WorldWideScience

Sample records for efficient membrane fusion

  1. Membrane fusion

    DEFF Research Database (Denmark)

    Bendix, Pól Martin

    2015-01-01

    At Stanford University, Boxer lab, I worked on membrane fusion of small unilamellar lipid vesicles to flat membranes tethered to glass surfaces. This geometry closely resembles biological systems in which liposomes fuse to plasma membranes. The fusion mechanism was studied using DNA zippering...... between complementary strands linked to the two apposing membranes closely mimicking the zippering mechanism of SNARE fusion complexes....

  2. Bacteriophage membrane protein P9 as a fusion partner for the efficient expression of membrane proteins in Escherichia coli.

    Science.gov (United States)

    Jung, Yuna; Jung, Hyeim; Lim, Dongbin

    2015-12-01

    Despite their important roles and economic values, studies of membrane proteins have been hampered by the difficulties associated with obtaining sufficient amounts of protein. Here, we report a novel membrane protein expression system that uses the major envelope protein (P9) of phage φ6 as an N-terminal fusion partner. Phage membrane protein P9 facilitated the synthesis of target proteins and their integration into the Escherichia coli cell membrane. This system was used to produce various multi-pass transmembrane proteins, including G-protein-coupled receptors, transporters, and ion channels of human origin. Green fluorescent protein fusion was used to confirm the correct folding of the expressed proteins. Of the 14 membrane proteins tested, eight were highly expressed, three were moderately expressed, and three were barely expressed in E. coli. Seven of the eight highly expressed proteins could be purified after extraction with the mild detergent lauryldimethylamine-oxide. Although a few proteins have previously been developed as fusion partners to augment membrane protein production, we believe that the major envelope protein P9 described here is better suited to the efficient expression of eukaryotic transmembrane proteins in E. coli. Copyright © 2015 Elsevier Inc. All rights reserved.

  3. Viral membrane fusion

    International Nuclear Information System (INIS)

    Harrison, Stephen C.

    2015-01-01

    Membrane fusion is an essential step when enveloped viruses enter cells. Lipid bilayer fusion requires catalysis to overcome a high kinetic barrier; viral fusion proteins are the agents that fulfill this catalytic function. Despite a variety of molecular architectures, these proteins facilitate fusion by essentially the same generic mechanism. Stimulated by a signal associated with arrival at the cell to be infected (e.g., receptor or co-receptor binding, proton binding in an endosome), they undergo a series of conformational changes. A hydrophobic segment (a “fusion loop” or “fusion peptide”) engages the target-cell membrane and collapse of the bridging intermediate thus formed draws the two membranes (virus and cell) together. We know of three structural classes for viral fusion proteins. Structures for both pre- and postfusion conformations of illustrate the beginning and end points of a process that can be probed by single-virion measurements of fusion kinetics. - Highlights: • Viral fusion proteins overcome the high energy barrier to lipid bilayer merger. • Different molecular structures but the same catalytic mechanism. • Review describes properties of three known fusion-protein structural classes. • Single-virion fusion experiments elucidate mechanism

  4. Fusion of biological membranes

    Indian Academy of Sciences (India)

    small hemifusion diaphragm. To obtain a direct view of the fusion process, we have carried out extensive simulations of two bilayers, composed of block copolymers, which are immersed in a solvent which favors one of the blocks. As in the biological case, the membranes are placed under tension. This is essential as fusion ...

  5. Fusion of biological membranes

    Indian Academy of Sciences (India)

    Logo of the Indian Academy of Sciences. Indian Academy of ... Home; Journals; Pramana – Journal of Physics; Volume 64; Issue 6. Fusion of biological ... The process of membrane fusion has been examined by Monte Carlo simulation, and is found to be very different than the conventional picture. The differences in ...

  6. Fusion of biological membranes

    Indian Academy of Sciences (India)

    Abstract. The process of membrane fusion has been examined by Monte Carlo simu- lation, and is found to be very different than the conventional picture. The differences in mechanism lead to several predictions, in particular that fusion is accompanied by tran- sient leakage. This prediction has recently been verified.

  7. Fusion of biological membranes

    Indian Academy of Sciences (India)

    The process of membrane fusion has been examined by Monte Carlo simulation, and is found to be very different than the conventional picture. The differences in mechanism lead to several predictions, in particular that fusion is accompanied by transient leakage. This prediction has recently been verified. Self-consistent ...

  8. A Model for Membrane Fusion

    Science.gov (United States)

    Ngatchou, Annita

    2010-01-01

    Pheochromocytoma is a tumor of the adrenal gland which originates from chromaffin cells and is characterized by the secretion of excessive amounts of neurotransmitter which lead to high blood pressure and palpitations. Pheochromocytoma contain membrane bound granules that store neurotransmitter. The release of these stored molecules into the extracellular space occurs by fusion of the granule membrane with the cell plasma membrane, a process called exocytosis. The molecular mechanism of this membrane fusion is not well understood. It is proposed that the so called SNARE proteins [1] are the pillar of vesicle fusion as their cleavage by clostridial toxin notably, Botulinum neurotoxin and Tetanus toxin abrogate the secretion of neurotransmitter [2]. Here, I describe how physical principles are applied to a biological cell to explore the role of the vesicle SNARE protein synaptobrevin-2 in easing granule fusion. The data presented here suggest a paradigm according to which the movement of the C-terminal of synaptobrevin-2 disrupts the lipid bilayer to form a fusion pore through which molecules can exit.

  9. Herpes Simplex Virus 1 Glycoprotein M and the Membrane-Associated Protein UL11 Are Required for Virus-Induced Cell Fusion and Efficient Virus Entry

    Science.gov (United States)

    Kim, In-Joong; Chouljenko, Vladimir N.; Walker, Jason D.

    2013-01-01

    Herpes simplex virus 1 (HSV-1) facilitates virus entry into cells and cell-to-cell spread by mediating fusion of the viral envelope with cellular membranes and fusion of adjacent cellular membranes. Although virus strains isolated from herpetic lesions cause limited cell fusion in cell culture, clinical herpetic lesions typically contain large syncytia, underscoring the importance of cell-to-cell fusion in virus spread in infected tissues. Certain mutations in glycoprotein B (gB), gK, UL20, and other viral genes drastically enhance virus-induced cell fusion in vitro and in vivo. Recent work has suggested that gB is the sole fusogenic glycoprotein, regulated by interactions with the viral glycoproteins gD, gH/gL, and gK, membrane protein UL20, and cellular receptors. Recombinant viruses were constructed to abolish either gM or UL11 expression in the presence of strong syncytial mutations in either gB or gK. Virus-induced cell fusion caused by deletion of the carboxyl-terminal 28 amino acids of gB or the dominant syncytial mutation in gK (Ala to Val at amino acid 40) was drastically reduced in the absence of gM. Similarly, syncytial mutations in either gB or gK did not cause cell fusion in the absence of UL11. Neither the gM nor UL11 gene deletion substantially affected gB, gC, gD, gE, and gH glycoprotein synthesis and expression on infected cell surfaces. Two-way immunoprecipitation experiments revealed that the membrane protein UL20, which is found as a protein complex with gK, interacted with gM while gM did not interact with other viral glycoproteins. Viruses produced in the absence of gM or UL11 entered into cells more slowly than their parental wild-type virus strain. Collectively, these results indicate that gM and UL11 are required for efficient membrane fusion events during virus entry and virus spread. PMID:23678175

  10. Membrane Trafficking and Vesicle Fusion

    Indian Academy of Sciences (India)

    Home; Journals; Resonance – Journal of Science Education; Volume 19; Issue 5. Membrane Trafficking and Vesicle Fusion: Post-Palade Era Researchers Win the Nobel Prize. Riddhi Atul Jani Subba Rao Gangi Setty. General Article Volume 19 Issue 5 May 2014 pp 421-445 ...

  11. The Yeast ATP-binding Cassette (ABC) Transporter Ycf1p Enhances the Recruitment of the Soluble SNARE Vam7p to Vacuoles for Efficient Membrane Fusion*

    Science.gov (United States)

    Sasser, Terry L.; Lawrence, Gus; Karunakaran, Surya; Brown, Christopher; Fratti, Rutilio A.

    2013-01-01

    The Saccharomyces cerevisiae vacuole contains five ATP-binding cassette class C (ABCC) transporters, including Ycf1p, a family member that was originally characterized as a Cd2+ transporter. Ycf1p has also been found to physically interact with a wide array of proteins, including factors that regulate vacuole homeostasis. In this study, we examined the role of Ycf1p and other ABCC transporters in the regulation of vacuole homotypic fusion. We found that deletion of YCF1 attenuated in vitro vacuole fusion by up to 40% relative to wild-type vacuoles. Plasmid-expressed wild-type Ycf1p rescued the deletion phenotype; however, Ycf1p containing a mutation of the conserved Lys-669 to Met in the Walker A box of the first nucleotide-binding domain (Ycf1pK669M) was unable to complement the fusion defect of ycf1Δ vacuoles. This indicates that the ATPase activity of Ycf1p is required for its function in regulating fusion. In addition, we found that deleting YCF1 caused a striking decrease in vacuolar levels of the soluble SNARE Vam7p, whereas total cellular levels were not altered. The attenuated fusion of ycf1Δ vacuoles was rescued by the addition of recombinant Vam7p to in vitro experiments. Thus, Ycf1p contributes in the recruitment of Vam7p to the vacuole for efficient membrane fusion. PMID:23658021

  12. Lipid Acrobatics in the Membrane Fusion Arena

    NARCIS (Netherlands)

    Markvoort, Albert J.; Marrink, Siewert J.; Chernomordik, Leonid V.; Kozlov, Michael M.

    2011-01-01

    In this review, we describe the recent contribution of computer simulation approaches to unravel the molecular details of membrane fusion. Over the past decade, fusion between apposed membranes and vesicles has been studied using a large variety of simulation methods and systems. Despite the variety

  13. Coiled coil driven membrane fusion between cyclodextrin vesicles and liposomes

    NARCIS (Netherlands)

    Versluis, Frank; Voskuhl, Jens; Vos, Jan; Friedrich, Heiner; Ravoo, Bart Jan; Bomans, Paul H H; Stuart, Marc C A; Sommerdijk, Nico A J M; Kros, Alexander

    2014-01-01

    Controlled fusion events between natural membranes composed of phospholipids with synthetic unnatural membranes will yield valuable fundamental information on the mechanism of membrane fusion. Here, fusion between vastly different phospholipid liposomes and cyclodextrin amphiphile based vesicles

  14. The dengue virus type 2 envelope protein fusion peptide is essential for membrane fusion

    International Nuclear Information System (INIS)

    Huang, Claire Y.-H.; Butrapet, Siritorn; Moss, Kelly J.; Childers, Thomas; Erb, Steven M.; Calvert, Amanda E.; Silengo, Shawn J.; Kinney, Richard M.; Blair, Carol D.; Roehrig, John T.

    2010-01-01

    The flaviviral envelope (E) protein directs virus-mediated membrane fusion. To investigate membrane fusion as a requirement for virus growth, we introduced 27 unique mutations into the fusion peptide of an infectious cDNA clone of dengue 2 virus and recovered seven stable mutant viruses. The fusion efficiency of the mutants was impaired, demonstrating for the first time the requirement for specific FP AAs in optimal fusion. Mutant viruses exhibited different growth kinetics and/or genetic stabilities in different cell types and adult mosquitoes. Virus particles could be recovered following RNA transfection of cells with four lethal mutants; however, recovered viruses could not re-infect cells. These viruses could enter cells, but internalized virus appeared to be retained in endosomal compartments of infected cells, thus suggesting a fusion blockade. Mutations of the FP also resulted in reduced virus reactivity with flavivirus group-reactive antibodies, confirming earlier reports using virus-like particles.

  15. Assembly of intermediates for rapid membrane fusion.

    Science.gov (United States)

    Harner, Max; Wickner, William

    2018-01-26

    Membrane fusion is essential for intracellular protein sorting, cell growth, hormone secretion, and neurotransmission. Rapid membrane fusion requires tethering and Sec1-Munc18 (SM) function to catalyze R-, Qa-, Qb-, and Qc-SNARE complex assembly in trans , as well as SNARE engagement by the SNARE-binding chaperone Sec17/αSNAP. The hexameric vacuolar HOPS ( ho motypic fusion and vacuole p rotein s orting) complex in the yeast Saccharomyces cerevisiae tethers membranes through its affinities for the membrane Rab GTPase Ypt7. HOPS also has specific affinities for the vacuolar SNAREs and catalyzes SNARE complex assembly, but the order of their assembly into a 4-SNARE complex is unclear. We now report defined assembly intermediates on the path to membrane fusion. We found that a prefusion intermediate will assemble with HOPS and the R, Qa, and Qc SNAREs, and that this assembly undergoes rapid fusion upon addition of Qb and Sec17. HOPS-tethered membranes and all four vacuolar SNAREs formed a complex that underwent an even more dramatic burst of fusion upon Sec17p addition. These findings provide initial insights into an ordered fusion pathway consisting of the following intermediates and events: 1) Rab- and HOPS-tethered membranes, 2) a HOPS:R:Qa:Qc trans -complex, 3) a HOPS:4-SNARE trans -complex, 4) an engagement with Sec17, and 5) the rapid lipid rearrangements during fusion. In conclusion, our results indicate that the R:Qa:Qc complex forms in the context of membrane, Ypt7, HOPS, and trans -SNARE assembly and serves as a functional intermediate for rapid fusion after addition of the Qb-SNARE and Sec17 proteins. © 2018 by The American Society for Biochemistry and Molecular Biology, Inc.

  16. Efficient liposome fusion mediated by lipid-nucleic acid conjugates

    DEFF Research Database (Denmark)

    Ries, O; Löffler, P M G; Rabe, A

    2017-01-01

    The fusion of biomembranes with release of encapsulated content in a controlled way is crucial for cell signaling, endo- and exocytosis and intracellular trafficking. Programmable fusion of liposomes and an efficient mixing of their contents have the potential to enable the study of chemical...... and enzymatic processes in a confined environment and under crowded conditions outside biological systems. We report on DNA-controlled fusion of lipid bilayer membranes using lipid-nucleic acid conjugates (LiNAs) to mediate lipid and content mixing of liposomes. Screening of different membrane anchor and linker...

  17. Membrane Trafficking and Vesicle Fusion

    Indian Academy of Sciences (India)

    IAS Admin

    hemophagocytic syndrome) and metabolic (diabe- tes) disorders [2, 23, 33]. Mutations in the genes of the basic secretory protein machinery lead to a number of membrane trafficking diseases such as Charcot–Marie–Tooth disease, Cohen.

  18. Membrane Trafficking and Vesicle Fusion

    Indian Academy of Sciences (India)

    IAS Admin

    investigators were awarded the Nobel Prize in Physiology or. Medicine in 2013. Introduction. Membrane Transport: In the eukaryotic cell, a majority of proteins are made in the cytosol. But the transmembrane and secretory proteins are synthesized in an organelle called the rough endoplasmic reticulum (ER). These proteins ...

  19. Fusion of Sendai virus with vesicles of oligomerizable lipids: a microcalorimetric analysis of membrane fusion.

    Science.gov (United States)

    Ravoo, B J; Weringa, W D; Engberts, J B

    2000-01-01

    Sendai virus fuses efficiently with small and large unilamellar vesicles of the lipid 1,2-di-n-hexadecyloxypropyl-4- (beta-nitrostyryl) phosphate (DHPBNS) at pH 7.4 and 37 degrees C, as shown by lipid mixing assays and electron microscopy. However, fusion is strongly inhibited by oligomerization of the head groups of DHPBNS in the bilayer vesicles. The enthalpy associated with fusion of Sendai virus with DHPBNS vesicles was measured by isothermal titration microcalorimetry, comparing titrations of Sendai virus into (i) solutions of DHPBNS vesicles (which fuse with the virus) and (ii) oligomerized DHPBNS vesicles (which do not fuse with the virus), respectively. The observed heat effect of fusion of Sendai virus with DHPBNS vesicles is strongly dependent on the buffer medium, reflecting a partial charge neutralization of the Sendai F and HN proteins upon insertion into the negatively-charged vesicle membrane. No buffer effect was observed for the titration of Sendai virus into oligomerized DHPBNS vesicles, indicating that inhibition of fusion is a result of inhibition of insertion of the fusion protein into the target membrane. Fusion of Sendai virus with DHPBNS vesicles is endothermic and entropy-driven. The positive enthalpy term is dominated by heat effects resulting from merging of the protein-rich viral envelope with the lipid vesicle bilayers rather than by the fusion of the viral with the vesicle bilayers per se. Copyright 2000 Academic Press.

  20. Visualization and Sequencing of Membrane Remodeling Leading to Influenza Virus Fusion

    Science.gov (United States)

    Gui, Long; Ebner, Jamie L.; Mileant, Alexander; Williams, James A.

    2016-01-01

    ABSTRACT Protein-mediated membrane fusion is an essential step in many fundamental biological events, including enveloped virus infection. The nature of protein and membrane intermediates and the sequence of membrane remodeling during these essential processes remain poorly understood. Here we used cryo-electron tomography (cryo-ET) to image the interplay between influenza virus and vesicles with a range of lipid compositions. By following the population kinetics of membrane fusion intermediates imaged by cryo-ET, we found that membrane remodeling commenced with the hemagglutinin fusion protein spikes grappling onto the target membrane, followed by localized target membrane dimpling as local clusters of hemagglutinin started to undergo conformational refolding. The local dimples then transitioned to extended, tightly apposed contact zones where the two proximal membrane leaflets were in most cases indistinguishable from each other, suggesting significant dehydration and possible intermingling of the lipid head groups. Increasing the content of fusion-enhancing cholesterol or bis-monoacylglycerophosphate in the target membrane led to an increase in extended contact zone formation. Interestingly, hemifused intermediates were found to be extremely rare in the influenza virus fusion system studied here, most likely reflecting the instability of this state and its rapid conversion to postfusion complexes, which increased in population over time. By tracking the populations of fusion complexes over time, the architecture and sequence of membrane reorganization leading to efficient enveloped virus fusion were thus resolved. IMPORTANCE Enveloped viruses employ specialized surface proteins to mediate fusion of cellular and viral membranes that results in the formation of pores through which the viral genetic material is delivered to the cell. For influenza virus, the trimeric hemagglutinin (HA) glycoprotein spike mediates host cell attachment and membrane fusion. While

  1. Peptides and membrane fusion : Towards an understanding of the molecular mechanism of protein-induced fusion

    NARCIS (Netherlands)

    Pecheur, EI; Sainte-Marie, J; Bienvenue, A; Hoekstra, D

    1999-01-01

    Processes such as endo- or exocytosis, membrane recycling, fertilization and enveloped viruses infection require one or more critical membrane fusion reactions. A key feature in viral and cellular fusion phenomena is the involvement of specific fusion proteins. Among the few well-characterized

  2. Dissipative Particle Dynamics of tension-induced membrane fusion

    DEFF Research Database (Denmark)

    Shillcock, Julian C.

    2009-01-01

    Recent studies of tension-induced membrane fusion using dissipative particle dynamics (DPD) simulations are briefly reviewed. The stochastic nature of the fusion process makes it necessary to simulate a large number of fusion attempts in order to obtain reliable fusion statistics and to extract...... meaningful values for the fusion probability and the average fusion times. All successful fusion events follow the same pathway. In this fusion pathway, configurations of individual lipids play an important role. Fusion starts with individual lipids assuming a splayed tail configuration with one tail......, three sub-processes have been identified in the fusion pathway. Their energy barriers are estimated to lie in the range 8-15kBT. The fusion probability is found to possess a maximum at intermediate tension values. As one decreases the tension, the fusion probability seems to vanish before...

  3. Coiled-coil driven membrane fusion: zipper-like vs. non-zipper-like peptide orientation.

    Science.gov (United States)

    Versluis, Frank; Dominguez, Juan; Voskuhl, Jens; Kros, Alexander

    2013-01-01

    Membrane fusion plays a central role in biological processes such as neurotransmission and exocytosis. An important class of proteins that induce membrane fusion are called SNARE (soluble N-ethyl malemeide sensitive factor attachment protein receptors) proteins. To induce membrane fusion, two SNARE proteins embedded in opposing membranes form a four-helix coiled-coil motif together with a third, cytoplasmic, SNARE protein. Coiled-coil formation brings the two membranes into close proximity allowing fusion to occur. Importantly, structural investigations have demonstrated that native membrane fusion only occurs when the orientation of the coiled-coil motif resembles that of a zipper. The zipper orientation arises when parallel coiled-coil formation takes place between peptides that are anchored into apposing membranes at identical termini, thereby forcing the membranes into close contact. Recently, we have designed a synthetic model for membrane fusion, which is based on a set of lipidated coiled-coil forming peptide pairs which are denoted E-K. When incorporated into liposomal membranes, coiled-coil formation between these lipidated peptides induces targeted and efficient membrane fusion of liposomes. Our model system mimics SNARE-driven membrane fusion, as it contains a coiled-coil motif which has a zipper-like orientation, similar to that of the SNARE proteins. Here we investigate whether the zipper-like orientation of the coiled-coil motifs is a prerequisite for membrane fusion in our model system. Our strategy is based on conjugation of the transmembrane anchor to either the N- or the C-terminus of peptides E and K. Whereas the use of a set of complementary peptides with the membrane anchor on identical peptide termini yields the zipper-like orientation of the coiled-coil complex, membrane anchors on opposite peptide termini results in a non-zipper-like coiled-coil orientation. Surprisingly, it was observed that efficient and targeted membrane fusion was

  4. The TIP30 protein complex, arachidonic acid and coenzyme A are required for vesicle membrane fusion.

    Directory of Open Access Journals (Sweden)

    Chengliang Zhang

    Full Text Available Efficient membrane fusion has been successfully mimicked in vitro using artificial membranes and a number of cellular proteins that are currently known to participate in membrane fusion. However, these proteins are not sufficient to promote efficient fusion between biological membranes, indicating that critical fusogenic factors remain unidentified. We have recently identified a TIP30 protein complex containing TIP30, acyl-CoA synthetase long-chain family member 4 (ACSL4 and Endophilin B1 (Endo B1 that promotes the fusion of endocytic vesicles with Rab5a vesicles, which transport endosomal acidification enzymes vacuolar (H⁺-ATPases (V-ATPases to the early endosomes in vivo. Here, we demonstrate that the TIP30 protein complex facilitates the fusion of endocytic vesicles with Rab5a vesicles in vitro. Fusion of the two vesicles also depends on arachidonic acid, coenzyme A and the synthesis of arachidonyl-CoA by ACSL4. Moreover, the TIP30 complex is able to transfer arachidonyl groups onto phosphatidic acid (PA, producing a new lipid species that is capable of inducing close contact between membranes. Together, our data suggest that the TIP30 complex facilitates biological membrane fusion through modification of PA on membranes.

  5. Fusion Pore Diameter Regulation by Cations Modulating Local Membrane Anisotropy

    Directory of Open Access Journals (Sweden)

    Doron Kabaso

    2012-01-01

    Full Text Available The fusion pore is an aqueous channel that is formed upon the fusion of the vesicle membrane with the plasma membrane. Once the pore is open, it may close again (transient fusion or widen completely (full fusion to permit vesicle cargo discharge. While repetitive transient fusion pore openings of the vesicle with the plasma membrane have been observed in the absence of stimulation, their frequency can be further increased using a cAMP-increasing agent that drives the opening of nonspecific cation channels. Our model hypothesis is that the openings and closings of the fusion pore are driven by changes in the local concentration of cations in the connected vesicle. The proposed mechanism of fusion pore dynamics is considered as follows: when the fusion pore is closed or is extremely narrow, the accumulation of cations in the vesicle (increased cation concentration likely leads to lipid demixing at the fusion pore. This process may affect local membrane anisotropy, which reduces the spontaneous curvature and thus leads to the opening of the fusion pore. Based on the theory of membrane elasticity, we used a continuum model to explain the rhythmic opening and closing of the fusion pore.

  6. On the mechanism of intracellular membrane fusion : In search of the genuine fusion factor

    NARCIS (Netherlands)

    Pecheur, EI; Maier, O; Hoekstra, D

    2000-01-01

    Intracellular membrane Fusion events require a general protein machinery that functions in vesicular traffic and in assembly and maintenance of organelles. An array of cytosolic and integral membrane proteins are currently identified, and in conjunction with ongoing detailed structural studies,

  7. Direct observation of intermediate states in model membrane fusion

    Science.gov (United States)

    Keidel, Andrea; Bartsch, Tobias F.; Florin, Ernst-Ludwig

    2016-01-01

    We introduce a novel assay for membrane fusion of solid supported membranes on silica beads and on coverslips. Fusion of the lipid bilayers is induced by bringing an optically trapped bead in contact with the coverslip surface while observing the bead’s thermal motion with microsecond temporal and nanometer spatial resolution using a three-dimensional position detector. The probability of fusion is controlled by the membrane tension on the particle. We show that the progression of fusion can be monitored by changes in the three-dimensional position histograms of the bead and in its rate of diffusion. We were able to observe all fusion intermediates including transient fusion, formation of a stalk, hemifusion and the completion of a fusion pore. Fusion intermediates are characterized by axial but not lateral confinement of the motion of the bead and independently by the change of its rate of diffusion due to the additional drag from the stalk-like connection between the two membranes. The detailed information provided by this assay makes it ideally suited for studies of early events in pure lipid bilayer fusion or fusion assisted by fusogenic molecules. PMID:27029285

  8. Tension-induced fusion of bilayer membranes and vesicles

    Science.gov (United States)

    Shillcock, Julian C.; Lipowsky, Reinhard

    2005-03-01

    Maintaining the integrity of their protective plasma membrane is a primary requirement of cells. Accordingly, cellular events that breach the membrane are tightly regulated. Artificial vesicles used in drug delivery must also stay intact until they have reached the desired target. In both cases, the intrinsic resistance of the membrane to rupture must be overcome to allow the efflux of the vesicle's contents. Here, we use mesoscopic simulations to study the fusion of 28-nm-diameter vesicles to 50 × 50 nm2 planar membrane patches over 2 μs. We monitor the time evolution of 93 different fusion attempts. This allows us to construct a global morphology diagram, using the initial tensions of the vesicle and the planar membrane patch as control parameters, and to determine the corresponding fusion statistics. All successful fusion events are observed to occur within 350 ns, which reflects the presence of alternative pathways for the tension relaxation.

  9. Multi-layered nanoparticles for penetrating the endosome and nuclear membrane via a step-wise membrane fusion process.

    Science.gov (United States)

    Akita, Hidetaka; Kudo, Asako; Minoura, Arisa; Yamaguti, Masaya; Khalil, Ikramy A; Moriguchi, Rumiko; Masuda, Tomoya; Danev, Radostin; Nagayama, Kuniaki; Kogure, Kentaro; Harashima, Hideyoshi

    2009-05-01

    Efficient targeting of DNA to the nucleus is a prerequisite for effective gene therapy. The gene-delivery vehicle must penetrate through the plasma membrane, and the DNA-impermeable double-membraned nuclear envelope, and deposit its DNA cargo in a form ready for transcription. Here we introduce a concept for overcoming intracellular membrane barriers that involves step-wise membrane fusion. To achieve this, a nanotechnology was developed that creates a multi-layered nanoparticle, which we refer to as a Tetra-lamellar Multi-functional Envelope-type Nano Device (T-MEND). The critical structural elements of the T-MEND are a DNA-polycation condensed core coated with two nuclear membrane-fusogenic inner envelopes and two endosome-fusogenic outer envelopes, which are shed in stepwise fashion. A double-lamellar membrane structure is required for nuclear delivery via the stepwise fusion of double layered nuclear membrane structure. Intracellular membrane fusions to endosomes and nuclear membranes were verified by spectral imaging of fluorescence resonance energy transfer (FRET) between donor and acceptor fluorophores that had been dually labeled on the liposome surface. Coating the core with the minimum number of nucleus-fusogenic lipid envelopes (i.e., 2) is essential to facilitate transcription. As a result, the T-MEND achieves dramatic levels of transgene expression in non-dividing cells.

  10. Efficient Activation of Human T Cells of Both CD4 and CD8 Subsets by Urease-Deficient Recombinant Mycobacterium bovis BCG That Produced a Heat Shock Protein 70-M. tuberculosis-Derived Major Membrane Protein II Fusion Protein

    Science.gov (United States)

    Mukai, Tetsu; Tsukamoto, Yumiko; Maeda, Yumi; Tamura, Toshiki

    2014-01-01

    For the purpose of obtaining Mycobacterium bovis bacillus Calmette-Guérin (BCG) capable of activating human naive T cells, urease-deficient BCG expressing a fusion protein composed of Mycobacterium tuberculosis-derived major membrane protein II (MMP-II) and heat shock protein 70 (HSP70) of BCG (BCG-DHTM) was produced. BCG-DHTM secreted the HSP70-MMP-II fusion protein and effectively activated human monocyte-derived dendritic cells (DCs) by inducing phenotypic changes and enhanced cytokine production. BCG-DHTM-infected DCs activated naive T cells of both CD4 and naive CD8 subsets, in an antigen (Ag)-dependent manner. The T cell activation induced by BCG-DHTM was inhibited by the pretreatment of DCs with chloroquine. The naive CD8+ T cell activation was mediated by the transporter associated with antigen presentation (TAP) and the proteosome-dependent cytosolic cross-priming pathway. Memory CD8+ T cells and perforin-producing effector CD8+ T cells were efficiently produced from the naive T cell population by BCG-DHTM stimulation. Single primary infection with BCG-DHTM in C57BL/6 mice efficiently produced T cells responsive to in vitro secondary stimulation with HSP70, MMP-II, and M. tuberculosis-derived cytosolic protein and inhibited the multiplication of subsequently aerosol-challenged M. tuberculosis more efficiently than did vector control BCG. These results indicate that the introduction of MMP-II and HSP70 into urease-deficient BCG may be useful for improving BCG for control of tuberculosis. PMID:24152387

  11. Projected thermodynamic efficiencies of fusion power plants

    International Nuclear Information System (INIS)

    McKinnon, M.A.

    1976-09-01

    Estimated thermal efficiencies of proposed fusion power plant concepts are compared to the efficiencies of nonfusion power plants. Present trends in electrical power generation are also discussed. The fusion reactor system designs will have about the same thermal efficiencies as present day power plants using steam if these designs require the collection of thermal energy at the blanket and the transfer of that energy to a heat exchanger or boiler using the current technology. Two general methods should be pursued for increasing the thermal efficiencies of fusion power plants and thereby reducing the amount of waste heat. Methods should be developed for increasing the temperatures of the reactor coolants since the maximum attainable thermal efficiency of systems using coolants can be increased only by increasing the coolant temperatures. Second, advanced power recovery systems such as potassium topping turbines, MHD, and direct conversion should be developed since such systems avoid the limits on steam systems due to excessive operating pressures at high temperatures. Direct conversion is particularly attractive because it avoids the theoretical Carnot limit on thermal efficiency when heat is converted to electrical energy

  12. 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.

