Energy Technology Data Exchange (ETDEWEB)

Pinto, Fabrizio, E-mail: fpinto@jazanu.edu.sa [Laboratory for Quantum Vacuum Applications, Department of Physics, Faculty of Science, Jazan University, P.O. Box 114, Gizan 45142 (Saudi Arabia)

2016-06-10

The measurement of Casimir forces by means of condenser microphones has been shown to be quite promising since its early introduction almost half-a-century ago. However, unlike the remarkable progress achieved in characterizing the vibrating membrane in the dynamical case, the accurate determination of the membrane static figure under electrostatic bias remains a challenge. In this paper, we discuss our first data obtained by measuring the centroid shift of an optical beam with subpixel accuracy by charge coupled device (CCD) and by an extensive analysis of noise sources present in the experimental setup.

Retention of Silica Nanoparticles in a Lab-Scale Membrane Bioreactor: Implications for Process Performance and Membrane Fouling

Directory of Open Access Journals (Sweden)

Mark Larracas Sibag

2016-07-01

Full Text Available In conventional activated sludge (CAS involving aerobic biological processes, the retention of silica nanoparticles (SiO2 NPs has no detrimental effect on chemical oxygen demand (COD and ammonia nitrogen (NH3–N removal. However, for the membrane bioreactor (MBR system, which is also based on the activated sludge process in addition to the membrane separation process, it has implications not only on the process performance but also on membrane fouling. To investigate these two implications in lab-scale experiments, we continuously operated a control MBR and two experimental MBRs, in which the 28 nm SiO2 NPs and 144 nm SiO2 NPs were added separately to the influent at a final concentration of 100 mg/L. Although the retention of SiO2 NPs in the MBR, as confirmed by dynamic light scattering (DLS analysis, did not compromise the COD and NH3–N removal, it resulted in substantial increases in the transmembrane pressure (TMP suggesting the onset of membrane fouling. Analyses by batch-dead end filtration revealed the same fouling trend as observed during the continuous MBR experiments; membrane fouling is aggravated in the presence of SiO2 NPs. This was evident from permeate flux decline of between 30% and 74% at very low TMP (5 kPa and the further increases in the total resistance.

Calcineurin signaling and membrane lipid homeostasis regulates iron mediated multidrug resistance mechanisms in Candida albicans.

Directory of Open Access Journals (Sweden)

Saif Hameed

2011-04-01

Full Text Available We previously demonstrated that iron deprivation enhances drug susceptibility of Candida albicans by increasing membrane fluidity which correlated with the lower expression of ERG11 transcript and ergosterol levels. The iron restriction dependent membrane perturbations led to an increase in passive diffusion and drug susceptibility. The mechanisms underlying iron homeostasis and multidrug resistance (MDR, however, are not yet resolved. To evaluate the potential mechanisms, we used whole genome transcriptome and electrospray ionization tandem mass spectrometry (ESI-MS/MS based lipidome analyses of iron deprived Candida cells to examine the new cellular circuitry of the MDR of this pathogen. Our transcriptome data revealed a link between calcineurin signaling and iron homeostasis. Among the several categories of iron deprivation responsive genes, the down regulation of calcineurin signaling genes including HSP90, CMP1 and CRZ1 was noteworthy. Interestingly, iron deprived Candida cells as well as iron acquisition defective mutants phenocopied molecular chaperone HSP90 and calcineurin mutants and thus were sensitive to alkaline pH, salinity and membrane perturbations. In contrast, sensitivity to above stresses did not change in iron deprived DSY2146 strain with a hyperactive allele of calcineurin. Although, iron deprivation phenocopied compromised HSP90 and calcineurin, it was independent of protein kinase C signaling cascade. Notably, the phenotypes associated with iron deprivation in genetically impaired calcineurin and HSP90 could be reversed with iron supplementation. The observed down regulation of ergosterol (ERG1, ERG2, ERG11 and ERG25 and sphingolipid biosynthesis (AUR1 and SCS7 genes followed by lipidome analysis confirmed that iron deprivation not only disrupted ergosterol biosynthesis, but it also affected sphingolipid homeostasis in Candida cells. These lipid compositional changes suggested extensive remodeling of the membranes in iron

Characterization of retrokeratoprosthetic membranes in the Boston type 1 keratoprosthesis.

Science.gov (United States)

Stacy, Rebecca C; Jakobiec, Frederick A; Michaud, Norman A; Dohlman, Claes H; Colby, Kathryn A

2011-03-01

To evaluate retroprosthetic membranes that can occur in 25% to 65% of patients with the Boston type 1 keratoprosthesis (KPro). Two patients with Peter anomaly and 2 with neurotrophic scarred corneas underwent revisions of their type 1 KPros because of visually compromising retroprosthetic membranes. The excised membranes were studied by light microscopy with hematoxylin-eosin, periodic acid-Schiff, and toluidine blue stains. Immunohistochemical and transmission electron microscopic examination were also used. Light microscopic examination revealed that the retro-KPro fibrous membranes originated from the host's corneal stroma. These mildly to moderately vascularized membranes grew through gaps in the Descemet membrane to reach behind the KPro back plate and adhere to the anterior iris surface, which had undergone partial lysis. In 2 cases, the fibrous membranes merged at the pupil with matrical portions of metaplastic lens epithelium, forming a bilayered structure that crossed the optical axis. Retro-KPro membranes stained positively for α-smooth muscle actin but negatively for pancytokeratin. Electron microscopy confirmed the presence of actin filaments within myofibroblasts and small surviving clusters of metaplastic lens epithelial cells. Stromal downgrowth, rather than epithelial downgrowth, was the major element of the retro-KPro membranes in this series. Metaplastic lens epithelium also contributed to opacification of the visual axis. Florid membranous inflammation was not a prominent finding and thus probably not a requisite stimulus for membrane development. Further advances in prosthetic design and newer antifibroproliferative agents may reduce membrane formation.

Metastatic Adenocarcinoma Presenting as Extensive Cavoatrial Tumor Thrombus

International Nuclear Information System (INIS)

Johari, Bushra; Abdul Aziz, Yang Faridah; Krishnasamy, Sivakumar; Looi, Lai Meng; Hashim, Shahrul Amry; Raja Mokhtar, Raja Amin

2015-01-01

The presence of tumor thrombus in the right atrium is frequently the result of direct intraluminal extension of infra-diaphragmatic malignancy into the inferior vena cava (IVC) or supradiaphragmatic carcinoma into the superior vena cava (SVC). Right atrial tumor thrombus with extension into both SVC and IVC has not been reported in the literature. We present a patient who presented with symptoms of right atrial and SVC obstruction. Imaging revealed presence of a thrombus in the right atrium, extending to the SVC and IVC, with the additional findings of a left adrenal mass and multiple liver lesions. The histopathological examination of the right atrial mass revealed metastatic adenocarcinoma cells. The patient was given a presumptive diagnosis of metastatic adenocarcinoma, most likely adrenal in origin, with multiple hepatic lesions suspicious for metastasis. The clinical outcome of the patient was not favorable; the patient succumbed before the adrenal mass could be confirmed to be the primary tumor. This case highlights that in patients manifesting with extensive cavoatrial thrombus as, the existence of primary carcinoma should be considered especially in the adrenal cortex or in the lung

Expression analysis of two gene subfamilies encoding the plasma membrane H+-ATPase in Nicotiana plumbaginifolia reveals the major transport functions of this enzyme.

Science.gov (United States)

Moriau, L; Michelet, B; Bogaerts, P; Lambert, L; Michel, A; Oufattole, M; Boutry, M

1999-07-01

The plasma membrane H+-ATPase couples ATP hydrolysis to proton transport, thereby establishing the driving force for solute transport across the plasma membrane. In Nicotiana plumbaginifolia, this enzyme is encoded by at least nine pma (plasma membrane H+-ATPase) genes. Four of these are classified into two gene subfamilies, pma1-2-3 and pma4, which are the most highly expressed in plant species. We have isolated genomic clones for pma2 and pma4. Mapping of their transcript 5' end revealed the presence of a long leader that contained small open reading frames, regulatory features typical of other pma genes. The gusA reporter gene was then used to determine the expression of pma2, pma3 and pma4 in N. tabacum. These data, together with those obtained previously for pma1, led to the following conclusions. (i) The four pma-gusA genes were all expressed in root, stem, leaf and flower organs, but each in a cell-type specific manner. Expression in these organs was confirmed at the protein level, using subfamily-specific antibodies. (ii) pma4-gusA was expressed in many cell types and notably in root hair and epidermis, in companion cells, and in guard cells, indicating that in N. plumbaginifolia the same H+-ATPase isoform might be involved in mineral nutrition, phloem loading and control of stomata aperture. (iii) The second gene subfamily is composed, in N. plumbaginifolia, of a single gene (pma4) with a wide expression pattern and, in Arabidopsis thaliana, of three genes (aha1, aha2, aha3), at least two of them having a more restrictive expression pattern. (iv) Some cell types expressed pma2 and pma4 at the same time, which encode H+-ATPases with different enzymatic properties.

SEPARATION OF HAZARDOUS ORGANICS BY LOW PRESSURE REVERSE OSMOSIS MEMBRANES - PHASE II FINAL REPORT

Science.gov (United States)

Extensive experimental studies showed that thin-film, composite membranes can be used effectively for the separation of selected hazardous organic compounds. This waste treatment technique offers definite advantages in terms of high solute separations at low pressures (<2MPa) and...

Membrane fouling mechanism of biofilm-membrane bioreactor (BF-MBR): Pore blocking model and membrane cleaning.

Science.gov (United States)

Zheng, Yi; Zhang, Wenxiang; Tang, Bing; Ding, Jie; Zheng, Yi; Zhang, Zhien

2018-02-01

Biofilm membrane bioreactor (BF-MBR) is considered as an important wastewater treatment technology that incorporates advantages of both biofilm and MBR process, as well as can alleviate membrane fouling, with respect to the conventional activated sludge MBR. But, to be efficient, it necessitates the establishment of proper methods for the assessment of membrane fouling. Four Hermia membrane blocking models were adopted to quantify and evaluate the membrane fouling of BF-MBR. The experiments were conducted with various operational conditions, including membrane types, agitation speeds and transmembrane pressure (TMP). Good agreement between cake formation model and experimental data was found, confirming the validity of the Hermia models for assessing the membrane fouling of BF-MBR and that cake layer deposits on membrane. Moreover, the influences of membrane types, agitation speeds and transmembrane pressure on the Hermia pore blocking coefficient of cake layer were investigated. In addition, the permeability recovery after membrane cleaning at various operational conditions was studied. This work confirms that, unlike conventional activated sludge MBR, BF-MBR possesses a low degree of membrane fouling and a higher membrane permeability recovery after cleaning. Copyright © 2017 Elsevier Ltd. All rights reserved.

Microfluidic systems with ion-selective membranes.

Science.gov (United States)

Slouka, Zdenek; Senapati, Satyajyoti; Chang, Hsueh-Chia

2014-01-01

When integrated into microfluidic chips, ion-selective nanoporous polymer and solid-state membranes can be used for on-chip pumping, pH actuation, analyte concentration, molecular separation, reactive mixing, and molecular sensing. They offer numerous functionalities and are hence superior to paper-based devices for point-of-care biochips, with only slightly more investment in fabrication and material costs required. In this review, we first discuss the fundamentals of several nonequilibrium ion current phenomena associated with ion-selective membranes, many of them revealed by studies with fabricated single nanochannels/nanopores. We then focus on how the plethora of phenomena has been applied for transport, separation, concentration, and detection of biomolecules on biochips.

Sulfonated polyimides containing triphenylphosphine oxide for proton exchange membranes

Energy Technology Data Exchange (ETDEWEB)

Mandal, Arun Kumar; Bera, Debaditya; Banerjee, Susanta, E-mail: susanta@matsc.iitkgp.ernet.in

2016-09-15

A series of sulfonated co-polyimides (co-SPI) were prepared by one pot polycondensation reaction of a combination of diamines namely; 4,4′-diaminostilbene-2,2′-disulfonic acid (DSDSA) and prepared non-sulfonated diamine (DATPPO) containing triphenylphosphine oxide with 1,4,5,8-naphthalenetetracarboxylic dianhydride (NTDA). All these soluble co-SPI gave flexible membranes with high thermal stability and showed good mechanical property. Transmission electron microscopy (TEM) analysis revealed the microphase separated morphology with well-dispersed hydrophilic (cluster size in the range of 5–55 nm) domains. The co-SPI membranes showed high oxidative and hydrolytic stability with higher proton conductivity. All these co-SPI membranes exhibited low water uptake and swelling ratio. The co-SPI membrane TPPO-60 (60% degree of sulfonation) with IEC{sub W} = 1.84 mequiv g{sup −1} showed high proton conductivity (99 mS cm{sup −1} at 80 °C and 107 mS cm{sup −1} at 90 °C) in water with high oxidative (20 h) and hydrolytic stability (only 5% degradation in 24 h). - Highlights: • Triphenylphosphine oxide containing sulfonated polyimides (SPIs) was synthesized. • The SPIs showed good oxidative and hydrolytic stability and high proton conductivity. • TEM analysis revealed well separated morphology of the SPIs.

Spectral studies of Lanthanide interactions with membrane surfaces

Energy Technology Data Exchange (ETDEWEB)

Karukstis, K.K.; Kao, M.Y.; Savin, D.A.; Bittker, R.A.; Kaphengst, K.J.; Emetarom, C.M.; Naito, N.R.; Takamoto, D.Y. [Harvey Mudd College, Claremont, CA (United States)

1995-03-23

We have monitored the interactions of the series of trivalent lanthanide cations with the thylakoid membrane surface of spinach chloroplasts using two complementary spectral techniques. Measurements of the fluorescence emission of the extrinsic probe 2-p-toluidinonaphthalene-6-sulfonate (TNS) and the absorbance of the intrinsic chromophore chlorophyll provide two sensitive means of characterizing the dependence of the cation-membrane interaction on the nature of the cation. In these systems, added lanthanide cations adsorb onto the membrane surface to neutralize exposed segments of membrane-embedded protein complexes. The lanthanide-induced charge neutralization increases the proximity of added TNS anion to the membrane surface as evidenced by variations in the TNS fluorescence level and wavelength of maximum emission. Our results reveal a strong dependence of TNS fluorescence parameters on both lanthanide size and total orbital angular momentum L value. Lanthanides with greater charge density (small size and/or low L value) enhance the TNS fluorescence level to a greater extent. A possible origin for the lanthanide-dependent TNS fluorescence levels is suggested in terms of a heterogeneity in the number and type of TNS binding sites. The data are consistent with the proposal that larger lanthanides with smaller enthalpies of hydration induce more significant membrane appression. 59 refs., 9 figs., 2 tabs.

Elastic properties of ultrathin diamond/AlN membranes

International Nuclear Information System (INIS)

Zuerbig, V.; Hees, J.; Pletschen, W.; Sah, R.E.; Wolfer, M.; Kirste, L.; Heidrich, N.; Nebel, C.E.; Ambacher, O.; Lebedev, V.

2014-01-01

Nanocrystalline diamond- (NCD) and AlN-based ultrathin single layer and bilayer membranes are investigated towards their mechanical properties. It is shown that chemo-mechanical polishing and heavy boron doping of NCD thin films do not impact the elastic properties of NCD layers as revealed by negligible variations of the NCD Young's modulus (E). In addition, it is demonstrated that the combination of NCD elastic layer and AlN piezo-actuator is highly suitable for the fabrication of mechanically stable ultrathin membranes in comparison to AlN single layer membranes. The elastic parameters of NCD/AlN heterostructures are mainly determined by the outstanding high Young's modulus of NCD (E = 1019 ± 19 GPa). Such ultrathin unimorph membranes allow for fabrication of piezo-actuated AlN/NCD microlenses with tunable focus length. - Highlights: • Mechanical properties of nanocrystalline diamond (NCD) and AlN circular membranes • No influence of polishing of NCD thin films on the mechanical properties of NCD • No influence of heavy boron-doping on the mechanical properties of NCD • Demonstration of mechanically stable piezo-actuated NCD/AlN membranes • Reported performance of AlN/NCD microlenses with adjustable focus length

[Ultrastructural organization of cytoplasmatic membrane of Anaerobacter polyendosporus studied by electron microscopic cryofractography].