  13. Membrane fusion triggers rapid degradation of two gamete-specific, fusion-essential proteins in a membrane block to polygamy in Chlamydomonas

    OpenAIRE

    Liu, Yanjie; Misamore, Michael J.; Snell, William J.

    2010-01-01

    The plasma membranes of gametes are specialized for fusion, yet, once fusion occurs, in many organisms the new zygote becomes incapable of further membrane fusion reactions. The molecular mechanisms that underlie this loss of fusion capacity (block to polygamy) remain unknown. During fertilization in the green alga Chlamydomonas, the plus gamete-specific membrane protein FUS1 is required for adhesion between the apically localized sites on the plasma membranes of plus and minus gametes that a...

  14. MEMBRANE-FUSION OF SEMLIKI FOREST VIRUS INVOLVES HOMOTRIMERS OF THE FUSION PROTEIN

    NARCIS (Netherlands)

    WAHLBERG, JM; WILSCHUT, J; GAROFF, H

    1992-01-01

    Infection of cells with enveloped viruses is accomplished through membrane fusion. The binding and fusion Processes are mediated by the spike proteins in the envelope of the virus particle and usually involve a series of conformational changes in these proteins. We have studied the low-pH-mediated

  15. Molecular View of the Role of Fusion Peptides in Promoting Positive Membrane Curvature

    NARCIS (Netherlands)

    Fuhrmans, Marc; Marrink, Siewert J.

    2012-01-01

    Fusion peptides are moderately hydrophobic segments of viral and nonviral membrane fusion proteins that enable these proteins to fuse two closely apposed biological membranes. In vitro assays furthermore show that even isolated fusion peptides alone can support membrane fusion in model systems. In

  16. Inner membrane fusion mediates spatial distribution of axonal mitochondria

    Science.gov (United States)

    Yu, Yiyi; Lee, Hao-Chih; Chen, Kuan-Chieh; Suhan, Joseph; Qiu, Minhua; Ba, Qinle; Yang, Ge

    2016-01-01

    In eukaryotic cells, mitochondria form a dynamic interconnected network to respond to changing needs at different subcellular locations. A fundamental yet unanswered question regarding this network is whether, and if so how, local fusion and fission of individual mitochondria affect their global distribution. To address this question, we developed high-resolution computational image analysis techniques to examine the relations between mitochondrial fusion/fission and spatial distribution within the axon of Drosophila larval neurons. We found that stationary and moving mitochondria underwent fusion and fission regularly but followed different spatial distribution patterns and exhibited different morphology. Disruption of inner membrane fusion by knockdown of dOpa1, Drosophila Optic Atrophy 1, not only increased the spatial density of stationary and moving mitochondria but also changed their spatial distributions and morphology differentially. Knockdown of dOpa1 also impaired axonal transport of mitochondria. But the changed spatial distributions of mitochondria resulted primarily from disruption of inner membrane fusion because knockdown of Milton, a mitochondrial kinesin-1 adapter, caused similar transport velocity impairment but different spatial distributions. Together, our data reveals that stationary mitochondria within the axon interconnect with moving mitochondria through fusion and fission and that local inner membrane fusion between individual mitochondria mediates their global distribution. PMID:26742817

  17. Amphiphilic gold nanoparticles as modulators of lipid membrane fusion

    Science.gov (United States)

    Tahir, Mukarram; Alexander-Katz, Alfredo

    The fusion of lipid membranes is central to biological functions like inter-cellular transport and signaling and is coordinated by proteins of the soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs) superfamily. We utilize molecular dynamics simulations to demonstrate that gold nanoparticles functionalized with a mixed-monolayer of hydrophobic and hydrophilic alkanethiol ligands can act as synthetic analogues of these fusion proteins and mediate lipid membrane fusion by catalyzing the formation of a toroidal stalk between adjacent membranes and enabling the formation of a fusion pore upon influx of Ca2+ into the exterior solvent. The fusion pathway enabled by these synthetic nanostructures is analogous to the regulated fast fusion pathway observed during synaptic vesicle fusion; it therefore provides novel physical insights into this important biological process while also being relevant in a number of single-cell therapeutic applications. Computational resources from NSF XSEDE contract TG-DMR130042. Financial support from DOE CSGF fellowship DE-FG02-97ER25308.

  18. Vesicle fusion with bilayer lipid membrane controlled by electrostatic interaction

    Directory of Open Access Journals (Sweden)

    Azusa Oshima

    2017-09-01

    Full Text Available The fusion of proteoliposomes is a promising approach for incorporating membrane proteins in artificial lipid membranes. In this study, we employed an electrostatic interaction between vesicles and supported bilayer lipid membranes (s-BLMs to control the fusion process. We combined large unilamellar vesicles (LUVs containing anionic lipids, which we used instead of proteoliposomes, and s-BLMs containing cationic lipids to control electrostatic interaction. Anionic LUVs were never adsorbed or ruptured on the SiO2 substrate with a slight negative charge, and selectively fused with cationic s-BLMs. The LUVs can be fused effectively to the target position. Furthermore, as the vesicle fusion proceeds and some of the positive charges are neutralized, the attractive interaction weakens and finally the vesicle fusion saturates. In other words, we can control the number of LUVs fused with s-BLMs by controlling the concentration of the cationic lipids in the s-BLMs. The fluidity of the s-BLMs after vesicle fusion was confirmed to be sufficiently high. This indicates that the LUVs attached to the s-BLMs were almost completely fused, and there were few intermediate state vesicles in the fusion process. We could control the position and amount of vesicle fusion with the s-BLMs by employing an electrostatic interaction.

  19. Autographa californica Multicapsid Nucleopolyhedrovirus efficiently infects Sf9 cells and transduces mammalian cells via direct fusion with the plasma membrane at low pH

    NARCIS (Netherlands)

    Dong, S.; Wang, M.; Qiu, Z.; Deng, F.; Vlak, J.M.; Hu, Z.H.; Wang, H.L.

    2010-01-01

    The budded virus (BV) of the Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) infects insect cells and transduces mammalian cells mainly through the endocytosis pathway. However, this study revealed that the treatment of the virus bound to Sf9 cells at low pH could efficiently rescue

  20. The Effect of Multispectral Image Fusion Enhancement on Human Efficiency

    Science.gov (United States)

    2017-03-20

    5a. CONTRACT NUMBER In-House The effect of multispectral image fusion enhancement on human efficiency 5b. GRANT NUMBER 5c. PROGRAM ELEMENT...fusion across its associated problem space of application. 15. SUBJECT TERMS Ideal observer analysis, Efficiency, Image fusion, Multispectral imagery...Implications (2017) 2:19 DOI 10.1186/s41235-016-0045-0 ORIGINAL ARTICLE Open Access The effect of multispectral image fusion enhancement on human

  1. Green fluorescence protein-based content-mixing assay of SNARE-driven membrane fusion.

    Science.gov (United States)

    Heo, Paul; Kong, Byoungjae; Jung, Young-Hun; Park, Joon-Bum; Shin, Jonghyeok; Park, Myungseo; Kweon, Dae-Hyuk

    2017-06-17

    Soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) proteins mediate intracellular membrane fusion by forming a ternary SNARE complex. A minimalist approach utilizing proteoliposomes with reconstituted SNARE proteins yielded a wealth of information pinpointing the molecular mechanism of SNARE-mediated fusion and its regulation by accessory proteins. Two important attributes of a membrane fusion are lipid-mixing and the formation of an aqueous passage between apposing membranes. These two attributes are typically observed by using various fluorescent dyes. Currently available in vitro assay systems for observing fusion pore opening have several weaknesses such as cargo-bleeding, incomplete removal of unencapsulated dyes, and inadequate information regarding the size of the fusion pore, limiting measurements of the final stage of membrane fusion. In the present study, we used a biotinylated green fluorescence protein and streptavidin conjugated with Dylight 594 (DyStrp) as a Föster resonance energy transfer (FRET) donor and acceptor, respectively. This FRET pair encapsulated in each v-vesicle containing synaptobrevin and t-vesicle containing a binary acceptor complex of syntaxin 1a and synaptosomal-associated protein 25 revealed the opening of a large fusion pore of more than 5 nm, without the unwanted signals from unencapsulated dyes or leakage. This system enabled determination of the stoichiometry of the merging vesicles because the FRET efficiency of the FRET pair depended on the molar ratio between dyes. Here, we report a robust and informative assay for SNARE-mediated fusion pore opening. Copyright © 2017 Elsevier Inc. All rights reserved.

  2. Conflicting views on the membrane fusion machinery and the fusion pore

    DEFF Research Database (Denmark)

    Sørensen, Jakob B

    2009-01-01

    of the assembly of the fusogenic SNARE-complex. Here, I review conflicting views on the function of the core fusion machinery consisting of the SNAREs, Munc18, complexin, and synaptotagmin. Munc18 controls docking of vesicles to the plasma membrane and initial SNARE-complex assembly, whereas complexin...... of the water-soluble vesicle content. The fusion pore remains the most elusive part of the exocytosis pathway, owing to its short lifetime....

  3. Slow fusion of liposomes composed of membrane-spanning lipids

    NARCIS (Netherlands)

    Elferink, MGL; vanBreemen, J; Konings, WN; Driessen, AJM; Wilschut, J; Elferink, Marieke G.L.

    1997-01-01

    The fusion characteristics of large unilamellar liposomes composed of bipolar tetraether lipids extracted from the thermophilic archaeon Sulfolobus acidocaldarius, was investigated. These lipids span the entire membrane and form single monolayer liposomes in aqueous media [Elferink, M.G.L., de Wit,

  4. A Cell-Cell Fusion Assay to Assess Arenavirus Envelope Glycoprotein Membrane-Fusion Activity.

    Science.gov (United States)

    York, Joanne; Nunberg, Jack H

    2018-01-01

    For many viruses that enter their target cells through pH-dependent fusion of the viral and endosomal membranes, cell-cell fusion assays can provide an experimental platform for investigating the structure-function relationships that promote envelope glycoprotein membrane-fusion activity. Typically, these assays employ effector cells expressing the recombinant envelope glycoprotein on the cell surface and target cells engineered to quantitatively report fusion with the effector cell. In the protocol described here, Vero cells are transfected with a plasmid encoding the arenavirus envelope glycoprotein complex GPC and infected with the vTF7-3 vaccinia virus expressing the bacteriophage T7 RNA polymerase. These effector cells are mixed with target cells infected with the vCB21R-lacZ vaccinia virus encoding a β-galactosidase reporter under the control of the T7 promoter. Cell-cell fusion is induced upon exposure to low-pH medium (pH 5.0), and the resultant expression of the β-galactosidase reporter is quantitated using a chemiluminescent substrate. We have utilized this robust microplate cell-cell fusion assay extensively to study arenavirus entry and its inhibition by small-molecule fusion inhibitors.

  5. Shear-Induced Membrane Fusion in Viscous Solutions

    KAUST Repository

    Kogan, Maxim

    2014-05-06

    Large unilamellar lipid vesicles do not normally fuse under fluid shear stress. They might deform and open pores to relax the tension to which they are exposed, but membrane fusion occurring solely due to shear stress has not yet been reported. We present evidence that shear forces in a viscous solution can induce lipid bilayer fusion. The fusion of 1,2-dioleoyl-sn-glycero-3- phosphocholine (DOPC) liposomes is observed in Couette flow with shear rates above 3000 s-1 provided that the medium is viscous enough. Liposome samples, prepared at different viscosities using a 0-50 wt % range of sucrose concentration, were studied by dynamic light scattering, lipid fusion assays using Förster resonance energy transfer (FRET), and linear dichroism (LD) spectroscopy. Liposomes in solutions with 40 wt % (or more) sucrose showed lipid fusion under shear forces. These results support the hypothesis that under suitable conditions lipid membranes may fuse in response to mechanical-force- induced stress. © 2014 American Chemical Society.

  6. Energy efficiency in membrane bioreactors.

    Science.gov (United States)

    Barillon, B; Martin Ruel, S; Langlais, C; Lazarova, V

    2013-01-01

    Energy consumption remains the key factor for the optimisation of the performance of membrane bioreactors (MBRs). This paper presents the results of the detailed energy audits of six full-scale MBRs operated by Suez Environnement in France, Spain and the USA based on on-site energy measurement and analysis of plant operation parameters and treatment performance. Specific energy consumption is compared for two different MBR configurations (flat sheet and hollow fibre membranes) and for plants with different design, loads and operation parameters. The aim of this project was to understand how the energy is consumed in MBR facilities and under which operating conditions, in order to finally provide guidelines and recommended practices for optimisation of MBR operation and design to reduce energy consumption and environmental impacts.

  7. Zeolite membranes for efficient synthesis of biofuels

    OpenAIRE

    Sandström, Linda

    2009-01-01

    The greenhouse effect and the limited fossil oil resources have increased the demand of renewable fuels. Zeolite membranes have potential applications in numerous separation processes, and could be useful in the development of efficient processes for renewable fuel production. Methanol synthesis from synthesis gas is equilibrium limited, and continuous removal of products in a zeolite membrane reactor could improve the productivity of a conventional methanol synthesis process. In this work, m...

  8. The coronavirus spike protein : mechanisms of membrane fusion and virion incorporation

    NARCIS (Netherlands)

    Bosch, B.J.

    2004-01-01

    The coronavirus spike protein is a membrane-anchored glycoprotein responsible for virus-cell attachment and membrane fusion, prerequisites for a successful virus infection. In this thesis, two aspects are described regarding the molecular biology of the coronavirus spike protein: its membrane fusion

  9. A compensatory mutation provides resistance to disparate HIV fusion inhibitor peptides and enhances membrane fusion.

    Directory of Open Access Journals (Sweden)

    Matthew P Wood

    Full Text Available Fusion inhibitors are a class of antiretroviral drugs used to prevent entry of HIV into host cells. Many of the fusion inhibitors being developed, including the drug enfuvirtide, are peptides designed to competitively inhibit the viral fusion protein gp41. With the emergence of drug resistance, there is an increased need for effective and unique alternatives within this class of antivirals. One such alternative is a class of cyclic, cationic, antimicrobial peptides known as θ-defensins, which are produced by many non-human primates and exhibit broad-spectrum antiviral and antibacterial activity. Currently, the θ-defensin analog RC-101 is being developed as a microbicide due to its specific antiviral activity, lack of toxicity to cells and tissues, and safety in animals. Understanding potential RC-101 resistance, and how resistance to other fusion inhibitors affects RC-101 susceptibility, is critical for future development. In previous studies, we identified a mutant, R5-tropic virus that had evolved partial resistance to RC-101 during in vitro selection. Here, we report that a secondary mutation in gp41 was found to restore replicative fitness, membrane fusion, and the rate of viral entry, which were compromised by an initial mutation providing partial RC-101 resistance. Interestingly, we show that RC-101 is effective against two enfuvirtide-resistant mutants, demonstrating the clinical importance of RC-101 as a unique fusion inhibitor. These findings both expand our understanding of HIV drug-resistance to diverse peptide fusion inhibitors and emphasize the significance of compensatory gp41 mutations.

  10. In vitro assay using engineered yeast vacuoles for neuronal SNARE-mediated membrane fusion

    Science.gov (United States)

    Ko, Young-Joon; Lee, Miriam; Kang, KyeongJin; Song, Woo Keun; Jun, Youngsoo

    2014-01-01

    Intracellular membrane fusion requires not only SNARE proteins but also other regulatory proteins such as the Rab and Sec1/Munc18 (SM) family proteins. Although neuronal SNARE proteins alone can drive the fusion between synthetic liposomes, it remains unclear whether they are also sufficient to induce the fusion of biological membranes. Here, through the use of engineered yeast vacuoles bearing neuronal SNARE proteins, we show that neuronal SNAREs can induce membrane fusion between yeast vacuoles and that this fusion does not require the function of the Rab protein Ypt7p or the SM family protein Vps33p, both of which are essential for normal yeast vacuole fusion. Although excess vacuolar SNARE proteins were also shown to mediate Rab-bypass fusion, this fusion required homotypic fusion and vacuole protein sorting complex, which bears Vps33p and was accompanied by extensive membrane lysis. We also show that this neuronal SNARE-driven vacuole fusion can be stimulated by the neuronal SM protein Munc18 and blocked by botulinum neurotoxin serotype E, a well-known inhibitor of synaptic vesicle fusion. Taken together, our results suggest that neuronal SNARE proteins are sufficient to induce biological membrane fusion, and that this new assay can be used as a simple and complementary method for investigating synaptic vesicle fusion mechanisms. PMID:24821814

  11. Fusion of Sendai Virus with Vesicles of Oligomerizable Lipids : A Micro Calorimetric Analysis of Membrane Fusion

    NARCIS (Netherlands)

    Ravoo, Bart Jan; Weringa, Wilke D.; Engberts, Jan B.F.N.

    2000-01-01

    Sendai virus fuses efficiently with small and large unilamellar vesicles of the lipid 1,2-di-n-hexadecyloxypropyl-4-(beta-nitrostyryl) phosphate (DHPBNS) at pH 7.4 and 37°C, as shown by lipid mixing assays and electron microscopy. However, fusion is strongly inhibited by oligomerization of the head

  12. Fusion of Sendai Virus with Vesicles of Oligomerizable Lipids: A Micro Calorimetric Analysis of Membrane Fusion

    OpenAIRE

    Ravoo, Bart Jan; Weringa, Wilke D.; Engberts, Jan B.F.N.

    2000-01-01

    Sendai virus fuses efficiently with small and large unilamellar vesicles of the lipid 1,2-di-n-hexadecyloxypropyl-4-(beta-nitrostyryl) phosphate (DHPBNS) at pH 7.4 and 37°C, as shown by lipid mixing assays and electron microscopy. However, fusion is strongly inhibited by oligomerization of the head groups of DHPBNS in the bilayer vesicles. The enthalpy associated with fusion of Sendai virus with DHPBNS vesicles was measured by isothermal titration microcalorimetry, comparing titrations of Sen...

  13. Henipavirus Mediated Membrane Fusion, Virus Entry and Targeted Therapeutics

    Directory of Open Access Journals (Sweden)

    Dimitar B. Nikolov

    2012-02-01

    Full Text Available The Paramyxoviridae genus Henipavirus is presently represented by the type species Hendra and Nipah viruses which are both recently emerged zoonotic viral pathogens responsible for repeated outbreaks associated with high morbidity and mortality in Australia, Southeast Asia, India and Bangladesh. These enveloped viruses bind and enter host target cells through the coordinated activities of their attachment (G and class I fusion (F envelope glycoproteins. The henipavirus G glycoprotein interacts with host cellular B class ephrins, triggering conformational alterations in G that lead to the activation of the F glycoprotein, which facilitates the membrane fusion process. Using the recently published structures of HeV-G and NiV-G and other paramyxovirus glycoproteins, we review the features of the henipavirus envelope glycoproteins that appear essential for mediating the viral fusion process, including receptor binding, G-F interaction, F activation, with an emphasis on G and the mutations that disrupt viral infectivity. Finally, recent candidate therapeutics for henipavirus-mediated disease are summarized in light of their ability to inhibit HeV and NiV entry by targeting their G and F glycoproteins.

  14. FUSION OF ARTIFICIAL MEMBRANES WITH MAMMALIAN SPERMATOZOA - SPECIFIC INVOLVEMENT OF THE EQUATORIAL SEGMENT AFTER ACROSOME REACTION

    NARCIS (Netherlands)

    ARTS, EGJM; KUIKEN, J; JAGER, S; HOEKSTRA, D

    1993-01-01

    The fusogenic properties of bovine and human spermatozoa membranes were investigated, using phospholipid bilayers (liposomes) as target membranes. Fusion was monitored by following lipid mixing, as revealed by an assay based on resonance-energy transfer. In addition, fusion was visualized by

  15. Paramyxovirus membrane fusion: Lessons from the F and HN atomic structures

    International Nuclear Information System (INIS)

    Lamb, Robert A.; Paterson, Reay G.; Jardetzky, Theodore S.

    2006-01-01

    Paramyxoviruses enter cells by fusion of their lipid envelope with the target cell plasma membrane. Fusion of the viral membrane with the plasma membrane allows entry of the viral genome into the cytoplasm. For paramyxoviruses, membrane fusion occurs at neutral pH, but the trigger mechanism that controls the viral entry machinery such that it occurs at the right time and in the right place remains to be elucidated. Two viral glycoproteins are key to the infection process-an attachment protein that varies among different paramyxoviruses and the fusion (F) protein, which is found in all paramyxoviruses. For many of the paramyxoviruses (parainfluenza viruses 1-5, mumps virus, Newcastle disease virus and others), the attachment protein is the hemagglutinin/neuraminidase (HN) protein. In the last 5 years, atomic structures of paramyxovirus F and HN proteins have been reported. The knowledge gained from these structures towards understanding the mechanism of viral membrane fusion is described

  16. Inhibition of HIV-1 endocytosis allows lipid mixing at the plasma membrane, but not complete fusion

    Directory of Open Access Journals (Sweden)

    de la Vega Michelle

    2011-12-01

    Full Text Available Abstract Background We recently provided evidence that HIV-1 enters HeLa-derived TZM-bl and lymphoid CEMss cells by fusing with endosomes, whereas its fusion with the plasma membrane does not proceed beyond the lipid mixing step. The mechanism of restriction of HIV-1 fusion at the cell surface and/or the factors that aid the virus entry from endosomes remain unclear. Results We examined HIV-1 fusion with a panel of target cells lines and with primary CD4+ T cells. Kinetic measurements of fusion combined with time-resolved imaging of single viruses further reinforced the notion that HIV-1 enters the cells via endocytosis and fusion with endosomes. Furthermore, we attempted to deliberately redirect virus fusion to the plasma membrane, using two experimental strategies. First, the fusion reaction was synchronized by pre-incubating the viruses with cells at reduced temperature to allow CD4 and coreceptors engagement, but not the virus uptake or fusion. Subsequent shift to a physiological temperature triggered accelerated virus uptake followed by entry from endosomes, but did not permit fusion at the cell surface. Second, blocking HIV-1 endocytosis by a small-molecule dynamin inhibitor, dynasore, resulted in transfer of viral lipids to the plasma membrane without any detectable release of the viral content into the cytosol. We also found that a higher concentration of dynasore is required to block the HIV-endosome fusion compared to virus internalization. Conclusions Our results further support the notion that HIV-1 enters disparate cell types through fusion with endosomes. The block of HIV-1 fusion with the plasma membrane at a post-lipid mixing stage shows that this membrane is not conducive to fusion pore formation and/or enlargement. The ability of dynasore to interfere with the virus-endosome fusion suggests that dynamin could be involved in two distinct steps of HIV-1 entry - endocytosis and fusion within intracellular compartments.

  17. A new combination of membranes and membrane reactors for improved tritium management in breeder blanket of fusion machines

    International Nuclear Information System (INIS)

    Demange, D.; Staemmler, S.; Kind, M.

    2011-01-01

    Tritium used as fuel in future fusion machines will be produced within the breeder blanket. The tritium extraction system recovers the tritium to be routed into the inner-fuel cycle of the machine. Accurate and precise tritium accountancy between both systems is mandatory to ensure a reliable operation. Handling in the blanket huge helium flow rates containing tritium as traces in molecular and oxide forms is challenging both for the process and the accountancy. Alternative tritium processes based on combinations of membranes and membrane reactors are proposed to facilitate the tritium management. The PERMCAT process is based on counter-current isotope swamping in a palladium membrane reactor. It allows recovering tritium efficiently from any chemical species. It produces a pure hydrogen stream enriched in tritium of advantage for integration upstream of the accountancy stage. A pre-separation and pre-concentration stage using new zeolite membranes has been studied to optimize the whole process. Such a combination could improve the tritium processes and facilitate accountancy in DEMO.

  18. Sphingolipids activate membrane fusion of Semliki Forest virus in a stereospecific manner

    DEFF Research Database (Denmark)

    Moesby, Lise; Corver, J; Erukulla, R K

    1995-01-01

    on degradation of the viral capsid protein by trypsin encapsulated in the target liposomes. Fusion mediated by D-erythro-ceramide was not affected by the additional presence in the target liposomes of ceramide stereoisomers incapable of fusion activation. Binding of the virus to the liposomes, as assessed...... by flotation on sucrose density gradients, was not dependent on the presence of fusion-competent or fusion-incompetent sphingolipids in the liposomes. The results of this study support the notion that a stereospecific interaction of the viral fusion protein with D-erythro sphingolipids in the target membrane......The alphavirus Semliki Forest virus (SFV) enters cells through receptor-mediated endocytosis. Subsequently, triggered by the acid pH in endosomes, the viral envelope fuses with the endosomal membrane. Membrane fusion of SFV has been shown previously to be dependent on the presence of cholesterol...

  19. A tethering complex drives the terminal stage of SNARE-dependent membrane fusion

    Science.gov (United States)

    D'Agostino, Massimo; Risselada, Herre Jelger; Lürick, Anna; Ungermann, Christian; Mayer, Andreas

    2017-11-01

    Membrane fusion in eukaryotic cells mediates the biogenesis of organelles, vesicular traffic between them, and exo- and endocytosis of important signalling molecules, such as hormones and neurotransmitters. Distinct tasks in intracellular membrane fusion have been assigned to conserved protein systems. Tethering proteins mediate the initial recognition and attachment of membranes, whereas SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor) protein complexes are considered as the core fusion engine. SNARE complexes provide mechanical energy to distort membranes and drive them through a hemifusion intermediate towards the formation of a fusion pore. This last step is highly energy-demanding. Here we combine the in vivo and in vitro fusion of yeast vacuoles with molecular simulations to show that tethering proteins are critical for overcoming the final energy barrier to fusion pore formation. SNAREs alone drive vacuoles only into the hemifused state. Tethering proteins greatly increase the volume of SNARE complexes and deform the site of hemifusion, which lowers the energy barrier for pore opening and provides the driving force. Thereby, tethering proteins assume a crucial mechanical role in the terminal stage of membrane fusion that is likely to be conserved at multiple steps of vesicular traffic. We therefore propose that SNAREs and tethering proteins should be considered as a single, non-dissociable device that drives fusion. The core fusion machinery may then be larger and more complex than previously thought.

  20. Protein-induced fusion can be modulated by target membrane lipids through a structural switch at the level of the fusion peptide

    NARCIS (Netherlands)

    Pecheur, EI; Martin, [No Value; Bienvenue, A; Ruysschaert, JM; Hoekstra, D

    2000-01-01

    Regulatory features of protein-induced membrane fusion are largely unclear, particularly at the level of the fusion peptide. Fusion peptides being part of larger protein complexes, such investigations are met with technical limitations. Here, we show that the fusion activity of influenza virus or

  1. Binding and Fusion of Extracellular Vesicles to the Plasma Membrane of Their Cell Targets.

    Science.gov (United States)

    Prada, Ilaria; Meldolesi, Jacopo

    2016-08-09

    Exosomes and ectosomes, extracellular vesicles of two types generated by all cells at multivesicular bodies and the plasma membrane, respectively, play critical roles in physiology and pathology. A key mechanism of their function, analogous for both types of vesicles, is the fusion of their membrane to the plasma membrane of specific target cells, followed by discharge to the cytoplasm of their luminal cargo containing proteins, RNAs, and DNA. Here we summarize the present knowledge about the interactions, binding and fusions of vesicles with the cell plasma membrane. The sequence initiates with dynamic interactions, during which vesicles roll over the plasma membrane, followed by the binding of specific membrane proteins to their cell receptors. Membrane binding is then converted rapidly into fusion by mechanisms analogous to those of retroviruses. Specifically, proteins of the extracellular vesicle membranes are structurally rearranged, and their hydrophobic sequences insert into the target cell plasma membrane which undergoes lipid reorganization, protein restructuring and membrane dimpling. Single fusions are not the only process of vesicle/cell interactions. Upon intracellular reassembly of their luminal cargoes, vesicles can be regenerated, released and fused horizontally to other target cells. Fusions of extracellular vesicles are relevant also for specific therapy processes, now intensely investigated.

  2. High efficiency fusion reactor based on bootstrap current

    International Nuclear Information System (INIS)

    Kikuchi, Mitsuru

    1990-01-01

    The establishment of the steady operation technology which has been the largest subject of the research on the nuclear fusion reactors utilizing tokamak type confinement principle advanced largely by the recent research, and the concept of the power reactor which can be made steady with high efficiency was established. This is to utilize positively the bootstrap current naturally flowing in tokamak plasma. This power reactor can be materialized with the technologies in near future, and there is the possibility that it can become the clear target of developing nuclear fusion reactors as electric power generation plants. In this report, explanation is made centering around the high efficiency nuclear fusion reactor SSTR, for which Japan Atomic Energy Research Institute advances the conceptual design as the prototype power reactor. In nuclear fusion energy CO 2 gas is not generated, essentially nuclear runaway never occurs, and radioactive wastes can be relatively reduced, therefore, it is expected to become a powerful substitute energy source. The main parameters and features of the steady state tokamak nuclear fusion power reactor (SSTR) are reported. (K.I.)

  3. Membrane fusion inducers, chloroquine and spermidine increase lipoplex-mediated gene transfection

    International Nuclear Information System (INIS)

    Wong-Baeza, Carlos; Bustos, Israel; Serna, Manuel; Tescucano, Alonso; Alcantara-Farfan, Veronica; Ibanez, Miguel; Montanez, Cecilia; Wong, Carlos; Baeza, Isabel

    2010-01-01

    Gene transfection into mammalian cells can be achieved with viral and non-viral vectors. Non-viral vectors, such as cationic lipids that form lipoplexes with DNA, are safer and more stable than viral vectors, but their transfection efficiencies are lower. Here we describe that the simultaneous treatment with a membrane fusion inducer (chlorpromazine or procainamide) plus the lysosomotropic agent chloroquine increases lipoplex-mediated gene transfection in human (HEK293 and C-33 A) and rat (PC12) cell lines (up to 9.2-fold), as well as in situ in BALB/c mice spleens and livers (up to 6-fold); and that the polyamine spermidine increases lipoplex-mediated gene transfection and expression in cell cultures. The use of these four drugs provides a novel, safe and relatively inexpensive way to considerably increase lipoplex-mediated gene transfection efficiency.

  4. 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.