Science.gov (United States)

Duda, V I; Suzina, N E; Dmitriev, V V

2001-01-01

Anaerobacter polyendosporus cells do not have typical mesosomes. However, the analysis of this anaerobic multispore bacterium by electron microscopic cryofractography showed that its cytoplasmic membrane contains specific intramembrane structures in the form of flat lamellar inverted lipid membranes tenths of nanometers to several microns in size. It was found that these structures are located in the hydrophobic interior between the outer and inner leaflets of the cytoplasmic membrane and do not contain intramembrane particles that are commonly present on freeze-fracture replicas. The flat inverted lipid membranes were revealed in bacterial cells cultivated under normal growth conditions, indicating the existence of a complex-type compartmentalization in biological membranes, which manifests itself in the formation of intramembrane compartments having the appearance of vesicles and inverted lipid membranes.

Coordination of membrane and actin cytoskeleton dynamics during filopodia protrusion.

Directory of Open Access Journals (Sweden)

Changsong Yang

2009-05-01

Full Text Available Leading edge protrusion of migrating cells involves tightly coordinated changes in the plasma membrane and actin cytoskeleton. It remains unclear whether polymerizing actin filaments push and deform the membrane, or membrane deformation occurs independently and is subsequently stabilized by actin filaments. To address this question, we employed an ability of the membrane-binding I-BAR domain of IRSp53 to uncouple the membrane and actin dynamics and to induce filopodia in expressing cells. Using time-lapse imaging and electron microscopy of IRSp53-I-BAR-expressing B16F1 melanoma cells, we demonstrate that cells are not able to protrude or maintain durable long extensions without actin filaments in their interior, but I-BAR-dependent membrane deformation can create a small and transient space at filopodial tips that is subsequently filled with actin filaments. Moreover, the expressed I-BAR domain forms a submembranous coat that may structurally support these transient actin-free protrusions until they are further stabilized by the actin cytoskeleton. Actin filaments in the I-BAR-induced filopodia, in contrast to normal filopodia, do not have a uniform length, are less abundant, poorly bundled, and display erratic dynamics. Such unconventional structural organization and dynamics of actin in I-BAR-induced filopodia suggests that a typical bundle of parallel actin filaments is not necessary for generation and mechanical support of the highly asymmetric filopodial geometry. Together, our data suggest that actin filaments may not directly drive the protrusion, but only stabilize the space generated by the membrane deformation; yet, such stabilization is necessary for efficient protrusion.

Flapping Tail Membrane in Bats Produces Potentially Important Thrust during Horizontal Takeoffs and Very Slow Flight

Science.gov (United States)

Adams, Rick A.; Snode, Emily R.; Shaw, Jason B.

2012-01-01

Historically, studies concerning bat flight have focused primarily on the wings. By analyzing high-speed video taken on 48 individuals of five species of vespertilionid bats, we show that the capacity to flap the tail-membrane (uropatagium) in order to generate thrust and lift during takeoffs and minimal-speed flight (ecomorphology. The extensive fanning motion of the tail membrane by vespertilionid bats has not been reported for other flying vertebrates. PMID:22393378

Examination of Amine-Functionalised Anion-Exchange Membranes for Possible Use in the All-Vanadium Redox Flow Battery

International Nuclear Information System (INIS)

Mallinson, Sarah L.; Varcoe, John R.; Slade, Robert C.T.

2014-01-01

The applicability of amine-functionalised anion-exchange membranes (AEMs) for use in the all-vanadium redox flow battery has been studied. A selection of radiation-grafted aminated membranes functionalised with dimethylamine, trimethylamine or diazabicyclo(2,2,2)octane were extensively tested. The success of each grafting process was confirmed by Raman and infrared spectroscopies, titrimetry and ionic conductivity measurements. The amine-functionalised membranes were found to have poor thermo-oxidative stability and high vanadium cation permeabilities. The results highlight the importance of balancing ionic conductivity with vanadium cation permeability and indicate that amine-based functional groups may not be suitably stable for the membranes to remain true AEMs when in use in the all-vanadium redox flow battery

Heat stress causes spatially-distinct membrane re-modelling in K562 leukemia cells.

Directory of Open Access Journals (Sweden)

Gábor Balogh

Full Text Available Cellular membranes respond rapidly to various environmental perturbations. Previously we showed that modulations in membrane fluidity achieved by heat stress (HS resulted in pronounced membrane organization alterations which could be intimately linked to the expression and cellular distribution of heat shock proteins. Here we examine heat-induced membrane changes using several visualisation methods. With Laurdan two-photon microscopy we demonstrate that, in contrast to the enhanced formation of ordered domains in surface membranes, the molecular disorder is significantly elevated within the internal membranes of cells preexposed to mild HS. These results were compared with those obtained by anisotropy, fluorescence lifetime and electron paramagnetic resonance measurements. All probes detected membrane changes upon HS. However, the structurally different probes revealed substantially distinct alterations in membrane heterogeneity. These data call attention to the careful interpretation of results obtained with only a single label. Subtle changes in membrane microstructure in the decision-making of thermal cell killing could have potential application in cancer therapy.

Medulloblastoma with extensive nodularity: US, CT and MRI findings

International Nuclear Information System (INIS)

Bekiesinska-Figatowska, M.; Uliasz, M.; Roszkowski, M.

2008-01-01

Medulloblastoma accounts for up to 25% of all paediatric CNS tumours. According to WHO classification (2007) medulloblastoma with extensive nodularity (MBEN) is a separate rare entity associated with younger age and better prognosis. A 9-month-old girl was admitted and examined because of macrocephaly and disturbed psychomotor development. Transfontanel ultrasound revealed dilated ventricular system and hyperechoic mass in the posterior cranial fossa. Computed tomography showed hyperdense mass in the cerebellum. Magnetic resonance imaging revealed a mass with gyriform pattern and strong contrast enhancement after gadolinium administration. Differential diagnosis included dysplastic heterotopic cortex, Lhermitte-Duclos disease, atypical teratoid/rhabdoid tumour (AT/RT), and medulloblastoma (MB). The patient was operated on. Medulloblastoma with extensive nodularity (MBEN) was finally diagnosed. The authors present and discuss three other cases of this rare entity.Transfontanel sonografic examination is capable of detection of the posterior fossa tumour as a cause of hydrocephalus and macrocephaly. The mass in a child's posterior cranial fossa that is hyperdense on unenhanced CT and gyriform, nodular, and markedly enhancing on MRI may strongly suggest medulloblastoma with extensive nodularity (MBEN). (authors)

Membrane dynamics

DEFF Research Database (Denmark)

Bendix, Pól Martin

2015-01-01

Current topics include membrane-protein interactions with regard to membrane deformation or curvature sensing by BAR domains. Also, we study the dynamics of membrane tubes of both cells and simple model membrane tubes. Finally, we study membrane phase behavior which has important implications...... for the lateral organization of membranes as wells as for physical properties like bending, permeability and elasticity...

Inverse colloidal crystal membranes for hydrophobic interaction membrane chromatography.

Science.gov (United States)

Vu, Anh T; Wang, Xinying; Wickramasinghe, S Ranil; Yu, Bing; Yuan, Hua; Cong, Hailin; Luo, Yongli; Tang, Jianguo

2015-08-01

Hydrophobic interaction membrane chromatography has gained interest due to its excellent performance in the purification of humanized monoclonal antibodies. The membrane material used in hydrophobic interaction membrane chromatography has typically been commercially available polyvinylidene fluoride. In this contribution, newly developed inverse colloidal crystal membranes that have uniform pores, high porosity and, therefore, high surface area for protein binding are used as hydrophobic interaction membrane chromatography membranes for humanized monoclonal antibody immunoglobulin G purification. The capacity of the inverse colloidal crystal membranes developed here is up to ten times greater than commercially available polyvinylidene fluoride membranes with a similar pore size. This work highlights the importance of developing uniform pore size high porosity membranes in order to maximize the capacity of hydrophobic interaction membrane chromatography. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Annexins are instrumental for efficient plasma membrane repair in cancer cells.

Science.gov (United States)

Lauritzen, Stine Prehn; Boye, Theresa Louise; Nylandsted, Jesper

2015-09-01

Plasma membrane stress can cause damage to the plasma membrane, both when imposed by the extracellular environment and by enhanced oxidative stress. Cells cope with these injuries by rapidly activating their plasma membrane repair system, which is triggered by Ca(2+) influx at the wound site. The repair system is highly dynamic, depends on both lipid and protein components, and include cytoskeletal reorganization, membrane replacements, and membrane fusion events. Cancer cells experience enhanced membrane stress when navigating through dense extracellular matrix, which increases the frequency of membrane injuries. In addition, increased motility and oxidative stress further increase the risk of plasma membrane lesions. Cancer cells compensate by overexpressing Annexin proteins including Annexin A2 (ANXA2). Annexin family members can facilitate membrane fusion events and wound healing by binding to negatively charged phospholipids in the plasma membrane. Plasma membrane repair in cancer cells depends on ANXA2 protein, which is recruited to the wound site and forms a complex with the Ca(2+)-binding EF-hand protein S100A11. Here they regulate actin accumulation around the wound perimeter, which is required for wound closure. In this review, we will discuss the requirement for Annexins, S100 proteins and actin cytoskeleton in the plasma membrane repair response of cancer cells, which reveals a novel avenue for targeting metastatic cancers. Copyright © 2015 Elsevier Ltd. All rights reserved.

Prostate-specific membrane antigen-directed nanoparticle targeting for extreme nearfield ablation of prostate cancer cells.

Science.gov (United States)

Lee, Seung S; Roche, Philip Jr; Giannopoulos, Paresa N; Mitmaker, Elliot J; Tamilia, Michael; Paliouras, Miltiadis; Trifiro, Mark A

2017-03-01

Almost all biological therapeutic interventions cannot overcome neoplastic heterogeneity. Physical ablation therapy is immune to tumor heterogeneity, but nearby tissue damage is the limiting factor in delivering lethal doses. Multi-walled carbon nanotubes offer a number of unique properties: chemical stability, photonic properties including efficient light absorption, thermal conductivity, and extensive surface area availability for covalent chemical ligation. When combined together with a targeting moiety such as an antibody or small molecule, one can deliver highly localized temperature increases and cause extensive cellular damage. We have functionalized multi-walled carbon nanotubes by conjugating an antibody against prostate-specific membrane antigen. In our in vitro studies using prostate-specific membrane antigen-positive LNCaP prostate cancer cells, we have effectively demonstrated cell ablation of >80% with a single 30-s exposure to a 2.7-W, 532-nm laser for the first time without bulk heating. We also confirmed the specificity and selectivity of prostate-specific membrane antigen targeting by assessing prostate-specific membrane antigen-null PC3 cell lines under the same conditions (<10% cell ablation). This suggests that we can achieve an extreme nearfield cell ablation effect, thus restricting potential tissue damage when transferred to in vivo clinical applications. Developing this new platform will introduce novel approaches toward current therapeutic modalities and will usher in a new age of effective cancer treatment squarely addressing tumoral heterogeneity.

CO2-Switchable Membranes Prepared by Immobilization of CO2-Breathing Microgels.

Science.gov (United States)

Zhang, Qi; Wang, Zhenwu; Lei, Lei; Tang, Jun; Wang, Jianli; Zhu, Shiping

2017-12-20

Herein, we report the development of a novel CO 2 -responsive membrane system through immobilization of CO 2 -responsive microgels into commercially available microfiltration membranes using a method of dynamic adsorption. The microgels, prepared from soap-free emulsion polymerization of CO 2 -responsive monomer 2-(diethylamino)ethyl methacrylate (DEA), can be reversibly expanded and shrunken upon CO 2 /N 2 alternation. When incorporated into the membranes, this switching behavior was preserved and further led to transformation between microfiltration and ultrafiltration membranes, as indicated from the dramatic changes on water flux and BSA rejection results. This CO 2 -regulated performance switching of membranes was caused by the changes of water transportation channel, as revealed from the dynamic water contact angle tests and SEM observation. This work represents a simple yet versatile strategy for making CO 2 -responsive membranes.

Nanoclay embedded mixed matrix PVDF nanocomposite membrane: Preparation, characterization and biofouling resistance

International Nuclear Information System (INIS)

Rajabi, Hamid; Ghaemi, Negin; Madaeni, Sayed S.; Daraei, Parisa; Khadivi, Mohammad Ali; Falsafi, Monir

2014-01-01

Highlights: • Nanocomposite membranes were prepared by addition of OMMT to PVDF membrane. • Addition of nanoclay considerably increased the hydrophilicity of PVDF membrane. • Nanocomposite membranes had higher water flux and antifouling properties. • Fouling of membranes blended with nanoclay (<4 wt.%) reduced. - Abstract: In this paper, nanocomposite PVDF/nanoclay membranes were prepared with addition of different concentrations of organically modified montmorillonite (OMMT) into the polymeric casting solution using combination of solution dispersion and phase inversion methods. Membranes were characterized by use of X-ray diffraction (XRD), water contact angle, scanning electron microscopy (SEM) and atomic force microscopy (AFM), and their performances were evaluated in terms of pure water flux and fouling parameters. The surface hydrophilicity of all nanocomposites markedly improved compared to nascent PVDF. In addition, XRD patterns revealed the formation of intercalated layers of mineral clays in PVDF matrix. SEM and AFM images showed that addition of OMMT resulted in nanocomposite membranes with thinner skin layer and higher porosity rather than PVDF membranes. Pure water flux of PVDF/OMMT membranes increased significantly (particularly for fabricated membranes by 4 and 6 wt.% OMMT) compared to that of PVDF membrane. Moreover, nanocomposite membranes showed the elevated antifouling properties, and flux recovery of nascent PVDF membranes increased from 51 to 72% with addition of 2 wt.% OMMT nanoparticles. These nanocomposite membranes also offered a remarkable reusability and durability against biofouling

Nanoclay embedded mixed matrix PVDF nanocomposite membrane: Preparation, characterization and biofouling resistance

Energy Technology Data Exchange (ETDEWEB)

Rajabi, Hamid [Membrane Research Centre, Department of Chemical Engineering, Razi University, Tagh Bostan, 67149 Kermanshah (Iran, Islamic Republic of); Department of Civil Engineering, Razi University, 67149 Kermanshah (Iran, Islamic Republic of); Ghaemi, Negin, E-mail: negin_ghaemi@kut.ac.ir [Department of Chemical Engineering, Kermanshah University of Technology, 67178 Kermanshah (Iran, Islamic Republic of); Madaeni, Sayed S. [Membrane Research Centre, Department of Chemical Engineering, Razi University, Tagh Bostan, 67149 Kermanshah (Iran, Islamic Republic of); Daraei, Parisa [Department of Chemical Engineering, Kermanshah University of Technology, 67178 Kermanshah (Iran, Islamic Republic of); Khadivi, Mohammad Ali [Friedrich-Alexander University, Erlangen-Nuremberg, Egerland Strasse 3, D-91058 Erlangen (Germany); Falsafi, Monir [Department of Chemistry, Faculty of Science, Razi University, 67149 Kermanshah (Iran, Islamic Republic of)

2014-09-15

Highlights: • Nanocomposite membranes were prepared by addition of OMMT to PVDF membrane. • Addition of nanoclay considerably increased the hydrophilicity of PVDF membrane. • Nanocomposite membranes had higher water flux and antifouling properties. • Fouling of membranes blended with nanoclay (<4 wt.%) reduced. - Abstract: In this paper, nanocomposite PVDF/nanoclay membranes were prepared with addition of different concentrations of organically modified montmorillonite (OMMT) into the polymeric casting solution using combination of solution dispersion and phase inversion methods. Membranes were characterized by use of X-ray diffraction (XRD), water contact angle, scanning electron microscopy (SEM) and atomic force microscopy (AFM), and their performances were evaluated in terms of pure water flux and fouling parameters. The surface hydrophilicity of all nanocomposites markedly improved compared to nascent PVDF. In addition, XRD patterns revealed the formation of intercalated layers of mineral clays in PVDF matrix. SEM and AFM images showed that addition of OMMT resulted in nanocomposite membranes with thinner skin layer and higher porosity rather than PVDF membranes. Pure water flux of PVDF/OMMT membranes increased significantly (particularly for fabricated membranes by 4 and 6 wt.% OMMT) compared to that of PVDF membrane. Moreover, nanocomposite membranes showed the elevated antifouling properties, and flux recovery of nascent PVDF membranes increased from 51 to 72% with addition of 2 wt.% OMMT nanoparticles. These nanocomposite membranes also offered a remarkable reusability and durability against biofouling.