  5. Mitochondrial DNA mutations provoke dominant inhibition of mitochondrial inner membrane fusion.

    Directory of Open Access Journals (Sweden)

    Cécile Sauvanet

    Full Text Available Mitochondria are highly dynamic organelles that continuously move, fuse and divide. Mitochondrial dynamics modulate overall mitochondrial morphology and are essential for the proper function, maintenance and transmission of mitochondria and mitochondrial DNA (mtDNA. We have investigated mitochondrial fusion in yeast cells with severe defects in oxidative phosphorylation (OXPHOS due to removal or various specific mutations of mtDNA. We find that, under fermentative conditions, OXPHOS deficient cells maintain normal levels of cellular ATP and ADP but display a reduced mitochondrial inner membrane potential. We demonstrate that, despite metabolic compensation by glycolysis, OXPHOS defects are associated to a selective inhibition of inner but not outer membrane fusion. Fusion inhibition was dominant and hampered the fusion of mutant mitochondria with wild-type mitochondria. Inhibition of inner membrane fusion was not systematically associated to changes of mitochondrial distribution and morphology, nor to changes in the isoform pattern of Mgm1, the major fusion factor of the inner membrane. However, inhibition of inner membrane fusion correlated with specific alterations of mitochondrial ultrastructure, notably with the presence of aligned and unfused inner membranes that are connected to two mitochondrial boundaries. The fusion inhibition observed upon deletion of OXPHOS related genes or upon removal of the entire mtDNA was similar to that observed upon introduction of point mutations in the mitochondrial ATP6 gene that are associated to neurogenic ataxia and retinitis pigmentosa (NARP or to maternally inherited Leigh Syndrome (MILS in humans. Our findings indicate that the consequences of mtDNA mutations may not be limited to OXPHOS defects but may also include alterations in mitochondrial fusion. Our results further imply that, in healthy cells, the dominant inhibition of fusion could mediate the exclusion of OXPHOS-deficient mitochondria from

  6. Effect of lipid molecule headgroup mismatch on non steroidal anti-inflammatory drugs induced membrane fusion.

    Science.gov (United States)

    Mondal Roy, Sutapa; Sarkar, Munna

    2011-12-20

    Membrane fusion is an essential process guiding many important biological events, which most commonly requires the aid of proteins and peptides as fusogenic agents. Small drug induced fusion at low drug concentration is a rare event. Only three drugs, namely, meloxicam (Mx), piroxicam (Px), and tenoxicam (Tx), belonging to the oxicam group of non steroidal anti-inflammatory drugs (NSAIDs) have been shown by us to induce membrane fusion successfully at low drug concentration. A better elucidation of the mechanism and the effect of different parameters in modulating the fusion process will allow the use of these common drugs to induce and control membrane fusion in various biochemical processes. In this study, we monitor the effect of lipid headgroup size mismatch in the bilayer on oxicam NSAIDs induced membrane fusion, by introducing dimyristoylphosphatidylethanolamine (DMPE) in dimyristoylphosphatidylcholine (DMPC) small unilamellar vesicles (SUVs). Such headgroup mismatch affects various lipid parameters which includes inhibition of trans-bilayer motion, domain formation, decrease in curvature, etc. Changes in various lipidic parameters introduce defects in the membrane bilayer and thereby modulate membrane fusion. SUVs formed by DMPC with increasing DMPE content (10, 20, and 30 mol %) were used as simple model membranes. Transmission electron microscopy (TEM) and differential scanning calorimetry (DSC) were used to characterize the DMPC-DMPE mixed vesicles. Fluorescence assays were used to probe the time dependence of lipid mixing, content mixing, and leakage and also used to determine the partitioning of the drugs in the membrane bilayer. How the inhibition of trans-bilayer motion, heterogeneous distribution of lipids, decrease in vesicle curvature, etc., arising due to headgroup mismatch affect the fusion process has been isolated and identified here. Mx amplifies these effects maximally followed by Px and Tx. This has been correlated to the enhanced

  7. The fusion of membranes and vesicles: pathway and energy barriers from Dissipative Particle Dynamics

    DEFF Research Database (Denmark)

    Shillcock, Julian C.

    2009-01-01

    are simulated using the optimized parameter set. In the observed fusion pathway, configurations of individual lipids play an important role. Fusion starts with individual lipids assuming a splayed tail configuration with one tail inserted in each membrane. To determine the corresponding energy barrier, we...

  8. Influenza-virus membrane fusion by cooperative fold-back of stochastically induced hemagglutinin intermediates

    NARCIS (Netherlands)

    Ivanovic, Tijana; Choi, Jason L.; Whelan, Sean P.; Oijen, Antoine M. van; Harrison, Stephen C.

    2013-01-01

    Influenza virus penetrates cells by fusion of viral and endosomal membranes catalyzed by the viral hemagglutinin (HA). Structures of the initial and final states of the HA trimer define the fusion endpoints, but do not specify intermediates. We have characterized these transitions by analyzing

  9. Switching between Successful and Dead-End Intermediates in Membrane Fusion.

    Science.gov (United States)

    Molotkovsky, Rodion J; Galimzyanov, Timur R; Jiménez-Munguía, Irene; Pavlov, Konstantin V; Batishchev, Oleg V; Akimov, Sergey A

    2017-12-02

    Fusion of cellular membranes during normal biological processes, including proliferation, or synaptic transmission, is mediated and controlled by sophisticated protein machinery ensuring the preservation of the vital barrier function of the membrane throughout the process. Fusion of virus particles with host cell membranes is more sparingly arranged and often mediated by a single fusion protein, and the virus can afford to be less discriminative towards the possible different outcomes of fusion attempts. Formation of leaky intermediates was recently observed in some fusion processes, and an alternative trajectory of the process involving formation of π-shaped structures was suggested. In this study, we apply the methods of elasticity theory and Lagrangian formalism augmented by phenomenological and molecular geometry constraints and boundary conditions to investigate the traits of this trajectory and the drivers behind the choice of one of the possible scenarios depending on the properties of the system. The alternative pathway proved to be a dead end, and, depending on the parameters of the participating membranes and fusion proteins, the system can either reversibly enter the corresponding "leaky" configuration or be trapped in it. A parametric study in the biologically relevant range of variables emphasized the fusion protein properties crucial for the choice of the fusion scenario.

  10. Switching between Successful and Dead-End Intermediates in Membrane Fusion

    Directory of Open Access Journals (Sweden)

    Rodion J. Molotkovsky

    2017-12-01

    Full Text Available Fusion of cellular membranes during normal biological processes, including proliferation, or synaptic transmission, is mediated and controlled by sophisticated protein machinery ensuring the preservation of the vital barrier function of the membrane throughout the process. Fusion of virus particles with host cell membranes is more sparingly arranged and often mediated by a single fusion protein, and the virus can afford to be less discriminative towards the possible different outcomes of fusion attempts. Formation of leaky intermediates was recently observed in some fusion processes, and an alternative trajectory of the process involving formation of π-shaped structures was suggested. In this study, we apply the methods of elasticity theory and Lagrangian formalism augmented by phenomenological and molecular geometry constraints and boundary conditions to investigate the traits of this trajectory and the drivers behind the choice of one of the possible scenarios depending on the properties of the system. The alternative pathway proved to be a dead end, and, depending on the parameters of the participating membranes and fusion proteins, the system can either reversibly enter the corresponding “leaky” configuration or be trapped in it. A parametric study in the biologically relevant range of variables emphasized the fusion protein properties crucial for the choice of the fusion scenario.

  11. Fusion blankets for high-efficiency power cycles

    International Nuclear Information System (INIS)

    Usher, J.L.; Lazareth, O.W.; Fillo, J.A.; Horn, F.L.; Powell, J.R.

    1980-01-01

    The efficiencies of blankets for fusion reactors are usually in the range of 30 to 40%, limited by the operating temperatures (500 0 C) of conventional structural materials such as stainless steels. In this project two-zone blankets are proposed; these blankets consist of a low-temperature shell surrounding a high-temperature interior zone. A survey of nucleonics and thermal hydraulic parameters has led to a reference blanket design consisting of a water-cooled stainless steel shell around a BeO, ZrO 2 interior (cooled by argon) utilizing Li 2 O for tritium breeding. In this design, approximately 60% of the fusion energy is deposited in the high-temperature interior. The maximum argon temperature is 2230 0 C leading to an overall efficiency estimate of 55 to 60% for this reference case

  12. Blanket options for high-efficiency fusion power

    International Nuclear Information System (INIS)

    Usher, J.L.; Lazareth, O.W.; Fillo, J.A.; Horn, F.L.; Powell, J.R.

    1980-01-01

    The efficiencies of blankets for fusion reactors are usually in the range of 30 to 40%, limited by the operating temperatures (500 0 C) of conventional structural materials such as stainless steels. In this project two-zone blankets are proposed; these blankets consist of a low-temperature shell surrounding a high-temperature interior zone. A survey of nucleonics and thermal hydraulic parameters has led to a reference blanket design consisting of a water-cooled stainless steel shell around a BeO, ZrO 2 interior (cooled by argon) utilizing Li 2 O for tritium breeding. In this design, approximately 60% of the fusion energy is deposited in the high-temperature interior. The maximum argon temperature is 2230 0 C leading to an overall efficiency estimate of 55 to 60% for this reference case

  13. Fusion blanket for high-efficiency power cycles

    International Nuclear Information System (INIS)

    Usher, J.L.; Powell, J.R.; Fillo, J.A.; Horn, F.L.; Lazareth, O.W.; Taussig, R.

    1980-01-01

    The efficiencies of blankets for fusion reactors are usually in the range of 30 to 40%, limited by the operating temperature (500 0 C) of conventional structural materials such as stainless steels. In this project two-zone blankets are proposed; these blankets consist of a low-temperature shell surrounding a high-temperature interior zone. A survey of nucleonics and thermal hydraulic parameters has led to a reference blanket design consisting of a water-cooled stainless steel shell around a BeO, ZrO 2 interior (cooled by Ar) utilizing Li 2 O for tritium breeding. In this design, approx. 60% of the fusion energy is deposited in the high-temperature interior. The maximum Ar temperature is 2230 0 C leading to an overall efficiency estimate of 55 to 60% for this reference case

  14. Fusion blankets for high-efficiency power cycles

    International Nuclear Information System (INIS)

    Usher, J.L.; Lazareth, O.W.; Fillo, J.A.; Horn, F.L.; Powell, J.R.

    1981-01-01

    The efficiencies of blankets for fusion reactors are usually in the range of 30 to 40%, limited by the operating temperatures (500 deg C) of conventional structural materials such as stainless steels. In this project 'two-zone' blankets are proposed; these blankets consist of a low-temperature shell surrounding a high-temperature interior zone. A survey of nucleonics and thermal hydraulic parameters has led to a reference blanket design consisting of a water-cooled stainless steel shell around a BeO, ZrO 2 interior (cooled by argon) utilizing Li 2 O for tritium breeding. In this design, approximately 60% of the fusion energy is deposited in the high-temperature interior. The maximum argon temperature is 2230 deg C leading to an overall efficiency estimate of 55 to 60% for this reference case. (author)

  15. Mitotic phosphorylation of VCIP135 blocks p97ATPase-mediated Golgi membrane fusion

    International Nuclear Information System (INIS)

    Totsukawa, Go; Matsuo, Ayaka; Kubota, Ayano; Taguchi, Yuya; Kondo, Hisao

    2013-01-01

    Highlights: •VCIP135 is mitotically phosphorylated on Threonine-760 and Serine-767 by Cdc2. •Phosphorylated VCIP135 does not bind to p97ATPase. •The phosphorylation of VCIP135 inhibits p97ATPase-mediated Golgi membrane fusion. -- Abstract: In mammals, the Golgi apparatus is disassembled early mitosis and reassembled at the end of mitosis. For Golgi disassembly, membrane fusion needs to be blocked. Golgi biogenesis requires two distinct p97ATPase-mediated membrane fusion, the p97/p47 and p97/p37 pathways. We previously reported that p47 phosphorylation on Serine-140 and p37 phosphorylation on Serine-56 and Threonine-59 result in mitotic inhibition of the p97/p47 and the p97/p37 pathways, respectively [11,14]. In this study, we show another mechanism of mitotic inhibition of p97-mediated Golgi membrane fusion. We clarified that VCIP135, an essential factor in both p97 membrane fusion pathways, is phosphorylated on Threonine-760 and Serine-767 by Cdc2 at mitosis and that this phosphorylated VCIP135 does not bind to p97. An in vitro Golgi reassembly assay revealed that VCIP135(T760E, S767E), which mimics mitotic phosphorylation, caused no cisternal regrowth. Our results indicate that the phosphorylation of VCIP135 on Threonine-760 and Serine-767 inhibits p97-mediated Golgi membrane fusion at mitosis

  16. Mitotic phosphorylation of VCIP135 blocks p97ATPase-mediated Golgi membrane fusion

    Energy Technology Data Exchange (ETDEWEB)

    Totsukawa, Go; Matsuo, Ayaka; Kubota, Ayano; Taguchi, Yuya; Kondo, Hisao, E-mail: hk228@med.kyushu-u.ac.jp

    2013-04-05

    Highlights: •VCIP135 is mitotically phosphorylated on Threonine-760 and Serine-767 by Cdc2. •Phosphorylated VCIP135 does not bind to p97ATPase. •The phosphorylation of VCIP135 inhibits p97ATPase-mediated Golgi membrane fusion. -- Abstract: In mammals, the Golgi apparatus is disassembled early mitosis and reassembled at the end of mitosis. For Golgi disassembly, membrane fusion needs to be blocked. Golgi biogenesis requires two distinct p97ATPase-mediated membrane fusion, the p97/p47 and p97/p37 pathways. We previously reported that p47 phosphorylation on Serine-140 and p37 phosphorylation on Serine-56 and Threonine-59 result in mitotic inhibition of the p97/p47 and the p97/p37 pathways, respectively [11,14]. In this study, we show another mechanism of mitotic inhibition of p97-mediated Golgi membrane fusion. We clarified that VCIP135, an essential factor in both p97 membrane fusion pathways, is phosphorylated on Threonine-760 and Serine-767 by Cdc2 at mitosis and that this phosphorylated VCIP135 does not bind to p97. An in vitro Golgi reassembly assay revealed that VCIP135(T760E, S767E), which mimics mitotic phosphorylation, caused no cisternal regrowth. Our results indicate that the phosphorylation of VCIP135 on Threonine-760 and Serine-767 inhibits p97-mediated Golgi membrane fusion at mitosis.

  17. SNARE motif: A common motif used by pathogens to manipulate membrane fusion

    Science.gov (United States)

    Wesolowski, Jordan

    2010-01-01

    To penetrate host cells through their membranes, pathogens use a variety of molecular components in which the presence of heptad repeat motifs seems to be a prevailing element. Heptad repeats are characterized by a pattern of seven, generally hydrophobic, residues. In order to initiate membrane fusion, viruses use glycoproteins-containing heptad repeats. These proteins are structurally and functionally similar to the SNARE proteins known to be involved in eukaryotic membrane fusion. SNAREs also display a heptad repeat motif called the “SNARE motif”. As bacterial genomes are being sequenced, microorganisms also appear to be carrying membrane proteins resembling eukaryotic SNAREs. This category of SNARE-like proteins might share similar functions and could be used by microorganisms to either promote or block membrane fusion. Such a recurrence across pathogenic organisms suggests that this architectural motif was evolutionarily selected because it most effectively ensures the survival of pathogens within the eukaryotic environment. PMID:21178463

  18. Measuring the strength of interaction between the Ebola fusion peptide and lipid rafts: implications for membrane fusion and virus infection.

    Directory of Open Access Journals (Sweden)

    Mônica S Freitas

    Full Text Available The Ebola fusion peptide (EBO₁₆ is a hydrophobic domain that belongs to the GP2 membrane fusion protein of the Ebola virus. It adopts a helical structure in the presence of mimetic membranes that is stabilized by the presence of an aromatic-aromatic interaction established by Trp8 and Phe12. In spite of its infectious cycle becoming better understood recently, several steps still remain unclear, a lacuna that makes it difficult to develop strategies to block infection. In order to gain insight into the mechanism of membrane fusion, we probed the structure, function and energetics of EBO₁₆ and its mutant W8A, in the absence or presence of different lipid membranes, including isolated domain-resistant membranes (DRM, a good experimental model for lipid rafts. The depletion of cholesterol from living mammalian cells reduced the ability of EBO₁₆ to induce lipid mixing. On the other hand, EBO₁₆ was structurally sensitive to interaction with lipid rafts (DRMs, but the same was not observed for W8A mutant. In agreement with these data, W8A showed a poor ability to promote membrane aggregation in comparison to EBO₁₆. Single molecule AFM experiments showed a high affinity force pattern for the interaction of EBO₁₆ and DRM, which seems to be a complex energetic event as observed by the calorimetric profile. Our study is the first to show a strong correlation between the initial step of Ebola virus infection and cholesterol, thus providing a rationale for Ebola virus proteins being co-localized with lipid-raft domains. In all, the results show how small fusion peptide sequences have evolved to adopt highly specific and strong interactions with membrane domains. Such features suggest these processes are excellent targets for therapeutic and vaccine approaches to viral diseases.

  19. HIV gp41 fusion peptide increases membrane ordering in a cholesterol-dependent fashion.

    Science.gov (United States)

    Lai, Alex L; Freed, Jack H

    2014-01-07

    Fusion between viral envelopes and host cell membranes, which is mediated by special glycoproteins anchored on the viral membrane, is required for HIV viral entry and infection. The HIV gp41 fusion peptide (FP), which initiates membrane fusion, adopts either an α-helical or β-sheeted structure depending on the cholesterol concentration. We used phosphocholine spin labels on the lipid headgroup and different positions on the acyl chain to detect its perturbation on lipid bilayers containing different cholesterol concentrations by electron-spin resonance. Our findings were as follows. 1), gp41 FP affects the lipid order in the same manner as previously shown for influenza hemagglutinin FP, i.e., it has a cooperative effect versus the peptide/lipid ratio, supporting our hypothesis that membrane ordering is a common prerequisite for viral membrane fusion. 2), gp41 FP induces membrane ordering in all lipid compositions studied, whereas a nonfusion mutant FP perturbs lipid order to a significantly smaller extent. 3), In high-cholesterol-containing lipid bilayers, where gp41 FP is in the β-aggregation conformation, its effect on the lipid ordering reaches deeper into the bilayer. The different extent to which the two conformers perturb is correlated with their fusogenicity. The possible role of the two conformers in membrane fusion is discussed. Copyright © 2014 Biophysical Society. Published by Elsevier Inc. All rights reserved.

  20. The Structural Dynamics of the Flavivirus Fusion Peptide–Membrane Interaction

    Science.gov (United States)

    Souza, Theo L. F.; Sousa, Ivanildo P.; Bianconi, M. Lucia; Bernardi, Rafael C.; Pascutti, Pedro G.; Silva, Jerson L.; Gomes, Andre M. O.; Oliveira, Andréa C.

    2012-01-01

    Membrane fusion is a crucial step in flavivirus infections and a potential target for antiviral strategies. Lipids and proteins play cooperative roles in the fusion process, which is triggered by the acidic pH inside the endosome. This acidic environment induces many changes in glycoprotein conformation and allows the action of a highly conserved hydrophobic sequence, the fusion peptide (FP). Despite the large volume of information available on the virus-triggered fusion process, little is known regarding the mechanisms behind flavivirus–cell membrane fusion. Here, we evaluated the contribution of a natural single amino acid difference on two flavivirus FPs, FLAG (98DRGWGNGCGLFGK110) and FLAH (98DRGWGNHCGLFGK110), and investigated the role of the charge of the target membrane on the fusion process. We used an in silico approach to simulate the interaction of the FPs with a lipid bilayer in a complementary way and used spectroscopic approaches to collect conformation information. We found that both peptides interact with neutral and anionic micelles, and molecular dynamics (MD) simulations showed the interaction of the FPs with the lipid bilayer. The participation of the indole ring of Trp appeared to be important for the anchoring of both peptides in the membrane model, as indicated by MD simulations and spectroscopic analyses. Mild differences between FLAG and FLAH were observed according to the pH and the charge of the target membrane model. The MD simulations of the membrane showed that both peptides adopted a bend structure, and an interaction between the aromatic residues was strongly suggested, which was also observed by circular dichroism in the presence of micelles. As the FPs of viral fusion proteins play a key role in the mechanism of viral fusion, understanding the interactions between peptides and membranes is crucial for medical science and biology and may contribute to the design of new antiviral drugs. PMID:23094066

  1. Membrane fusion-competent virus-like proteoliposomes and proteinaceous supported bilayers made directly from cell plasma membranes.

    Science.gov (United States)

    Costello, Deirdre A; Hsia, Chih-Yun; Millet, Jean K; Porri, Teresa; Daniel, Susan

    2013-05-28

    Virus-like particles are useful materials for studying virus-host interactions in a safe manner. However, the standard production of pseudovirus based on the vesicular stomatitis virus (VSV) backbone is an intricate procedure that requires trained laboratory personnel. In this work, a new strategy for creating virus-like proteoliposomes (VLPLs) and virus-like supported bilayers (VLSBs) is presented. This strategy uses a cell blebbing technique to induce the formation of nanoscale vesicles from the plasma membrane of BHK cells expressing the hemagglutinin (HA) fusion protein of influenza X-31. These vesicles and supported bilayers contain HA and are used to carry out single particle membrane fusion events, monitored using total internal reflection fluorescence microscopy. The results of these studies show that the VLPLs and VLSBs contain HA proteins that are fully competent to carry out membrane fusion, including the formation of a fusion pore and the release of fluorophores loaded into vesicles. This new strategy for creating spherical and planar geometry virus-like membranes has many potential applications. VLPLs could be used to study fusion proteins of virulent viruses in a safe manner, or they could be used as therapeutic delivery particles to transport beneficial proteins coexpressed in the cells to a target cell. VLSBs could facilitate high throughput screening of antiviral drugs or pathogen-host cell interactions.

  2. Influenza-virus membrane fusion by cooperative fold-back of stochastically induced hemagglutinin intermediates.

    Science.gov (United States)

    Ivanovic, Tijana; Choi, Jason L; Whelan, Sean P; van Oijen, Antoine M; Harrison, Stephen C

    2013-02-19

    Influenza virus penetrates cells by fusion of viral and endosomal membranes catalyzed by the viral hemagglutinin (HA). Structures of the initial and final states of the HA trimer define the fusion endpoints, but do not specify intermediates. We have characterized these transitions by analyzing low-pH-induced fusion kinetics of individual virions and validated the analysis by computer simulation. We detect initial engagement with the target membrane of fusion peptides from independently triggered HAs within the larger virus-target contact patch; fusion then requires engagement of three or four neighboring HA trimers. Effects of mutations in HA indicate that withdrawal of the fusion peptide from a pocket in the pre-fusion trimer is rate-limiting for both events, but the requirement for cooperative action of several HAs to bring the fusing membranes together leads to a long-lived intermediate state for single, extended HA trimers. This intermediate is thus a fundamental aspect of the fusion mechanism. DOI:http://dx.doi.org/10.7554/eLife.00333.001.

  3. Sphingolipids activate membrane fusion of Semliki Forest virus in a stereospecific manner

    DEFF Research Database (Denmark)

    Moesby, Lise; Corver, J; Erukulla, R K

    1995-01-01

    The alphavirus Semliki Forest virus (SFV) enters cells through receptor-mediated endocytosis. Subsequently, triggered by the acid pH in endosomes, the viral envelope fuses with the endosomal membrane. Membrane fusion of SFV has been shown previously to be dependent on the presence of cholesterol ...

  4. Translocation of cell penetrating peptides and calcium-induced membrane fusion share same mechanism

    Czech Academy of Sciences Publication Activity Database

    Magarkar, Aniket; Allolio, Christoph; Jurkiewicz, Piotr; Baxová, Katarína; Šachl, Radek; Horinek, D.; Heinz, V.; Rachel, R.; Ziegler, C.; Jungwirth, Pavel

    2017-01-01

    Roč. 46, Suppl 1 (2017), S386 ISSN 0175-7571. [IUPAB congress /19./ and EBSA congress /11./. 16.07.2017-20.07.2017, Edinburgh] Institutional support: RVO:61388963 ; RVO:61388955 Keywords : membrane interactions * membrane fusion * cell penetration Subject RIV: BO - Biophysics

  5. Assessing the efficacy of vesicle fusion with planar membrane arrays using a mitochondrial porin as reporter

    International Nuclear Information System (INIS)

    Pszon-Bartosz, Kamila; Hansen, Jesper S.; Stibius, Karin B.; Groth, Jesper S.; Emneus, Jenny; Geschke, Oliver; Helix-Nielsen, Claus

    2011-01-01

    Research highlights: → We have established a vesicle fusion efficacy assay based on the major non-specific porin of Fusobacterium nucleatum (FomA). → Maximal fusion obtained was almost 150,000 porin insertions during 20 min. → Incorporation can be either first order or exponential kinetics which has implications for establishing protein delivery to biomimetic membranes. -- Abstract: Reconstitution of functionally active membrane protein into artificially made lipid bilayers is a challenge that must be overcome to create a membrane-based biomimetic sensor and separation device. In this study we address the efficacy of proteoliposome fusion with planar membrane arrays. We establish a protein incorporation efficacy assay using the major non-specific porin of Fusobacterium nucleatum (FomA) as reporter. We use electrical conductance measurements and fluorescence microscopy to characterize proteoliposome fusion with an array of planar membranes. We show that protein reconstitution in biomimetic membrane arrays may be quantified using the developed FomA assay. Specifically, we show that FomA vesicles are inherently fusigenic. Optimal FomA incorporation is obtained with a proteoliposome lipid-to-protein molar ratio (LPR) = 50 more than 10 5 FomA proteins could be incorporated in a bilayer array with a total membrane area of 2 mm 2 within 20 min. This novel assay for quantifying protein delivery into lipid bilayers may be a useful tool in developing biomimetic membrane applications.

  6. Assessing the efficacy of vesicle fusion with planar membrane arrays using a mitochondrial porin as reporter

    Energy Technology Data Exchange (ETDEWEB)

    Pszon-Bartosz, Kamila; Hansen, Jesper S. [Aquaporin A/S, Ole Maaloes Vej 3, DK-2200 Copenhagen N (Denmark); Technical University of Denmark, Department of Micro- and Nanotechnology, DK-2800 Kongens Lyngby (Denmark); Stibius, Karin B. [Aquaporin A/S, Ole Maaloes Vej 3, DK-2200 Copenhagen N (Denmark); Technical University of Denmark, Department of Physics, DTU Physics, DK-2800 Kongens Lyngby (Denmark); Groth, Jesper S. [Aquaporin A/S, Ole Maaloes Vej 3, DK-2200 Copenhagen N (Denmark); Emneus, Jenny; Geschke, Oliver [Technical University of Denmark, Department of Micro- and Nanotechnology, DK-2800 Kongens Lyngby (Denmark); Helix-Nielsen, Claus, E-mail: claus.helix.nielsen@fysik.dtu.dk [Aquaporin A/S, Ole Maaloes Vej 3, DK-2200 Copenhagen N (Denmark); Technical University of Denmark, Department of Physics, DTU Physics, DK-2800 Kongens Lyngby (Denmark)

    2011-03-04

    Research highlights: {yields} We have established a vesicle fusion efficacy assay based on the major non-specific porin of Fusobacterium nucleatum (FomA). {yields} Maximal fusion obtained was almost 150,000 porin insertions during 20 min. {yields} Incorporation can be either first order or exponential kinetics which has implications for establishing protein delivery to biomimetic membranes. -- Abstract: Reconstitution of functionally active membrane protein into artificially made lipid bilayers is a challenge that must be overcome to create a membrane-based biomimetic sensor and separation device. In this study we address the efficacy of proteoliposome fusion with planar membrane arrays. We establish a protein incorporation efficacy assay using the major non-specific porin of Fusobacterium nucleatum (FomA) as reporter. We use electrical conductance measurements and fluorescence microscopy to characterize proteoliposome fusion with an array of planar membranes. We show that protein reconstitution in biomimetic membrane arrays may be quantified using the developed FomA assay. Specifically, we show that FomA vesicles are inherently fusigenic. Optimal FomA incorporation is obtained with a proteoliposome lipid-to-protein molar ratio (LPR) = 50 more than 10{sup 5} FomA proteins could be incorporated in a bilayer array with a total membrane area of 2 mm{sup 2} within 20 min. This novel assay for quantifying protein delivery into lipid bilayers may be a useful tool in developing biomimetic membrane applications.

  7. Structure and orientation study of Ebola fusion peptide inserted in lipid membrane models.

    Science.gov (United States)

    Agopian, Audrey; Castano, Sabine

    2014-01-01

    The fusion peptide of Ebola virus comprises a highly hydrophobic sequence located downstream from the N-terminus of the glycoprotein GP2 responsible for virus-host membrane fusion. The internal fusion peptide of GP2 inserts into membranes of infected cell to mediate the viral and the host cell membrane fusion. Since the sequence length of Ebola fusion peptide is still not clear, we study in the present work the behavior of two fusion peptides of different lengths which were named EBO17 and EBO24 referring to their amino acid length. The secondary structure and orientation of both peptides in lipid model systems made of DMPC:DMPG:cholesterol:DMPE (6:2:5:3) were investigated using PMIRRAS and polarized ATR spectroscopy coupled with Brewster angle microscopy. The infrared results showed a structural flexibility of both fusion peptides which are able to transit reversibly from an α-helix to antiparallel β-sheets. Ellipsometry results corroborate together with isotherm measurements that EBO peptides interacting with lipid monolayer highly affected the lipid organization. When interacting with a single lipid bilayer, at low peptide content, EBO peptides insert as mostly α-helices mainly perpendicular into the lipid membrane thus tend to organize the lipid acyl chains. Inserted in multilamellar vesicles at higher peptide content, EBO peptides are mostly in β-sheet structures and induce a disorganization of the lipid chain order. In this paper, we show that the secondary structure of the Ebola fusion peptide is reversibly flexible between α-helical and β-sheet conformations, this feature being dependent on its concentration in lipids, eventually inducing membrane fusion. © 2013.