Smart membranes for monitoring membrane based desalination processes

KAUST Repository

Laleg-Kirati, Taous-Meriem; Karam, Ayman M.

2017-01-01

Various examples are related to smart membranes for monitoring membrane based process such as, e.g., membrane distillation processes. In one example, a membrane, includes a porous surface and a plurality of sensors (e.g., temperature, flow and

Reduced Graphene Oxide Membranes: Applications in Fog Collection and Water Purification

KAUST Repository

Tang, Bo

2017-05-01

Reduced graphene oxide (rGO) has attracted considerable interest recently as the low cost and chemical stable derivative of pristine graphene with application in many applications such as energy storage, water purification and electronic devices. This dissertation thoroughly investigated stacked rGO membrane fabrication process by vacuum-driven filtration, discovered asymmetry of the two surfaces of the rGO membrane, explored application perspectives of the asymmetric rGO membrane in fog collection and microstructure patterning, and disclosed membrane compaction issue during water filtration and species rejection. In more details, this dissertation revealed that, with suitable pore size, the filtration membrane substrate would leave its physical imprint on the bottom surface of the rGO membrane in the form of surface microstructures, which result in asymmetric dynamic water wettability properties of the two surfaces of the rGO membrane. The asymmetric wettability of the rGO membrane would lead to contrasting fog harvesting behavior of its two surfaces. The physical imprint mechanism was further extended to engineering pre-designed patterns selectively on the bottom surface of the rGO membrane. This dissertation, for the first time, reported the water flux and rejection kinetics, which was related to the compaction of the rGO membrane under pressure in the process of water filtration.

The development of zirconia membrane oxygen separation technology

International Nuclear Information System (INIS)

Chiacchi, F.T.; Badwal, S.P.S.; Velizko, V.

2000-01-01

The oxygen separation technology based on ceramic membranes constructed from stabilised zirconia is currently under development for applications ranging from oxygen generation or air enrichment for medical use to control of oxygen concentration or oxygen removal from gas streams and enclosures for semiconductor, food packaging and process control instrumentation industries. The technology is based on a rugged tubular design with extensive thermal cycling capability. Several single and three tube devices have been operated for periods up to 5000h. An eight tube module, as a building block for larger scale oxygen production or removal devices, has been constructed and is being evaluated. In this paper, the construction of the device, oxygen generating capacity, life time tests and performance of the ceramic membrane device under development at CSIRO will be discussed. Copyright (2000) The Australian Ceramic Society

Need for Methamphetamine Programming in Extension Education

Science.gov (United States)

Beaudreault, Amy R.; Miller, Larry E.

2011-01-01

The study reported sought to identify the prevention education needs involving methamphetamine through survey methodology. The study focused on a random sample of U.S. states and the Extension Directors within each state, resulting in a 70% response rate (n = 134). Findings revealed that 11% reported they had received methamphetamine user…

Identification of Current Proficiency Level of Extension Competencies and the Competencies Needed for Extension Agents to Be Successful in the 21st Century

Directory of Open Access Journals (Sweden)

Dona Lakai

2014-02-01

Full Text Available In this era of globalization, competency is an issue of concern to any field of professionals and their clients. Competency is an integrated set of skills, knowledge, and attitudes that allow one to effectively carry out the activities of a given work to the standards expected in the employment context. The purpose of this descriptive survey study was to determine the current proficiency level of North Carolina Cooperative Extension agents’ competencies and the other competencies they need to develop to be successful in Cooperative Extension. Findings indicate that the current proficiency level of competency for Extension agents in North Carolina Cooperative Extension varies from moderate to high in all 42 items listed in the survey. Multiple regression analysis confirmed that Extension agents’ years of Extension experience and age were major determinants of their overall proficiency level. Extension agents’ proficiency levels did not vary with gender, level of education, professional association affiliation, job position, or area of job responsibility. The research revealed that emotional intelligence, interpersonal skills, flexibility for adapting to changing environments, and ability to manage resources were the most significant other competencies needed for Extension agents to be successful in current context.

Acyl transfer from membrane lipids to peptides is a generic process.

Science.gov (United States)

Dods, Robert H; Bechinger, Burkhard; Mosely, Jackie A; Sanderson, John M

2013-11-15

The generality of acyl transfer from phospholipids to membrane-active peptides has been probed using liquid chromatography-mass spectrometry analysis of peptide-lipid mixtures. The peptides examined include melittin, magainin II, PGLa, LAK1, LAK3 and penetratin. Peptides were added to liposomes with membrane lipid compositions ranging from pure phosphatidylcholine (PC) to mixtures of PC with phosphatidylethanolamine, phosphatidylserine or phosphatidylglycerol. Experiments were typically conducted at pH7.4 at modest salt concentrations (90 mM NaCl). In favorable cases, lipidated peptides were further characterized by tandem mass spectrometry methods to determine the sites of acylation. Melittin and magainin II were the most reactive peptides, with significant acyl transfer detected under all conditions and membrane compositions. Both peptides were lipidated at the N-terminus by transfer from PC, phosphatidylethanolamine, phosphatidylserine or phosphatidylglycerol, as well as at internal sites: lysine for melittin; serine and lysine for magainin II. Acyl transfer could be detected within 3h of melittin addition to negatively charged membranes. The other peptides were less reactive, but for each peptide, acylation was found to occur in at least one of the conditions examined. The data demonstrate that acyl transfer is a generic process for peptides bound to membranes composed of diacylglycerophospholipids. Phospholipid membranes cannot therefore be considered as chemically inert toward peptides and by extension proteins. © 2013. Published by Elsevier Ltd. All rights reserved.

Pigmented villonodular synovitis of the temporomandibular joint with intracranial extension: A case series and systematic review.

Science.gov (United States)

Safaee, Michael; Oh, Taemin; Sun, Matthew Z; Parsa, Andrew T; McDermott, Michael W; El-Sayed, Ivan H; Bloch, Orin

2015-08-01

Pigmented villonodular synovitis (PVNS) is a rare proliferative disorder of the synovial membrane. PVNS generally affects large joints but occasionally involves the temporomandibular joint (TMJ), with occasional extension into the middle cranial fossa. The purpose of this study was to report our experience with PVNS along with a focused literature review. Patients with PVNS of the TMJ treated at the University of California - San Francisco from 2007 to 2013 were reviewed. A PubMed search was performed to identify additional cases. Five patients underwent surgical resection, with 1 recurrence at 61 months. A literature review identified 58 patients, 19 of which had intracranial involvement. Interestingly, intracranial extension was more common in men. Intracranial extension was not associated with an increased rate of recurrence. PVNS of the TMJ is a rare entity associated with excellent outcomes, even with intracranial extension. Management should consist of maximal resection, with radiotherapy reserved for extensive or recurrent lesions. © 2014 Wiley Periodicals, Inc.

Biomimetic membranes and methods of making biomimetic membranes

Science.gov (United States)

Rempe, Susan; Brinker, Jeffrey C.; Rogers, David Michael; Jiang, Ying-Bing; Yang, Shaorong

2016-11-08

The present disclosure is directed to biomimetic membranes and methods of manufacturing such membranes that include structural features that mimic the structures of cellular membrane channels and produce membrane designs capable of high selectivity and high permeability or adsorptivity. The membrane structure, material and chemistry can be selected to perform liquid separations, gas separation and capture, ion transport and adsorption for a variety of applications.

Gel layer formation on membranes in Membrane Bioreactors

NARCIS (Netherlands)

Van den Brink, P.F.H.

2014-01-01

The widespread application of membrane bioreactors (MBRs) for municipal wastewater treatment is hampered by membrane fouling. Fouling increases energy demand, reduces process performance and creates the need for more frequent (chemical) membrane cleaning or replacement. Membrane fouling in MBRs is

Surface patterning of polymeric separation membranes and its influence on the filtration performance

Science.gov (United States)

Maruf, Sajjad

Polymeric membrane based separation technologies are crucial for addressing the global issues such as water purification. However, continuous operations of these processes are often hindered by fouling which increases mass transport resistance of the membrane to permeation and thus the energy cost, and eventually replacement of the membrane in the system. In comparison to other anti-fouling strategies, the use of controlled surface topography to mitigate fouling has not been realized mainly due to the lack of methods to create targeted topography on the porous membrane surface. This thesis aims to develop a new methodology to create surface-patterned polymeric separation membrane to improve their anti-fouling characteristics during filtration. First, successful fabrication of sub-micron surface patterns directly on a commercial ultrafiltration (UF) membrane surface using nanoimprint lithographic (NIL) technique was demonstrated. Comprehensive filtration studies revealed that the presence of these sub-micron surface patterns mitigates not only the onset of colloidal particle deposition, but also lowers the rate of growth of cake layer after initial deposition, in comparison with un-patterned membranes. The anti-fouling effects were also observed for model protein solutions. Staged filtration experiments, with backwash cleaning, revealed that the permeate flux of the patterned membrane after protein fouling was considerably higher than that of the pristine or un-patterned membrane. In addition to the surface-patterning of UF membranes, successful fabrication of a surface-patterned thin film composite (TFC) membrane was shown for the first time. A two-step fabrication process was carried out by (1) nanoimprinting a polyethersulfone (PES) support using NIL, and (2) forming a thin dense film atop the PES support via interfacial polymerization (IP). Fouling experiments suggest that the surface patterns alter the hydrodynamics at the membrane-feed interface, which is

Bright and photostable push-pull pyrene dye visualizes lipid order variation between plasma and intracellular membranes.

Science.gov (United States)

Niko, Yosuke; Didier, Pascal; Mely, Yves; Konishi, Gen-ichi; Klymchenko, Andrey S

2016-01-11

Imaging lipid organization in cell membranes requires advanced fluorescent probes. Here, we show that a recently synthesized push-pull pyrene (PA), similarly to popular probe Laurdan, changes the emission maximum as a function of lipid order, but outperforms it by spectroscopic properties. In addition to red-shifted absorption compatible with common 405 nm diode laser, PA shows higher brightness and much higher photostability than Laurdan in apolar membrane environments. Moreover, PA is compatible with two-photon excitation at wavelengths >800 nm, which was successfully used for ratiometric imaging of coexisting liquid ordered and disordered phases in giant unilamellar vesicles. Fluorescence confocal microscopy in Hela cells revealed that PA efficiently stains the plasma membrane and the intracellular membranes at >20-fold lower concentrations, as compared to Laurdan. Finally, ratiometric imaging using PA reveals variation of lipid order within different cellular compartments: plasma membranes are close to liquid ordered phase of model membranes composed of sphingomyelin and cholesterol, while intracellular membranes are much less ordered, matching well membranes composed of unsaturated phospholipids without cholesterol. These differences in the lipid order were confirmed by fluorescence lifetime imaging (FLIM) at the blue edge of PA emission band. PA probe constitutes thus a new powerful tool for biomembrane research.

Cell membranes in radiation injury

International Nuclear Information System (INIS)

Koeteles, G.J.

1986-01-01

Cell membrane-related phenomena caused by low linear energy transfer radiation with doses lower than those producing cell killing are outlined. Micromorphological alterations as well as functional activities appearing with the receptors and in binding sites render it possible to reveal early and temporary changes. The cell injuries are suggested to transfer damaging conditions to surviving cells and to contribute to further development of non-stochastic effects in tissues

Functionalized membranes for environmental remediation and selective separation

Science.gov (United States)

Xiao, Li

Membrane process including microfiltration (MF), ultrafiltration (UF), nanofiltration (NF) and reverse osmosis (RO) have provided numerous successful applications ranging from drinking water purification, wastewater treatment, to material recovery. The addition of functional moiety in the membranes pores allows such membranes to be used in challenging areas including tunable separations, toxic metal capture, and catalysis. In this work, polyvinylidene fluoride (PVDF) MF membrane was functionalized with temperature responsive (poly(N-isopropylacrylamide), PNIPAAm) and pH responsive (polyacrylic acid, PAA) polymers. It's revealed that the permeation of various molecules (water, salt and dextran) through the membrane can be thermally or pH controlled. The introduction of PAA as a polyelectrolyte offers an excellent platform for the immobilization of metal nanoparticles (NPs) applied for degradation of toxic chlorinated organics with significantly increased longevity and stability. The advantage of using temperature and pH responsive polymers/hydrogels also includes the high reactivity and effectiveness in dechlorination. Further advancement on the PVDF functionalization involved the alkaline treatment to create partially defluorinated membrane (Def-PVDF) with conjugated double bounds allowing for the covalent attachment of different polymers. The PAA-Def-PVDF membrane shows pH responsive behavior on both the hydraulic permeability and solute retention. The sponge-like PVDF (SPVDF) membranes by phase inversion were developed through casting PVDF solution on polyester backing. The SPVDF membrane was demonstrated to have 4 times more surface area than commercial PVDF MF membrane, allowing for enhanced nanoparticles loading for chloro-organics degradation. The advanced functionalization method and process were also validated to be able to be scaled-up through the evaluation of full-scale functionalized membrane provided by Ultura Inc. California, USA. Nanofiltration (NF

Automated builder and database of protein/membrane complexes for molecular dynamics simulations.

Directory of Open Access Journals (Sweden)

Sunhwan Jo

2007-09-01

Full Text Available Molecular dynamics simulations of membrane proteins have provided deeper insights into their functions and interactions with surrounding environments at the atomic level. However, compared to solvation of globular proteins, building a realistic protein/membrane complex is still challenging and requires considerable experience with simulation software. Membrane Builder in the CHARMM-GUI website (http://www.charmm-gui.org helps users to build such a complex system using a web browser with a graphical user interface. Through a generalized and automated building process including system size determination as well as generation of lipid bilayer, pore water, bulk water, and ions, a realistic membrane system with virtually any kinds and shapes of membrane proteins can be generated in 5 minutes to 2 hours depending on the system size. Default values that were elaborated and tested extensively are given in each step to provide reasonable options and starting points for both non-expert and expert users. The efficacy of Membrane Builder is illustrated by its applications to 12 transmembrane and 3 interfacial membrane proteins, whose fully equilibrated systems with three different types of lipid molecules (DMPC, DPPC, and POPC and two types of system shapes (rectangular and hexagonal are freely available on the CHARMM-GUI website. One of the most significant advantages of using the web environment is that, if a problem is found, users can go back and re-generate the whole system again before quitting the browser. Therefore, Membrane Builder provides the intuitive and easy way to build and simulate the biologically important membrane system.

Organic/inorganic composite membranes based on polybenzimidazole and nano-SiO2

International Nuclear Information System (INIS)

Pu Hongting; Liu Lu; Chang Zhihong; Yuan Junjie

2009-01-01

Organic/inorganic composite membranes based on polybenzimidazole (PBI) and nano-SiO 2 were prepared in this work. However, the preparation of PBI/SiO 2 composite membrane is not easy since PBI is insoluble in water, while nano-SiO 2 is hydrophilic due to the hydrophilicity of nano-SiO 2 and water-insolubility of PBI. Thus, a solvent-exchange method was employed to prepare the composite membrane. The morphology of the composite membranes was studied by scanning electron microscopy (SEM). It was revealed that inorganic particles were dispersed homogenously in the PBI matrix. The thermal stability of the composite membrane is higher than that of pure PBI, both for doped and undoped membranes. PBI/SiO 2 composite membranes with up to 15 wt% SiO 2 exhibited improved mechanical properties compared with PBI membranes. The proton conductivity of the composite membranes containing phosphoric acid was studied. The nano-SiO 2 in the composite membranes enhanced the ability to trap phosphoric acid, which improved the proton conductivity of the composite membranes. The membrane with 15 wt% of inorganic material is oxidatively stable and has a proton conductivity of 3.9 x 10 -3 S/cm at 180 deg. C.

Preparation of catalyst coated membrane by modified decal transfer method for proton exchange membrane fuel cell

Science.gov (United States)

Indriyati; Irmawati, Y.; Prihandoko, B.