  8. Design, construction, and characterization of high-performance membrane fusion devices with target-selectivity.

    Science.gov (United States)

    Kashiwada, Ayumi; Yamane, Iori; Tsuboi, Mana; Ando, Shun; Matsuda, Kiyomi

    2012-01-31

    Membrane fusion proteins such as the hemagglutinin glycoprotein have target recognition and fusion accelerative domains, where some synergistically working elements are essential for target-selective and highly effective native membrane fusion systems. In this work, novel membrane fusion devices bearing such domains were designed and constructed. We selected a phenylboronic acid derivative as a recognition domain for a sugar-like target and a transmembrane-peptide (Leu-Ala sequence) domain interacting with the target membrane, forming a stable hydrophobic α-helix and accelerating the fusion process. Artificial membrane fusion behavior between the synthetic devices in which pilot and target liposomes were incorporated was characterized by lipid-mixing and inner-leaflet lipid-mixing assays. Consequently, the devices bearing both the recognition and transmembrane domains brought about a remarkable increase in the initial rate for the membrane fusion compared with the devices containing the recognition domain alone. In addition, a weakly acidic pH-responsive device was also constructed by replacing three Leu residues in the transmembrane-peptide domain by Glu residues. The presence of Glu residues made the acidic pH-dependent hydrophobic α-helix formation possible as expected. The target-selective liposome-liposome fusion was accelerated in a weakly acidic pH range when the Glu-substituted device was incorporated in pilot liposomes. The use of this pH-responsive device seems to be a potential strategy for novel applications in a liposome-based delivery system. © 2011 American Chemical Society

  9. Study on Efficiency of Fusion Techniques for IKONOS Images

    International Nuclear Information System (INIS)

    Liu, Yanmei; Yu, Haiyang; Guijun, Yang; Nie, Chenwei; Yang, Xiaodong; Ren, Dong

    2014-01-01

    Many image fusion techniques have been proposed to achieve optimal resolution in the spatial and spectral domains. Six different merging methods were listed in this paper and the efficiency of fusion techniques was assessed in qualitative and quantitative aspect. Both local and global evaluation parameters were used in the spectral quality and a Laplace filter method was used in spatial quality assessment. By simulation, the spectral quality of the images merged by Brovery was demonstrated to be the worst. In contrast, GS and PCA algorithms, especially the Pansharpening provided higher spectral quality than the standard Brovery, wavelet and CN methods. In spatial quality assessment, the CN method represented best compared with that of others, while the Brovery algorithm was worst. The wavelet parameters that performed best achieved acceptable spectral and spatial quality compared to the others

  10. Cytosol-dependent membrane fusion in ER, nuclear envelope and nuclear pore assembly: biological implications.

    Science.gov (United States)

    Rafikova, Elvira R; Melikov, Kamran; Chernomordik, Leonid V

    2010-01-01

    Endoplasmic reticulum and nuclear envelope rearrangements after mitosis are often studied in the reconstitution system based on Xenopus egg extract. In our recent work we partially replaced the membrane vesicles in the reconstitution mix with protein-free liposomes to explore the relative contributions of cytosolic and transmembrane proteins. Here we discuss our finding that cytosolic proteins mediate fusion between membranes lacking functional transmembrane proteins and the role of membrane fusion in endoplasmic reticulum and nuclear envelope reorganization. Cytosol-dependent liposome fusion has allowed us to restore, without adding transmembrane nucleoporins, functionality of nuclear pores, their spatial distribution and chromatin decondensation in nuclei formed at insufficient amounts of membrane material and characterized by only partial decondensation of chromatin and lack of nuclear transport. Both the mechanisms and the biological implications of the discovered coupling between spatial distribution of nuclear pores, chromatin decondensation and nuclear transport are discussed.

  11. Complementation between avirulent Newcastle disease virus and a fusion protein gene expressed from a retrovirus vector: requirements for membrane fusion.

    OpenAIRE

    Morrison, T; McQuain, C; McGinnes, L

    1991-01-01

    The cDNA derived from the fusion gene of the virulent AV strain of Newcastle disease virus (NDV) was expressed in chicken embryo cells by using a retrovirus vector. The fusion protein expressed in this system was transported to the cell surface and was efficiently cleaved into the disulfide-linked F1-F2 form found in infectious virions. The cells expressing the fusion gene grew normally and could be passaged many times. Monolayers of these cells would plaque, in the absence of trypsin, avirul...

  12. Susceptibility to virus-cell fusion at the plasma membrane is reduced through expression of HIV gp41 cytoplasmic domains

    International Nuclear Information System (INIS)

    Malinowsky, Katharina; Luksza, Julia; Dittmar, Matthias T.

    2008-01-01

    The cytoplasmic tail of the HIV transmembrane protein plays an important role in viral infection. In this study we analyzed the role of retroviral cytoplasmic tails in modulating the cytoskeleton and interfering with virus-cell fusion. HeLaP4 cells expressing different HIV cytoplasmic tail constructs showed reduced acetylated tubulin levels whereas the cytoplasmic tail of MLV did not alter microtubule stability indicating a unique function for the lentiviral cytoplasmic tail. The effect on tubulin is mediated through the membrane proximal region of the HIV cytoplasmic tail and was independent of membrane localization. Site-directed mutagenesis identified three motifs in the HIV-2 cytoplasmic tail required to effect the reduction in acetylated tubulin. Both the YxxΦ domain and amino acids 21 to 45 of the HIV-2 cytoplasmic tail need to be present to change the level of acetylated tubulin in transfected cells. T-cells stably expressing one HIV-2 cytoplasmic tail derived construct showed also a reduction in acetylated tubulin thus confirming the importance of this effect not only for HeLaP4 and 293T cells. Challenge experiments using transiently transfected HeLaP4 cells and T cells stably expressing an HIV cytoplasmic tail construct revealed both reduced virus-cell fusion and replication of HIV-1 NL4.3 compared to control cells. In the virus-cell fusion assay only virions pseudotyped with either HIV or MLV envelopes showed reduced fusion efficiency, whereas VSV-G pseudotyped virions where not affected by the expression of HIV derived cytoplasmic tail constructs, indicating that fusion at the plasma but not endosomal membrane is affected. Overexpression of human histone-deacetylase 6 (HDAC6) and constitutively active RhoA resulted in a reduction of acetylated tubulin and reduced virus-cell fusion as significant as that observed following expression of HIV cytoplasmic tail constructs. Inhibition of HDAC6 showed a strong increase in acetylated tubulin and increase of

  13. Rational design of a fusion partner for membrane protein expression in E. coli.

    Science.gov (United States)

    Luo, Jianying; Choulet, Julie; Samuelson, James C

    2009-08-01

    We have designed a novel protein fusion partner (P8CBD) to utilize the co-translational SRP pathway in order to target heterologous proteins to the E. coli inner membrane. SRP-dependence was demonstrated by analyzing the membrane translocation of P8CBD-PhoA fusion proteins in wt and SRP-ffh77 mutant cells. We also demonstrate that the P8CBD N-terminal fusion partner promotes over-expression of a Thermotoga maritima polytopic membrane protein by replacement of the native signal anchor sequence. Furthermore, the yeast mitochondrial inner membrane protein Oxa1p was expressed as a P8CBD fusion and shown to function within the E. coli inner membrane. In this example, the mitochondrial targeting peptide was replaced by P8CBD. Several practical features were incorporated into the P8CBD expression system to aid in protein detection, purification, and optional in vitro processing by enterokinase. The basis of membrane protein over-expression toxicity is discussed and solutions to this problem are presented. We anticipate that this optimized expression system will aid in the isolation and study of various recombinant forms of membrane-associated protein.

  14. Physiological and molecular triggers for SARS-CoV membrane fusion and entry into host cells.

    Science.gov (United States)

    Millet, Jean Kaoru; Whittaker, Gary R

    2018-04-01

    During viral entry, enveloped viruses require the fusion of their lipid envelope with host cell membranes. For coronaviruses, this critical step is governed by the virally-encoded spike (S) protein, a class I viral fusion protein that has several unique features. Coronavirus entry is unusual in that it is often biphasic in nature, and can occur at or near the cell surface or in late endosomes. Recent advances in structural, biochemical and molecular biology of the coronavirus S protein has shed light on the intricacies of coronavirus entry, in particular the molecular triggers of coronavirus S-mediated membrane fusion. Furthermore, characterization of the coronavirus fusion peptide (FP), the segment of the fusion protein that inserts to a target lipid bilayer during membrane fusion, has revealed its particular attributes which imparts some of the unusual properties of the S protein, such as Ca 2+ -dependency. These unusual characteristics can explain at least in part the biphasic nature of coronavirus entry. In this review, using severe acute respiratory syndrome coronavirus (SARS-CoV) as model virus, we give an overview of advances in research on the coronavirus fusion peptide with an emphasis on its role and properties within the biological context of host cell entry. Copyright © 2017 Elsevier Inc. All rights reserved.

  15. Structure and interaction with lipid membrane models of Semliki Forest virus fusion peptide.

    Science.gov (United States)

    Agopian, A; Quetin, M; Castano, S

    2016-11-01

    Semliki Forest virus (SFV) is a well-characterized alphavirus that infects cells via endocytosis and an acid-triggered fusion step using class II fusion proteins. Membrane fusion is mediated by the viral spike protein, a heterotrimer of two transmembrane subunits, E1 and E2, and a peripheral protein, E3. Sequence analysis of the E1 ectodomain of a number of alphaviruses demonstrated the presence of a highly conserved hydrophobic domain on the E1 ectodomain. This sequence was proposed to be the fusion peptide of SFV and is believed to be the domain of E1 that interacts with the target membrane and triggers fusion. Here, we investigate the structure and the interaction with lipid membrane models of 76 YQCKVYTGVYPFMWGGAYCFC 96 sequence from SFV, named SFV21, using optical method (ellipsometry) and vibrational spectroscopiy approaches (Polarization Modulation infra-Red Reflection Absorption Spectroscopy, PMIRRAS, and polarized ATR-FTIR). We demonstrate a structural flexibility of SFV21 sequence whether the lateral pressure and the lipid environment. In a lipid environment that mimics eukaryotic cell membranes, a conformational transition from an α-helix to a β-sheet is induced in the presence of lipid by increasing the peptide to lipid ratio, which leads to important perturbations in the membrane organisation. Copyright © 2016 Elsevier B.V. All rights reserved.

  16. Efficient preparation and analysis of membrane and membrane protein systems

    Czech Academy of Sciences Publication Activity Database

    Javanainen, M.; Martinez-Seara, Hector

    2016-01-01

    Roč. 1858, č. 10 (2016), s. 2468-2482 ISSN 0005-2736 Institutional support: RVO:61388963 Keywords : tools and software * membrane building * protein insertion * molecular dynamics * lipid bilayer Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 3.498, year: 2016

  17. Assessing the efficacy of vesicle fusion with planar membrane arrays using a mitochondrial porin as reporter

    DEFF Research Database (Denmark)

    Pszon-Bartosz, Kamila Justyna; Hansen, Jesper S.; Stibius, Karin B.

    2011-01-01

    Reconstitution of functionally active membrane protein into artificially made lipid bilayers is a challenge that must be overcome to create a membrane-based biomimetic sensor and separation device. In this study we address the efficacy of proteoliposome fusion with planar membrane arrays. We...... establish a protein incorporation efficacy assay using the major non-specific porin of Fusobacterium nucleatum (FomA) as reporter. We use electrical conductance measurements and fluorescence microscopy to characterize proteoliposome fusion with an array of planar membranes. We show that protein...... reconstitution in biomimetic membrane arrays may be quantified using the developed FomA assay. Specifically, we show that FomA vesicles are inherently fusigenic. Optimal FomA incorporation is obtained with a proteoliposome lipid-to-protein molar ratio (LPR)=50 more than 105 FomA proteins could be incorporated...

  18. Inner/Outer nuclear membrane fusion in nuclear pore assembly: biochemical demonstration and molecular analysis.

    Science.gov (United States)

    Fichtman, Boris; Ramos, Corinne; Rasala, Beth; Harel, Amnon; Forbes, Douglass J

    2010-12-01

    Nuclear pore complexes (NPCs) are large proteinaceous channels embedded in double nuclear membranes, which carry out nucleocytoplasmic exchange. The mechanism of nuclear pore assembly involves a unique challenge, as it requires creation of a long-lived membrane-lined channel connecting the inner and outer nuclear membranes. This stabilized membrane channel has little evolutionary precedent. Here we mapped inner/outer nuclear membrane fusion in NPC assembly biochemically by using novel assembly intermediates and membrane fusion inhibitors. Incubation of a Xenopus in vitro nuclear assembly system at 14°C revealed an early pore intermediate where nucleoporin subunits POM121 and the Nup107-160 complex were organized in a punctate pattern on the inner nuclear membrane. With time, this intermediate progressed to diffusion channel formation and finally to complete nuclear pore assembly. Correct channel formation was blocked by the hemifusion inhibitor lysophosphatidylcholine (LPC), but not if a complementary-shaped lipid, oleic acid (OA), was simultaneously added, as determined with a novel fluorescent dextran-quenching assay. Importantly, recruitment of the bulk of FG nucleoporins, characteristic of mature nuclear pores, was not observed before diffusion channel formation and was prevented by LPC or OA, but not by LPC+OA. These results map the crucial inner/outer nuclear membrane fusion event of NPC assembly downstream of POM121/Nup107-160 complex interaction and upstream or at the time of FG nucleoporin recruitment.

  19. The destructive effect of botulinum neurotoxins on the SNARE protein: SNAP-25 and synaptic membrane fusion

    Directory of Open Access Journals (Sweden)

    Bin Lu

    2015-06-01

    Full Text Available Synaptic exocytosis requires the assembly of syntaxin 1A and SNAP-25 on the plasma membrane and synaptobrevin 2 (VAMP2 on the vesicular membrane to bridge the two opposite membranes. It is believed that the three SNARE proteins assemble in steps along the dynamic assembly pathway. The C-terminus of SNAP-25 is known to be the target of botulinum neurotoxins (BoNT/A and BoNT/E that block neurotransmitters release in vivo. In this study, we employed electron paramagnetic resonance (EPR spectroscopy to investigate the conformation of the SNAP-25 C-terminus in binary and ternary SNARE complexes. The fluorescence lipid mixing assay shows that the C-terminal of SNAP-25 is essential for membrane fusion, and that the truncated SNAP-25 mutants cleaved by BoNT/A and BoNT/E display different inhibition effects on membrane fusion: SNAP-25E (Δ26 abolishes the fusion activity of the SNARE complex, while SNAP-25A (Δ9 loses most of its function, although it can still form a SDS-resistant SNARE complex as the wild-type SNAP-25. CW-EPR spectra validate the unstable structures of the SNARE complex formed by SNAP-25 mutants. We propose that the truncated SNAP-25 mutants will disrupt the assembly of the SNARE core complex, and then inhibit the synaptic membrane fusion accordingly.

  20. The Fusion Loops of the Initial Prefusion Conformation of Herpes Simplex Virus 1 Fusion Protein Point Toward the Membrane

    Directory of Open Access Journals (Sweden)

    Juan Fontana

    2017-08-01

    Full Text Available All enveloped viruses, including herpesviruses, must fuse their envelope with the host membrane to deliver their genomes into target cells, making this essential step subject to interference by antibodies and drugs. Viral fusion is mediated by a viral surface protein that transits from an initial prefusion conformation to a final postfusion conformation. Strikingly, the prefusion conformation of the herpesvirus fusion protein, gB, is poorly understood. Herpes simplex virus (HSV, a model system for herpesviruses, causes diseases ranging from mild skin lesions to serious encephalitis and neonatal infections. Using cryo-electron tomography and subtomogram averaging, we have characterized the structure of the prefusion conformation and fusion intermediates of HSV-1 gB. To this end, we have set up a system that generates microvesicles displaying full-length gB on their envelope. We confirmed proper folding of gB by nondenaturing electrophoresis-Western blotting with a panel of monoclonal antibodies (MAbs covering all gB domains. To elucidate the arrangement of gB domains, we labeled them by using (i mutagenesis to insert fluorescent proteins at specific positions, (ii coexpression of gB with Fabs for a neutralizing MAb with known binding sites, and (iii incubation of gB with an antibody directed against the fusion loops. Our results show that gB starts in a compact prefusion conformation with the fusion loops pointing toward the viral membrane and suggest, for the first time, a model for gB’s conformational rearrangements during fusion. These experiments further illustrate how neutralizing antibodies can interfere with the essential gB structural transitions that mediate viral entry and therefore infectivity.

  1. The fusion-related hydrophobic domain of Sendai F protein can be moved through the cytoplasmic membrane of Escherichia coli.

    OpenAIRE

    Davis, N G; Hsu, M C

    1986-01-01

    Recent work on a prokaryotic membrane protein, gene III protein (pIII) of coliphage f1, showed that polypeptide segments of sufficient hydrophobicity functioned to stop transfer of the polypeptide across the cell membrane: strings of 16 or more hydrophobic amino acids sufficed. A fusion-related hydrophobic domain (FRHD) of Sendai F protein, a sequence of 26 consecutive uncharged residues, has been implicated in the fusion of the viral membrane envelope and the target-cell membrane through a h...

  2. Shielding efficiency of metal hydrides and borohydrides in fusion reactors

    DEFF Research Database (Denmark)

    Singh, Vishvanath P.; Badiger, Nagappa M.; Gerward, Leif

    2016-01-01

    at energies 0.015 MeV to15 MeV, and for penetration depths up to 40 mean free paths. Fast-neutron shielding efficiency has been characterized by the effective neutron removal cross-section. It is shown that ZrH2 and VH2 are very good shielding materials for gamma rays and fast neutrons due to their suitable......Mass attenuation coefficients, mean free paths and exposure buildup factors have been used to characterize the shielding efficiency of metal hydrides and borohydrides, with high density of hydrogen. Gamma ray exposure buildup factors were computed using five-parameter geometric progression fitting...... combination of low-and high-Z elements. The present work should be useful for the selection and design of blankets and shielding, and for dose evaluation for components in fusion reactors....

  3. Yeast Lipin 1 Orthologue Pah1p Regulates Vacuole Homeostasis and Membrane Fusion*

    Science.gov (United States)

    Sasser, Terry; Qiu, Quan-Sheng; Karunakaran, Surya; Padolina, Mark; Reyes, Anna; Flood, Blake; Smith, Sheena; Gonzales, Chad; Fratti, Rutilio A.

    2012-01-01

    Vacuole homotypic fusion requires a group of regulatory lipids that includes diacylglycerol, a fusogenic lipid that is produced through multiple metabolic pathways including the dephosphorylation of phosphatidic acid (PA). Here we examined the relationship between membrane fusion and PA phosphatase activity. Pah1p is the single yeast homologue of the Lipin family of PA phosphatases. Deletion of PAH1 was sufficient to cause marked vacuole fragmentation and abolish vacuole fusion. The function of Pah1p solely depended on its phosphatase activity as complementation studies showed that wild type Pah1p restored fusion, whereas the phosphatase dead mutant Pah1pD398E had no effect. We discovered that the lack of PA phosphatase activity blocked fusion by inhibiting the binding of SNAREs to Sec18p, an N-ethylmaleimide-sensitive factor homologue responsible for priming inactive cis-SNARE complexes. In addition, pah1Δ vacuoles were devoid of the late endosome/vacuolar Rab Ypt7p, the phosphatidylinositol 3-kinase Vps34p, and Vps39p, a subunit of the HOPS (homotypic fusion and vacuole protein sorting) tethering complex, all of which are required for vacuole fusion. The lack of Vps34p resulted in the absence of phosphatidylinositol 3-phosphate, a lipid required for SNARE activity and vacuole fusion. These findings demonstrate that Pah1p and PA phosphatase activity are critical for vacuole homeostasis and fusion. PMID:22121197

  4. Trans-complex formation by proteolipid channels in the terminal phase of membrane fusion

    DEFF Research Database (Denmark)

    Peters, C; Bayer, M J; Bühler, S

    2001-01-01

    -complex formation occurs downstream from trans-SNARE pairing, and depends on both the Rab-GTPase Ypt7 and calmodulin. The maintenance of existing complexes and completion of fusion are independent of trans-SNARE pairs. Reconstituted proteolipids form sealed channels, which can expand to form aqueous pores in a Ca2......+/calmodulin-dependent fashion. V0 trans-complexes may therefore form a continuous, proteolipid-lined channel at the fusion site. We propose that radial expansion of such a protein pore may be a mechanism for intracellular membrane fusion....

  5. The Heptad Repeat C Domain of the Respiratory Syncytial Virus Fusion Protein Plays a Key Role in Membrane Fusion.

    Science.gov (United States)

    Bermingham, Imogen M; Chappell, Keith J; Watterson, Daniel; Young, Paul R

    2018-02-15

    Respiratory syncytial virus (RSV) mediates host cell entry through the fusion (F) protein, which undergoes a conformational change to facilitate the merger of viral and host lipid membrane envelopes. The RSV F protein comprises a trimer of disulfide-bonded F 1 and F 2 subunits that is present on the virion surface in a metastable prefusion state. This prefusion form is readily triggered to undergo refolding to bring two heptad repeats (heptad repeat A [HRA] and HRB) into close proximity to form a six-helix bundle that stabilizes the postfusion form and provides the free energy required for membrane fusion. This process can be triggered independently of other proteins. Here, we have performed a comprehensive analysis of a third heptad repeat region, HRC (amino acids 75 to 97), an amphipathic α-helix that lies at the interface of the prefusion F trimer and is a major structural feature of the F 2 subunit. We performed alanine scanning mutagenesis from Lys-75 to Met-97 and assessed all mutations in transient cell culture for expression, proteolytic processing, cell surface localization, protein conformation, and membrane fusion. Functional characterization revealed a striking distribution of activity in which fusion-increasing mutations localized to one side of the helical face, while fusion-decreasing mutations clustered on the opposing face. Here, we propose a model in which HRC plays a stabilizing role within the globular head for the prefusion F trimer and is potentially involved in the early events of triggering, prompting fusion peptide release and transition into the postfusion state. IMPORTANCE RSV is recognized as the most important viral pathogen among pediatric populations worldwide, yet no vaccine or widely available therapeutic treatment is available. The F protein is critical for the viral replication process and is the major target for neutralizing antibodies. Recent years have seen the development of prefusion stabilized F protein-based approaches to

  6. Two coiled-coil domains of Chlamydia trachomatis IncA affect membrane fusion events during infection.

    Directory of Open Access Journals (Sweden)

    Erik Ronzone

    Full Text Available Chlamydia trachomatis replicates in a parasitophorous membrane-bound compartment called an inclusion. The inclusions corrupt host vesicle trafficking networks to avoid the degradative endolysosomal pathway but promote fusion with each other in order to sustain higher bacterial loads in a process known as homotypic fusion. The Chlamydia protein IncA (Inclusion protein A appears to play central roles in both these processes as it participates to homotypic fusion and inhibits endocytic SNARE-mediated membrane fusion. How IncA selectively inhibits or activates membrane fusion remains poorly understood. In this study, we analyzed the spatial and molecular determinants of IncA's fusogenic and inhibitory functions. Using a cell-free membrane fusion assay, we found that inhibition of SNARE-mediated fusion requires IncA to be on the same membrane as the endocytic SNARE proteins. IncA displays two coiled-coil domains showing high homology with SNARE proteins. Domain swap and deletion experiments revealed that although both these domains are capable of independently inhibiting SNARE-mediated fusion, these two coiled-coil domains cooperate in mediating IncA multimerization and homotypic membrane interaction. Our results support the hypothesis that Chlamydia employs SNARE-like virulence factors that positively and negatively affect membrane fusion and promote infection.

  7. Two coiled-coil domains of Chlamydia trachomatis IncA affect membrane fusion events during infection.

    Science.gov (United States)

    Ronzone, Erik; Paumet, Fabienne

    2013-01-01

    Chlamydia trachomatis replicates in a parasitophorous membrane-bound compartment called an inclusion. The inclusions corrupt host vesicle trafficking networks to avoid the degradative endolysosomal pathway but promote fusion with each other in order to sustain higher bacterial loads in a process known as homotypic fusion. The Chlamydia protein IncA (Inclusion protein A) appears to play central roles in both these processes as it participates to homotypic fusion and inhibits endocytic SNARE-mediated membrane fusion. How IncA selectively inhibits or activates membrane fusion remains poorly understood. In this study, we analyzed the spatial and molecular determinants of IncA's fusogenic and inhibitory functions. Using a cell-free membrane fusion assay, we found that inhibition of SNARE-mediated fusion requires IncA to be on the same membrane as the endocytic SNARE proteins. IncA displays two coiled-coil domains showing high homology with SNARE proteins. Domain swap and deletion experiments revealed that although both these domains are capable of independently inhibiting SNARE-mediated fusion, these two coiled-coil domains cooperate in mediating IncA multimerization and homotypic membrane interaction. Our results support the hypothesis that Chlamydia employs SNARE-like virulence factors that positively and negatively affect membrane fusion and promote infection.

  8. Reverse flow catalytic membrane reactors for energy efficient syngas production

    OpenAIRE

    Smit, Joris

    2006-01-01

    To improve the recuperative heat exchange, a Reverse Flow Catalytic Membrane Reactor (RFCMR) with porous membranes is proposed in this thesis, in which very efficient heat exchange between the feed and product streams is achieved by using the reverse flow concept (i.e. periodic alternation of the flow direction of the gas through a fixed catalyst bed).

  9. Reverse flow catalytic membrane reactors for energy efficient syngas production

    NARCIS (Netherlands)

    Smit, Joris

    2006-01-01

    To improve the recuperative heat exchange, a Reverse Flow Catalytic Membrane Reactor (RFCMR) with porous membranes is proposed in this thesis, in which very efficient heat exchange between the feed and product streams is achieved by using the reverse flow concept (i.e. periodic alternation of the

  10. Novel thermal efficiency-based model for determination of thermal conductivity of membrane distillation membranes

    International Nuclear Information System (INIS)

    Vanneste, Johan; Bush, John A.; Hickenbottom, Kerri L.; Marks, Christopher A.; Jassby, David

    2017-01-01

    Development and selection of membranes for membrane distillation (MD) could be accelerated if all performance-determining characteristics of the membrane could be obtained during MD operation without the need to recur to specialized or cumbersome porosity or thermal conductivity measurement techniques. By redefining the thermal efficiency, the Schofield method could be adapted to describe the flux without prior knowledge of membrane porosity, thickness, or thermal conductivity. A total of 17 commercially available membranes were analyzed in terms of flux and thermal efficiency to assess their suitability for application in MD. The thermal-efficiency based model described the flux with an average %RMSE of 4.5%, which was in the same range as the standard deviation on the measured flux. The redefinition of the thermal efficiency also enabled MD to be used as a novel thermal conductivity measurement device for thin porous hydrophobic films that cannot be measured with the conventional laser flash diffusivity technique.

  11. High-yield membrane protein expression from E. coli using an engineered outer membrane protein F fusion.

    Science.gov (United States)

    Su, Pin-Chuan; Si, William; Baker, Deidre L; Berger, Bryan W

    2013-04-01

    Obtaining high yields of membrane proteins necessary to perform detailed structural study is difficult due to poor solubility and variability in yields from heterologous expression systems. To address this issue, an Escherichia coli-based membrane protein overexpression system utilizing an engineered bacterial outer membrane protein F (pOmpF) fusion has been developed. Full-length human receptor activity-modifying protein 1 (RAMP1) was expressed using pOmpF, solubilized in FC15 and purified to homogeneity. Using circular dichroism and fluorescence spectroscopy, purified full-length RAMP1 is composed of approximately 90% α-helix, and retains its solubility and structure in FC15 over a wide range of temperatures (20-60°C). Thus, our approach provides a useful, complementary approach to achieve high-yield, full-length membrane protein overexpression for biophysical studies. Copyright © 2013 The Protein Society.

  12. Membrane fusion is induced by a distinct peptide sequence of the sea urchin fertilization protein bindin

    NARCIS (Netherlands)

    Ulrich, AS; Glabe, CG; Hoekstra, D

    1998-01-01

    Fertilization in the sea urchin is mediated by the membrane-associated acrosomal protein bindin, which plays a key role in the adhesion and fusion between sperm and egg. We have investigated the structure/function relationship of an 18-amino acid peptide fragment "B18," which represents the minimal

  13. Towards fully automated Identification of Vesicle-Membrane Fusion Events in TIRF Microscopy

    Science.gov (United States)

    Vallotton, Pascal; James, David E.; Hughes, William E.

    2007-11-01

    Total Internal Reflection Fluorescence Microscopy (TIRFM) is imposing itself as the tool of choice for studying biological activity in close proximity to the plasma membrane. For example, the exquisite selectivity of TIRFM allows monitoring the diffusion of GFP-phogrin vesicles and their recruitment to the plasma membrane in pancreatic β-cells. We present a novel computer vision system for automatically identifying the elusive fusion events of GFP-phogrin vesicles with the plasma membrane. Our method is based on robust object tracking and matched filtering. It should accelerate the quantification of TIRFM data and allow the extraction of more biological information from image data to support research in diabetes and obesity.

  14. Influenza Hemifusion Phenotype Depends on Membrane Context: Differences in Cell-Cell and Virus-Cell Fusion.

    Science.gov (United States)

    Zawada, Katarzyna E; Okamoto, Kenta; Kasson, Peter M

    2018-03-02

    Influenza viral entry into the host cell cytoplasm is accomplished by a process of membrane fusion mediated by the viral hemagglutinin protein. Hemagglutinin acts in a pH-triggered fashion, inserting a short fusion peptide into the host membrane followed by refolding of a coiled-coil structure to draw the viral envelope and host membranes together. Mutations to this fusion peptide provide an important window into viral fusion mechanisms and protein-membrane interactions. Here, we show that a well-described fusion peptide mutant, G1S, has a phenotype that depends strongly on the viral membrane context. The G1S mutant is well known to cause a "hemifusion" phenotype based on experiments in transfected cells, where cells expressing G1S hemagglutinin can undergo lipid mixing in a pH-triggered fashion similar to virus but will not support fusion pores. We compare fusion by the G1S hemagglutinin mutant expressed either in cells or in influenza virions and show that this hemifusion phenotype occurs in transfected cells but that native virions are able to support full fusion, albeit at a slower rate and 10-100× reduced infectious titer. We explain this with a quantitative model where the G1S mutant, instead of causing an absolute block of fusion, alters the protein stoichiometry required for fusion. This change slightly slows fusion at high hemagglutinin density, as on the viral surface, but at lower hemagglutinin density produces a hemifusion phenotype. The quantitative model thus reproduces the observed virus-cell and cell-cell fusion phenotypes, yielding a unified explanation where membrane context can control the observed viral fusion phenotype. Copyright © 2018 Elsevier Ltd. All rights reserved.

  15. Study on a multi-component palladium alloy membrane for the fusion fuel cycle

    International Nuclear Information System (INIS)

    Yoshida, Hiroshi; Okuno, Kenji; Nagasaki, Takanori; Noda, Kenji; Ishii, Yoshinobu; Takeshita, Hidefumi.

    1985-11-01

    A feasibility study on the material integrity with respect to the hydride formation and helium damage of the palladium alloy membrane was performed for an application of the palladium diffuser to a fusion fuel cleanup process. This study was conducted under the Japan/US Fusion Cooperation Program. Experimental works on the crystallography, hydrogen solubility and 3 He release characteristics were carried out with a multi-component palladium alloy(Pd-25Ag.Au.Ru). The excellent hydrogen permeability and mechanical properties of the membrane made of this alloy had been confirmed by authors' previous study. Based on the present study, this alloy membrane has high resistivity to the hydrogen embrittlement, and swelling and fracture due to the helium bubble formation under the practical operating conditions of the diffuser. (author)

  16. Membrane support of accelerated fuel capsules for inertial fusion energy reactors

    International Nuclear Information System (INIS)

    Petzoldt, R.W.; Moir, R.W.

    1993-01-01

    The use of a thin membrane to suspend an (inertial fusion energy) fuel capsule in a holder for injection into a reactor chamber is investigated. Capsule displacement and membrane deformation angle are calculated for an axisymmetric geometry for a range of membrane strain and capsule size. This information is used to calculate maximum target accelerations. Membranes must be thin (perhaps of order one micron) to minimize their effect on capsule implosion symmetry. For example, a 5 μm thick cryogenic mylar membrane is calculated to allow 1,000 m/s 2 acceleration of a 3 mm radius, 100 mg capsule. Vibration analysis (for a single membrane support) shows that if membrane vibration is not deliberately minimized, allowed acceleration may be reduced by a factor of four. A two membrane alternative geometry would allow several times greater acceleration. Therefore, alternative membrane geometry's should be used to provide greater target acceleration potential and reduce capsule displacement within the holder (for a given membrane thickness)

  17. Carbon membranes for efficient water-ethanol separation

    Science.gov (United States)

    Gravelle, Simon; Yoshida, Hiroaki; Joly, Laurent; Ybert, Christophe; Bocquet, Lydéric

    2016-09-01

    We demonstrate, on the basis of molecular dynamics simulations, the possibility of an efficient water-ethanol separation using nanoporous carbon membranes, namely, carbon nanotube membranes, nanoporous graphene sheets, and multilayer graphene membranes. While these carbon membranes are in general permeable to both pure liquids, they exhibit a counter-intuitive "self-semi-permeability" to water in the presence of water-ethanol mixtures. This originates in a preferred ethanol adsorption in nanoconfinement that prevents water molecules from entering the carbon nanopores. An osmotic pressure is accordingly expressed across the carbon membranes for the water-ethanol mixture, which agrees with the classic van't Hoff type expression. This suggests a robust and versatile membrane-based separation, built on a pressure-driven reverse-osmosis process across these carbon-based membranes. In particular, the recent development of large-scale "graphene-oxide" like membranes then opens an avenue for a versatile and efficient ethanol dehydration using this separation process, with possible application for bio-ethanol fabrication.

  18. High-efficiency technology for lithium isotope separation using an ionic-liquid impregnated organic membrane

    International Nuclear Information System (INIS)

    Hoshino, Tsuyoshi; Terai, Takayuki

    2011-01-01

    The tritium needed as a fuel for fusion reactors is produced by the neutron capture reaction of lithium-6 ( 6 Li) in tritium breeding materials. New lithium isotope separation technique using ionic-liquid impregnated organic membranes (Ionic-Liquid-i-OMs) have been developed. Lithium ions are able to move by electrodialysis through certain Ionic-Liquid-i-OMs between the cathode and the anode in lithium solutions. In this report, the effects of protection cover and membrane thickness on the durability of membrane and the efficiency of isotope separation were evaluated. In order to improve the durability of the Ionic-Liquid-i-OM, we developed highly-durable Ionic-Liquid-i-OM. Both surfaces of the Ionic-Liquid-i-OM were covered by a nafion 324 overcoat or a cation exchange membrane (SELEMION TM CMD) to prevent the outflow of the ionic liquid. It was observed that the durability of the Ionic-Liquid-i-OM was improved by a nafion 324 overcoat. On the other hand, the organic membrane selected was 1, 2 or 3 mm highly-porous Teflon film, in order to efficiently impregnate the ionic liquid. The 6 Li isotope separation factor by electrodialysis using highly-porous Teflon film of 3 mm thickness was larger than using that of 1 or 2 mm thickness.

  19. Sequential analysis of trans-SNARE formation in intracellular membrane fusion.

    Directory of Open Access Journals (Sweden)

    Kannan Alpadi

    2012-01-01

    Full Text Available SNARE complexes are required for membrane fusion in the endomembrane system. They contain coiled-coil bundles of four helices, three (Q(a, Q(b, and Q(c from target (t-SNAREs and one (R from the vesicular (v-SNARE. NSF/Sec18 disrupts these cis-SNARE complexes, allowing reassembly of their subunits into trans-SNARE complexes and subsequent fusion. Studying these reactions in native yeast vacuoles, we found that NSF/Sec18 activates the vacuolar cis-SNARE complex by selectively displacing the vacuolar Q(a SNARE, leaving behind a Q(bcR subcomplex. This subcomplex serves as an acceptor for a Q(a SNARE from the opposite membrane, leading to Q(a-Q(bcR trans-complexes. Activity tests of vacuoles with diagnostic distributions of inactivating mutations over the two fusion partners confirm that this distribution accounts for a major share of the fusion activity. The persistence of the Q(bcR cis-complex and the formation of the Q(a-Q(bcR trans-complex are both sensitive to the Rab-GTPase inhibitor, GDI, and to mutations in the vacuolar tether complex, HOPS (HOmotypic fusion and vacuolar Protein Sorting complex. This suggests that the vacuolar Rab-GTPase, Ypt7, and HOPS restrict cis-SNARE disassembly and thereby bias trans-SNARE assembly into a preferred topology.

  20. Sequential Analysis of Trans-SNARE Formation in Intracellular Membrane Fusion

    Science.gov (United States)

    Alpadi, Kannan; Kulkarni, Aditya; Comte, Veronique; Reinhardt, Monique; Schmidt, Andrea; Namjoshi, Sarita; Mayer, Andreas; Peters, Christopher

    2012-01-01

    SNARE complexes are required for membrane fusion in the endomembrane system. They contain coiled-coil bundles of four helices, three (Qa, Qb, and Qc) from target (t)-SNAREs and one (R) from the vesicular (v)-SNARE. NSF/Sec18 disrupts these cis-SNARE complexes, allowing reassembly of their subunits into trans-SNARE complexes and subsequent fusion. Studying these reactions in native yeast vacuoles, we found that NSF/Sec18 activates the vacuolar cis-SNARE complex by selectively displacing the vacuolar Qa SNARE, leaving behind a QbcR subcomplex. This subcomplex serves as an acceptor for a Qa SNARE from the opposite membrane, leading to Qa-QbcR trans-complexes. Activity tests of vacuoles with diagnostic distributions of inactivating mutations over the two fusion partners confirm that this distribution accounts for a major share of the fusion activity. The persistence of the QbcR cis-complex and the formation of the Qa-QbcR trans-complex are both sensitive to the Rab-GTPase inhibitor, GDI, and to mutations in the vacuolar tether complex, HOPS (HOmotypic fusion and vacuolar Protein Sorting complex). This suggests that the vacuolar Rab-GTPase, Ypt7, and HOPS restrict cis-SNARE disassembly and thereby bias trans-SNARE assembly into a preferred topology. PMID:22272185

  1. Autographa californica multiple nucleopolyhedrovirus GP64 protein: Analysis of domain I and V amino acid interactions and membrane fusion activity

    Energy Technology Data Exchange (ETDEWEB)

    Yu, Qianlong [State Key Laboratory of Crop Stress Biology for Arid Areas, Key Laboratory of Northwest Loess Plateau Crop Pest Management of Ministry of Agriculture, College of Plant Protection, Northwest A& F University, Yangling, Shaanxi 712100 (China); Blissard, Gary W. [Boyce Thompson Institute, Cornell University, Ithaca, NY 14853, United State (United States); Liu, Tong-Xian [State Key Laboratory of Crop Stress Biology for Arid Areas, Key Laboratory of Northwest Loess Plateau Crop Pest Management of Ministry of Agriculture, College of Plant Protection, Northwest A& F University, Yangling, Shaanxi 712100 (China); Li, Zhaofei, E-mail: zhaofeili73@outlook.com [State Key Laboratory of Crop Stress Biology for Arid Areas, Key Laboratory of Northwest Loess Plateau Crop Pest Management of Ministry of Agriculture, College of Plant Protection, Northwest A& F University, Yangling, Shaanxi 712100 (China)

    2016-01-15

    The Autographa californica multiple nucleopolyhedrovirus GP64 is a class III viral fusion protein. Although the post-fusion structure of GP64 has been solved, its pre-fusion structure and the detailed mechanism of conformational change are unknown. In GP64, domain V is predicted to interact with two domain I segments that flank fusion loop 2. To evaluate the significance of the amino acids involved in these interactions, we examined 24 amino acid positions that represent interacting and conserved residues within domains I and V. In several cases, substitution of a single amino acid involved in a predicted interaction disrupted membrane fusion activity, but no single amino acid pair appears to be absolutely required. We identified 4 critical residues in domain V (G438, W439, T452, and T456) that are important for membrane fusion, and two residues (G438 and W439) that appear to be important for formation or stability of the pre-fusion conformation of GP64. - Highlights: • The baculovirus envelope glycoprotein GP64 is a class III viral fusion protein. • The detailed mechanism of conformational change of GP64 is unknown. • We analyzed 24 positions that might stabilize the post-fusion structure of GP64. • We identified 4 residues in domain V that were critical for membrane fusion. • Two residues are critical for formation of the pre-fusion conformation of GP64.

  2. Autographa californica multiple nucleopolyhedrovirus GP64 protein: Analysis of domain I and V amino acid interactions and membrane fusion activity

    International Nuclear Information System (INIS)

    Yu, Qianlong; Blissard, Gary W.; Liu, Tong-Xian; Li, Zhaofei

    2016-01-01

    The Autographa californica multiple nucleopolyhedrovirus GP64 is a class III viral fusion protein. Although the post-fusion structure of GP64 has been solved, its pre-fusion structure and the detailed mechanism of conformational change are unknown. In GP64, domain V is predicted to interact with two domain I segments that flank fusion loop 2. To evaluate the significance of the amino acids involved in these interactions, we examined 24 amino acid positions that represent interacting and conserved residues within domains I and V. In several cases, substitution of a single amino acid involved in a predicted interaction disrupted membrane fusion activity, but no single amino acid pair appears to be absolutely required. We identified 4 critical residues in domain V (G438, W439, T452, and T456) that are important for membrane fusion, and two residues (G438 and W439) that appear to be important for formation or stability of the pre-fusion conformation of GP64. - Highlights: • The baculovirus envelope glycoprotein GP64 is a class III viral fusion protein. • The detailed mechanism of conformational change of GP64 is unknown. • We analyzed 24 positions that might stabilize the post-fusion structure of GP64. • We identified 4 residues in domain V that were critical for membrane fusion. • Two residues are critical for formation of the pre-fusion conformation of GP64.

  3. Regulation of membrane fusion and secretory events in the sea urchin embryo

    Energy Technology Data Exchange (ETDEWEB)

    Roe, J.L.

    1990-01-01

    Membrane fusion and secretory events play a key role in fertilization and early development in the sea urchin embryo. To investigate the mechanism of membrane fusion, the effect of inhibitors of metalloendoprotease activity was studied on two model systems of cell fusion; fertilization and spiculogenesis by primary mesenchyme cells in the embryo. Both the zinc chelator, 1,10-phenanthroline, and peptide metalloprotease substrates were found to inhibit both fertilization and gamete fusion, while peptides that are not substrates of metalloproteases did not affect either process. Primary mesenchyme cells form the larval skeleton in the embryo by deposition of mineral and an organic matrix into a syncytial cavity formed by fusion of filopodia of these cells. Metalloprotease inhibitors were found to inhibit spiculogenesis both in vivo and in cultures of isolated primary mesenchyme cells, and the activity of a metalloprotease of the appropriate specificity was found in the primary mesenchyme cells. These two studies implicate the activity of a metalloprotease in a necessary step in membrane fusion. Following fertilization, exocytosis of the cortical granules results in the formation of the fertilization envelope and the hyaline layer, that surround the developing embryo. The hatching enzyme is secreted by the blastula stage sea urchin embryo, which proteolyzes the fertilization envelope surrounding the embryo, allowing the embryo to hatch. Using an assay that measures {sup 125}I-fertilization envelope degradation, the hatching enzyme was identified as a 33 kDa metalloprotease, and was purified by ion-exchange and affinity chromatography from the hatching media of Strongylocentrotus purpuratus embryos. The hatching enzyme showed a substrate preference for only a minor subset of fertilization envelope proteins.

  4. Reciprocal complementation of bovine parainfluenza virus type 3 lacking either the membrane or fusion gene.

    Science.gov (United States)

    Takada, Marina; Matsuura, Ryosuke; Kokuho, Takehiro; Tsuboi, Takamitsu; Kameyama, Ken-Ichiro; Takeuchi, Kaoru

    2017-11-01

    Two defective bovine parainfluenza virus type 3 (BPIV3) strains were generated, one lacking the membrane (M) protein gene and expressing EGFP (ΔM-EGFP) and the other lacking the fusion (F) protein gene and expressing mStrawberry (ΔF-mSB), by supplying deficient proteins in trans. When Madin-Darby bovine kidney (MDBK) cells were co-infected with ΔM-EGFP and ΔF-mSB at a multiplicity of infection (MOI) of 0.1, complemented viruses were easily obtained. Complemented viruses grew as efficiently as wild-type BPIV3 and could be passaged in MDBK cell cultures even at an MOI of 0.01, possibly due to multiploid virus particles containing genomes of both ΔM-EGFP and ΔF-mSB. This reciprocal complementation method using two defective viruses would be useful to express large or multiple proteins in cell cultures using paramyxovirus vectors. Copyright © 2017 Elsevier B.V. All rights reserved.

  5. Tri-membrane nanoparticles produced by combining liposome fusion and a novel patchwork of bicelles to overcome endosomal and nuclear membrane barriers to cargo delivery.

    Science.gov (United States)

    Yamada, Asako; Mitsueda, Asako; Hasan, Mahadi; Ueda, Miho; Hama, Susumu; Warashina, Shota; Nakamura, Takashi; Harashima, Hideyoshi; Kogure, Kentaro

    2016-03-01

    Membrane fusion is a rational strategy for crossing intracellular membranes that present barriers to liposomal nanocarrier-mediated delivery of plasmid DNA into the nucleus of non-dividing cells, such as dendritic cells. Based on this strategy, we previously developed nanocarriers consisting of a nucleic acid core particle coated with four lipid membranes [Akita, et al., Biomaterials, 2009, 30, 2940-2949]. However, including the endosomal membrane and two nuclear membranes, cells possess three intracellular membranous barriers. Thus, after entering the nucleus, nanoparticles coated with four membranes would still have one lipid membrane remaining, and could impede cargo delivery. Until now, coating a core particle with an odd number of lipid membranes was challenging. To produce nanocarriers with an odd number of lipid membranes, we developed a novel coating method involving lipid nano-discs, also known as bicelles, as a material for packaging DNA in a carrier with an odd number of lipid membranes. In this procedure, bicelles fuse to form an outer coating that resembles a patchwork quilt, which allows the preparation of nanoparticles coated with only three lipid membranes. Moreover, the transfection activity of dendritic cells with these three-membrane nanoparticles was higher than that for nanoparticles coated with four lipid membranes. In summary, we developed novel nanoparticles coated with an odd number of lipid membranes using the novel "patchwork-packaging method" to deliver plasmid DNA into the nucleus via membrane fusion.

  6. Sequential Conformational Changes in the Morbillivirus Attachment Protein Initiate the Membrane Fusion Process

    Science.gov (United States)

    Ader-Ebert, Nadine; Khosravi, Mojtaba; Herren, Michael; Avila, Mislay; Alves, Lisa; Bringolf, Fanny; Örvell, Claes; Langedijk, Johannes P.; Zurbriggen, Andreas; Plemper, Richard K.; Plattet, Philippe

    2015-01-01

    Despite large vaccination campaigns, measles virus (MeV) and canine distemper virus (CDV) cause major morbidity and mortality in humans and animals, respectively. The MeV and CDV cell entry system relies on two interacting envelope glycoproteins: the attachment protein (H), consisting of stalk and head domains, co-operates with the fusion protein (F) to mediate membrane fusion. However, how receptor-binding by the H-protein leads to F-triggering is not fully understood. Here, we report that an anti-CDV-H monoclonal antibody (mAb-1347), which targets the linear H-stalk segment 126-133, potently inhibits membrane fusion without interfering with H receptor-binding or F-interaction. Rather, mAb-1347 blocked the F-triggering function of H-proteins regardless of the presence or absence of the head domains. Remarkably, mAb-1347 binding to headless CDV H, as well as standard and engineered bioactive stalk-elongated CDV H-constructs treated with cells expressing the SLAM receptor, was enhanced. Despite proper cell surface expression, fusion promotion by most H-stalk mutants harboring alanine substitutions in the 126-138 “spacer” section was substantially impaired, consistent with deficient receptor-induced mAb-1347 binding enhancement. However, a previously reported F-triggering defective H-I98A variant still exhibited the receptor-induced “head-stalk” rearrangement. Collectively, our data spotlight a distinct mechanism for morbillivirus membrane fusion activation: prior to receptor contact, at least one of the morbillivirus H-head domains interacts with the membrane-distal “spacer” domain in the H-stalk, leaving the F-binding site located further membrane-proximal in the stalk fully accessible. This “head-to-spacer” interaction conformationally stabilizes H in an auto-repressed state, which enables intracellular H-stalk/F engagement while preventing the inherent H-stalk’s bioactivity that may prematurely activate F. Receptor-contact disrupts the

  7. An efficient multiple exposure image fusion in JPEG domain

    Science.gov (United States)

    Hebbalaguppe, Ramya; Kakarala, Ramakrishna

    2012-01-01

    In this paper, we describe a method to fuse multiple images taken with varying exposure times in the JPEG domain. The proposed algorithm finds its application in HDR image acquisition and image stabilization for hand-held devices like mobile phones, music players with cameras, digital cameras etc. Image acquisition at low light typically results in blurry and noisy images for hand-held camera's. Altering camera settings like ISO sensitivity, exposure times and aperture for low light image capture results in noise amplification, motion blur and reduction of depth-of-field respectively. The purpose of fusing multiple exposures is to combine the sharp details of the shorter exposure images with high signal-to-noise-ratio (SNR) of the longer exposure images. The algorithm requires only a single pass over all images, making it efficient. It comprises of - sigmoidal boosting of shorter exposed images, image fusion, artifact removal and saturation detection. Algorithm does not need more memory than a single JPEG macro block to be kept in memory making it feasible to be implemented as the part of a digital cameras hardware image processing engine. The Artifact removal step reuses the JPEGs built-in frequency analysis and hence benefits from the considerable optimization and design experience that is available for JPEG.

  8. Pilot demonstration of energy-efficient membrane bioreactor (MBR) using reciprocating submerged membrane.

    Science.gov (United States)

    Ho, Jaeho; Smith, Shaleena; Patamasank, Jaren; Tontcheva, Petia; Kim, Gyu Dong; Roh, Hyung Keun

    2015-03-01

    Membrane bioreactor (MBR) is becoming popular for advanced wastewater treatment and water reuse. Air scouring to "shake" the membrane fibers is most suitable and applicable to maintain filtration without severe and rapidfouling. However, membrane fouling mitigating technologies are energy intensive. The goal of this research is to develop an alternative energy-saving MBR system to reduce energy consumption; a revolutionary system that will directly compete with air scouring technologies currently in the membrane water reuse market. The innovative MBR system, called reciprocation MBR (rMBR), prevents membrane fouling without the use of air scouring blowers. The mechanism featured is a mechanical reciprocating membrane frame that uses inertia to prevent fouling. Direct strong agitation of the fiber is also beneficial for the constant removal of solids built up on the membrane surface. The rMBR pilot consumes less energy than conventional coarse air scouring MBR systems. Specific energy consumption for membrane reciprocation for the pilot rMBR system was 0.072 kWh/m3 permeate produced at 40 LMH, which is 75% less than the conventional air scouring in an MBR system (0.29 kWh/m3). Reciprocation of the hollow-fiber membrane can overcome the hydrodynamic limitations of air scouring or cross-flow membrane systems with less energy consumption and/or higher energy efficiency.

  9. Nuclear inner membrane fusion facilitated by yeast Jem1p is required for spindle pole body fusion but not for the first mitotic nuclear division during yeast mating.

    Science.gov (United States)

    Nishikawa, Shuh-ichi; Hirata, Aiko; Endo, Toshiya

    2008-11-01

    During mating of budding yeast, Saccharomyces cerevisiae, two haploid nuclei fuse to produce a diploid nucleus. The process of nuclear fusion requires two J proteins, Jem1p in the endoplasmic reticulum (ER) lumen and Sec63p, which forms a complex with Sec71p and Sec72p, in the ER membrane. Zygotes of mutants defective in the functions of Jem1p or Sec63p contain two haploid nuclei that were closely apposed but failed to fuse. Here we analyzed the ultrastructure of nuclei in jem1 Delta and sec71 Delta mutant zygotes using electron microscope with the freeze-substituted fixation method. Three-dimensional reconstitution of nuclear structures from electron microscope serial sections revealed that Jem1p facilitates nuclear inner-membrane fusion and spindle pole body (SPB) fusion while Sec71p facilitates nuclear outer-membrane fusion. Two haploid SPBs that failed to fuse could duplicate, and mitotic nuclear division of the unfused haploid nuclei started in jem1 Delta and sec71 Delta mutant zygotes. This observation suggests that nuclear inner-membrane fusion is required for SPB fusion, but not for SPB duplication in the first mitotic cell division.

  10. 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.

  11. MXene molecular sieving membranes for highly efficient gas separation.

    Science.gov (United States)

    Ding, Li; Wei, Yanying; Li, Libo; Zhang, Tao; Wang, Haihui; Xue, Jian; Ding, Liang-Xin; Wang, Suqing; Caro, Jürgen; Gogotsi, Yury

    2018-01-11

    Molecular sieving membranes with sufficient and uniform nanochannels that break the permeability-selectivity trade-off are desirable for energy-efficient gas separation, and the arising two-dimensional (2D) materials provide new routes for membrane development. However, for 2D lamellar membranes, disordered interlayer nanochannels for mass transport are usually formed between randomly stacked neighboring nanosheets, which is obstructive for highly efficient separation. Therefore, manufacturing lamellar membranes with highly ordered nanochannel structures for fast and precise molecular sieving is still challenging. Here, we report on lamellar stacked MXene membranes with aligned and regular subnanometer channels, taking advantage of the abundant surface-terminating groups on the MXene nanosheets, which exhibit excellent gas separation performance with H 2 permeability >2200 Barrer and H 2 /CO 2 selectivity >160, superior to the state-of-the-art membranes. The results of molecular dynamics simulations quantitatively support the experiments, confirming the subnanometer interlayer spacing between the neighboring MXene nanosheets as molecular sieving channels for gas separation.

  12. Efficiency and temperature dependence of water removal by membrane dryers

    Science.gov (United States)

    Leckrone, K. J.; Hayes, J. M.

    1997-01-01

    The vapor pressure of water in equilibrium with sorption sites within a Nafion membrane is given by log P(WN) = -3580/T + 10.01, where P(WN) is expressed in Torr and T is the membrane temperature, in kelvin. The efficiency of dryers based on selective permeation of water through Nafion can thus be enhanced by cooling the membrane. Residual water in effluents exceeds equilibrium levels if insufficient time is allowed for water to diffuse to the membrane surface as gas passes through the dryer. For tubular configurations, this limitation can be avoided if L > or = Fc(10(3.8)/120 pi D), where L is the length of the tubular membrane, in centimeters, Fc is the gas flow rate, in mL/ min, and D is the diffusion coefficient for water in the carrier gas at the operating temperature of the dryer, in cm2/s. An efficient dryer that at room temperature dries gas to a dew point of -61 degrees C is described; the same dryer maintained at 0 degrees C yields a dew point of -80 degrees C and removes water as effectively as Mg(ClO4)2 or a dry ice/acetone slush. The use of Nafion membranes to construct devices capable of delivering gas streams with low but precisely controlled humidities is discussed.

  13. Outer nuclear membrane fusion of adjacent nuclei in varicella-zoster virus-induced syncytia.

    Science.gov (United States)

    Wang, Wei; Yang, Lianwei; Huang, Xiumin; Fu, Wenkun; Pan, Dequan; Cai, Linli; Ye, Jianghui; Liu, Jian; Xia, Ningshao; Cheng, Tong; Zhu, Hua

    2017-12-01

    Syncytia formation has been considered important for cell-to-cell spread and pathogenesis of many viruses. As a syncytium forms, individual nuclei often congregate together, allowing close contact of nuclear membranes and possibly fusion to occur. However, there is currently no reported evidence of nuclear membrane fusion between adjacent nuclei in wild-type virus-induced syncytia. Varicella-zoster virus (VZV) is one typical syncytia-inducing virus that causes chickenpox and shingles in humans. Here, we report, for the first time, an interesting observation of apparent fusion of the outer nuclear membranes from juxtaposed nuclei that comprise VZV syncytia both in ARPE-19 human epithelial cells in vitro and in human skin xenografts in the SCID-hu mouse model in vivo. This work reveals a novel aspect of VZV-related cytopathic effect in the context of multinucleated syncytia. Additionally, the information provided by this study could be helpful for future studies on interactions of viruses with host cell nuclei. Copyright © 2017 Elsevier Inc. All rights reserved.

  14. A molecular model for membrane fusion based on solution studies of an amphiphilic peptide from HIV gp41.

    OpenAIRE

    Fujii, G.; Horvath, S.; Woodward, S.; Eiserling, F.; Eisenberg, D.

    1992-01-01

    The mechanism of protein-mediated membrane fusion and lysis has been investigated by solution-state studies of the effects of peptides on liposomes. A peptide (SI) corresponding to a highly amphiphilic C-terminal segment from the envelope protein (gp41) of the human immunodeficiency virus (HIV) was synthesized and tested for its ability to cause lipid membranes to fuse together (fusion) or to break open (lysis). These effects were compared to those produced by the lytic and fusogenic peptide ...

  15. Semliki Forest virus produced in the absence of the 6K protein has an altered spike structure as revealed by decreased membrane fusion capacity

    NARCIS (Netherlands)

    McInerney, GM; Smit, JM; Liljestrom, P; Wilschut, J

    2004-01-01

    We examined the kinetics of membrane fusion of wild type (wt) and Delta6K mutant Semliki Forest virus in a liposomal model system. The final extent of membrane fusion of the mutant (at pH 5.5) was approximately one third that of the wt virus, although the level of E1 (fusion protein) trimerization

  16. High Efficiency RF Heating for Small Nuclear Fusion Rocket Engines, Phase I

    Data.gov (United States)

    National Aeronautics and Space Administration — High power nuclear fusion propulsion systems will require high efficiency radio-frequency heating systems in the MHz range for plasma heating. This proposal is for a...

  17. Review of Membrane Oxygen Enrichment for Efficient Combustion

    Science.gov (United States)

    Ariono, Danu; Kusuma Wardani, Anita

    2017-07-01

    Oxygen enrichment from air is a simple way of increasing the efficiency of combustion process, as in oxy-combustion. Oxy-combustion has become one of the most attracting combustion technologies because of its potential to address both pollutant reduction and CO2 capture. In oxy-combustion, the fuel and recycled flue gas are combusted with oxygen enriched air (OEA). By using OEA, many benefits can be obtained, such as increasing available heat, improving ignition characteristics, flue gas reduction, increasing productivity, energy efficiency, turndown ratio, and flame stability. Membrane-based gas separation for OEA production becomes an attractive technology over the conventional technology due to the some advantages, including low capital cost, low energy consumption, compact size, and modularity. A single pass through membrane usually can enrich O2 concentration in the air up to 35% and a 50% concentration can be achieved with a double pass of membrane. The use of OEA in the combustion process eliminates the presence of nitrogen in the flue gas. Hence, the flue gas is mainly composed of CO2 and condensable water that can be easily separated. This paper gives an overview of oxy-combustion with membrane technology for oxygen enrichment process. Special attention is given to OEA production and the effect of OEA to the efficiency of combustion.

  18. SNARE-fusion mediated insertion of membrane proteins into native and artificial membranes.

    Science.gov (United States)

    Nordlund, Gustav; Brzezinski, Peter; von Ballmoos, Christoph

    2014-07-02

    Membrane proteins carry out functions such as nutrient uptake, ATP synthesis or transmembrane signal transduction. An increasing number of reports indicate that cellular processes are underpinned by regulated interactions between these proteins. Consequently, functional studies of these networks at a molecular level require co-reconstitution of the interacting components. Here, we report a SNARE protein-based method for incorporation of multiple membrane proteins into artificial membrane vesicles of well-defined composition, and for delivery of large water-soluble substrates into these vesicles. The approach is used for in vitro reconstruction of a fully functional bacterial respiratory chain from purified components. Furthermore, the method is used for functional incorporation of the entire F1F0 ATP synthase complex into native bacterial membranes from which this component had been genetically removed. The novel methodology offers a tool to investigate complex interaction networks between membrane-bound proteins at a molecular level, which is expected to generate functional insights into key cellular functions.

  19. An efficient algorithm for multisensory data fusion under uncertainty condition

    Directory of Open Access Journals (Sweden)

    Mohammad M. Alyannezhadi

    2017-05-01

    Full Text Available This paper addresses multisensory data fusion for unknown systems. The main focus is on identifying and dealing with uncertainty and inconsistent conditions. Most data fusion methods depends on system behavior, which do not allow to easily deal with unknown systems. This method, works based on a clustering technique followed by an MLP predictor. It is specifically designed for unknown systems in uncertain and inconsistent conditions. However, it can also be applied for known and exact sources. When the sources contain uncertainty and inconsistency, data fusion may fail. The proposed method can recognize and remove the inconsistent data points and produces reliable results. The experimental results on both synthetic and real data confirm the effectiveness of the proposed approach.

  20. Spatiotemporal dynamics of membrane remodeling and fusion proteins during endocytic transport.

    Science.gov (United States)

    Arlt, Henning; Auffarth, Kathrin; Kurre, Rainer; Lisse, Dominik; Piehler, Jacob; Ungermann, Christian

    2015-04-01

    Organelles of the endolysosomal system undergo multiple fission and fusion events to combine sorting of selected proteins to the vacuole with endosomal recycling. This sorting requires a consecutive remodeling of the organelle surface in the course of endosomal maturation. Here we dissect the remodeling and fusion machinery on endosomes during the process of endocytosis. We traced selected GFP-tagged endosomal proteins relative to exogenously added fluorescently labeled α-factor on its way from the plasma membrane to the vacuole. Our data reveal that the machinery of endosomal fusion and ESCRT proteins has similar temporal localization on endosomes, whereas they precede the retromer cargo recognition complex. Neither deletion of retromer nor the fusion machinery with the vacuole affects this maturation process, although the kinetics seems to be delayed due to ESCRT deletion. Of importance, in strains lacking the active Rab7-like Ypt7 or the vacuolar SNARE fusion machinery, α-factor still proceeds to late endosomes with the same kinetics. This indicates that endosomal maturation is mainly controlled by the early endosomal fusion and remodeling machinery but not the downstream Rab Ypt7 or the SNARE machinery. Our data thus provide important further understanding of endosomal biogenesis in the context of cargo sorting. © 2015 Arlt et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

  1. Membrane cholesterol regulates lysosome-plasma membrane fusion events and modulates Trypanosoma cruzi invasion of host cells.