2017-07-01

A new catalyst coated membrane (CCM) was prepared by modified decal transfer method. A structure of ionomer/catalyst/carbon/substrate was used to facilitate the transfer of catalyst layer from decal substrate to the membrane at quite low hot-pressing temperature (120 °C) for 8 min. Several decal substrates were tested to select a proper substrate, namely PTFE cloth, PTFE film, aluminium foil, and OHP transparent sheet. The transfer degree of catalyst layer was estimated. Elemental analysis and SEM-mapping were performed to evaluate the residue, whereas contact angle measurement was conducted to characterize the hydrophobicity of decal substrates. The results showed that PTFE cloth and PFTE film transferred approximately 90% of catalyst layer onto the membrane, while the other two substrates were around 70%. Furthermore, the elemental analysis of the residue on the substrate revealed that it was mainly composed of carbon and fluorine for PTFE cloth and PTFE film. This result supports other findings that PTFE cloth and PTFE film are suitable as decal substrate at low temperature hot pressing for fabricating CCM.

Hybrid membranes of polyamide applied in treatment of waste water

International Nuclear Information System (INIS)

Medeiros, Keila Machado de; Araujo, Edcleide Maria; Lira, Helio de Lucena; Lima, Diego de Farias; Lima, Carlos Antonio Pereira de

2017-01-01

In this work, it was prepared hybrid membranes of polyamide6 (PA6) with montmorillonite (MMT) and porogenic agent (CaCl 2 ). The hybrid membranes with CaCl 2 were characterized by X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), differential scanning calorimetry (DSC), scanning electron microscopy (SEM), porosimetry by mercury intrusion (PMI), flux measurements and rejection. By means of X-ray diffraction, it was revealed that the hybrid membranes with CaCl 2 have an exfoliated and/or partially exfoliated structure. For FTIR and DSC of hybrid membranes with CaCl 2 , it was found that the spectra and the crystalline melting temperature remained virtually unchanged compared to PA6 membrane. From the SEM images, it was observed that the addition of the MMT and the CaCl 2 in the membrane of PA6 caused an increase in the amount of pores the surface and cross section of these membranes. By PMI, it was observed that the presence of MMT and CaCl 2 in the membrane caused an increase in the average diameters of pores. The water-oil separation tests, indicated a significant reduction of oil in the permeate, allowing treatment of wastewater contaminated with oil. (author)

Hybrid membranes of polyamide applied in treatment of waste water

Energy Technology Data Exchange (ETDEWEB)

Medeiros, Keila Machado de; Araujo, Edcleide Maria; Lira, Helio de Lucena, E-mail: keilamachadodemedeiros@gmail.com [Universidade Federal de Campina Grande (UFCG), PB (Brazil). Departamento de Engenharia de Materiais; Lima, Diego de Farias; Lima, Carlos Antonio Pereira de [Universidade Estadual da Paraiba (UEPB), Campina Grande, PB (Brazil). Departamento de Engenharia Sanitaria e Ambiental

2017-03-15

In this work, it was prepared hybrid membranes of polyamide6 (PA6) with montmorillonite (MMT) and porogenic agent (CaCl{sub 2} ). The hybrid membranes with CaCl{sub 2} were characterized by X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), differential scanning calorimetry (DSC), scanning electron microscopy (SEM), porosimetry by mercury intrusion (PMI), flux measurements and rejection. By means of X-ray diffraction, it was revealed that the hybrid membranes with CaCl{sub 2} have an exfoliated and/or partially exfoliated structure. For FTIR and DSC of hybrid membranes with CaCl{sub 2} , it was found that the spectra and the crystalline melting temperature remained virtually unchanged compared to PA6 membrane. From the SEM images, it was observed that the addition of the MMT and the CaCl{sub 2} in the membrane of PA6 caused an increase in the amount of pores the surface and cross section of these membranes. By PMI, it was observed that the presence of MMT and CaCl{sub 2} in the membrane caused an increase in the average diameters of pores. The water-oil separation tests, indicated a significant reduction of oil in the permeate, allowing treatment of wastewater contaminated with oil. (author)

Dynamic Membrane Formation in Anaerobic Dynamic Membrane Bioreactors: Role of Extracellular Polymeric Substances.

Directory of Open Access Journals (Sweden)

Hongguang Yu

Full Text Available Dynamic membrane (DM formation in dynamic membrane bioreactors plays an important role in achieving efficient solid-liquid separation. In order to study the contribution of extracellular polymeric substances (EPS to DM formation in anaerobic dynamic membrane bioreactor (AnDMBR processes, EPS extraction from and re-addition to bulk sludge were carried out in short-term filtration tests. DM formation behaviors could be well simulated by cake filtration model, and sludge with EPS re-addition showed the highest resistance coefficient, followed by sludge after EPS extraction. The DM layers exhibited a higher resistance and a lower porosity for the sludge sample after EPS extraction and for the sludge with EPS re-addition. Particle size of sludge flocs decreased after EPS extraction, and changed little with EPS re-addition, which was confirmed by interaction energy analysis. Further investigations by confocal laser scanning microscopy (CLSM analysis and batch tests suggested that the removal of in-situ EPS stimulated release of soluble EPS, and re-added EPS were present as soluble EPS rather than bound EPS, which thus improved the formation of DM. The present work revealed the role of EPS in anaerobic DM formation, and could facilitate the operation of AnDMBR processes.

Development of polyelectrolyte multilayer thin film composite membrane for water desalination application

KAUST Repository

Fadhillah, F.; Zaidi, S.M.J.; Khan, Z.; Khaled, M.M.; Rahman, F.; Hammond, P.T.

2013-01-01

Thin film composite membranes were fabricated via spin assisted layer by layer (SA-LbL) assembly by depositing alternate layers of poly(allyl amine hydrochloride) (PAH) and poly(acrylic acid) (PAA) on a polysulfone (PSF) ultrafiltration membrane as support. The suitability of these membranes for potential water purification applications was explored by testing the stability of the deposited thin films and their permeation characteristic using cross-flow permeation cell. Permeation test conducted at a pressure of 40bar, temperature of 25°C, pH of 6 and feed water concentration of 2000ppm NaCl demonstrated that the PAH/PAA multilayer film deposited on polysulfone support remained stable and intact under long-term test conditions. The 120 bilayers of PAH/PAA membrane tested at the above condition showed flux of 15L/m2.h and salt rejection of 65%. The membrane performance evaluation also revealed that SA-LbL PAH/PAA membrane follows the characteristics of the solution diffusion membrane. © 2013 Elsevier B.V.

Development of polyelectrolyte multilayer thin film composite membrane for water desalination application

KAUST Repository

Fadhillah, F.

2013-06-01

Thin film composite membranes were fabricated via spin assisted layer by layer (SA-LbL) assembly by depositing alternate layers of poly(allyl amine hydrochloride) (PAH) and poly(acrylic acid) (PAA) on a polysulfone (PSF) ultrafiltration membrane as support. The suitability of these membranes for potential water purification applications was explored by testing the stability of the deposited thin films and their permeation characteristic using cross-flow permeation cell. Permeation test conducted at a pressure of 40bar, temperature of 25°C, pH of 6 and feed water concentration of 2000ppm NaCl demonstrated that the PAH/PAA multilayer film deposited on polysulfone support remained stable and intact under long-term test conditions. The 120 bilayers of PAH/PAA membrane tested at the above condition showed flux of 15L/m2.h and salt rejection of 65%. The membrane performance evaluation also revealed that SA-LbL PAH/PAA membrane follows the characteristics of the solution diffusion membrane. © 2013 Elsevier B.V.

A protein anomaly in erythrocyte membranes of patients with Duchenne muscular dystrophy

Science.gov (United States)

1983-01-01

Raman spectroscopic comparisons of erythrocyte membranes from 20 patients with Duchenne muscular dystrophy and 8 age-matched controls indicate a prominent and consistent protein anomaly in the patient samples. This was apparent in the following: (a) CH-stretching signals from control membranes reveal a thermotropic transition at 15.6 degrees C, attributable to a protein/lipid phase that is lacking in dystrophic membranes. (b) CH-stretching signals from control membranes also show a protein transition at 39 degrees C [pH 7.4] that is shifted to 45 degrees in dystrophic membranes. (c) A reduction in pH to 5.7 shifts this transition from 39 degrees C to 7 degrees C in normal membranes and from 45 degrees C to 24 degrees C in dystrophic membranes. (d) The Amide I/Amide III regions indicate a significant proportion of beta- structured peptide in dystrophic but not normal membranes. (e) Analysis of tyrosine signals indicates greater polar exposure of tyrosine hydroxyl groups in dystrophic vs normal membranes. All of the differences between dystrophic and normal membranes are highly significant (P less than 0.001). PMID:6854213

Assessment of extension agents' use of communication methods ...

African Journals Online (AJOL)

Findings from correlation analysis revealed that there was significant relationship between linkage and communication methods between institute (r =-0.377), linkage and communication methods among extension agents (r =0.379). However, the relationship between communication within and between institute was highly ...

Nanodisc-solubilized membrane protein library reflects the membrane proteome.

Science.gov (United States)

Marty, Michael T; Wilcox, Kyle C; Klein, William L; Sligar, Stephen G

2013-05-01

The isolation and identification of unknown membrane proteins offers the prospect of discovering new pharmaceutical targets and identifying key biochemical receptors. However, interactions between membrane protein targets and soluble ligands are difficult to study in vitro due to the insolubility of membrane proteins in non-detergent systems. Nanodiscs, nanoscale discoidal lipid bilayers encircled by a membrane scaffold protein belt, have proven to be an effective platform to solubilize membrane proteins and have been used to study a wide variety of purified membrane proteins. This report details the incorporation of an unbiased population of membrane proteins from Escherichia coli membranes into Nanodiscs. This solubilized membrane protein library (SMPL) forms a soluble in vitro model of the membrane proteome. Since Nanodiscs contain isolated proteins or small complexes, the SMPL is an ideal platform for interactomics studies and pull-down assays of membrane proteins. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of the protein population before and after formation of the Nanodisc library indicates that a large percentage of the proteins are incorporated into the library. Proteomic identification of several prominent bands demonstrates the successful incorporation of outer and inner membrane proteins into the Nanodisc library.

Regulation of multispanning membrane protein topology via post-translational annealing.

Science.gov (United States)

Van Lehn, Reid C; Zhang, Bin; Miller, Thomas F

2015-09-26

The canonical mechanism for multispanning membrane protein topogenesis suggests that protein topology is established during cotranslational membrane integration. However, this mechanism is inconsistent with the behavior of EmrE, a dual-topology protein for which the mutation of positively charged loop residues, even close to the C-terminus, leads to dramatic shifts in its topology. We use coarse-grained simulations to investigate the Sec-facilitated membrane integration of EmrE and its mutants on realistic biological timescales. This work reveals a mechanism for regulating membrane-protein topogenesis, in which initially misintegrated configurations of the proteins undergo post-translational annealing to reach fully integrated multispanning topologies. The energetic barriers associated with this post-translational annealing process enforce kinetic pathways that dictate the topology of the fully integrated proteins. The proposed mechanism agrees well with the experimentally observed features of EmrE topogenesis and provides a range of experimentally testable predictions regarding the effect of translocon mutations on membrane protein topogenesis.

Bmi-1 Regulates Extensive Erythroid Self-Renewal

Directory of Open Access Journals (Sweden)

Ah Ram Kim

2015-06-01

Full Text Available Red blood cells (RBCs, responsible for oxygen delivery and carbon dioxide exchange, are essential for our well-being. Alternative RBC sources are needed to meet the increased demand for RBC transfusions projected to occur as our population ages. We previously have discovered that erythroblasts derived from the early mouse embryo can self-renew extensively ex vivo for many months. To better understand the mechanisms regulating extensive erythroid self-renewal, global gene expression data sets from self-renewing and differentiating erythroblasts were analyzed and revealed the differential expression of Bmi-1. Bmi-1 overexpression conferred extensive self-renewal capacity upon adult bone-marrow-derived self-renewing erythroblasts, which normally have limited proliferative potential. Importantly, Bmi-1 transduction did not interfere with the ability of extensively self-renewing erythroblasts (ESREs to terminally mature either in vitro or in vivo. Bmi-1-induced ESREs can serve to generate in vitro models of erythroid-intrinsic disorders and ultimately may serve as a source of cultured RBCs for transfusion therapy.

Membranous nephropathy with Neuromyelitis optica spectrum disorder.

Science.gov (United States)

Li, Xiangling; Wang, YanQiang

2017-07-01

Membranous nephropathy (MN) accompanying Neuromyelitis optica spectrum disorders (NMOSD) has rarely been described previously. We recently presented a 45-year-old Chinese male presenting with recurrent lower extremity pitting edema, or eyelid edema, proteinuria and hyperlipidemia. especially intractable hiccup and vomiting, painful tonic spasm (PTS) as the revealing symptom of a demyelinating disorder of central nervous system. The kindey biopsy specimen showed MN stage 2. Serological testing revealed antibodies AQP4, MRI head and spine revealed medulla oblongata and C1-C2 cervical vertebra lesions. Treatment with methylprednisolone, cyclophosphamide, azathioprine resulted in consistent clinical improvement. Copyright © 2017 Elsevier B.V. All rights reserved.

Flux Enhancement in Membrane Distillation Using Nanofiber Membranes

Directory of Open Access Journals (Sweden)

T. Jiříček

2016-01-01

Full Text Available Membrane distillation (MD is an emerging separation technology, whose largest application potential lies in the desalination of highly concentrated solutions, which are out of the scope of reverse osmosis. Despite many attractive features, this technology is still awaiting large industrial application. The main reason is the lack of commercially available membranes with fluxes comparable to reverse osmosis. MD is a thermal separation process driven by a partial vapour pressure difference. Flux, distillate purity, and thermal efficiency are always in conflict, all three being strictly connected with pore size, membrane hydrophobicity, and thickness. The world has not seen the ideal membrane yet, but nanofibers may offer a solution to these contradictory requirements. Membranes of electrospun PVDF were tested under various conditions on a direct contact (DCMD unit, in order to determine the optimum conditions for maximum flux. In addition, their performance was compared to commonly available PTFE, PE, and PES membranes. It was confirmed that thinner membranes have higher fluxes and a lower distillate purity and also higher energy losses via conduction across the membrane. As both mass and heat transfer are connected, it is best to develop new membranes with a target application in mind, for the specific membrane module and operational conditions.

Synthesis of ion exchange membrane by radiation grafting of acrylic acid onto polyethylene

International Nuclear Information System (INIS)

Ishigaki, I.; Sugo, T.; Senoo, K.; Takayama, T.; Machi, S.; Okamoto, J.; Okada, T.

1981-01-01

Radiation grafting of vinyl monomers onto polymer films has been extensively studied by many workers. In the preirradiation method of grafting a polymer substrate is activated by irradiation (either in the presence or absence of oxygen) and subsequently allowed to react with a monomer. The preirradiation method was utilized in this study to synthesize an ion exchange membrane useful for a battery separator by grafting acrylic acid onto polyethylene film. The battery separator should be chemically and thermally stable, sufficiently durable in electrolyte as well as highly electrically conductive. Membranes made from regenerated cellulose, e.g., cellophane, have long been used as a separator in the batteries with alkaline electrolyte, such as silver oxide primary cell. However, it has poor durability, as short as one year, due to breakdown of the membrane during operation or storing. The acrylic acid-grafted polyethylene film was found to be quite useful for a separator in the alkaline batteries. This membrane has a high electric conductivity and an excellent durability. (author)

Development of Ultrafiltration Membrane-Separation Technology for Energy-Efficient Water Treatment and Desalination Process

Energy Technology Data Exchange (ETDEWEB)

Yim, Woosoon [Univ. of Nevada, Las Vegas, NV (United States); Bae, Chulsung [Rensselaer Polytechnic Inst., Troy, NY (United States)

2016-10-28

. Recently, sulfonated SEBS became commercially available and has been extensively explored for membrane-mediated water purification technology. The sulfonated block copolymer creates a well developed nano-sale phase-separated morphologies composed of hydrophilic domains (sulfonated polystyrene) and hydrophobic domains (polyethylene/polybutylene). The hydrophilic domains determines transport properties (water transport, salt and/or ion rejection, etc) and the hydrophobic domains provides mechanical stability of the membrane. Unfortunately, a high degree of sulfonation of SEBS induces excessive swelling and deterioration of mechanical stability of the membrane. In an effort to develop robust polymeric membrane materials for water purification technology, phosphonic acid-functionalized SEBS membranes are investigated during this report period. In compare to sulfonated polymers, the corresponding phosphonated polymers are known to swell less because of the formation of extensive hydrogen bonding networks between phosphonates. In addition to the expected better mechanical stability, phosphonated polymers has another advantage over sulfonated polymers for the use water purification membrane; each phosphonate can accommodate two ions while each sulfonate accommodates only one ion. Membrane properties (ion type, ionic density, etc) of new membranes will be studied and their separation performance will be evaluated in water purification and desalination process. Through systematic study of the relationship of chemical structure–surface property–membrane performance, we aim to better understand the nature of membrane fouling and develop more fouling-resistant water purification membranes. The basic understanding of this relationship will lead to the development of advanced membrane materials which can offer a solution to environmentally sustainable production of fresh water.