    Directory of Open Access Journals (Sweden)

    Bárbara Hissa

    Full Text Available BACKGROUND: Trypomastigotes of Trypanosoma cruzi are able to invade several types of non-phagocytic cells through a lysosomal dependent mechanism. It has been shown that, during invasion, parasites trigger host cell lysosome exocytosis, which initially occurs at the parasite-host contact site. Acid sphingomyelinase released from lysosomes then induces endocytosis and parasite internalization. Lysosomes continue to fuse with the newly formed parasitophorous vacuole until the parasite is completely enclosed by lysosomal membrane, a process indispensable for a stable infection. Previous work has shown that host membrane cholesterol is also important for the T. cruzi invasion process in both professional (macrophages and non-professional (epithelial phagocytic cells. However, the mechanism by which cholesterol-enriched microdomains participate in this process has remained unclear. METHODOLOGY/PRINCIPAL FINDING: In the present work we show that cardiomyocytes treated with MβCD, a drug able to sequester cholesterol from cell membranes, leads to a 50% reduction in invasion by T. cruzi trypomastigotes, as well as a decrease in the number of recently internalized parasites co-localizing with lysosomal markers. Cholesterol depletion from host membranes was accompanied by a decrease in the labeling of host membrane lipid rafts, as well as excessive lysosome exocytic events during the earlier stages of treatment. Precocious lysosomal exocytosis in MβCD treated cells led to a change in lysosomal distribution, with a reduction in the number of these organelles at the cell periphery, and probably compromises the intracellular pool of lysosomes necessary for T. cruzi invasion. CONCLUSION/SIGNIFICANCE: Based on these results, we propose that cholesterol depletion leads to unregulated exocytic events, reducing lysosome availability at the cell cortex and consequently compromise T. cruzi entry into host cells. The results also suggest that two different pools of

  2. Coiled-coils on lipid membranes : a new perspective on membrane fusion

    NARCIS (Netherlands)

    Rabe, Martin

    2014-01-01

    For decades a large amount of research has dealt with membrane interactions of peptides and proteins as well as peptide-peptide interactions to understand the mechanisms of essential biological processes such as protein-driven vesicle budding and fission, cell penetration and lysis by peptides, and

  3. High flux zeolite membranes for efficient production of biofuels

    OpenAIRE

    Sandström, Linda

    2012-01-01

    The greenhouse effect and the limited fossil oil resources have increased the demand for renewable fuels. Zeolite membranes have potential applications in numerous separation processes, and could be useful in the development of efficient processes for renewable fuel production.Synthesis gas is a gas mixture containing mainly carbon monoxide, hydrogen and carbon dioxide. Synthesis gas produced from biomass usually contains more carbon dioxide than desired, and it also contains hydrogen sulphid...

  4. Mechanism of HIV-1 Resistance to an Electronically Constrained α-Helical Peptide Membrane Fusion Inhibitor.

    Science.gov (United States)

    Wu, Xiyuan; Liu, Zixuan; Ding, Xiaohui; Yu, Danwei; Wei, Huamian; Qin, Bo; Zhu, Yuanmei; Chong, Huihui; Cui, Sheng; He, Yuxian

    2018-04-01

    SC29EK is an electronically constrained α-helical peptide HIV-1 fusion inhibitor that is highly effective against both wild-type and enfuvirtide (T20)-resistant viruses. In this study, we focused on investigating the mechanism of HIV-1 resistance to SC29EK by two approaches. First, SC29EK-escaping HIV-1 variants were selected and characterized. Three mutant viruses, which possessed two (N43K/E49A) or three (Q39R/N43K/N126K and N43K/E49A/N126K) amino acid substitutions in the N- and C-terminal repeat regions of gp41 were identified as conferring high resistance to SC29EK and cross-resistance to the first-generation (T20 and C34) and newly designed (sifuvirtide, MT-SC29EK, and 2P23) fusion inhibitors. The resistance mutations could reduce the binding stability of SC29EK, impair viral Env-mediated cell fusion and entry, and change the conformation of the gp41 core structure. Further, we determined the crystal structure of SC29EK in complex with a target mimic peptide, which revealed the critical intra- and interhelical interactions underlying the mode of action of SC29EK and the genetic pathway to HIV-1 resistance. Taken together, the present data provide new insights into the structure and function of gp41 and the structure-activity relationship (SAR) of viral fusion inhibitors. IMPORTANCE T20 is the only membrane fusion inhibitor available for treatment of viral infection, but it has relatively low anti-HIV activity and genetic barriers for resistance, thus calling for new drugs blocking the viral fusion process. As an electronically constrained α-helical peptide, SC29EK is highly potent against both wild-type and T20-resistant HIV-1 strains. Here, we report the characterization of HIV-1 variants resistant to SC29EK and the crystal structure of SC29EK. The key mutations mediating high resistance to SC29EK and cross-resistance to the first and new generations of fusion inhibitors as well as the underlying mechanisms were identified. The crystal structure of SC29EK

  5. Ebola virus glycoprotein needs an additional trigger, beyond proteolytic priming for membrane fusion.

    Directory of Open Access Journals (Sweden)

    Shridhar Bale

    2011-11-01

    Full Text Available Ebolavirus belongs to the family filoviridae and causes severe hemorrhagic fever in humans with 50-90% lethality. Detailed understanding of how the viruses attach to and enter new host cells is critical to development of medical interventions. The virus displays a trimeric glycoprotein (GP(1,2 on its surface that is solely responsible for membrane attachment, virus internalization and fusion. GP(1,2 is expressed as a single peptide and is cleaved by furin in the host cells to yield two disulphide-linked fragments termed GP1 and GP2 that remain associated in a GP(1,2 trimeric, viral surface spike. After entry into host endosomes, GP(1,2 is enzymatically cleaved by endosomal cathepsins B and L, a necessary step in infection. However, the functional effects of the cleavage on the glycoprotein are unknown.We demonstrate by antibody binding and Hydrogen-Deuterium Exchange Mass Spectrometry (DXMS of glycoproteins from two different ebolaviruses that although enzymatic priming of GP(1,2 is required for fusion, the priming itself does not initiate the required conformational changes in the ectodomain of GP(1,2. Further, ELISA binding data of primed GP(1,2 to conformational antibody KZ52 suggests that the low pH inside the endosomes also does not trigger dissociation of GP1 from GP2 to effect membrane fusion.The results reveal that the ebolavirus GP(1,2 ectodomain remains in the prefusion conformation upon enzymatic cleavage in low pH and removal of the glycan cap. The results also suggest that an additional endosomal trigger is necessary to induce the conformational changes in GP(1,2 and effect fusion. Identification of this trigger will provide further mechanistic insights into ebolavirus infection.

  6. Fusion

    International Nuclear Information System (INIS)

    Naraghi, M.

    1976-01-01

    It is proposed that Iran as a world's potential supplier of fossile fuel should participate in fusion research and gain experience in this new field. Fusion, as an ultimate source of energy in future, and the problems concerned with the fusion reactors are reviewed. Furthermore; plasma heating, magnetic and inertial confinement in a fusion reactor are discussed. A brief description of tokamak, theta pinch and magnetic mirror reactors is also included

  7. Inhibition of EBV-mediated membrane fusion by anti-gHgL antibodies

    Energy Technology Data Exchange (ETDEWEB)

    Sathiyamoorthy, Karthik; Jiang, Jiansen; Möhl, Britta S.; Chen, Jia; Zhou, Z. Hong; Longnecker, Richard; Jardetzky, Theodore S. (UCLA); (Stanford-MED); (NWU)

    2017-09-22

    Herpesvirus entry into cells requires the coordinated action of multiple virus envelope glycoproteins, including gH, gL, and gB. For EBV, the gp42 protein assembles into complexes with gHgL heterodimers and binds HLA class II to activate gB-mediated membrane fusion with B cells. EBV tropism is dictated by gp42 levels in the virion, as it inhibits entry into epithelial cells while promoting entry into B cells. The gHgL and gB proteins are targets of neutralizing antibodies and potential candidates for subunit vaccine development, but our understanding of their neutralizing epitopes and the mechanisms of inhibition remain relatively unexplored. Here we studied the structures and mechanisms of two anti-gHgL antibodies, CL40 and CL59, that block membrane fusion with both B cells and epithelial cells. We determined the structures of the CL40 and CL59 complexes with gHgL using X-ray crystallography and EM to identify their epitope locations. CL59 binds to the C-terminal domain IV of gH, while CL40 binds to a site occupied by the gp42 receptor binding domain. CL40 binding to gHgL/gp42 complexes is not blocked by gp42 and does not interfere with gp42 binding to HLA class II, indicating that its ability to block membrane fusion with B cells represents a defect in gB activation. These data indicate that anti-gHgL neutralizing antibodies can block gHgL-mediated activation of gB through different surface epitopes and mechanisms.

  8. Preliminary design of fusion reactor fuel cleanup system by palladium alloy membrane method

    International Nuclear Information System (INIS)

    Yoshida, Hiroshi; Konishi, Satoshi; Naruse, Yuji

    1981-10-01

    A design of palladium diffuser and Fuel Cleanup System (FCU) for D-T fusion reactor is proposed. Feasibility of palladium alloy membrane method is discussed based on the early studies by the authors. Operating conditions of the palladium diffuser are determined experimentally. Dimensions of the diffuser are estimated from computer simulation. FCU system is designed under the feed conditions of Tritium Systems Test Assembly (TSTA) at Los Alamos Scientific Laboratory. The system is composed of Pd-diffusers, catalytic oxidizer, freezer and zink beds, and has some advantages in system layout and operation. This design can readily be extended to other conditions of plasma exhaust gases. (author)

  9. Extended Synaptotagmin 1 Interacts with Herpes Simplex Virus 1 Glycoprotein M and Negatively Modulates Virus-Induced Membrane Fusion.

    Science.gov (United States)

    El Kasmi, Imane; Khadivjam, Bita; Lackman, Miki; Duron, Johanne; Bonneil, Eric; Thibault, Pierre; Lippé, Roger

    2018-01-01

    Enveloped viruses typically encode their own fusion machinery to enter cells. Herpesviruses are unusual, as they fuse with a number of cellular compartments throughout their life cycles. As uncontrolled fusion of the host membranes should be avoided in these events, tight regulation of the viral fusion machinery is critical. While studying herpes simplex virus 1 (HSV-1) glycoprotein gM, we identified the cellular protein E-Syt1 (extended synaptotagmin 1) as an interaction partner. The interaction took place in both infected and transfected cells, suggesting other viral proteins were not required for the interaction. Most interestingly, E-Syt1 is a member of the synaptotagmin family of membrane fusion regulators. However, the protein is known to promote the tethering of the endoplasmic reticulum (ER) to the plasma membrane. We now show that E-Syt1, along with the related E-Syt3, negatively modulates viral release into the extracellular milieu, cell-to-cell viral spread, and viral entry, all processes that implicate membrane fusion events. Similarly, these E-Syt proteins impacted the formation of virus-induced syncytia. Altogether, these findings hint at the modulation of the viral fusion machinery by the E-Syt family of proteins. IMPORTANCE Viruses typically encode their own fusion apparatus to enable them to enter cells. For many viruses, this means a single fusogenic protein. However, herpesviruses are large entities that express several accessory viral proteins to regulate their fusogenic activity. The present study hints at the additional participation of cellular proteins in this process, suggesting the host can also modulate viral fusion to some extent. Hence E-Syt proteins 1 and 3 seem to negatively modulate the different viral fusion events that take place during the HSV-1 life cycle. This could represent yet another innate immunity response to the virus. Copyright © 2017 American Society for Microbiology.

  10. Fusion

    CERN Document Server

    Mahaffey, James A

    2012-01-01

    As energy problems of the world grow, work toward fusion power continues at a greater pace than ever before. The topic of fusion is one that is often met with the most recognition and interest in the nuclear power arena. Written in clear and jargon-free prose, Fusion explores the big bang of creation to the blackout death of worn-out stars. A brief history of fusion research, beginning with the first tentative theories in the early 20th century, is also discussed, as well as the race for fusion power. This brand-new, full-color resource examines the various programs currently being funded or p

  11. Reassessment of the lineage fusion hypothesis for the origin of double membrane bacteria.

    Science.gov (United States)

    Swithers, Kristen S; Fournier, Gregory P; Green, Anna G; Gogarten, J Peter; Lapierre, Pascal

    2011-01-01

    In 2009, James Lake introduced a new hypothesis in which reticulate phylogeny reconstruction is used to elucidate the origin of gram-negative bacteria (Nature 460: 967-971). The presented data supported the gram-negative bacteria originating from an ancient endosymbiosis between the Actinobacteria and Clostridia. His conclusion was based on a presence-absence analysis of protein families that divided all prokaryotes into five groups: Actinobacteria, Double Membrane bacteria (DM), Clostridia, Archaea and Bacilli. Of these five groups, the DM are by far the largest and most diverse group compared to the other groupings. While the fusion hypothesis for the origin of double membrane bacteria is enticing, we show that the signal supporting an ancient symbiosis is lost when the DM group is broken down into smaller subgroups. We conclude that the signal detected in James Lake's analysis in part results from a systematic artifact due to group size and diversity combined with low levels of horizontal gene transfer.

  12. A molecular model for membrane fusion based on solution studies of an amphiphilic peptide from HIV gp41.

    Science.gov (United States)

    Fujii, G; Horvath, S; Woodward, S; Eiserling, F; Eisenberg, D

    1992-11-01

    The mechanism of protein-mediated membrane fusion and lysis has been investigated by solution-state studies of the effects of peptides on liposomes. A peptide (SI) corresponding to a highly amphiphilic C-terminal segment from the envelope protein (gp41) of the human immunodeficiency virus (HIV) was synthesized and tested for its ability to cause lipid membranes to fuse together (fusion) or to break open (lysis). These effects were compared to those produced by the lytic and fusogenic peptide from bee venom, melittin. Other properties studied included the changes in visible absorbance and mean particle size, and the secondary structure of peptides as judged by CD spectroscopy. Taken together, the observations suggest that protein-mediated membrane fusion is dependent not only on hydrophobic and electrostatic forces but also on the spatial arrangement of the amino acid residues to form an amphiphilic structure that promotes the mixing of the lipids between membranes. A speculative molecular model is proposed for membrane fusion by alpha-helical peptides, and its relationship to the forces involved in protein-membrane interactions is discussed.

  13. An insight into fusion technology aiding efficient recombinant protein production for functional proteomics.

    Science.gov (United States)

    Yadav, Dinesh K; Yadav, Neelam; Yadav, Sarika; Haque, Shafiul; Tuteja, Narendra

    2016-12-15

    Advancements in peptide fusion technologies to maximize the protein production has taken a big leap to fulfill the demands of post-genomics era targeting elucidation of structure/function of the proteome and its therapeutic applications, by over-expression in heterologous expression systems. Despite being most preferred protein expression system armed with variety of cardinal fusion tags, expression of the functionally active recombinant protein in E. coli remains plagued. The present review critically analyses the aptness of well-characterized fusion tags utilized for over-expression of recombinant proteins with improved solubility and their compatibility with downstream purification procedures. The combinatorial tandem affinity strategies have shown to provide more versatile options. Solubility decreasing fusion tags have proved to facilitate the overproduction of antimicrobial peptides. Efficient removal of fusion tags prior to final usage is of utmost importance and has been summarized discussing the efficiency of various enzymatic and chemical methods of tag removal. Unfortunately, no single fusion tag works as a magic bullet to completely fulfill the requirements of protein expression and purification in active form. The information provided might help in selection and development of a successful protocol for efficient recombinant protein production for functional proteomics. Copyright © 2016 Elsevier Inc. All rights reserved.

  14. Enhanced human somatic cell reprogramming efficiency by fusion of the MYC transactivation domain and OCT4

    Directory of Open Access Journals (Sweden)

    Ling Wang

    2017-12-01

    Full Text Available The development of human induced pluripotent stem cells (iPSCs holds great promise for regenerative medicine. However the iPSC induction efficiency is still very low and with lengthy reprogramming process. We utilized the highly potent transactivation domain (TAD of MYC protein to engineer the human OCT4 fusion proteins. Applying the MYC-TAD-OCT4 fusion proteins in mouse iPSC generation leads to shorter reprogramming dynamics, with earlier activation of pluripotent markers in reprogrammed cells than wild type OCT4 (wt-OCT4. Dramatic enhancement of iPSC colony induction efficiency and shortened reprogramming dynamics were observed when these MYC-TAD-OCT4 fusion proteins were used to reprogram primary human cells. The OCT4 fusion proteins induced human iPSCs are pluripotent. We further show that the MYC Box I (MBI is dispensable while both MBII and the linking region between MBI/II are essential for the enhanced reprogramming activity of MYC-TAD-OCT4 fusion protein. Consistent with an enhanced transcription activity, the engineered OCT4 significantly stimulated the expression of genes specifically targeted by OCT4-alone, OCT4/SOX2, and OCT4/SOX2/KLF4 during human iPSC induction, compared with the wt-OCT4. The MYC-TAD-OCT4 fusion proteins we generated will be valuable tools for studying the reprogramming mechanisms and for efficient iPSC generation for humans as well as for other species.

  15. Enhancement of the Computational Efficiency of Membrane Computing Models

    National Research Council Canada - National Science Library

    Das, Digendra K

    2007-01-01

    .... Membrane computing consists of cell-like membranes placed inside a unique skin membrane. In regions delimited by a membrane structure, cells are placed in multisets of objects which evolve according to evolution rules associated with the regions...

  16. Performance of a palladium membrane reactor using a Ni catalyst for fusion fuel impurities processing

    International Nuclear Information System (INIS)

    Willm, R.S.; Okuno, K.

    1994-01-01

    The palladium membrane reactor (PMR) provides a means to recover hydrogen isotopes from impurities expected to be present in fusion reactor exhaust. This recovery is based on reactions such as water gas shift and steam reforming for which conversion is equilibrium limited. By including a selectively permeable membrane such as Pd/Ag in the catalyst bed, hydrogen isotopes can be removed from the reacting environment, thus promoting the reaction to complete conversion. Such a device has been built and operated at the Tritium Systems Test Assembly (TSTA) at Los Alamos National Laboratory (LANL). For the reactions listed above, earlier study with this unit has shown that hydrogen single-pass recoveries approaching 100% can be achieved. It was also determined that a nickel catalyst is a feasible choice for use with a PMR appropriate for fusion fuel impurities processing. The purpose of this study is to systematically assess the performance of the PMR using a nickel catalyst. Reactions which will be studied include the water gas shift reaction, steam reforming and methane cracking. Extended operation will be performed to detect performance degradation if it exists. The use of methane in these tests may lead to the formation of coke on the catalyst. Methods of removing the coke such as treatment with carbon dioxide or diluted oxygen will be examined

  17. On the efficiency of conical targets for laser thermonuclear fusion

    International Nuclear Information System (INIS)

    Borovskij, A.V.; Korobkin, V.V.

    1981-01-01

    Advantages and drawbacks of conical targets (CT) for laser fusion (LF) are discussed. Possibility of the laser power reduction, laser pulse lengthening and neutron yield increase are analyzed for an ideal conical target with absolutely rigid and heat-proof walls as compared to a spherical target of the same mass. A simple theory is suggested which makes it possible to take into account an effect of walls on the fusion process in the conical target with gaseous fuel and heavy shell. Energy losses due to wall deformations and heat conduction are estimated. An influence of these effects on the neutron yield is estimated. CT used in the LF experiments are found to have serious drawbacks in comparison with spherical ones. These drawbacks are connected with the effect of walls on the processes taking place in CT. Analysis of CT, for which the effect of walls is not significant, points up some definite advantages of CT as compared with spherical one. These advantages are the possibility of laser pulse lengthening and laser power reduction in comparison with the irradiation of a sphere of an equal mass. These two positive qualities are connected with the fact that CT has large linear dimensions [ru

  18. Cdc42p and Rho1p are sequentially activated and mechanistically linked to vacuole membrane fusion

    International Nuclear Information System (INIS)

    Logan, Michael R.; Jones, Lynden; Eitzen, Gary

    2010-01-01

    Small monomeric GTPases act as molecular switches, regulating many biological functions via activation of membrane localized signaling cascades. Activation of their switch function is controlled by GTP binding and hydrolysis. Two Rho GTPases, Cdc42p and Rho1p, are localized to the yeast vacuole where they regulate membrane fusion. Here, we define a method to directly examine vacuole membrane Cdc42p and Rho1p activation based on their affinity to probes derived from effectors. Cdc42p and Rho1p showed unique temporal activation which aligned with distinct subreactions of in vitro vacuole fusion. Cdc42p was rapidly activated in an ATP-independent manner while Rho1p activation was kinetically slower and required ATP. Inhibitors that are known to block vacuole membrane fusion were examined for their effect on Cdc42p and Rho1p activation. Rdi1p, which inhibits the dissociation of GDP from Rho proteins, blocked both Cdc42p and Rho1p activation. Ligands of PI(4,5)P 2 specifically inhibited Rho1p activation while pre-incubation with U73122, which targets Plc1p function, increased Rho1p activation. These results define unique activation mechanisms for Cdc42p and Rho1p, which may be linked to the vacuole membrane fusion mechanism.

  19. Coiled-coil formation of the membrane-fusion K/E peptides viewed by electron paramagnetic resonance.

    Directory of Open Access Journals (Sweden)

    Pravin Kumar

    Full Text Available The interaction of the complementary K (Ac-(KIAALKE3-GW-NH2 and E (Ac-(EIAALEK3-GY-NH2 peptides, components of the zipper of an artificial membrane fusion system (Robson Marsden H. et al. Angew Chemie Int Ed. 2009 is investigated by electron paramagnetic resonance (EPR. By frozen solution continuous-wave EPR and double electron-electron resonance (DEER, the distance between spin labels attached to the K- and to the E-peptide is measured. Three constructs of spin-labelled K- and E-peptides are used in five combinations for low temperature investigations. The K/E heterodimers are found to be parallel, in agreement with previous studies. Also, K homodimers in parallel orientation were observed, a finding that was not reported before. Comparison to room-temperature, solution EPR shows that the latter method is less specific to detect this peptide-peptide interaction. Combining frozen solution cw-EPR for short distances (1.8 nm to 2.0 nm and DEER for longer distances thus proves versatile to detect the zipper interaction in membrane fusion. As the methodology can be applied to membrane samples, the approach presented suggests itself for in-situ studies of the complete membrane fusion process, opening up new avenues for the study of membrane fusion.

  20. Porous germanene as a highly efficient gas separation membrane.

    Science.gov (United States)

    Bian, Ang; Dai, Yafei; Yang, Jinlong

    2017-11-16

    Using a gas separation membrane as a simple gas separation device has an obvious advantage because of the low energy consumption and pollution-free manufacturing. The first-principles calculations used in this work show that germanene with its divacancy is an excellent material for use as a hydrogen (H 2 ) and helium (He) separation membrane, and that it displays an even better competitive advantage than porous graphene and porous silicene. Porous germanene with its divacancy is chemically inert to gas molecules, because it lacks additional atoms to protect the edged dangling germanium atoms in defects, and thus shows great advantages for gas separation over previously prepared graphene. The energy barriers to H 2 and He penetrating porous germanene are quite low, and the permeabilities to H 2 and He are high. Furthermore, the selectivities of porous germanene for H 2 and He relative to other gas molecules are high, up to 10 31 and 10 27 , respectively, which are superior to those of porous graphene (10 23 ) and porous silicene (10 13 ); thus the separation efficiency of porous germanene is much higher than that of porous graphene and porous silicene. Therefore, germanene is a favorable candidate as a gas separation membrane material. At the same time, the successful synthesis of germanene in the laboratory means that it is possible to use it in real applications.

  1. Dual phase oxygen transport membrane for efficient oxyfuel combustion

    International Nuclear Information System (INIS)

    Ramasamy, Madhumidha

    2016-01-01

    Oxygen transport membranes (OTMs) are attracting great interest for the separation of oxygen from air in an energy efficient way. A variety of solid oxide ceramic materials that possess mixed ionic and electronic conductivity (MIEC) are being investigated for efficient oxygen separation (Betz '10, Skinner '03). Unfortunately these materials do not exhibit high degradation stability under harsh ambient conditions such as flue gas containing CO 2 , SO x , H 2 O and dust, pressure gradients and high temperatures that are typical in fossil fuel power plants. For this reason, dual phase composite membranes are developed to combine the best characteristics of different compounds to achieve high oxygen permeability and sufficient chemical and mechanical stability at elevated temperatures. In this thesis, the dual phase membrane Ce 0.8 Gd 0.2 O 2-δ - FeCo 2 O 4 (CGO-FCO) was developed after systematic investigation of various combinations of ionic and electronic conductors. The phase distribution of the composite was investigated in detail using electron microscopes and this analysis revealed the phase interaction leading to grain boundary rock salt phase and formation of perovskite secondary phase. A systematic study explored the onset of phase interactions to form perovskite phase and the role of this unintended phase as pure electronic conductor was identified. Additionally optimization of conventional sintering process to eliminate spinel phase decomposition into rock salt was identified. An elaborate study on the absolute minimum electronic conductor requirement for efficient percolation network was carried out and its influence on oxygen flux value was measured. Oxygen permeation measurements in the temperature range of 600 C - 1000 C under partial pressure gradient provided by air and argon as feed and sweep gases are used to identify limiting transport processes. The dual phase membranes are much more prone to surface exchange limitations because of the

  2. H1N1 Swine Influenza Viruses Differ from Avian Precursors by a Higher pH Optimum of Membrane Fusion

    Science.gov (United States)

    Baumann, Jan; Kouassi, Nancy Mounogou; Foni, Emanuela; Klenk, Hans-Dieter

    2015-01-01

    ABSTRACT The H1N1 Eurasian avian-like swine (EAsw) influenza viruses originated from an avian H1N1 virus. To characterize potential changes in the membrane fusion activity of the hemagglutinin (HA) during avian-to-swine adaptation of the virus, we studied EAsw viruses isolated in the first years of their circulation in pigs and closely related contemporary H1N1 viruses of wild aquatic birds. Compared to the avian viruses, the swine viruses were less sensitive to neutralization by lysosomotropic agent NH4Cl in MDCK cells, had a higher pH optimum of hemolytic activity, and were less stable at acidic pH. Eight amino acid substitutions in the HA were found to separate the EAsw viruses from their putative avian precursor; four substitutions—T492S, N722D, R752K, and S1132F—were located in the structural regions of the HA2 subunit known to play a role in acid-induced conformational transition of the HA. We also studied low-pH-induced syncytium formation by cell-expressed HA proteins and found that the HAs of the 1918, 1957, 1968, and 2009 pandemic viruses required a lower pH for fusion induction than did the HA of a representative EAsw virus. Our data show that transmission of an avian H1N1 virus to pigs was accompanied by changes in conformational stability and fusion promotion activity of the HA. We conclude that distinctive host-determined fusion characteristics of the HA may represent a barrier for avian-to-swine and swine-to-human transmission of influenza viruses. IMPORTANCE Continuing cases of human infections with zoonotic influenza viruses highlight the necessity to understand which viral properties contribute to interspecies transmission. Efficient binding of the HA to cellular receptors in a new host species is known to be essential for the transmission. Less is known about required adaptive changes in the membrane fusion activity of the HA. Here we show that adaptation of an avian influenza virus to pigs in Europe in 1980s was accompanied by mutations in

  3. Recombinant fusion proteins TAT-Mu, Mu and Mu-Mu mediate efficient non-viral gene delivery.

    Science.gov (United States)

    Rajagopalan, Rukkumani; Xavier, Jennifer; Rangaraj, Nandini; Rao, Nalam Madhusudhana; Gopal, Vijaya

    2007-04-01

    The inherent ability of certain peptides or proteins of viral, prokaryotic and eukaryotic origin to bind DNA was used to generate novel peptide-based DNA delivery protocols. We have developed a recombinant approach to make fusion proteins with motifs for DNA-binding ability, Mu and membrane transduction domains, TAT, and tested them for their DNA-binding, uptake and transfection efficiencies. In one of the constructs, the recombinant plasmid was designed to encode the Mu moiety of sequence MRRAHHRRRRASHRRMRGG in-frame with TAT of sequence YGRKKRRQRRR to generate TAT-Mu, while the other two constructs, Mu and Mu-Mu, harbor a single copy or two copies of the Mu moiety. Recombinant his-tag fusion proteins TAT-Mu, Mu and Mu-Mu were purified by overexpression of plasmid constructs using cobalt-based affinity resins. The peptides were characterized for their size and interaction with DNA, complexed with plasmid pCMVbeta-gal, and shown to transfect MCF-7, COS and CHOK-1 cells efficiently. Recombinant fusion proteins TAT-Mu, Mu and Mu-Mu were cloned and overexpressed in BL21(DE3)pLysS with greater than 95% purity. The molecular weight of TAT-Mu was determined by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) to be 11.34 kDa while those of Mu and Mu-Mu were 7.78 and 9.83 kDa, respectively. Live uptake analysis of TAT-Mu, Mu and Mu-Mu as DP (DNA+peptide) or DPL (DNA+peptide+lipid) complexes into MCF-7 cells, followed by immunostaining and laser scanning confocal microscopy, demonstrated that the complexes are internalized very efficiently and localized in the nucleus. DNA:peptide complexes (DP) transfect MCF-7, COS and CHOK-1 cells. The addition of cationic liposomes enhances the uptake of the ternary complexes (DPL) further and also brings about 3-7-fold enhancement in reporter gene expression compared to DP alone. Recombinant proteins that are heterologous fusions, having DNA-binding domains and nuclear localization epitopes

  4. A soluble form of Epstein-Barr virus gH/gL inhibits EBV-induced membrane fusion and does not function in fusion

    Energy Technology Data Exchange (ETDEWEB)

    Rowe, Cynthia L. [Department of Microbiology and Immunology, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611 (United States); Connolly, Sarah A. [Department of Health Sciences, DePaul University, Chicago, IL 60614 (United States); Chen, Jia [Department of Microbiology and Immunology, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611 (United States); Jardetzky, Theodore S. [Department of Structural Biology, Stanford University School of Medicine, 371 Serra Mall, Stanford, CA 94305 (United States); Longnecker, Richard, E-mail: r-longnecker@northwestern.edu [Department of Microbiology and Immunology, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611 (United States)

    2013-02-05

    We investigated whether soluble EBV gH/gL (sgH/gL) functions in fusion and made a series of truncations of gH/gL domains based on the gH/gL crystal structure. We found sgH/gL failed to mediate cell-cell fusion both when co-expressed with the other entry glycoproteins and when added exogenously to fusion assays. Interestingly, sgH/gL inhibited cell-cell fusion in a dose dependent manner when co-expressed. sgH/gL from HSV was unable to inhibit EBV fusion, suggesting the inhibition was specific to EBV gH/gL. sgH/gL stably binds gp42, but not gB nor gH/gL. The domain mutants, DI/gL, DI-II/gL and DI-II-III/gL were unable to bind gp42. Instead, DI-II/gL, DI-II-III/gL and sgH/gL but not DI/gL decreased the expression of gp42, resulting in decreased overall fusion. Overall, our results suggest that domain IV may be required for proper folding and the transmembrane domain and cytoplasmic tail of EBV gH/gL are required for the most efficient fusion.