APPLICATION OF MEMBRANE SORPTION REACTOR TECHNOLOGY FOR LRW MANAGEMENT

International Nuclear Information System (INIS)

Glagolenko, Yuri; Dzekun, Evgeny; Myasoedovg, Boris; Gelis, Vladimir; Kozlitin, Evgeny; Milyutin, Vitaly; Trusov, Lev; Rengel, Mike; Mackay, Stewart M.; Johnson, Michael E.

2003-01-01

A new membrane-sorption technology has been recently developed and industrially implemented in Russia for the treatment of the Liquid (Low-Level) Radioactive Waste (LRW). The first step of the technology is a precipitation of the radionuclides and/or their adsorption onto sorbents of small particle size. The second step is filtration of the precipitate/sorbent through the metal-ceramic membrane, Trumem.. The unique feature of the technology is a Membrane-Sorption Reactor (MSR), in which the precipitation / sorption and the filtration of the radionuclides occur simultaneously, in one stage. This results in high efficiency, high productivity and compactness of the equipment, which are the obvious advantages of the developed technology. Two types of MSR based on Flat Membranes device and Centrifugal Membrane device were developed. The advantages and disadvantages of application of each type of the reactors are discussed. The MSR technology has been extensively tested and efficiently implemented at ''Mayak '' nuclear facility near Chelyabinsk, Russia as well as at other Russian sites. The results of this and other applications of the MSR technology at the different Russian nuclear facilities are discussed. The results of the first industrial applications of the MSR technology for radioactive waste treatment in Russia and analysis of the available information about LRW accumulated in other countries imply that this technology can be successfully used for the Low Level Radioactive Waste treatment in the USA and in other nuclear countries

Effects of transmembrane hydraulic pressure on performance of forward osmosis membranes.

Science.gov (United States)

Coday, Bryan D; Heil, Dean M; Xu, Pei; Cath, Tzahi Y

2013-03-05

Forward osmosis (FO) is an emerging membrane separation process that continues to be tested and implemented in various industrial water and wastewater treatment applications. The growing interests in the technology have prompted laboratories and manufacturers to adopt standard testing methods to ensure accurate comparison of membrane performance under laboratory-controlled conditions; however, standardized methods might not capture specific operating conditions unique to industrial applications. Experiments with cellulose triacetate (CTA) and polyamide thin-film composite (TFC) FO membranes demonstrated that hydraulic transmembrane pressure (TMP), common in industrial operation of FO membrane elements, could affect membrane performance. Experiments were conducted with three FO membranes and with increasing TMP up to a maximum of 50 psi (3.45 bar). The feed solution was a mixture of salts and the draw solution was either a NaCl solution or concentrated seawater at similar osmotic pressure. Results revealed that TMP minimally affected water flux, reverse salt flux (RSF), and solute rejection of the CTA membrane. However, water flux through TFC membranes might slightly increase with increasing TMP, and RSF substantially declines with increasing TMP. It was observed that rejection of feed constituents was influenced by TMP and RSF.

Effect of high carbon dioxide storage and gamma irradiation on membrane deterioration in cauliflower florets

International Nuclear Information System (INIS)

Voisine, R.; Hombourger, C.; Willemot, C.; Castaigne, F.; Makhlouf, J.

1993-01-01

Controlled atmospheres and gamma irradiation are technologies which extend storage-life of fruits and vegetables. Separate and combined effects of high CO 2 storage and gamma irradiation on cell membranes from cauliflower florets (Brassica oleracea L., Botrytis group) were investigated. Storage of the florets for 8 days at 13°C, either under 15% carbon dioxide or in air after irradiation at 2 kGy, accelerated the deterioration of microsomal membranes during storage. Both treatments caused an early loss in lipid phosphate. Irradiation enhanced the free fatty acid content of the membranes during storage and caused an extensive protein loss. When irradiation and high CO 2 storage were combined, electrolyte leakage significantly increased while protein loss was considerably reduced. The results indicate that high CO 2 and irradiation accelerate membrane degradation through different mechanisms. The combined effects of the treatments were not additive, but membrane yield was apparently reduced. CO 2 protected the membranes from protein loss induced by irradiation. The apparent increase in electrolyte leakage after irradiation may be caused by the release of ions following cell wall deterioration

Studies by radioiodination of normal adult, fetal and leukemic cell membranes

Energy Technology Data Exchange (ETDEWEB)

Kannourakis, G; Cauchi, M N [Department of Pathology and Immunology, Monash Medical School, Melbourne, Australia

1978-01-01

A comparison was made between cord blood lymphocytes, normal adult lymphocytes and leukemic cells after membrane iodination with lactoperoxidase. A double-labeling technique using lactoperoxidase iodination with /sup 125/I and /sup 131/I followed by analysis on polyacrylamide gel electrophoresis revealed a number of membrane differences between leukemic, normal and fetal cells. There was a reduction in the 70,000 molecular weight component in cord blood cells compared to adult lymphocytes, and an increase in membrane peptides with molecular weights of 35,000, 20,000, 9,000 and 4,000. Although smaller molecular weight peptides were also present in chronic lymphatic leukemia as well as acute myeloid leukemia, these were shown to be distinct from fetal type membrane components.

Characterisation of detergent-insoluble membranes in pollen tubes of Nicotiana tabacum (L.

Directory of Open Access Journals (Sweden)

Alessandra Moscatelli

2015-02-01

Full Text Available Pollen tubes are the vehicle for sperm cell delivery to the embryo sac during fertilisation of Angiosperms. They provide an intriguing model for unravelling mechanisms of growing to extremes. The asymmetric distribution of lipids and proteins in the pollen tube plasma membrane modulates ion fluxes and actin dynamics and is maintained by a delicate equilibrium between exocytosis and endocytosis. The structural constraints regulating polarised secretion and asymmetric protein distribution on the plasma membrane are mostly unknown. To address this problem, we investigated whether ordered membrane microdomains, namely membrane rafts, might contribute to sperm cell delivery. Detergent insoluble membranes, rich in sterols and sphingolipids, were isolated from tobacco pollen tubes. MALDI TOF/MS analysis revealed that actin, prohibitins and proteins involved in methylation reactions and in phosphoinositide pattern regulation are specifically present in pollen tube detergent insoluble membranes. Tubulins, voltage-dependent anion channels and proteins involved in membrane trafficking and signalling were also present. This paper reports the first evidence of membrane rafts in Angiosperm pollen tubes, opening new perspectives on the coordination of signal transduction, cytoskeleton dynamics and polarised secretion.

Membrane Structure Studies by Means of Small-Angle Neutron Scattering (SANS)

International Nuclear Information System (INIS)

Knott, R. B.

2008-01-01

The basic model for membrane structure--a lipid bilayer with imbedded proteins--was formulated 35 years ago, however the detailed structure is still under active investigation using a variety of physical, chemical and computational techniques. Every biologically active cell is encapsulated by a plasma membrane with most cells also equipped with an extensive intracellular membrane system. The plasma membrane is an important boundary between the cytoplasm of the cell and the external environment, and selectively isolates the cell from that environment. Passive diffusion and/or active transport mechanisms are provided for water, ions, substrates etc. which are vital for cell metabolism and viability. Membranes also facilitate excretion of substances either as useful cellular products or as waste. Despite their complexity and diverse function, plasma membranes from quite different cells have surprisingly similar compositions. A typical membrane structure consists of a phospholipid bilayer with a number of proteins scattered throughout, along with carbohydrates (glycoproteins), glycolipids and sterols. The plasma membranes of most eukaryotic cells contain approximately equal weights of lipid and protein, which corresponds to about 100 lipid molecules per protein molecule. Clearly, lipids are a major constituent and the study of their structure and function in isolation provides valuable insight into the more complex intact multicomponent membrane. The membrane bound protein is the other major constituent and is a very active area of research for a number of reasons including the fact that over 60% of modern drugs act on their receptor sites. The interaction between the protein and the supporting lipid bilayer is clearly of major importance. Neutron scattering is a powerful technique for exploring the structure of membranes, either as reconstituted membranes formed from well characterised lipids, or as intact membranes isolated from selected biological systems. A brief

Membraneous stenosis of the upper oesophagus ('webs')

International Nuclear Information System (INIS)

Koch, H.L.; Kurtz, B.

1981-01-01

Webs of the upper oesophagus are sail-like mucosal folds of unknown aetiology. Small, transverse webs on the anterior wall of the oesophagus are not uncommon incidental findings which are easily overlooked on routine examination. Extensive, circular membranes in the upper oesophagus, on the other hand, are rare; these may lead to severe difficulty with swallowing and may be associated with regurgitation. One example of a transverse, and three cases of circular webs are described, which caused stenosis and dysphagia and which, in some cases, were multiple. The aetiology is discussed. (orig.) [de

Characterization of Membrane Foulants in Full-scale and Lab-scale Membrane Bioreactors for Wastewater Treatment and Reuse

KAUST Repository

Matar, Gerald

2015-12-01

Membrane bioreactors (MBRs) offer promising solution for wastewater treatment and reuse to address the problem of water scarcity. Nevertheless, this technology is still facing challenges associated with membrane biofouling. This phenomenon has been mainly investigated in lab-scale MBRs with little or no insight on biofouling in full-scale MBR plants. Furthermore, the temporal dynamics of biofouling microbial communities and their extracellular polymeric substances (EPS) are less studied. Herein, a multidisciplinary approach was adopted to address the above knowledge gaps in lab- and full-scale MBRs. In the full-scale MBR study, 16S rRNA gene pyrosequencing with multivariate statistical analysis revealed that the early and mature biofilm communities from five full-scale MBRs differed significantly from the source community (i.e. activated sludge), and random immigration of species from the source community was unlikely to shape the community structure of biofilms. Also, a core biofouling community was shared between the five MBR plants sampled despite differences in their operating conditions. In the lab-scale MBR studies, temporal dynamics of microbial communities and their EPS products were monitored on different hydrophobic and hydrophilic membranes during 30 days. At the early stages of filtration (1 d), the same early colonizers belonging to the class Betaproteobacteria were identified on all the membranes. However, their relative abundance decreased on day 20 and 30, and sequence reads belonging to the phylum Firmicutes and Chlorobi became dominant on all the membranes on day 20 and 30. In addition, the intrinsic membrane characteristic did not select any specific EPS fractions at the initial stages of filtration and the same EPS foulants developed with time on the hydrophobic and hydrophilic membranes. Our results indicated that the membrane surface characteristics did not select for specific biofouling communities or EPS foulants, and the same early

Surface expression, single-channel analysis and membrane topology of recombinant Chlamydia trachomatis Major Outer Membrane Protein

Directory of Open Access Journals (Sweden)

McClafferty Heather

2005-01-01

Full Text Available Abstract Background Chlamydial bacteria are obligate intracellular pathogens containing a cysteine-rich porin (Major Outer Membrane Protein, MOMP with important structural and, in many species, immunity-related roles. MOMP forms extensive disulphide bonds with other chlamydial proteins, and is difficult to purify. Leaderless, recombinant MOMPs expressed in E. coli have yet to be refolded from inclusion bodies, and although leadered MOMP can be expressed in E. coli cells, it often misfolds and aggregates. We aimed to improve the surface expression of correctly folded MOMP to investigate the membrane topology of the protein, and provide a system to display native and modified MOMP epitopes. Results C. trachomatis MOMP was expressed on the surface of E. coli cells (including "porin knockout" cells after optimizing leader sequence, temperature and medium composition, and the protein was functionally reconstituted at the single-channel level to confirm it was folded correctly. Recombinant MOMP formed oligomers even in the absence of its 9 cysteine residues, and the unmodified protein also formed inter- and intra-subunit disulphide bonds. Its topology was modeled as a (16-stranded β-barrel, and specific structural predictions were tested by removing each of the four putative surface-exposed loops corresponding to highly immunogenic variable sequence (VS domains, and one or two of the putative transmembrane strands. The deletion of predicted external loops did not prevent folding and incorporation of MOMP into the E. coli outer membrane, in contrast to the removal of predicted transmembrane strands. Conclusions C. trachomatis MOMP was functionally expressed on the surface of E. coli cells under newly optimized conditions. Tests of its predicted membrane topology were consistent with β-barrel oligomers in which major immunogenic regions are displayed on surface-exposed loops. Functional surface expression, coupled with improved understanding of MOMP

All-solid-state ion-selective silicone rubber membrane electrodes with a new conducting polymer

International Nuclear Information System (INIS)

Park, Eun Rang; Chung, Yeon Joon; Hwang, Sun Woo

2012-01-01

New conducting polymers containing heterocyclic rings with carbazole, ethylene dioxythiophene (EDOT) and benzobisthiazole were synthesized and the characterized by using organic spectroscopic methods. Potentiometric ion-selective membrane electrodes (ISMEs) have been extensively used for ion analysis in clinical, environmental, and industrial fields owing to their wide response range (4 to 7 orders of magnitude), no effect of sample turbidity, fast response time, and ease of miniaturization. Considerable attention has been given to alternative use of room-temperature vulcanizing (RTV)-type silicone rubber (SR) owing to its strong adhesion and high thermal durability. Unfortunately, the high membrane resistance of SR-based ion-selective membranes (ISMs) (2 to 3 higher orders of magnitude compared to those of poly(vinyl chloride)(PVC)-based ones) has significantly restricted their application. Herein, we demonstrate a new method to reduce the membrane resistance via addition of a new conducting polymer into the SR-based ISMs.

Polymer-inorganic hybrid proton conductive membranes: Effect of the interfacial transfer pathways

International Nuclear Information System (INIS)

Chen, Pingping; Hao, Lie; Wu, Wenjia; Li, Yifan; Wang, Jingtao

2016-01-01

Highlights: • A series of hybrid membranes are prepared using fillers with different structures. • The fillers (0-D, 1-D, and 2-D) are sulfonated to ensure close surface component. • The effect of filler’s structure on microstructure of hydrid membrane is explored. • For single-kind filler series, 2-D filler has the strongest conduction promotion. • The synergy effect of different kinds of fillers is systematacially investigated. - Abstract: For hybrid membrane, the polymer-inorganic interface along filler surface can be facilely created to be distinctive and controllable pathway for mass transfer. Herein, three kinds of fillers are used as inorganic additives including zero-dimensional silica (0-D, SiO_2), one-dimensional halloysite nanotube (1-D, HNT), and two-dimensional graphene oxide (2-D, GO), which are functionalized by sulfonated polymer layer to ensure close surface component. Then the fillers are incorporated into two types of polymer matrixes (phase-separated sulfonated poly(ether ether ketone) and non-phase-separated chitosan) to prepare three series of hybrid membranes with single-kind filler, double-kinds fillers, or triple-kinds fillers, respectively. The microstructures, physicochemical properties, and proton conduction properties (under hydrated and anhydrous conditions) of the membranes are extensively investigated. It is found that (i) for the single-kind filler-filled membranes, 2-D filler has the strongest promotion ability for proton conductivity of membrane due to the constructed wide and long-range pathways for proton transfer; (ii) while for the hybrid membranes with double-kinds fillers, instead of synergistic promotion effect, the fillers cause more tortuous transfer pathways within membranes and then decrease proton conductivity; (iii) the hybrid membranes with triple-kinds fillers exhibit similar behavior but a little higher conductivity than the membranes with double-kinds fillers.