  5. Early Events in Chikungunya Virus Infection—From Virus CellBinding to Membrane Fusion

    Directory of Open Access Journals (Sweden)

    Mareike K. S. van Duijl-Richter

    2015-07-01

    Full Text Available Chikungunya virus (CHIKV is a rapidly emerging mosquito-borne alphavirus causing millions of infections in the tropical and subtropical regions of the world. CHIKV infection often leads to an acute self-limited febrile illness with debilitating myalgia and arthralgia. A potential long-term complication of CHIKV infection is severe joint pain, which can last for months to years. There are no vaccines or specific therapeutics available to prevent or treat infection. This review describes the critical steps in CHIKV cell entry. We summarize the latest studies on the virus-cell tropism, virus-receptor binding, internalization, membrane fusion and review the molecules and compounds that have been described to interfere with virus cell entry. The aim of the review is to give the reader a state-of-the-art overview on CHIKV cell entry and to provide an outlook on potential new avenues in CHIKV research.

  6. Cysteines and N-Glycosylation Sites Conserved among All Alphaherpesviruses Regulate Membrane Fusion in Herpes Simplex Virus 1 Infection.

    Science.gov (United States)

    Rider, Paul J F; Naderi, Misagh; Bergeron, Scott; Chouljenko, Vladimir N; Brylinski, Michal; Kousoulas, Konstantin G

    2017-11-01

    Neurotropism is a defining characteristic of alphaherpesvirus pathogenicity. Glycoprotein K (gK) is a conserved virion glycoprotein of all alphaherpesviruses that is not found in other herpesvirus subfamilies. The extracellular amino terminus of gK has been shown to be important to the ability of the prototypic alphaherpesvirus herpes simplex virus 1 (HSV-1) to enter neurons via axonal termini. Here, we determined the role of the two conserved N-linked glycosylation (N48 and N58) sites of gK in virus-induced cell fusion and replication. We found that N-linked glycosylation is important to the regulation of HSV-1-induced membrane fusion since mutating N58 to alanine (N58A) caused extensive virus-induced cell fusion. Due to the known contributions of N-linked glycosylation to protein processing and correct disulfide bond formation, we investigated whether the conserved extracellular cysteine residues within the amino terminus of gK contributed to the regulation of HSV-1-induced membrane fusion. We found that mutation of C37 and C114 residues led to a gK-null phenotype characterized by very small plaque formation and drastic reduction in infectious virus production, while mutation of C82 and C243 caused extensive virus-induced cell fusion. Comparison of N-linked glycosylation and cysteine mutant replication kinetics identified disparate effects on infectious virion egress from infected cells. Specifically, cysteine mutations caused defects in the accumulation of infectious virus in both the cellular and supernatant fractions, while glycosylation site mutants did not adversely affect virion egress from infected cells. These results demonstrate a critical role for the N glycosylation sites and cysteines for the structure and function of the amino terminus of gK. IMPORTANCE We have previously identified important entry and neurotropic determinants in the amino terminus of HSV-1 glycoprotein K (gK). Alphaherpesvirus-mediated membrane fusion is a complex and highly

  7. Fusion between perinuclear virions and the outer nuclear membrane requires the fusogenic activity of herpes simplex virus gB.

    Science.gov (United States)

    Wright, Catherine C; Wisner, Todd W; Hannah, Brian P; Eisenberg, Roselyn J; Cohen, Gary H; Johnson, David C

    2009-11-01

    Herpesviruses cross nuclear membranes (NMs) in two steps, as follows: (i) capsids assemble and bud through the inner NM into the perinuclear space, producing enveloped virus particles, and (ii) the envelopes of these virus particles fuse with the outer NM. Two herpes simplex virus (HSV) glycoproteins, gB and gH (the latter, likely complexed as a heterodimer with gL), are necessary for the second step of this process. Mutants lacking both gB and gH accumulate in the perinuclear space or in herniations (membrane vesicles derived from the inner NM). Both gB and gH/gL are also known to act directly in fusing the virion envelope with host cell membranes during HSV entry into cells, i.e., both glycoproteins appear to function directly in different aspects of the membrane fusion process. We hypothesized that HSV gB and gH/gL also act directly in the membrane fusion that occurs during virus egress from the nucleus. Previous studies of the role of gB and gH/gL in nuclear egress involved HSV gB and gH null mutants that could potentially also possess gross defects in the virion envelope. Here, we produced recombinant HSV-expressing mutant forms of gB with single amino acid substitutions in the hydrophobic "fusion loops." These fusion loops are thought to play a direct role in membrane fusion by insertion into cellular membranes. HSV recombinants expressing gB with any one of four fusion loop mutations (W174R, W174Y, Y179K, and A261D) were unable to enter cells. Moreover, two of the mutants, W174Y and Y179K, displayed reduced abilities to mediate HSV cell-to-cell spread, and W174R and A261D exhibited no spread. All mutant viruses exhibited defects in nuclear egress, enveloped virions accumulated in herniations and in the perinuclear space, and fewer enveloped virions were detected on cell surfaces. These results support the hypothesis that gB functions directly to mediate the fusion between perinuclear virus particles and the outer NM.

  8. Different sets of ER-resident J-proteins regulate distinct polar nuclear-membrane fusion events in Arabidopsis thaliana.

    Science.gov (United States)

    Maruyama, Daisuke; Yamamoto, Masaya; Endo, Toshiya; Nishikawa, Shuh-ichi

    2014-11-01

    Angiosperm female gametophytes contain a central cell with two polar nuclei. In many species, including Arabidopsis thaliana, the polar nuclei fuse during female gametogenesis. We previously showed that BiP, an Hsp70 in the endoplasmic reticulum (ER), was essential for membrane fusion during female gametogenesis. Hsp70 function requires partner proteins for full activity. J-domain containing proteins (J-proteins) are the major Hsp70 functional partners. A. thaliana ER contains three soluble J-proteins, AtERdj3A, AtERdj3B, and AtP58(IPK). Here, we analyzed mutants of these proteins and determined that double-mutant ovules lacking AtP58(IPK) and AtERdj3A or AtERdj3B were defective in polar nuclear fusion. Electron microscopy analysis identified that polar nuclei were in close contact, but no membrane fusion occurred in mutant ovules lacking AtP58(IPK) and AtERdj3A. The polar nuclear outer membrane appeared to be connected via the ER remaining at the inner unfused membrane in mutant ovules lacking AtP58(IPK) and AtERdj3B. These results indicate that ER-resident J-proteins, AtP58(IPK)/AtERdj3A and AtP58(IPK)/AtERdj3B, function at distinct steps of polar nuclear-membrane fusion. Similar to the bip1 bip2 double mutant female gametophytes, the aterdj3a atp58(ipk) double mutant female gametophytes defective in fusion of the outer polar nuclear membrane displayed aberrant endosperm proliferation after fertilization with wild-type pollen. However, endosperm proliferated normally after fertilization of the aterdj3b atp58(ipk) double mutant female gametophytes defective in fusion of the inner membrane. Our results indicate that the polar nuclear fusion defect itself does not cause an endosperm proliferation defect. © The Author 2014. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.

  9. Fusion

    Science.gov (United States)

    Herman, Robin

    1990-10-01

    The book abounds with fascinating anecdotes about fusion's rocky path: the spurious claim by Argentine dictator Juan Peron in 1951 that his country had built a working fusion reactor, the rush by the United States to drop secrecy and publicize its fusion work as a propaganda offensive after the Russian success with Sputnik; the fortune Penthouse magazine publisher Bob Guccione sank into an unconventional fusion device, the skepticism that met an assertion by two University of Utah chemists in 1989 that they had created "cold fusion" in a bottle. Aimed at a general audience, the book describes the scientific basis of controlled fusion--the fusing of atomic nuclei, under conditions hotter than the sun, to release energy. Using personal recollections of scientists involved, it traces the history of this little-known international race that began during the Cold War in secret laboratories in the United States, Great Britain and the Soviet Union, and evolved into an astonishingly open collaboration between East and West.

  10. The efficiency of a membrane bioreactor in drinking water denitrification

    Directory of Open Access Journals (Sweden)

    Petrovič Aleksandra

    2015-01-01

    Full Text Available The membrane bioreactor (MBR system was investigated regarding its nitrate removal capacity from drinking water. The performance of a pilot-scale MBR was tested, depending on the operational parameters, using sucrose as a carbon source. Drinking water from the source was introduced into the reactor in order to study the influence of flow-rate on the nitrate removal and denitrification efficiency of drinking water. The content of the nitrate was around 70 mg/L and the C/N ratio was 3:1. Nitrate removal efficiencies above 90% were obtained by flow-rates lower than 4.8 L/h. The specific denitrification rates varied between 0.02 and 0.16 g/L NO3/ (g/L MLSS•d. The efficiencies and nitrate removal were noticeably affected by the flow-rate and hydraulic retention times. At the maximum flow-rate of 10.2 L/h still 68% of the nitrate had been removed, whilst the highest specific denitrification rate was achieved at 0.2738 g/L NO3/ (g/L MLSS•d. The maximum reactor removal capacity was calculated at 8.75 g NO3/m3•h.

  11. Using Haloarcula marismortui bacteriorhodopsin as a fusion tag for enhancing and visible expression of integral membrane proteins in Escherichia coli.

    Directory of Open Access Journals (Sweden)

    Min-Feng Hsu

    Full Text Available Membrane proteins are key targets for pharmacological intervention because of their vital functions. Structural and functional studies of membrane proteins have been severely hampered because of the difficulties in producing sufficient quantities of properly folded and biologically active proteins. Here we generate a high-level expression system of integral membrane proteins in Escherichia coli by using a mutated bacteriorhodopsin (BR from Haloarcula marismortui (HmBRI/D94N as a fusion partner. A purification strategy was designed by incorporating a His-tag on the target membrane protein for affinity purification and an appropriate protease cleavage site to generate the final products. The fusion system can be used to detect the intended target membrane proteins during overexpression and purification either with the naked eye or by directly monitoring their characteristic optical absorption. In this study, we applied this approach to produce two functional integral membrane proteins, undecaprenyl pyrophosphate phosphatase and carnitine/butyrobetaine antiporter with significant yield enhancement. This technology could facilitate the development of a high-throughput strategy to screen for conditions that improve the yield of correctly folded target membrane proteins. Other robust BRs can also be incorporated in this system.

  12. Reassessment of the lineage fusion hypothesis for the origin of double membrane bacteria.

    Directory of Open Access Journals (Sweden)

    Kristen S Swithers

    Full Text Available In 2009, James Lake introduced a new hypothesis in which reticulate phylogeny reconstruction is used to elucidate the origin of gram-negative bacteria (Nature 460: 967-971. The presented data supported the gram-negative bacteria originating from an ancient endosymbiosis between the Actinobacteria and Clostridia. His conclusion was based on a presence-absence analysis of protein families that divided all prokaryotes into five groups: Actinobacteria, Double Membrane bacteria (DM, Clostridia, Archaea and Bacilli. Of these five groups, the DM are by far the largest and most diverse group compared to the other groupings. While the fusion hypothesis for the origin of double membrane bacteria is enticing, we show that the signal supporting an ancient symbiosis is lost when the DM group is broken down into smaller subgroups. We conclude that the signal detected in James Lake's analysis in part results from a systematic artifact due to group size and diversity combined with low levels of horizontal gene transfer.

  13. Direct targeting of membrane fusion by SNARE mimicry: Convergent evolution of Legionella effectors.

    Science.gov (United States)

    Shi, Xingqi; Halder, Partho; Yavuz, Halenur; Jahn, Reinhard; Shuman, Howard A

    2016-08-02

    Legionella pneumophila, the Gram-negative pathogen causing Legionnaires' disease, infects host cells by hijacking endocytic pathways and forming a Legionella-containing vacuole (LCV) in which the bacteria replicate. To promote LCV expansion and prevent lysosomal targeting, effector proteins are translocated into the host cell where they alter membrane traffic. Here we show that three of these effectors [LegC2 (Legionella eukaryotic-like gene C2)/YlfB (yeast lethal factor B), LegC3, and LegC7/YlfA] functionally mimic glutamine (Q)-SNARE proteins. In infected cells, the three proteins selectively form complexes with the endosomal arginine (R)-SNARE vesicle-associated membrane protein 4 (VAMP4). When reconstituted in proteoliposomes, these proteins avidly fuse with liposomes containing VAMP4, resulting in a stable complex with properties resembling canonical SNARE complexes. Intriguingly, however, the LegC/SNARE hybrid complex cannot be disassembled by N-ethylmaleimide-sensitive factor. We conclude that LegCs use SNARE mimicry to divert VAMP4-containing vesicles for fusion with the LCV, thus promoting its expansion. In addition, the LegC/VAMP4 complex avoids the host's disassembly machinery, thus effectively trapping VAMP4 in an inactive state.

  14. Fusion of local and global features for efficient object detection

    Science.gov (United States)

    Le, Duy-Dinh; Satoh, Shin'ichi

    2005-02-01

    In this paper, we propose a two-stage method for efficient object detection that combines full advantages of AdaBoost and SVM to achieve a reasonable balance in both training and classification speed. In the first stage, we use Haar wavelet features and AdaBoost to train a cascade of classifiers for quick and efficient rejection. This cascade of classifiers consists of simple-to-complex classifiers that allow adapting to complexity of input patterns, rejects almost 90%-99% non-object patterns rapidly. Hard patterns, object-like patterns, which are passed through the first stage, will be classified by the second stage which uses a non linear SVM-based classifier with pixel-based global features. The nonlinear SVM classifier is robust enough in order to reach high performance. We have investigated our proposed method to detect different kinds of objects such as face and facial features like eye and mouth regions. In training, our system is roughly 25 times faster than the system trained by AdaBoost. In running, the experimental results show 1,000 times faster than SVM based method and slightly slower than AdaBoost based method with a comparable accuracy.

  15. A simple, flexible and efficient PCR-fusion/Gateway cloning procedure for gene fusion, site-directed mutagenesis, short sequence insertion and domain deletions and swaps

    Directory of Open Access Journals (Sweden)

    Etchells J Peter

    2009-10-01

    Full Text Available Abstract Background The progress and completion of various plant genome sequencing projects has paved the way for diverse functional genomic studies that involve cloning, modification and subsequent expression of target genes. This requires flexible and efficient procedures for generating binary vectors containing: gene fusions, variants from site-directed mutagenesis, addition of protein tags together with domain swaps and deletions. Furthermore, efficient cloning procedures, ideally high throughput, are essential for pyramiding of multiple gene constructs. Results Here, we present a simple, flexible and efficient PCR-fusion/Gateway cloning procedure for construction of binary vectors for a range of gene fusions or variants with single or multiple nucleotide substitutions, short sequence insertions, domain deletions and swaps. Results from selected applications of the procedure which include ORF fusion, introduction of Cys>Ser mutations, insertion of StrepII tag sequence and domain swaps for Arabidopsis secondary cell wall AtCesA genes are demonstrated. Conclusion The PCR-fusion/Gateway cloning procedure described provides an elegant, simple and efficient solution for a wide range of diverse and complicated cloning tasks. Through streamlined cloning of sets of gene fusions and modification variants into binary vectors for systematic functional studies of gene families, our method allows for efficient utilization of the growing sequence and expression data.

  16. Fusion proteins and select lipids cooperate as membrane receptors for the soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) Vam7p.

    Science.gov (United States)

    Karunakaran, Vidya; Wickner, William

    2013-10-04

    Vam7p, the vacuolar soluble Qc-SNARE, is essential for yeast vacuole fusion. The large tethering complex, homotypic fusion and vacuole protein sorting complex (HOPS), and phosphoinositides, which interact with the Vam7p PX domain, have each been proposed to serve as its membrane receptors. Studies with the isolated organelle cannot determine whether these receptor elements suffice and whether ligands or mutations act directly or indirectly on Vam7p binding to the membrane. Using pure components that are active in reconstituted vacuolar fusion, we now find that Vam7p binds to membranes through its combined affinities for several vacuolar membrane constituents: HOPS, phosphatidylinositol 3-phosphate, SNAREs, and acidic phospholipids. Acidic lipids allow low concentrations of Vam7p to suffice for fusion; without acidic lipids, the block to fusion is partially bypassed by high concentrations of Vam7p.

  17. High efficiency targets for high gain inertial confinement fusion

    International Nuclear Information System (INIS)

    Gardner, J.H.; Bodner, S.E.

    1986-01-01

    Rocket efficiencies as high as 15% are possible using short wavelength lasers and moderately high aspect ratio pellet designs. These designs are made possible by two recent breakthroughs in physics constraints. First is the development of the Induced Spatial Incoherence (ISI) technique which allows uniform illumination of the pellet and relaxes the constraint of thermal smoothing, permitting the use of short wavelength laser light. Second is the discovery that the Rayleigh-Taylor growth rate is considerably reduced at the short laser wavelengths. By taking advantage of the reduced constraints imposed by nonuniform laser illumination and Rayleigh-Taylor instability, pellets using 1/4 micron laser light and initial aspect ratios of about 10 (with in flight aspect ratios of about 150 to 200) may produce energy gains as high as 200 to 250

  18. Crystal Structure of the Membrane Fusion Protein CusB from Escherichia coli

    Energy Technology Data Exchange (ETDEWEB)

    Su, Chih-Chia; Yang, Feng; Long, Feng; Reyon, Deepak; Routh, Mathew D.; Kuo, Dennis W.; Mokhtari, Adam K.; Van Ornam, Jonathan D.; Rabe, Katherine L.; Hoy, Julie A.; Lee, Young Jin; Rajashankar, Kanagalaghatta R.; Yu, Edward W.; (Cornell); (Iowa State)

    2010-03-29

    Gram-negative bacteria, such as Escherichia coli, frequently utilize tripartite efflux complexes belonging to the resistance-nodulation-division family to expel diverse toxic compounds from the cell. These systems contain a periplasmic membrane fusion protein (MFP) that is critical for substrate transport. We here present the x-ray structures of the CusB MFP from the copper/silver efflux system of E. coli. This is the first structure of any MFPs associated with heavy-metal efflux transporters. CusB bridges the inner-membrane efflux pump CusA and outer-membrane channel CusC to mediate resistance to Cu{sup +} and Ag{sup +} ions. Two distinct structures of the elongated molecules of CusB were found in the asymmetric unit of a single crystal, which suggests the flexible nature of this protein. Each protomer of CusB can be divided into four different domains, whereby the first three domains are mostly {beta}-strands and the last domain adopts an entirely helical architecture. Unlike other known structures of MFPs, the {alpha}-helical domain of CusB is folded into a three-helix bundle. This three-helix bundle presumably interacts with the periplasmic domain of CusC. The N- and C-termini of CusB form the first {beta}-strand domain, which is found to interact with the periplasmic domain of the CusA efflux pump. Atomic details of how this efflux protein binds Cu{sup +} and Ag{sup +} were revealed by the crystals of the CusB-Cu(I) and CusB-Ag(I) complexes. The structures indicate that CusB consists of multiple binding sites for these metal ions. These findings reveal novel structural features of an MFP in the resistance-nodulation-division efflux system and provide direct evidence that this protein specifically interacts with transported substrates.

  19. Tunable C2N Membrane for High Efficient Water Desalination

    Science.gov (United States)

    Yang, Yanmei; Li, Weifeng; Zhou, Hongcai; Zhang, Xiaoming; Zhao, Mingwen

    2016-01-01

    Water scarcity represents one of the most serious global problems of our time and challenges the advancements in desalination techniques. Although water-filtering architectures based on graphene have greatly advanced the approach to high performance desalination membranes, the controlled-generation of nanopores with particular diameter is tricky and has stunted its wide applications. Here, through molecular dynamic simulations and first-principles calculations, we propose that the recently reported graphene-like carbon nitride (g-C2N) monolayer can serve as high efficient filters for water desalination. Taking the advantages of the intrisic nanoporous structure and excellent mechanical properties of g-C2N, high water transparency and strong salt filtering capability have been demonstrated in our simulations. More importantly, the “open” and “closed” states of the g-C2N filter can be precisely regulated by tensile strain. It is found that the water permeability of g-C2N is significantly higher than that reported for graphene filters by almost one order of magnitude. In the light of the abundant family of graphene-like carbon nitride monolayered materials, our results thus offer a promising approach to the design of high efficient filteration architectures. PMID:27384666

  20. Hirudin as a novel fusion tag for efficient production of lunasin in Escherichia coli.

    Science.gov (United States)

    Tian, Qinghua; Zhang, Ping; Gao, Zhan; Li, Hengli; Bai, Zhengli; Tan, Shuhua

    2017-07-03

    Fusion expression provides an effective means for the biosynthesis of longer peptides in Escherichia coli. However, the commonly used fusion tags are primarily suitable for laboratory scale applications due to the high cost of commercial affinity resins. Herein, a novel approach exploiting hirudin as a multipurpose fusion tag in combination with tobacco etch virus (TEV) protease cleavage has been developed for the efficient and cost-effective production of a 43-amino acid model peptide lunasin in E. coli at preparative scale. A fusion gene which allows for lunasin to be N-terminally fused to the C-terminus of hirudin through a flexible linker comprising a TEV protease cleavage site was designed and cloned in a secretion vector pTASH. By cultivation in a 7-L bioreactor, the fusion protein was excreted into the culture medium at a high yield of ~380 mg/L, which was conveniently recovered and purified by inexpensive HP20 hydrophobic chromatography at a recovery rate of ~80%. After polishing and cleavage with TEV protease, the finally purified lunasin was obtained with ≥95% purity and yield of ~86 mg/L culture medium. Conclusively, this hirudin tagging strategy is powerful in the production of lunasin and could be applicable for the production of other peptides at preparative scale.

  1. Effects of thermal efficiency in DCMD and the preparation of membranes with low thermal conductivity

    Science.gov (United States)

    Li, Zhehao; Peng, Yuelian; Dong, Yajun; Fan, Hongwei; Chen, Ping; Qiu, Lin; Jiang, Qi

    2014-10-01

    The effects of the membrane characteristics and operational conditions on the vapor flux and thermal efficiency in a direct contact membrane distillation (DCMD) process were studied with a mathematical simulation. The membrane temperature, driving force of vapor transfer, membrane distillation coefficient, etc. were used to analyze the effects. The operating conditions that increased the vapor flux improved the thermal efficiency. The membrane characteristics of four microporous membranes and their performances in DCMD were compared. A polysulfone (PSf) membrane prepared via vapor-induced phase separation exhibited the lowest thermal conductivity. The PSf and polyvinylidene difluoride (PVDF) membranes were modified using SiO2 aerogel blending and coating to reduce the thermal conductivity of the membrane. The coating process was more effective than the blending process toward this end. The changes in the structure of the modified membrane were observed with a scanning electron microscope. Si was found on the modified membrane surface with an energy spectrometer. The PVDF composite and support membranes were tested during the DCMD process; the composite membrane had a higher vapor flux and a better thermal efficiency than the support. A new method based on a 3ω technique was used to measure the thermal conductivity of the membranes.

  2. Computationally efficient visual–inertial sensor fusion for Global Positioning System–denied navigation on a small quadrotor

    Directory of Open Access Journals (Sweden)

    Chang Liu

    2016-03-01

    Full Text Available Because of the complementary nature of visual and inertial sensors, the combination of both is able to provide fast and accurate 6 degree-of-freedom state estimation, which is the fundamental requirement for robotic (especially, unmanned aerial vehicle navigation tasks in Global Positioning System–denied environments. This article presents a computationally efficient visual–inertial fusion algorithm, by separating orientation fusion from the position fusion process. The algorithm is designed to perform 6 degree-of-freedom state estimation, based on a gyroscope, an accelerometer and a monocular visual-based simultaneous localisation and mapping algorithm measurement. It also recovers the visual scale for the monocular visual-based simultaneous localisation and mapping. In particular, the fusion algorithm treats the orientation fusion and position fusion as two separate processes, where the orientation fusion is based on a very efficient gradient descent algorithm, whereas the position fusion is based on a 13-state linear Kalman filter. The elimination of the magnetometer sensor avoids the problem of magnetic distortion, which makes it a power-on-and-go system once the accelerometer is factory calibrated. The resulting algorithm shows a significant computational reduction over the conventional extended Kalman filter, with competitive accuracy. Moreover, the separation between orientation and position fusion processes enables the algorithm to be easily implemented onto two individual hardware elements and thus allows the two fusion processes to be executed concurrently.

  3. MITO-Porter: A liposome-based carrier system for delivery of macromolecules into mitochondria via membrane fusion.

    Science.gov (United States)

    Yamada, Yuma; Akita, Hidetaka; Kamiya, Hiroyuki; Kogure, Kentaro; Yamamoto, Takenori; Shinohara, Yasuo; Yamashita, Kikuji; Kobayashi, Hideo; Kikuchi, Hiroshi; Harashima, Hideyoshi

    2008-02-01

    Mitochondria are the principal producers of energy in higher cells. Mitochondrial dysfunction is implicated in a variety of human diseases, including cancer and neurodegenerative disorders. Effective medical therapies for such diseases will ultimately require targeted delivery of therapeutic proteins or nucleic acids to the mitochondria, which will be achieved through innovations in the nanotechnology of intracellular trafficking. Here we describe a liposome-based carrier that delivers its macromolecular cargo to the mitochondrial interior via membrane fusion. These liposome particles, which we call MITO-Porters, carry octaarginine surface modifications to stimulate their entry into cells as intact vesicles (via macropinocytosis). We identified lipid compositions for the MITO-Porter which promote both its fusion with the mitochondrial membrane and the release of its cargo to the intra-mitochondrial compartment in living cells. Thus, the MITO-Porter holds promise as an efficacious system for the delivery of both large and small therapeutic molecules into mitochondria.

  4. Anti-tumor effects of gene therapy with GALV membrane fusion glycoprotein in lung adenocarcinoma.

    Science.gov (United States)

    Zhu, Bing; Yang, Jian-ru; Fu, Xin-ping; Jiang, Yue-quan

    2014-07-01

    This study examined the efficacy of gene therapy of lung adenocarcinoma using specifically controlled type I herpes simplex virus recombinant vector expressing Gibbon ape leukemia virus membrane fusion glycoprotein gene (GALV.fus). Recombinant HSV-I plasmid carrying target transgene was constructed, and recombinant viral vector was generated in Vero cells using Lipofectamine transfection. Viral vector was introduced into lung adenocarcinoma A549 cells or human fetal fibroblast HFL-I GNHu 5 cells, or inoculated into human lung adenocarcinoma xenografts in nude mice. The anti-tumor and cytotoxic effects of GALV-FMG, the transgene, were examined in these cell and animal models. Expression of GALV-FMG in xenographs achieved 100 % tumorigenicity. Recombinant HSV-I viral vector also exhibited significant tumor cell killing effect in vitro. Relative survival rates of tumor cells treated with GALV-FMG or control vectors were, respectively, 20 and 70 %. GALV.fus has a potent anti-tumor effect against lung cancer both in vitro and in vivo. This anti-tumor potential provides foundation for further studies with this vector.

  5. Convenient synthesis and application of versatile nucleic acid lipid membrane anchors in the assembly and fusion of liposomes

    DEFF Research Database (Denmark)

    Ries, Oliver; Löffler, Philipp M. G.; Vogel, Stefan

    2015-01-01

    or the construction of DNA origami structures. We herein present the synthesis and applications of versatile lipid membrane anchor building blocks suitable for solid phase oligonucleotide synthesis. These are readily synthesized in bulk in five to seven steps from commercially available precursors and can...... be incorporated at any position within an oligonucleotide without significantly altering duplex stability and structure as proven by thermal denaturation experiments and circular dichroism. Furthermore, applicability could be demonstrated by assembly and fusion of liposomes mediated by lipid...

  6. Molecular dynamics simulations of T-2410 and T-2429 HIV fusion inhibitors interacting with model membranes: Insight into peptide behavior, structure and dynamics.

    Science.gov (United States)

    Mavioso, I C V C; de Andrade, V C R; Palace Carvalho, A J; Martins do Canto, A M T

    2017-09-01

    T-2410 and T-2429 are HIV fusion inhibitor peptides (FI) designed to present a higher efficiency even against HIV strains that developed resistance against other FIs. Similar peptides were shown to interact with model membranes both in the liquid disordered phase and in the liquid ordered state. Those results indicated that such interaction is important to function and could be correlated with their effectiveness. Extensive molecular dynamics simulations were carried out to investigate the interactions between both T-2410 and T-2429 with bilayers of pure 1-palmitoyl-2-oleoyl-phosphatidylcholine (POPC) and a mixture of POPC/cholesterol (Chol) (1:1). It was observed that both peptides interact strongly with both membrane systems, especially with the POPC/Chol systems, where these peptides show the highest number of H-bonds observed so far. T-2410 and T-2429 showed higher extent of interaction with bilayers when compared to T-20 or T-1249 in previous studies. This is most notable in POPC/Chol membranes where, although able to form H-bonds with Chol, they do so to a lesser extent than T-1249 does, the latter being the only FI peptide so far that was observed to form H-bonds with Chol. This behavior suggests that interaction of FI peptides with rigid Chol rich membranes may not be as dependent from peptide/Chol H-bond formation as previous results of T-1249 behavior led to believe. As in other similar peptides, the higher ability to interact with membranes shown by T-2410 and T2429 is probably correlated with its higher inhibitory efficiency. Copyright © 2017 Elsevier B.V. All rights reserved.

  7. Vesicular PtdIns(3,4,5)P3 and Rab7 are key effectors of sea urchin zygote nuclear membrane fusion.

    Science.gov (United States)

    Lete, Marta G; Byrne, Richard D; Alonso, Alicia; Poccia, Dominic; Larijani, Banafshé

    2017-01-15

    Regulation of nuclear envelope dynamics is an important example of the universal phenomena of membrane fusion. The signalling molecules involved in nuclear membrane fusion might also be conserved during the formation of both pronuclear and zygote nuclear envelopes in the fertilised egg. Here, we determine that class-I phosphoinositide 3-kinases (PI3Ks) are needed for in vitro nuclear envelope formation. We show that, in vivo, PtdIns(3,4,5)P 3 is transiently located in vesicles around the male pronucleus at the time of nuclear envelope formation, and around male and female pronuclei before membrane fusion. We illustrate that class-I PI3K activity is also necessary for fusion of the female and male pronuclear membranes. We demonstrate, using coincidence amplified Förster resonance energy transfer (FRET) monitored using fluorescence lifetime imaging microscopy (FLIM), a protein-lipid interaction of Rab7 GTPase and PtdIns(3,4,5)P 3 that occurs during pronuclear membrane fusion to create the zygote nuclear envelope. We present a working model, which includes several molecular steps in the pathways controlling fusion of nuclear envelope membranes. © 2017. Published by The Company of Biologists Ltd.