HAMLET interacts with lipid membranes and perturbs their structure and integrity.

Science.gov (United States)

Mossberg, Ann-Kristin; Puchades, Maja; Halskau, Øyvind; Baumann, Anne; Lanekoff, Ingela; Chao, Yinxia; Martinez, Aurora; Svanborg, Catharina; Karlsson, Roger

2010-02-23

Cell membrane interactions rely on lipid bilayer constituents and molecules inserted within the membrane, including specific receptors. HAMLET (human alpha-lactalbumin made lethal to tumor cells) is a tumoricidal complex of partially unfolded alpha-lactalbumin (HLA) and oleic acid that is internalized by tumor cells, suggesting that interactions with the phospholipid bilayer and/or specific receptors may be essential for the tumoricidal effect. This study examined whether HAMLET interacts with artificial membranes and alters membrane structure. We show by surface plasmon resonance that HAMLET binds with high affinity to surface adherent, unilamellar vesicles of lipids with varying acyl chain composition and net charge. Fluorescence imaging revealed that HAMLET accumulates in membranes of vesicles and perturbs their structure, resulting in increased membrane fluidity. Furthermore, HAMLET disrupted membrane integrity at neutral pH and physiological conditions, as shown by fluorophore leakage experiments. These effects did not occur with either native HLA or a constitutively unfolded Cys-Ala HLA mutant (rHLA(all-Ala)). HAMLET also bound to plasma membrane vesicles formed from intact tumor cells, with accumulation in certain membrane areas, but the complex was not internalized by these vesicles or by the synthetic membrane vesicles. The results illustrate the difference in membrane affinity between the fatty acid bound and fatty acid free forms of partially unfolded HLA and suggest that HAMLET engages membranes by a mechanism requiring both the protein and the fatty acid. Furthermore, HAMLET binding alters the morphology of the membrane and compromises its integrity, suggesting that membrane perturbation could be an initial step in inducing cell death.

Targeting membrane-bound viral RNA synthesis reveals potent inhibition of diverse coronaviruses including the middle East respiratory syndrome virus.

Directory of Open Access Journals (Sweden)

Anna Lundin

2014-05-01

Full Text Available Coronaviruses raise serious concerns as emerging zoonotic viruses without specific antiviral drugs available. Here we screened a collection of 16671 diverse compounds for anti-human coronavirus 229E activity and identified an inhibitor, designated K22, that specifically targets membrane-bound coronaviral RNA synthesis. K22 exerts most potent antiviral activity after virus entry during an early step of the viral life cycle. Specifically, the formation of double membrane vesicles (DMVs, a hallmark of coronavirus replication, was greatly impaired upon K22 treatment accompanied by near-complete inhibition of viral RNA synthesis. K22-resistant viruses contained substitutions in non-structural protein 6 (nsp6, a membrane-spanning integral component of the viral replication complex implicated in DMV formation, corroborating that K22 targets membrane bound viral RNA synthesis. Besides K22 resistance, the nsp6 mutants induced a reduced number of DMVs, displayed decreased specific infectivity, while RNA synthesis was not affected. Importantly, K22 inhibits a broad range of coronaviruses, including Middle East respiratory syndrome coronavirus (MERS-CoV, and efficient inhibition was achieved in primary human epithelia cultures representing the entry port of human coronavirus infection. Collectively, this study proposes an evolutionary conserved step in the life cycle of positive-stranded RNA viruses, the recruitment of cellular membranes for viral replication, as vulnerable and, most importantly, druggable target for antiviral intervention. We expect this mode of action to serve as a paradigm for the development of potent antiviral drugs to combat many animal and human virus infections.

Thermo-and pH-sensitive hydrogel membranes composed of poly(N-isopropylacrylamide)-hyaluronan for biomedical applications: Influence of hyaluronan incorporation on the membrane properties.

Science.gov (United States)

Kamoun, Elbadawy A; Fahmy, Alaa; Taha, Tarek H; El-Fakharany, Esmail M; Makram, Mohamed; Soliman, Hesham M A; Shehata, Hassan

2018-01-01

Interpenetrating hydrogel membranes consisting of pH-sensitive hyaluronan (HA) and thermo-sensitive poly(N-isopropylacrylamide) (PNIPAAM) were synthesized using redox polymerization, followed by N,N-methylenebisacrylamide (BIS) and epichlorohydrin (EPI) were added as chemical crosslinkers. The interaction between membrane compositions has been characterized by FTIR spectroscopy and discussed intensively. The result indicates that HA incorporation in membranes increase the gel fraction, swelling uptake, and the flexibility/elasticity of crosslinked membranes, however it reduced oppositely the mechanical elongation of membranes. PNIPAAm-HA hydrogels responded to both temperature and pH changes and the stimuli-responsiveness was reversible. However, in vitro bioevaluation results revealed that the released ampicillin during the burst release time was sharply influenced and increased with increasing HA contents in membranes; afterwards it became sustainable. Whereas, high HA contents in hydrogels unexpectedly impacted negatively on the cells viability, owing to the viscosity of cell culture media changed. A big resistance was observed against microbial growth of Staphylococcus aureus, Salmonella typhi, and Candida albicans in case of pure PNIPAAm hydrogel membranes without HA or ampicillin. However, HA incorporation or the loaded ampicillin in membranes showed unexpected easily microbial growth. The fast release performance with dual pH-thermo-sensitive hydrogels were suggested as promising materials for quick drug carrier in the biomedical field. Copyright © 2017 Elsevier B.V. All rights reserved.

A wrinkle in flight: the role of elastin fibres in the mechanical behaviour of bat wing membranes.

Science.gov (United States)

Cheney, Jorn A; Konow, Nicolai; Bearnot, Andrew; Swartz, Sharon M

2015-05-06

Bats fly using a thin wing membrane composed of compliant, anisotropic skin. Wing membrane skin deforms dramatically as bats fly, and its three-dimensional configurations depend, in large part, on the mechanical behaviour of the tissue. Large, macroscopic elastin fibres are an unusual mechanical element found in the skin of bat wings. We characterize the fibre orientation and demonstrate that elastin fibres are responsible for the distinctive wrinkles in the surrounding membrane matrix. Uniaxial mechanical testing of the wing membrane, both parallel and perpendicular to elastin fibres, is used to distinguish the contribution of elastin and the surrounding matrix to the overall membrane mechanical behaviour. We find that the matrix is isotropic within the plane of the membrane and responsible for bearing load at high stress; elastin fibres are responsible for membrane anisotropy and only contribute substantially to load bearing at very low stress. The architecture of elastin fibres provides the extreme extensibility and self-folding/self-packing of the wing membrane skin. We relate these findings to flight with membrane wings and discuss the aeromechanical significance of elastin fibre pre-stress, membrane excess length, and how these parameters may aid bats in resisting gusts and preventing membrane flutter. © 2015 The Author(s) Published by the Royal Society. All rights reserved.

Benign Mature Mediastinal Dysembryoma with Pulmonary Extension Revealed by Recurrent Hemoptysis in a Young Woman

International Nuclear Information System (INIS)

Filaire, M.; Michel-Letonturier, M.; Garcier, J. M.; Escande, G.; Boyer, L.

2006-01-01

We report one case of mature mediastinal teratoma with pulmonary extension surgically diagnosed in a 22-year-old woman complaining of recurrent hemoptyses for which no etiological explanation could be found. Thoracic surgery was only decided on after three embolizations proved ineffective

Transverse relaxation dispersion of the p7 membrane channel from hepatitis C virus reveals conformational breathing

International Nuclear Information System (INIS)

Dev, Jyoti; Brüschweiler, Sven; Ouyang, Bo; Chou, James J.

2015-01-01

The p7 membrane protein encoded by hepatitis C virus (HCV) assembles into a homo-hexamer that selectively conducts cations. An earlier solution NMR structure of the hexameric complex revealed a funnel-like architecture and suggests that a ring of conserved asparagines near the narrow end of the funnel are important for cation interaction. NMR based drug-binding experiments also suggest that rimantadine can allosterically inhibit ion conduction via a molecular wedge mechanism. These results suggest the presence of dilation and contraction of the funnel tip that are important for channel activity and that the action of the drug is attenuating this motion. Here, we determined the conformational dynamics and solvent accessibility of the p7 channel. The proton exchange measurements show that the cavity-lining residues are largely water accessible, consistent with the overall funnel shape of the channel. Our relaxation dispersion data show that residues Val7 and Leu8 near the asparagine ring are subject to large chemical exchange, suggesting significant intrinsic channel breathing at the tip of the funnel. Moreover, the hinge regions connecting the narrow and wide regions of the funnel show strong relaxation dispersion and these regions are the binding sites for rimantadine. Presence of rimantadine decreases the conformational dynamics near the asparagine ring and the hinge area. Our data provide direct observation of μs–ms dynamics of the p7 channel and support the molecular wedge mechanism of rimantadine inhibition of the HCV p7 channel

Transverse relaxation dispersion of the p7 membrane channel from hepatitis C virus reveals conformational breathing

Energy Technology Data Exchange (ETDEWEB)

Dev, Jyoti; Brüschweiler, Sven [Harvard Medical School, Department of Biological Chemistry and Molecular Pharmacology (United States); Ouyang, Bo [Chinese Academy of Sciences, State Key Laboratory of Molecular Biology, Shanghai Institute of Biochemistry and Cell Biology (China); Chou, James J., E-mail: james-chou@hms.harvard.edu [Harvard Medical School, Department of Biological Chemistry and Molecular Pharmacology (United States)

2015-04-15

The p7 membrane protein encoded by hepatitis C virus (HCV) assembles into a homo-hexamer that selectively conducts cations. An earlier solution NMR structure of the hexameric complex revealed a funnel-like architecture and suggests that a ring of conserved asparagines near the narrow end of the funnel are important for cation interaction. NMR based drug-binding experiments also suggest that rimantadine can allosterically inhibit ion conduction via a molecular wedge mechanism. These results suggest the presence of dilation and contraction of the funnel tip that are important for channel activity and that the action of the drug is attenuating this motion. Here, we determined the conformational dynamics and solvent accessibility of the p7 channel. The proton exchange measurements show that the cavity-lining residues are largely water accessible, consistent with the overall funnel shape of the channel. Our relaxation dispersion data show that residues Val7 and Leu8 near the asparagine ring are subject to large chemical exchange, suggesting significant intrinsic channel breathing at the tip of the funnel. Moreover, the hinge regions connecting the narrow and wide regions of the funnel show strong relaxation dispersion and these regions are the binding sites for rimantadine. Presence of rimantadine decreases the conformational dynamics near the asparagine ring and the hinge area. Our data provide direct observation of μs–ms dynamics of the p7 channel and support the molecular wedge mechanism of rimantadine inhibition of the HCV p7 channel.

Membrane fatty acid composition and radiation response of Bp8 sarcoma ascites tumour cells

International Nuclear Information System (INIS)

Harms-Ringdahl, M.

1987-01-01

Radiation responses of Bp8 sarcoma ascites tumour cells with differences in membrane fatty acid composition was studied. The cells were grown i.p. in NMRI mice and their membrane composition was changed in response to different dietary regimes provided to the hosts. Cell survival, varied insignificantly between the four dietary groups, while repair capacity differed significantly. Increased repair capacity was observed for ascites cells grown in animals on diets enriched in sunflower seed oil and coconut oil, compared with cells from mice fed the hydrogenated lard diet or from cells from the control animals. The membrane fatty acid composition of the cells from the two dietary groups with increased levels of repair capacity differed extensively, and in general there was no correlation between radiation response and the membrane fatty acid composition of the four groups. For coconut oil and control groups with marked differences in membrane fatty acid composition, the effects of irradiation on ascites tumour growth rate and cell cycle distribution were followed in vivo. For none of the parameters was an effect on membrane fatty acid composition on radiation response observed. (author)

Membrane topology analysis of HIV-1 envelope glycoprotein gp41

Directory of Open Access Journals (Sweden)

Xiao Dan

2010-11-01

Full Text Available Abstract Background The gp41 subunit of the HIV-1 envelope glycoprotein (Env has been widely regarded as a type I transmembrane protein with a single membrane-spanning domain (MSD. An alternative topology model suggested multiple MSDs. The major discrepancy between the two models is that the cytoplasmic Kennedy sequence in the single MSD model is assigned as the extracellular loop accessible to neutralizing antibodies in the other model. We examined the membrane topology of the gp41 subunit in both prokaryotic and mammalian systems. We attached topological markers to the C-termini of serially truncated gp41. In the prokaryotic system, we utilized a green fluorescent protein (GFP that is only active in the cytoplasm. The tag protein (HaloTag and a membrane-impermeable ligand specific to HaloTag was used in the mammalian system. Results In the absence of membrane fusion, both the prokaryotic and mammalian systems (293FT cells supported the single MSD model. In the presence of membrane fusion in mammalian cells (293CD4 cells, the data obtained seem to support the multiple MSD model. However, the region predicted to be a potential MSD is the highly hydrophilic Kennedy sequence and is least likely to become a MSD based on several algorithms. Further analysis revealed the induction of membrane permeability during membrane fusion, allowing the membrane-impermeable ligand and antibodies to cross the membrane. Therefore, we cannot completely rule out the possible artifacts. Addition of membrane fusion inhibitors or alterations of the MSD sequence decreased the induction of membrane permeability. Conclusions It is likely that a single MSD model for HIV-1 gp41 holds true even in the presence of membrane fusion. The degree of the augmentation of membrane permeability we observed was dependent on the membrane fusion and sequence of the MSD.

A Multiparameter Network Reveals Extensive Divergence between C. elegans bHLH Transcription Factors

DEFF Research Database (Denmark)

Grove, C.; De Masi, Federico; Newburger, Daniel

2009-01-01

parameters remain undetermined. We comprehensively identify dimerization partners, spatiotemporal expression patterns, and DNA-binding specificities for the C. elegans bHLH family of TFs, and model these data into an integrated network. This network displays both specificity and promiscuity, as some b......HLH proteins, DNA sequences, and tissues are highly connected, whereas others are not. By comparing all bHLH TFs, we find extensive divergence and that all three parameters contribute equally to bHLH divergence. Our approach provides a framework for examining divergence for other protein families in C. elegans...

Investigation of cell adhesion in chitosan membranes for peripheral nerve regeneration

Energy Technology Data Exchange (ETDEWEB)

Carvalho, Cristiana R.; López-Cebral, Rita; Silva-Correia, Joana; Silva, Joana M.; Mano, João F.; Silva, Tiago H. [3B' s Research Group - Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, AvePark – Parque de Ciência e Tecnologia, 4805-017, Barco, Guimarães (Portugal); ICVS/3B' s - PT Government Associate Laboratory, Braga, Guimarães (Portugal); Freier, Thomas [MEDOVENT GmbH, Friedrich-Koenig-Str. 3, D-55129 Mainz (Germany); Reis, Rui L. [3B' s Research Group - Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, AvePark – Parque de Ciência e Tecnologia, 4805-017, Barco, Guimarães (Portugal); ICVS/3B' s - PT Government Associate Laboratory, Braga, Guimarães (Portugal); Oliveira, Joaquim M., E-mail: miguel.oliveira@dep.uminho.pt [3B' s Research Group - Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, AvePark – Parque de Ciência e Tecnologia, 4805-017, Barco, Guimarães (Portugal); ICVS/3B' s - PT Government Associate Laboratory, Braga, Guimarães (Portugal)

2017-02-01

Peripheral nerve injuries have produced major concerns in regenerative medicine for several years, as the recovery of normal nerve function continues to be a significant clinical challenge. Chitosan (CHT), because of its good biocompatibility, biodegradability and physicochemical properties, has been widely used as a biomaterial in tissue engineering scaffolding. In this study, CHT membranes were produced with three different Degrees of Acetylation (DA), envisioning its application in peripheral nerve regeneration. The three CHT membranes (DA I: 1%, DA II: 2%, DA III: 5%) were extensively characterized and were found to have a smooth and flat surface, with DA III membrane having slightly higher roughness and surface energy. All the membranes presented suitable mechanical properties and did not show any signs of calcification after SBF test. Biodegradability was similar for all samples, and adequate to physically support neurite outgrowth. The in vitro cell culture results indicate selective cell adhesion. The CHT membranes favoured Schwann cells invasion and proliferation, with a display of appropriate cytoskeletal morphology. At the same time they presented low fibroblast infiltration. This fact may be greatly beneficial for the prevention of fibrotic tissue formation, a common phenomenon impairing peripheral nerve regeneration. The great deal of results obtained during this work permitted to select the formulation with the greatest potential for further biological tests. - Highlights: • Three chitosan membranes were produced with very specific degrees of acetylation (DA I: 1%, DA II: 2%, DA III: 5%). • Physicochemical characterization of the membranes showed their suitability for peripheral nerve regeneration purposes. • In vitro cellular tests confirmed the potential of the membranes as peripheral nerve regeneration systems. • The results indicated that DA III membrane should be the one considered for further peripheral nerve regeneration studies.

Investigation of cell adhesion in chitosan membranes for peripheral nerve regeneration

International Nuclear Information System (INIS)

Carvalho, Cristiana R.; López-Cebral, Rita; Silva-Correia, Joana; Silva, Joana M.; Mano, João F.; Silva, Tiago H.; Freier, Thomas; Reis, Rui L.; Oliveira, Joaquim M.

2017-01-01

Peripheral nerve injuries have produced major concerns in regenerative medicine for several years, as the recovery of normal nerve function continues to be a significant clinical challenge. Chitosan (CHT), because of its good biocompatibility, biodegradability and physicochemical properties, has been widely used as a biomaterial in tissue engineering scaffolding. In this study, CHT membranes were produced with three different Degrees of Acetylation (DA), envisioning its application in peripheral nerve regeneration. The three CHT membranes (DA I: 1%, DA II: 2%, DA III: 5%) were extensively characterized and were found to have a smooth and flat surface, with DA III membrane having slightly higher roughness and surface energy. All the membranes presented suitable mechanical properties and did not show any signs of calcification after SBF test. Biodegradability was similar for all samples, and adequate to physically support neurite outgrowth. The in vitro cell culture results indicate selective cell adhesion. The CHT membranes favoured Schwann cells invasion and proliferation, with a display of appropriate cytoskeletal morphology. At the same time they presented low fibroblast infiltration. This fact may be greatly beneficial for the prevention of fibrotic tissue formation, a common phenomenon impairing peripheral nerve regeneration. The great deal of results obtained during this work permitted to select the formulation with the greatest potential for further biological tests. - Highlights: • Three chitosan membranes were produced with very specific degrees of acetylation (DA I: 1%, DA II: 2%, DA III: 5%). • Physicochemical characterization of the membranes showed their suitability for peripheral nerve regeneration purposes. • In vitro cellular tests confirmed the potential of the membranes as peripheral nerve regeneration systems. • The results indicated that DA III membrane should be the one considered for further peripheral nerve regeneration studies.

Thermal stability of nafion membranes under mechanical stress

Energy Technology Data Exchange (ETDEWEB)

Quintilii, M; Struis, R [Paul Scherrer Inst. (PSI), Villigen (Switzerland)

1997-06-01

The feasibility of adequately modified fluoro-ionomer membranes (NAFION{sup R}) is demonstrated for the selective separation of methanol synthesis products from the raw reactor gas at temperatures around 200{sup o}C. For an economically relevant application of this concept on a technical scale the Nafion membranes should be thin ({approx_equal}10 {mu}m) and thermally stable over a long period of time (1-2 years). In cooperation with industry (Methanol Casale SA, Lugano (CH)), we test the thermal stability of Nafion hollow fibers and supported Nafion thin sheet membranes at temperatures between 160 and 200{sup o}C under mechanical stress by applying a gas pressure difference over the membrane surface ({Delta}P{<=} 40 bar). Tests with the hollow fibers revealed that Nafion has visco-elastic properties. Tests with 50 {mu}m thin Nafion sheets supported by a porous metal carrier at 200{sup o}C and {Delta}P=39 bar showed no mechanical defects over a period of 92 days. (author) 5 figs., 4 refs.

RNAi-mediated downregulation of poplar plasma membrane intrinsic proteins (PIPs) changes plasma membrane proteome composition and affects leaf physiology.

Science.gov (United States)

Bi, Zhen; Merl-Pham, Juliane; Uehlein, Norbert; Zimmer, Ina; Mühlhans, Stefanie; Aichler, Michaela; Walch, Axel Karl; Kaldenhoff, Ralf; Palme, Klaus; Schnitzler, Jörg-Peter; Block, Katja

2015-10-14

Plasma membrane intrinsic proteins (PIPs) are one subfamily of aquaporins that mediate the transmembrane transport of water. To reveal their function in poplar, we generated transgenic poplar plants in which the translation of PIP genes was downregulated by RNA interference investigated these plants with a comprehensive leaf plasma membrane proteome and physiome analysis. First, inhibition of PIP synthesis strongly altered the leaf plasma membrane protein composition. Strikingly, several signaling components and transporters involved in the regulation of stomatal movement were differentially regulated in transgenic poplars. Furthermore, hormonal crosstalk related to abscisic acid, auxin and brassinosteroids was altered, in addition to cell wall biosynthesis/cutinization, the organization of cellular structures and membrane trafficking. A physiological analysis confirmed the proteomic results. The leaves had wider opened stomata and higher net CO2 assimilation and transpiration rates as well as greater mesophyll conductance for CO2 (gm) and leaf hydraulic conductance (Kleaf). Based on these results, we conclude that PIP proteins not only play essential roles in whole leaf water and CO2 flux but have important roles in the regulation of stomatal movement. Copyright © 2015. Published by Elsevier B.V.

Mitochondrial shape governs BAX-induced membrane permeabilization and apoptosis.

Science.gov (United States)

Renault, Thibaud T; Floros, Konstantinos V; Elkholi, Rana; Corrigan, Kelly-Ann; Kushnareva, Yulia; Wieder, Shira Y; Lindtner, Claudia; Serasinghe, Madhavika N; Asciolla, James J; Buettner, Christoph; Newmeyer, Donald D; Chipuk, Jerry E

2015-01-08

Proapoptotic BCL-2 proteins converge upon the outer mitochondrial membrane (OMM) to promote mitochondrial outer membrane permeabilization (MOMP) and apoptosis. Here we investigated the mechanistic relationship between mitochondrial shape and MOMP and provide evidence that BAX requires a distinct mitochondrial size to induce MOMP. We utilized the terminal unfolded protein response pathway to systematically define proapoptotic BCL-2 protein composition after stress and then directly interrogated their requirement for a productive mitochondrial size. Complementary biochemical, cellular, in vivo, and ex vivo studies reveal that Mfn1, a GTPase involved in mitochondrial fusion, establishes a mitochondrial size that is permissive for proapoptotic BCL-2 family function. Cells with hyperfragmented mitochondria, along with size-restricted OMM model systems, fail to support BAX-dependent membrane association and permeabilization due to an inability to stabilize BAXα9·membrane interactions. This work identifies a mechanistic contribution of mitochondrial size in dictating BAX activation, MOMP, and apoptosis. Copyright © 2015 Elsevier Inc. All rights reserved.

Study of Aging ion exchange membranes used in separation processes

International Nuclear Information System (INIS)

Bellakhal, N.; Ghalloussi, R.; Dammak, L.

2009-01-01

capacities of these new membranes and worn. Indeed, there is a marked decrease of this magnitude in the case of membrane treatment. - Extensive use of these membranes in the presence of media loaded molecules organic causes plugging gaps left open by the chains macromolecular. Such gaps are space travel and dissemination of electrolytes. The comparative study of ionic permeability of new and used IEM showed that over the membrane permeability is worn over the electrolyte is low. This was attributed to inhibition of some functional sites, and the clogging of interstices by organic molecules. FTIR tests are underway to confirm the presence of these macromolecules in the membrane treatment.

Caloric Restriction-Induced Extension of Chronological Lifespan Requires Intact Respiration in Budding Yeast.

Science.gov (United States)

Kwon, Young-Yon; Lee, Sung-Keun; Lee, Cheol-Koo

2017-04-01

Caloric restriction (CR) has been shown to extend lifespan and prevent cellular senescence in various species ranging from yeast to humans. Many effects of CR may contribute to extend lifespan. Specifically, CR prevents oxidative damage from reactive oxygen species (ROS) by enhancing mitochondrial function. In this study, we characterized 33 single electron transport chain (ETC) gene-deletion strains to identify CR-induced chronological lifespan (CLS) extension mechanisms. Interestingly, defects in 17 of these 33 ETC gene-deleted strains showed loss of both respiratory function and CR-induced CLS extension. On the contrary, the other 16 respiration-capable mutants showed increased CLS upon CR along with increased mitochondrial membrane potential (MMP) and intracellular adenosine triphosphate (ATP) levels, with decreased mitochondrial superoxide generation. We measured the same parameters in the 17 non-respiratory mutants upon CR. CR simultaneously increased MMP and mitochondrial superoxide generation without altering intracellular ATP levels. In conclusion, respiration is essential for CLS extension by CR and is important for balancing MMP, ROS, and ATP levels.

Membrane properties involved in calcium-stimulated microparticle release from the plasma membranes of S49 lymphoma cells.

Science.gov (United States)

Campbell, Lauryl E; Nelson, Jennifer; Gibbons, Elizabeth; Judd, Allan M; Bell, John D

2014-01-01

This study answered the question of whether biophysical mechanisms for microparticle shedding discovered in platelets and erythrocytes also apply to nucleated cells: cytoskeletal disruption, potassium efflux, transbilayer phospholipid migration, and membrane disordering. The calcium ionophore, ionomycin, disrupted the actin cytoskeleton of S49 lymphoma cells and produced rapid release of microparticles. This release was significantly inhibited by interventions that impaired calcium-activated potassium current. Microparticle release was also greatly reduced in a lymphocyte cell line deficient in the expression of scramblase, the enzyme responsible for calcium-stimulated dismantling of the normal phospholipid transbilayer asymmetry. Rescue of the scrambling function at high ionophore concentration also resulted in enhanced particle shedding. The effect of membrane physical properties was addressed by varying the experimental temperature (32-42°C). A significant positive trend in the rate of microparticle release as a function of temperature was observed. Fluorescence experiments with trimethylammonium diphenylhexatriene and Patman revealed significant decrease in the level of apparent membrane order along that temperature range. These results demonstrated that biophysical mechanisms involved in microparticle release from platelets and erythrocytes apply also to lymphocytes.

Biogenesis of the demarcation membrane system (DMS) in megakaryocytes.

Science.gov (United States)

Eckly, Anita; Heijnen, Harry; Pertuy, Fabien; Geerts, Willie; Proamer, Fabienne; Rinckel, Jean-Yves; Léon, Catherine; Lanza, François; Gachet, Christian

2014-02-06

The demarcation membrane system (DMS) in megakaryocytes forms the plasma membrane (PM) of future platelets. Using confocal microscopy, electron tomography, and large volume focused ion beam/scanning electron microscopy (FIB/SEM), we determined the sequential steps of DMS formation. We identified a pre-DMS that initiated at the cell periphery and was precisely located between the nuclear lobes. At all developmental stages, the DMS remained continuous with the cell surface. The number of these connections correlated well with the nuclear lobulation, suggesting a relationship with cleavage furrow formation and abortive cytokinesis. On DMS expansion, Golgi complexes assembled around the pre-DMS, and fusion profiles between trans-golgi network-derived vesicles and the DMS were observed. Brefeldin-A reduced DMS expansion, indicating that the exocytic pathway is essential for DMS biogenesis. Close contacts between the endoplasmic reticulum (ER) and the DMS were detected, suggesting physical interaction between the 2 membrane systems. FIB/SEM revealed that the DMS forms an intertwined tubular membrane network resembling the platelet open canalicular system. We thus propose the following steps in DMS biogenesis: (1) focal membrane assembly at the cell periphery; (2) PM invagination and formation of a perinuclear pre-DMS; (3) expansion through membrane delivery from Golgi complexes; and (4) ER-mediated lipid transfer.

The Membrane Steps of Bacterial Cell Wall Synthesis as Antibiotic Targets

Directory of Open Access Journals (Sweden)

Yao Liu

2016-08-01

Full Text Available Peptidoglycan is the major component of the cell envelope of virtually all bacteria. It has structural roles and acts as a selective sieve for molecules from the outer environment. Peptidoglycan synthesis is therefore one of the most important biogenesis pathways in bacteria and has been studied extensively over the last twenty years. The pathway starts in the cytoplasm, continues in the cytoplasmic membrane and finishes in the periplasmic space, where the precursor is polymerized into the peptidoglycan layer. A number of proteins involved in this pathway, such as the Mur enzymes and the penicillin binding proteins (PBPs, have been studied and regarded as good targets for antibiotics. The present review focuses on the membrane steps of peptidoglycan synthesis that involve two enzymes, MraY and MurG, the inhibitors of these enzymes and the inhibition mechanisms. We also discuss the challenges of targeting these two cytoplasmic membrane (associated proteins in bacterial cells and the perspectives on how to overcome the issues.

N- and O-glycosylation Analysis of Human C1-inhibitor Reveals Extensive Mucin-type O-Glycosylation.

Science.gov (United States)

Stavenhagen, Kathrin; Kayili, H Mehmet; Holst, Stephanie; Koeleman, Carolien A M; Engel, Ruchira; Wouters, Diana; Zeerleder, Sacha; Salih, Bekir; Wuhrer, Manfred

2018-06-01

Human C1-inhibitor (C1-Inh) is a serine protease inhibitor and the major regulator of the contact activation pathway as well as the classical and lectin complement pathways. It is known to be a highly glycosylated plasma glycoprotein. However, both the structural features and biological role of C1-Inh glycosylation are largely unknown. Here, we performed for the first time an in-depth site-specific N - and O -glycosylation analysis of C1-Inh combining various mass spectrometric approaches, including C18-porous graphitized carbon (PGC)-LC-ESI-QTOF-MS/MS applying stepping-energy collision-induced dissociation (CID) and electron-transfer dissociation (ETD). Various proteases were applied, partly in combination with PNGase F and exoglycosidase treatment, in order to analyze the (glyco)peptides. The analysis revealed an extensively O -glycosylated N-terminal region. Five novel and five known O -glycosylation sites were identified, carrying mainly core1-type O -glycans. In addition, we detected a heavily O -glycosylated portion spanning from Thr 82 -Ser 121 with up to 16 O -glycans attached. Likewise, all known six N -glycosylation sites were covered and confirmed by this site-specific glycosylation analysis. The glycoforms were in accordance with results on released N -glycans by MALDI-TOF/TOF-MS/MS. The comprehensive characterization of C1-Inh glycosylation described in this study will form the basis for further functional studies on the role of these glycan modifications. © 2018 by The American Society for Biochemistry and Molecular Biology, Inc.

Scanning electron microscopy of the collodion membrane from a self-healing collodion baby*

Science.gov (United States)

de Almeida Jr., Hiram Larangeira; Isaacsson, Henrique; Guarenti, Isabelle Maffei; Silva, Ricardo Marques e; de Castro, Luis Antônio Suita

2015-01-01

Abstract Self-healing collodion baby is a well-established subtype of this condition. We examined a male newborn, who was covered by a collodion membrane. The shed membrane was examined with scanning electron microscopy. The outer surface showed a very compact keratin without the normal elimination of corneocytes. The lateral view of the specimen revealed a very thick, horny layer. The inner surface showed the structure of lower corneocytes with polygonal contour. With higher magnifications villous projections were seen in the cell membrane. PMID:26375232

Isolation and characterization of the plasma membrane from the yeast Pichia pastoris.

Science.gov (United States)

Grillitsch, Karlheinz; Tarazona, Pablo; Klug, Lisa; Wriessnegger, Tamara; Zellnig, Günther; Leitner, Erich; Feussner, Ivo; Daum, Günther

2014-07-01

Despite similarities of cellular membranes in all eukaryotes, every compartment displays characteristic and often unique features which are important for the functions of the specific organelles. In the present study, we biochemically characterized the plasma membrane of the methylotrophic yeast Pichia pastoris with emphasis on the lipids which form the matrix of this compartment. Prerequisite for this effort was the design of a standardized and reliable isolation protocol of the plasma membrane at high purity. Analysis of isolated plasma membrane samples from P. pastoris revealed an increase of phosphatidylserine and a decrease of phosphatidylcholine compared to bulk membranes. The amount of saturated fatty acids in the plasma membrane was higher than in total cell extracts. Ergosterol, the final product of the yeast sterol biosynthetic pathway, was found to be enriched in plasma membrane fractions, although markedly lower than in Saccharomyces cerevisiae. A further characteristic feature of the plasma membrane from P. pastoris was the enrichment of inositol phosphorylceramides over neutral sphingolipids, which accumulated in internal membranes. The detailed analysis of the P. pastoris plasma membrane is discussed in the light of cell biological features of this microorganism especially as a microbial cell factory for heterologous protein production. Copyright © 2014 Elsevier B.V. All rights reserved.

Characterization of phospholipid composition and its control in the plasma membrane of developing soybean root

International Nuclear Information System (INIS)

Whitman, C.E.

1985-01-01

The phospholipid composition of plasma membrane enriched fractions from developing soybean root and several mechanisms which may regulate it have been examined. Plasma membrane vesicles were isolated from meristematic and mature sections of four-day-old dark grown soybean roots (Glycine max [L.] Merr. Cult. Wells II). Analysis of lipid extracts revealed two major phospholipid classes: phosphatidylcholine and phosphatidylethanolamine. Minor phospholipid classes were phosphatidylinositol, phosphatidylserine, phosphatidylgylcerol and diphosphatidylgylcerol. Phospholipid composition was similar at each developmental stage. Fatty acids of phosphatidylcholine and phosphatidylethanolamine were 16:0, 18:0, 18:2, and 18:3. Fatty acid composition varied with both phospholipid class and the developmental stage of the root. The degradation of phosphatidylcholine by endogenous phospholipase D during membrane isolation indicated that this enzyme might be involved in phospholipid turnover within the membrane. Phospholipase D activity was heat labile and increasing the pH of the enzyme assay from 5.3 to 7.8 resulted in 90% inhibition of activity. The turnover of fatty acids within the phospholipids of the plasma membrane was studied. Mature root sections were incubated with [1- 14 C] acetate, 1 mM Na acetate and 50 μg/ml chloramphenicol. Membrane lipid extracts analyzed for phospholipid class and acyl chain composition revealed that the long incubation times did not alter the phospholipid composition of the plasma membrane enriched fraction

Giant plasma membrane vesicles: models for understanding membrane organization.

Science.gov (United States)

Levental, Kandice R; Levental, Ilya

2015-01-01

The organization of eukaryotic membranes into functional domains continues to fascinate and puzzle cell biologists and biophysicists. The lipid raft hypothesis proposes that collective lipid interactions compartmentalize the membrane into coexisting liquid domains that are central to membrane physiology. This hypothesis has proven controversial because such structures cannot be directly visualized in live cells by light microscopy. The recent observations of liquid-liquid phase separation in biological membranes are an important validation of the raft hypothesis and enable application of the experimental toolbox of membrane physics to a biologically complex phase-separated membrane. This review addresses the role of giant plasma membrane vesicles (GPMVs) in refining the raft hypothesis and expands on the application of GPMVs as an experimental model to answer some of key outstanding problems in membrane biology. Copyright © 2015 Elsevier Inc. All rights reserved.

A remote arene-binding site on prostate specific membrane antigen revealed by antibody-recruiting small molecules

Czech Academy of Sciences Publication Activity Database

Zhang, A.X.; Murelli, R.P.; Bařinka, Cyril; Michel, J.; Cocleaza, A.; Jorgensen, W.L.; Lubkowski, J.; Spiegel, D.A.

2010-01-01

Roč. 132, č. 36 (2010), s. 12711-12716 ISSN 0002-7863 Institutional research plan: CEZ:AV0Z50520701 Keywords : Prostate -specific membrane antigen * antibody recruiting molecules * Structure-activity relationship Subject RIV: CE - Biochemistry Impact factor: 9.019, year: 2010

A Review on Cold Start of Proton Exchange Membrane Fuel Cells

Directory of Open Access Journals (Sweden)

Zhongmin Wan

2014-05-01

Full Text Available Successful and rapid startup of proton exchange membrane fuel cells (PEMFCs at subfreezing temperatures (also called cold start is of great importance for their commercialization in automotive and portable devices. In order to maintain good proton conductivity, the water content in the membrane must be kept at a certain level to ensure that the membrane remains fully hydrated. However, the water in the pores of the catalyst layer (CL, gas diffusion layer (GDL and the membrane may freeze once the cell temperature decreases below the freezing point (Tf. Thus, methods which could enable the fuel cell startup without or with slight performance degradation at subfreezing temperature need to be studied. This paper presents an extensive review on cold start of PEMFCs, including the state and phase changes of water in PEMFCs, impacts of water freezing on PEMFCs, numerical and experimental studies on PEMFCs, and cold start strategies. The impacts on each component of the fuel cell are discussed in detail. Related numerical and experimental work is also discussed. It should be mentioned that the cold start strategies, especially the enumerated patents, are of great reference value on the practical cold start process.

Oriented Morphology and Anisotropic Transport in Uniaxially Stretched Perfluorosulfonate Ionomer Membranes

Energy Technology Data Exchange (ETDEWEB)

J Park; J Li; G Divoux; L Madsen; R Moore

2011-12-31

Relations between morphology and transport sensitively govern proton conductivity in perfluorsulfonate ionomers (PFSIs) and thus determine useful properties of these technologically important materials. In order to understand such relations, we have conducted a broad systematic study of H{sup +}-form PFSI membranes over a range of uniaxial extensions and water uptakes. On the basis of small-angle X-ray scattering (SAXS) and {sup 2}H NMR spectroscopy, uniaxial deformation induces a strong alignment of ionic domains along the stretching direction. We correlate ionic domain orientation to transport using pulsed-field-gradient {sup 1}H NMR measurements of water diffusion coefficients along the three orthogonal membrane directions. Intriguingly, we observe that uniaxial deformation enhances water transport in one direction (parallel-to-draw direction) while reducing it in the other two directions (two orthogonal directions relative to the stretching direction). We evaluate another important transport parameter, proton conductivity, along two orthogonal in-plane directions. In agreement with water diffusion experiments, orientation of ionic channels increases proton conduction along the stretching direction while decreasing it in the perpendicular direction. These findings provide valuable fodder for optimal application of PFSI membranes as well as for the design of next generation polymer electrolyte membranes.

Single-cell analysis of pyroptosis dynamics reveals conserved GSDMD-mediated subcellular events that precede plasma membrane rupture.

Science.gov (United States)

de Vasconcelos, Nathalia M; Van Opdenbosch, Nina; Van Gorp, Hanne; Parthoens, Eef; Lamkanfi, Mohamed

2018-04-17

Pyroptosis is rapidly emerging as a mechanism of anti-microbial host defense, and of extracellular release of the inflammasome-dependent cytokines interleukin (IL)-1β and IL-18, which contributes to autoinflammatory pathology. Caspases 1, 4, 5 and 11 trigger this regulated form of necrosis by cleaving the pyroptosis effector gasdermin D (GSDMD), causing its pore-forming amino-terminal domain to oligomerize and perforate the plasma membrane. However, the subcellular events that precede pyroptotic cell lysis are ill defined. In this study, we triggered primary macrophages to undergo pyroptosis from three inflammasome types and recorded their dynamics and morphology using high-resolution live-cell spinning disk confocal laser microscopy. Based on quantitative analysis of single-cell subcellular events, we propose a model of pyroptotic cell disintegration that is initiated by opening of GSDMD-dependent ion channels or pores that are more restrictive than recently proposed GSDMD pores, followed by osmotic cell swelling, commitment of mitochondria and other membrane-bound organelles prior to sudden rupture of the plasma membrane and full permeability to intracellular proteins. This study provides a dynamic framework for understanding cellular changes that occur during pyroptosis, and charts a chronological sequence of GSDMD-mediated subcellular events that define pyroptotic cell death at the single-cell level.

A robust mass spectrometry method for rapid profiling of erythrocyte ghost membrane proteomes.

Science.gov (United States)

Fye, Haddy K S; Mrosso, Paul; Bruce, Lesley; Thézénas, Marie-Laëtitia; Davis, Simon; Fischer, Roman; Rwegasira, Gration L; Makani, Julie; Kessler, Benedikt M

2018-01-01

Red blood cell (RBC) physiology is directly linked to many human disorders associated with low tissue oxygen levels or anemia including chronic obstructive pulmonary disease, congenital heart disease, sleep apnea and sickle cell anemia. Parasites such as Plasmodium spp. and phylum Apicomplexa directly target RBCs, and surface molecules within the RBC membrane are critical for pathogen interactions. Proteomics of RBC membrane 'ghost' fractions has therefore been of considerable interest, but protocols described to date are either suboptimal or too extensive to be applicable to a larger set of clinical cohorts. Here, we describe an optimised erythrocyte isolation protocol from blood, tested for various storage conditions and explored using different fractionation conditions for isolating ghost RBC membranes. Liquid chromatography mass spectrometry (LC-MS) analysis on a Q-Exactive Orbitrap instrument was used to profile proteins isolated from the comparative conditions. Data analysis was run on the MASCOT and MaxQuant platforms to assess their scope and diversity. The results obtained demonstrate a robust method for membrane enrichment enabling consistent MS based characterisation of > 900 RBC membrane proteins in single LC-MS/MS analyses. Non-detergent based membrane solubilisation methods using the tissue and supernatant fractions of isolated ghost membranes are shown to offer effective haemoglobin removal as well as diverse recovery including erythrocyte membrane proteins of high and low abundance. The methods described in this manuscript propose a medium to high throughput framework for membrane proteome profiling by LC-MS of potential applicability to larger clinical cohorts in a variety of disease contexts.

An AFM-based pit-measuring method for indirect measurements of cell-surface membrane vesicles

International Nuclear Information System (INIS)

Zhang, Xiaojun; Chen, Yuan; Chen, Yong

2014-01-01

Highlights: • Air drying induced the transformation of cell-surface membrane vesicles into pits. • An AFM-based pit-measuring method was developed to measure cell-surface vesicles. • Our method detected at least two populations of cell-surface membrane vesicles. - Abstract: Circulating membrane vesicles, which are shed from many cell types, have multiple functions and have been correlated with many diseases. Although circulating membrane vesicles have been extensively characterized, the status of cell-surface membrane vesicles prior to their release is less understood due to the lack of effective measurement methods. Recently, as a powerful, micro- or nano-scale imaging tool, atomic force microscopy (AFM) has been applied in measuring circulating membrane vesicles. However, it seems very difficult for AFM to directly image/identify and measure cell-bound membrane vesicles due to the similarity of surface morphology between membrane vesicles and cell surfaces. Therefore, until now no AFM studies on cell-surface membrane vesicles have been reported. In this study, we found that air drying can induce the transformation of most cell-surface membrane vesicles into pits that are more readily detectable by AFM. Based on this, we developed an AFM-based pit-measuring method and, for the first time, used AFM to indirectly measure cell-surface membrane vesicles on cultured endothelial cells. Using this approach, we observed and quantitatively measured at least two populations of cell-surface membrane vesicles, a nanoscale population (<500 nm in diameter peaking at ∼250 nm) and a microscale population (from 500 nm to ∼2 μm peaking at ∼0.8 μm), whereas confocal microscopy only detected the microscale population. The AFM-based pit-measuring method is potentially useful for studying cell-surface membrane vesicles and for investigating the mechanisms of membrane vesicle formation/release

Reconstitution of proapoptotic BAK function in liposomes reveals a dual role for mitochondrial lipids in the BAK-driven membrane permeabilization process.

Science.gov (United States)

Landeta, Olatz; Landajuela, Ane; Gil, David; Taneva, Stefka; Di Primo, Carmelo; Sot, Begoña; Valle, Mikel; Frolov, Vadim A; Basañez, Gorka

2011-03-11

BAK is a key effector of mitochondrial outer membrane permeabilization (MOMP) whose molecular mechanism of action remains to be fully dissected in intact cells, mainly due to the inherent complexity of the intracellular apoptotic machinery. Here we show that the core features of the BAK-driven MOMP pathway can be reproduced in a highly simplified in vitro system consisting of recombinant human BAK lacking the carboxyl-terminal 21 residues (BAKΔC) and tBID in combination with liposomes bearing an appropriate lipid environment. Using this minimalist reconstituted system we established that tBID suffices to trigger BAKΔC membrane insertion, oligomerization, and pore formation. Furthermore, we demonstrate that tBID-activated BAKΔC permeabilizes the membrane by forming structurally dynamic pores rather than a large proteinaceous channel of fixed size. We also identified two distinct roles played by mitochondrial lipids along the molecular pathway of BAKΔC-induced membrane permeabilization. First, using several independent approaches, we showed that cardiolipin directly interacts with BAKΔC, leading to a localized structural rearrangement in the protein that "primes" BAKΔC for interaction with tBID. Second, we provide evidence that selected curvature-inducing lipids present in mitochondrial membranes specifically modulate the energetic expenditure required to create the BAKΔC pore. Collectively, our results support the notion that BAK functions as a direct effector of MOMP akin to BAX and also adds significantly to the growing evidence indicating that mitochondrial membrane lipids are actively implicated in BCL-2 protein family function.

Large-scale atomistic and quantum-mechanical simulations of a Nafion membrane: Morphology, proton solvation and charge transport

Directory of Open Access Journals (Sweden)

Pavel V. Komarov

2013-09-01

Full Text Available Atomistic and first-principles molecular dynamics simulations are employed to investigate the structure formation in a hydrated Nafion membrane and the solvation and transport of protons in the water channel of the membrane. For the water/Nafion systems containing more than 4 million atoms, it is found that the observed microphase-segregated morphology can be classified as bicontinuous: both majority (hydrophobic and minority (hydrophilic subphases are 3D continuous and organized in an irregular ordered pattern, which is largely similar to that known for a bicontinuous double-diamond structure. The characteristic size of the connected hydrophilic channels is about 25–50 Å, depending on the water content. A thermodynamic decomposition of the potential of mean force and the calculated spectral densities of the hindered translational motions of cations reveal that ion association observed with decreasing temperature is largely an entropic effect related to the loss of low-frequency modes. Based on the results from the atomistic simulation of the morphology of Nafion, we developed a realistic model of ion-conducting hydrophilic channel within the Nafion membrane and studied it with quantum molecular dynamics. The extensive 120 ps-long density functional theory (DFT-based simulations of charge migration in the 1200-atom model of the nanochannel consisting of Nafion chains and water molecules allowed us to observe the bimodality of the van Hove autocorrelation function, which provides the direct evidence of the Grotthuss bond-exchange (hopping mechanism as a significant contributor to the proton conductivity.

Structure and physical properties of bio membranes and model membranes

International Nuclear Information System (INIS)

Tibor Hianik

2006-01-01

Bio membranes belong to the most important structures of the cell and the cell organelles. They play not only structural role of the barrier separating the external and internal part of the membrane but contain also various functional molecules, like receptors, ionic channels, carriers and enzymes. The cell membrane also preserves non-equilibrium state in a cell which is crucial for maintaining its excitability and other signaling functions. The growing interest to the bio membranes is also due to their unique physical properties. From physical point of view the bio membranes, that are composed of lipid bilayer into which are incorporated integral proteins and on their surface are anchored peripheral proteins and polysaccharides, represent liquid s crystal of smectic type. The bio membranes are characterized by anisotropy of structural and physical properties. The complex structure of bio membranes makes the study of their physical properties rather difficult. Therefore several model systems that mimic the structure of bio membranes were developed. Among them the lipid monolayers at an air-water interphase, bilayer lipid membranes, supported bilayer lipid membranes and liposomes are most known. This work is focused on the introduction into the physical word of the bio membranes and their models. After introduction to the membrane structure and the history of its establishment, the physical properties of the bio membranes and their models are stepwise presented. The most focus is on the properties of lipid monolayers, bilayer lipid membranes, supported bilayer lipid membranes and liposomes that were most detailed studied. This lecture has tutorial character that may be useful for undergraduate and graduate students in the area of biophysics, biochemistry, molecular biology and bioengineering, however it contains also original work of the author and his co-worker and PhD students, that may be useful also for specialists working in the field of bio membranes and model