  8. Rapid Biogas Production by Compact Multi-Layer Membrane Bioreactor: Efficiency of Synthetic Polymeric Membranes

    Directory of Open Access Journals (Sweden)

    Supansa Youngsukkasem

    2013-11-01

    Full Text Available Entrapment of methane-producing microorganisms between semi-permeable synthetic membranes in a multi-layer membrane bioreactor (MMBR was studied and compared to the digestion capacity of a free-cell digester, using a hydraulic retention time of one day and organic loading rates (OLR of 3.08, 6.16, and 8.16 g COD/L·day. The reactor was designed to retain bacterial cells with uprising plug flow through a narrow tunnel between membrane layers, in order to acquire maximal mass transfer in a compact bioreactor. Membranes of hydrophobic polyamide 46 (PA and hydroxyethylated polyamide 46 (HPA as well as a commercial membrane of polyvinylidene fluoride (PVDF were examined. While the bacteria in the free-cell digester were washed out, the membrane bioreactor succeeded in retaining them. Cross-flow of the liquid through the membrane surface and diffusion of the substrate through the membranes, using no extra driving force, allowed the bacteria to receive nutrients and to produce biogas. However, the choice of membrane type was crucial. Synthesized hydrophobic PA membrane was not effective for this purpose, producing 50–121 mL biogas/day, while developed HPA membrane and the reference PVDF were able to transfer the nutrients and metabolites while retaining the cells, producing 1102–1633 and 1016–1960 mL biogas/day, respectively.

  9. Coiled-coil formation on lipid bilayers--implications for docking and fusion efficiency.

    Science.gov (United States)

    Pähler, Gesa; Panse, Cornelia; Diederichsen, Ulf; Janshoff, Andreas

    2012-12-05

    Coiled-coil formation of four different oligopeptides was characterized in solution, on hydrogels, and on membranes by employing circular dichroism spectroscopy, surface plasmon resonance spectroscopy, attenuated total reflection infrared spectroscopy, and ellipsometry. Peptide sequences rich in either glutamic acid (E: E3Cys, i-E3Cys) or lysine (K: K3Cys, i-K3Cys) were used to represent minimal mimics of eukaryotic SNARE motifs. Half of the peptides were synthesized in reverse sequence, so that parallel and antiparallel heptad coiled-coil structures were formed. Either E-peptides or K-peptides were attached covalently to phospholipid anchors via maleimide chemistry, and served as receptors for the recognition of the corresponding binding partners added to solution. Attenuated total reflection infrared spectroscopy of single bilayers confirmed the formation of coiled-coil complexes at the membrane interface. Coiled-coil formation in solution, as compared with association at the membrane surface, displays considerably larger binding constants that are largely attributed to loss of translational entropy at the interface. Finally, the fusogenicity of the various coiled-coil motifs was explored, and the results provide clear evidence that hemifusion followed by full fusion requires a parallel orientation of α-helices, whereas antiparallel oriented coiled-coil motifs display only docking. Copyright © 2012 Biophysical Society. Published by Elsevier Inc. All rights reserved.

  10. Coiled-Coil Formation on Lipid Bilayers—Implications for Docking and Fusion Efficiency

    Science.gov (United States)

    Pähler, Gesa; Panse, Cornelia; Diederichsen, Ulf; Janshoff, Andreas

    2012-01-01

    Coiled-coil formation of four different oligopeptides was characterized in solution, on hydrogels, and on membranes by employing circular dichroism spectroscopy, surface plasmon resonance spectroscopy, attenuated total reflection infrared spectroscopy, and ellipsometry. Peptide sequences rich in either glutamic acid (E: E3Cys, i-E3Cys) or lysine (K: K3Cys, i-K3Cys) were used to represent minimal mimics of eukaryotic SNARE motifs. Half of the peptides were synthesized in reverse sequence, so that parallel and antiparallel heptad coiled-coil structures were formed. Either E-peptides or K-peptides were attached covalently to phospholipid anchors via maleimide chemistry, and served as receptors for the recognition of the corresponding binding partners added to solution. Attenuated total reflection infrared spectroscopy of single bilayers confirmed the formation of coiled-coil complexes at the membrane interface. Coiled-coil formation in solution, as compared with association at the membrane surface, displays considerably larger binding constants that are largely attributed to loss of translational entropy at the interface. Finally, the fusogenicity of the various coiled-coil motifs was explored, and the results provide clear evidence that hemifusion followed by full fusion requires a parallel orientation of α-helices, whereas antiparallel oriented coiled-coil motifs display only docking. PMID:23283228

  11. Multivalent interactions between streptavidin-based pretargeting fusion proteins and cell receptors impede efficient internalization of biotinylated nanoparticles.

    Science.gov (United States)

    Parker, Christina L; Yang, Qi; Yang, Bing; McCallen, Justin D; Park, Steven I; Lai, Samuel K

    2017-11-01

    Pretargeting represents a promising strategy to enhance delivery of nanoparticles. The strategy involves first introducing bispecific antibodies or fusion proteins (BFP) that can bind specific epitopes on target cells with one arm, and use the other arm to capture subsequently administered effector molecules, such as radionuclides or drug-loaded nanoparticles. Nevertheless, it remains unclear whether BFP that bind slowly- or non-internalizing epitopes on target cells can facilitate efficient intracellular delivery. Here, we investigated the cellular uptake of biotin-functionalized nanoparticles with streptavidin-scFv against TAG-72, a membrane protein on Jurkat T-cell leukemia cells. Unlike conventional active-targeted nanoparticles, we found that pretargeting resulted in preferential retention of ∼100nm nanoparticles at the plasma membrane rather than internalization into cells. We found no improvement in nanoparticle internalization by simply reducing nanoparticle concentration or surface biotin density. Interestingly, by adding both the BFP and a monoclonal antibody against TAG-72, we observed a twofold improvement in internalization of pretargeted nanoparticles. Our work illustrates that the cellular fate of pretargeted nanoparticles can be controlled by carefully tuning the interactions between pretargeting molecules and nanoparticles on the cell surface. Pretargeting is a multi-step strategy that utilizes bispecific proteins that recognize both cellular epitopes and subsequently administered therapeutic molecules. This approach has been extensively studied for radiotherapy of blood cancers; however, pretargeting remains largely underexplored for nanoparticle targeting, including whether pretargeting can facilitate efficient intracellular delivery. Here, we found that high density of targeting proteins on the cell surface can effectively limit internalization of pretargeted nanoparticles. Our work underscores the need to carefully assess specific cell

  12. Novel Membrane for Highly Efficient Fuel Cells, Phase I

    Data.gov (United States)

    National Aeronautics and Space Administration — Proton Exchange Membrane (PEM) fuel cells and electrolyzers are key technologies for NASA space systems utilizing hydrogen, oxygen, or water as reactants. In order...

  13. Fast and efficient protein purification using membrane adsorber systems.

    Science.gov (United States)

    Suck, Kirstin; Walter, Johanna; Menzel, Frauke; Tappe, Alexander; Kasper, Cornelia; Naumann, Claudia; Zeidler, Robert; Scheper, Thomas

    2006-02-10

    The purification of proteins from complex cell culture samples is an essential step in proteomic research. Traditional chromatographic methods often require several steps resulting in time consuming and costly procedures. In contrast, protein purification via membrane adsorbers offers the advantage of fast and gentle but still effective isolation. In this work, we present a new method for purification of proteins from crude cell extracts via membrane adsorber based devices. This isolation procedure utilises the membranes favourable pore structure allowing high flow rates without causing high back pressure. Therefore, shear stress to fragile structures is avoided. In addition, mass transfer takes place through convection rather than diffusion, thus allowing very rapid separation processes. Based on this membrane adsorber technology the separation of two model proteins, human serum albumin (HSA) and immungluboline G (IgG) is shown. The isolation of human growth hormone (hGH) from chinese hamster ovary (CHO) cell culture supernatant was performed using a cation exchange membrane. The isolation of the enzyme penicillin acylase from the crude Escherichia coli supernatant was achieved using an anion exchange spin column within one step at a considerable purity. In summary, the membrane adsorber devices have proven to be suitable tools for the purification of proteins from different complex cell culture samples.

  14. Determination of the topology of endoplasmic reticulum membrane proteins using redox-sensitive green-fluorescence protein fusions.

    Science.gov (United States)

    Tsachaki, Maria; Birk, Julia; Egert, Aurélie; Odermatt, Alex

    2015-07-01

    Membrane proteins of the endoplasmic reticulum (ER) are involved in a wide array of essential cellular functions. Identification of the topology of membrane proteins can provide significant insight into their mechanisms of action and biological roles. This is particularly important for membrane enzymes, since their topology determines the subcellular site where a biochemical reaction takes place and the dependence on luminal or cytosolic co-factor pools and substrates. The methods currently available for the determination of topology of proteins are rather laborious and require post-lysis or post-fixation manipulation of cells. In this work, we have developed a simple method for defining intracellular localization and topology of ER membrane proteins in living cells, based on the fusion of the respective protein with redox-sensitive green-fluorescent protein (roGFP). We validated the method and demonstrated that roGFP fusion proteins constitute a reliable tool for the study of ER membrane protein topology, using as control microsomal 11β-hydroxysteroid dehydrogenase (11β-HSD) proteins whose topology has been resolved, and comparing with an independent approach. We then implemented this method to determine the membrane topology of six microsomal members of the 17β-hydroxysteroid dehydrogenase (17β-HSD) family. The results revealed a luminal orientation of the catalytic site for three enzymes, i.e. 17β-HSD6, 7 and 12. Knowledge of the intracellular location of the catalytic site of these enzymes will enable future studies on their biological functions and on the role of the luminal co-factor pool. Copyright © 2015 Elsevier B.V. All rights reserved.

  15. The Structure of Herpesvirus Fusion Glycoprotein B-Bilayer Complex Reveals the Protein-Membrane and Lateral Protein-Protein Interaction

    NARCIS (Netherlands)

    Maurer, Ulrike E.; Zeev-Ben-Mordehai, Tzviya; Pandurangan, Arun Prasad; Cairns, Tina M.; Hannah, Brian P.; Whitbeck, J. Charles; Eisenberg, Roselyn J.; Cohen, Gary H.; Topf, Maya; Huiskonen, Juha T.; Gruenewald, Kay

    2013-01-01

    Glycoprotein B (gB) is a key component of the complex herpesvirus fusion machinery. We studied membrane interaction of two gB ectodomain forms and present an electron cryotomography structure of the gB-bilayer complex. The two forms differed in presence or absence of the membrane proximal region

  16. Molecular dynamics analysis of conformational change of paramyxovirus F protein during the initial steps of membrane fusion

    International Nuclear Information System (INIS)

    Martín-García, Fernando; Mendieta-Moreno, Jesús Ignacio; Mendieta, Jesús; Gómez-Puertas, Paulino

    2012-01-01

    Highlights: ► Initial conformational change of paramyxovirus F protein is caused only by mechanical forces. ► HRA region undergoes a structural change from a beta + alpha conformation to an extended coil and then to an all-alpha conformation. ► HRS domains of F protein form three single α-helices prior to generation of the coiled coil. -- Abstract: The fusion of paramyxovirus to the cell membrane is mediated by fusion protein (F protein) present in the virus envelope, which undergoes a dramatic conformational change during the process. Unlike hemagglutinin in orthomyxovirus, this change is not mediated by an alteration of environmental pH, and its cause remains unknown. Steered molecular dynamics analysis leads us to suggest that the conformational modification is mediated only by stretching mechanical forces once the transmembrane fusion peptide of the protein is anchored to the cell membrane. Such elongating forces will generate major secondary structure rearrangement in the heptad repeat A region of the F protein; from β-sheet conformation to an elongated coil and then spontaneously to an α-helix. In addition, it is proposed that the heptad repeat A region adopts a final three-helix coiled coil and that this structure appears after the formation of individual helices in each monomer.

  17. K-alpha conversion efficiency measurments for x-ray scattering in inertial confinement fusion plasmas

    Energy Technology Data Exchange (ETDEWEB)

    Kritcher, A L; Neumayer, P; Urry, M K; Robey, H; Niemann, C; Landen, O L; Morse, E; Glenzer, S H

    2006-11-21

    The conversion efficiency of ultra short-pulse laser radiation to K-{alpha} x-rays has been measured for various chlorine-containing targets to be used as x-ray scattering probes of dense plasmas. The spectral and temporal properties of these sources will allow spectrally-resolved x-ray scattering probing with picosecond temporal resolution required for measuring the plasma conditions in inertial confinement fusion experiments. Simulations of x-ray scattering spectra from these plasmas show that fuel capsule density, capsule ablator density, and shock timing information may be inferred.

  18. Conservation of proteo-lipid nuclear membrane fusion machinery during early embryogenesis.

    Science.gov (United States)

    Byrne, Richard D; Veeriah, Selvaraju; Applebee, Christopher J; Larijani, Banafshé

    2014-01-01

    The fusogenic lipid diacylglycerol is essential for remodeling gamete and zygote nuclear envelopes (NE) during early embryogenesis. It is unclear whether upstream signaling molecules are likewise conserved. Here we demonstrate PLCγ and its activator SFK1, which co-operate during male pronuclear envelope formation, also promote the subsequent male and female pronuclear fusion. PLCγ and SFK1 interact directly at the fusion site leading to PLCγ activation. This is accompanied by a spatially restricted reduction of PtdIns(4,5)P2. Consequently, pronuclear fusion is blocked by PLCγ or SFK1 inhibition. These findings identify new regulators of events in the early embryo and suggest a conserved "toolkit" of fusion machinery drives successive NE fusion events during embryogenesis.

  19. Targeting of a chimeric human histone fusion mRNA to membrane-bound polysomes in HeLa cells

    International Nuclear Information System (INIS)

    Zambetti, G.; Stein, J.; Stein, G.

    1987-01-01

    The subcellular location of histone mRNA-containing polysomes may play a key role in the posttranscriptional events that mediate histone mRNA turnover following inhibition of DNA synthesis. Previously, it has been shown that histone mRNA is found primarily on free polysomes that are associated with the cytoskeleton. The authors report here the construction of an Escherichia coli pBR322 β-lactamase signal peptide-human H3 histone fusion gene. The fusion transcript is targeted to membrane-bound polysomes and remains stable following interruption of DNA replication. Relocating mRNA within the cell may provide a procedure for studying the posttranscriptional regulation of gene expression

  20. Highly efficient retrograde gene transfer into motor neurons by a lentiviral vector pseudotyped with fusion glycoprotein.

    Directory of Open Access Journals (Sweden)

    Miyabi Hirano

    Full Text Available The development of gene therapy techniques to introduce transgenes that promote neuronal survival and protection provides effective therapeutic approaches for neurological and neurodegenerative diseases. Intramuscular injection of adenoviral and adeno-associated viral vectors, as well as lentiviral vectors pseudotyped with rabies virus glycoprotein (RV-G, permits gene delivery into motor neurons in animal models for motor neuron diseases. Recently, we developed a vector with highly efficient retrograde gene transfer (HiRet by pseudotyping a human immunodeficiency virus type 1 (HIV-1-based vector with fusion glycoprotein B type (FuG-B or a variant of FuG-B (FuG-B2, in which the cytoplasmic domain of RV-G was replaced by the corresponding part of vesicular stomatitis virus glycoprotein (VSV-G. We have also developed another vector showing neuron-specific retrograde gene transfer (NeuRet with fusion glycoprotein C type, in which the short C-terminal segment of the extracellular domain and transmembrane/cytoplasmic domains of RV-G was substituted with the corresponding regions of VSV-G. These two vectors afford the high efficiency of retrograde gene transfer into different neuronal populations in the brain. Here we investigated the efficiency of the HiRet (with FuG-B2 and NeuRet vectors for retrograde gene transfer into motor neurons in the spinal cord and hindbrain in mice after intramuscular injection and compared it with the efficiency of the RV-G pseudotype of the HIV-1-based vector. The main highlight of our results is that the HiRet vector shows the most efficient retrograde gene transfer into both spinal cord and hindbrain motor neurons, offering its promising use as a gene therapeutic approach for the treatment of motor neuron diseases.

  1. Mechanics of post-fusion exocytotic vesicle

    Science.gov (United States)

    Stephens, Thomas; Wu, Zhanghan; Liu, Jian

    2017-06-01

    Exocytosis is an important cellular process controlled by metabolic signaling. It involves vesicle fusion to the plasma membrane, followed by the opening of a fusion pore, and the subsequent release of the vesicular lumen content into the extracellular space. While most modeling efforts focus on the events leading to membrane fusion, how the vesicular membrane remodels after fusing to plasma membrane remains unclear. This latter event dictates the nature and the efficiency of exocytotic vesicular secretions, and is thus critical for exocytotic function. We provide a generic membrane mechanical model to systematically study the fate of post-fusion vesicles. We show that while membrane stiffness favors full-collapse vesicle fusion into the plasma membrane, the intravesicular pressure swells the vesicle and causes the fusion pore to shrink. Dimensions of the vesicle and its associated fusion pore further modulate this mechanical antagonism. We systematically define the mechanical conditions that account for the full spectrum of the observed vesicular secretion modes. Our model therefore can serve as a unified theoretical framework that sheds light on the elaborate control mechanism of exocytosis.

  2. Incorporation of VSV-G produces fusogenic plasma membrane vesicles capable of efficient transfer of bioactive macromolecules and mitochondria.

    Science.gov (United States)

    Lin, Hao-Peng; Zheng, De-Jin; Li, Yun-Pan; Wang, Na; Chen, Shao-Jun; Fu, Yu-Cai; Xu, Wen-Can; Wei, Chi-Ju

    2016-06-01

    generate fusogenic plasma membrane vesicles (fPMVs), which contain bioactive molecules and organelles but not the nucleus. fPMVs can be endocytosed by target cells, while the cargo is released due to low-pH induced membrane fusion. These nucleus-free fPMVs are efficient at delivery of cytoplasmic proteins and mitochondria, leading to recovery of mitochondrial biogenesis and proliferative ability in mitochondria-deficient cells.

  3. Efficiency expectant management in women with premature rupture of membranes

    OpenAIRE

    ISMAILOVA SAVRINISА SULTANOVNA

    2016-01-01

    The aim of this study is to evaluate the effectiveness of the monitoring of pregnant women with preterm rupture of membranes, including the determination of the level of white blood cells, ESR in the blood, assessment of vaginal flora and the presence of elements of amniotic fluid in vaginal discharge (every 12 hours), thermometry (every 3 hours). Also assessed the condition of the fetus: Doppler and cardiotocography utero-placental and fetal blood flow.

  4. Enhancement of the Computational Efficiency of Membrane Computing Models

    Science.gov (United States)

    2007-04-01

    Information Retrieval, SPIRE 2000, La Coruna, Spain, 64-74. [9] S.N. Krishna, R. Rama: P Systems with Replicated Rewriting. Journal of Automata Languages...Paraschiv. Membrane software. A P system simulator. Fundamental Informaticae , 49(13):61-66, 2002. [25] P. Fitzgibbons, D. Das, and T. Renz, “Bio...systems with worm-objects, IEEE 7th International Conference on String Processing and Information Retrieval, SPIRE, La Coruna, Spain, 2000, pp. 64

  5. Efficient3He/4He separation in a nanoporous graphenylene membrane.

    Science.gov (United States)

    Qu, Yuanyuan; Li, Feng; Zhao, Mingwen

    2017-08-16

    Helium-3 is a precious noble gas, which is essential in many advanced technologies such as cryogenics, isotope labeling and nuclear weapons. The current imbalance of 3 He demand and supply shortage leads to the search for an efficient membrane with high performance for 3 He separation. In this study, based on first-principles calculations, we demonstrated that highly efficient 3 He harvesting can be achieved in a nanoporous graphenylene membrane with industrially-acceptable selectivity and permeance. The quantum tunneling effect leads to 3 He harvesting with high efficiency via kinetic sieving. Both the quantum tunneling effect and zero-point energy (ZPE) determine the 3 He/ 4 He separation via thermally-driven equilibrium sieving, where the ZPE effect dominates efficient 3 He/ 4 He separation between two reservoirs. The quantum effects revealed in this work suggest that the nanoporous graphenylene membrane is promising for efficient 3 He harvesting that can be exploited for industrial applications.

  6. Fusion of blastomeres in mouse embryos under the action of femtosecond laser radiation. Efficiency of blastocyst formation and embryo development

    Energy Technology Data Exchange (ETDEWEB)

    Osychenko, A A; Zalesskii, A D; Krivokharchenko, A S; Zhakhbazyan, A K; Nadtochenko, V A [N N Semenov Institute of Chemical Physics, Russian Academy of Sciences, Moscow (Russian Federation); Ryabova, A V [A M Prokhorov General Physics Institute, Russian Academy of Sciences, Moscow (Russian Federation)

    2015-05-31

    Using the method of femtosecond laser surgery we study the fusion of two-cell mouse embryos under the action of tightly focused femtosecond laser radiation with the fusion efficiency reaching 60%. The detailed statistical analysis of the efficiency of blastomere fusion and development of the embryo up to the blastocyst stage after exposure of the embryos from different mice to a femtosecond pulse is presented. It is shown that the efficiency of blastocyst formation essentially depends on the biological characteristics of the embryo, namely, the strain and age of the donor mouse. The possibility of obtaining hexaploid embryonal cells using the methods of femtosecond laser surgery is demonstrated. (extreme light fields and their applications)

  7. Hot Fusion: An Efficient Method to Clone Multiple DNA Fragments as Well as Inverted Repeats without Ligase

    Science.gov (United States)

    Fu, Changlin; Donovan, William P.; Shikapwashya-Hasser, Olga; Ye, Xudong; Cole, Robert H.

    2014-01-01

    Molecular cloning is utilized in nearly every facet of biological and medical research. We have developed a method, termed Hot Fusion, to efficiently clone one or multiple DNA fragments into plasmid vectors without the use of ligase. The method is directional, produces seamless junctions and is not dependent on the availability of restriction sites for inserts. Fragments are assembled based on shared homology regions of 17–30 bp at the junctions, which greatly simplifies the construct design. Hot Fusion is carried out in a one-step, single tube reaction at 50°C for one hour followed by cooling to room temperature. In addition to its utility for multi-fragment assembly Hot Fusion provides a highly efficient method for cloning DNA fragments containing inverted repeats for applications such as RNAi. The overall cloning efficiency is in the order of 90–95%. PMID:25551825

  8. Bioinspired Bifunctional Membrane for Efficient Clean Water Generation.

    Science.gov (United States)

    Liu, Yang; Lou, Jinwei; Ni, Mengtian; Song, Chengyi; Wu, Jianbo; Dasgupta, Neil P; Tao, Peng; Shang, Wen; Deng, Tao

    2016-01-13

    Solving the problems of water pollution and water shortage is an urgent need for the sustainable development of modern society. Different approaches, including distillation, filtration, and photocatalytic degradation, have been developed for the purification of contaminated water and the generation of clean water. In this study, we explored a new approach that uses solar light for both water purification and clean water generation. A bifunctional membrane consisting of a top layer of TiO2 nanoparticles (NPs), a middle layer of Au NPs, and a bottom layer of anodized aluminum oxide (AAO) was designed and fabricated through multiple filtration processes. Such a design enables both TiO2 NP-based photocatalytic function and Au NP-based solar-driven plasmonic evaporation. With the integration of these two functions into a single membrane, both the purification of contaminated water through photocatalytic degradation and the generation of clean water through evaporation were demonstrated using simulated solar illumination. Such a demonstration should also help open up a new strategy for maximizing solar energy conversion and utilization.

  9. Efficient One-Step Fusion PCR Based on Dual-Asymmetric Primers and Two-Step Annealing

    DEFF Research Database (Denmark)

    Liu, Yilan; Chen, Jinjin; Thygesen, Anders

    2018-01-01

    Gene splicing by fusion PCR is a versatile and widely used methodology, especially in synthetic biology. We here describe a rapid method for splicing two fragments by one-round fusion PCR with a dual-asymmetric primers and two-step annealing (ODT) method. During the process, the asymmetric...... intermediate fragments were generated in the early stage. Thereafter, they were hybridized in the subsequent cycles to serve as template for the target full-length product. The process parameters such as primer ratio, elongation temperature and cycle numbers were optimized. In addition, the fusion products...... produced with this method were successfully applied in seamless genome editing. The fusion of two fragments by this method takes less than 0.5 day. The method is expected to facilitate various kinds of complex genetic engineering projects with enhanced efficiency....

  10. Efficient One-Step Fusion PCR Based on Dual-Asymmetric Primers and Two-Step Annealing.

    Science.gov (United States)

    Liu, Yilan; Chen, Jinjin; Thygesen, Anders

    2018-02-01

    Gene splicing by fusion PCR is a versatile and widely used methodology, especially in synthetic biology. We here describe a rapid method for splicing two fragments by one-round fusion PCR with a dual-asymmetric primers and two-step annealing (ODT) method. During the process, the asymmetric intermediate fragments were generated in the early stage. Thereafter, they were hybridized in the subsequent cycles to serve as template for the target full-length product. The process parameters such as primer ratio, elongation temperature and cycle numbers were optimized. In addition, the fusion products produced with this method were successfully applied in seamless genome editing. The fusion of two fragments by this method takes less than 0.5 day. The method is expected to facilitate various kinds of complex genetic engineering projects with enhanced efficiency.

  11. NSF- and SNARE-mediated membrane fusion is required for nuclear envelope formation and completion of nuclear pore complex assembly in Xenopus laevis egg extracts.

    Science.gov (United States)

    Baur, Tina; Ramadan, Kristijan; Schlundt, Andreas; Kartenbeck, Jürgen; Meyer, Hemmo H

    2007-08-15

    Despite the progress in understanding nuclear envelope (NE) reformation after mitosis, it has remained unclear what drives the required membrane fusion and how exactly this is coordinated with nuclear pore complex (NPC) assembly. Here, we show that, like other intracellular fusion reactions, NE fusion in Xenopus laevis egg extracts is mediated by SNARE proteins that require activation by NSF. Antibodies against Xenopus NSF, depletion of NSF or the dominant-negative NSF(E329Q) variant specifically inhibited NE formation. Staging experiments further revealed that NSF was required until sealing of the envelope was completed. Moreover, excess exogenous alpha-SNAP that blocks SNARE function prevented membrane fusion and caused accumulation of non-flattened vesicles on the chromatin surface. Under these conditions, the nucleoporins Nup107 and gp210 were fully recruited, whereas assembly of FxFG-repeat-containing nucleoporins was blocked. Together, we define NSF- and SNARE-mediated membrane fusion events as essential steps during NE formation downstream of Nup107 recruitment, and upstream of membrane flattening and completion of NPC assembly.

  12. Efficient purification of arsenic-contaminated water using amyloid-carbon hybrid membranes.

    Science.gov (United States)

    Bolisetty, Sreenath; Reinhold, Noemi; Zeder, Christophe; Orozco, Monica N; Mezzenga, Raffaele

    2017-05-23

    We show the purification of arsenic-contaminated water using amyloid fibril-based membranes, which adsorb both the arsenate (+5) and arsenite (+3) oxidation forms at efficiencies of ∼99%. Binding isotherms indicate that amyloid fibrils possess multiple binding residues capable of strongly adsorbing arsenic ions via metal-ligand interactions, delaying the saturation of the membrane. We also show that these membranes can be reused for several cycles without any efficiency drop, and validate our technology in purifying real contaminated ground water by removing arsenic with an efficiency as high as 99.6%. These results make this technology promising for inexpensive, efficient and low-energy removal of arsenic from contaminated water.

  13. Submerged anaerobic membrane bioreactor (SAnMBR) performance on sewage treatment: removal efficiencies, biogas production and membrane fouling.

    Science.gov (United States)

    Chen, Rong; Nie, Yulun; Ji, Jiayuan; Utashiro, Tetsuya; Li, Qian; Komori, Daisuke; Li, Yu-You

    2017-09-01

    A submerged anaerobic membrane reactor (SAnMBR) was employed for comprehensive evaluation of sewage treatment at 25 °C and its performance in removal efficiency, biogas production and membrane fouling. Average 89% methanogenic degradation efficiency as well as 90%, 94% and 96% removal of total chemical oxygen demand (TCOD), biochemical oxygen demand (BOD) and nonionic surfactant were obtained, while nitrogen and phosphorus were only subjected to small removals. Results suggest that SAnMBRs can effectively decouple organic degradation and nutrients disposal, and reserve all the nitrogen and phosphorus in the effluent for further possible recovery. Small biomass yields of 0.11 g mixed liquor volatile suspended solids (MLVSS)/gCOD were achieved, coupled to excellent methane production efficiencies of 0.338 NLCH 4 /gCOD, making SAnMBR an attractive technology characterized by low excess sludge production and high bioenergy recovery. Batch tests revealed the SAnMBR appeared to have the potential to bear a high food-to-microorganism ratio (F/M) of 1.54 gCOD/gMLVSS without any inhibition effect, and maximum methane production rate occurred at F/M 0.7 gCOD/gMLVSS. Pore blocking dominated the membrane fouling behaviour at a relative long hydraulic retention time (HRT), i.e. >12 hours, while cake layer dominated significantly at shorter HRTs, i.e. <8 hours.

  14. Membrane fusion activity of Semliki forest virus in a liposomal model system : Specific inhibition by Zn2+ ions

    NARCIS (Netherlands)

    Corver, J; Snippe, H; Kraaijeveld, C; Wilschut, J

    1997-01-01

    Semliki Forest virus (SFV) has been shown previously to fuse efficiently with cholesterol-and sphingolipid-containing liposomal model membranes in a low-pH-dependent manner. Several steps can be distinguished in this process, including low-pH-induced irreversible binding of the virus to the

  15. Membrane pumping technology, helium and hydrogen isotopes separation in the fusion hydrogen

    International Nuclear Information System (INIS)

    Pigarov, A.Yu.; Pistunovich, V.I.; Busnyuk, A.O.

    1994-01-01

    A gas pumping system for the ITER, improved by implementation of superpermeable membranes for selective hydrogen isotope exhaust, is considered. The study of the pumping capability of a niobium membrane for a hydrogen-helium mixture has been fulfilled. The membrane superpermeability can be only realized for atomic hydrogen. Helium does not pass through the membrane, and its presence does not affect the hydrogen pumping. A detailed Monte Carlo simulation of gas behavior for the experimental facility has been done. The probability of permeation for a hydrogen atom for one collision with the membrane is ∼0.1; the same probability of molecule permeation is ∼10 -5 . The probability for atomization, i.e. re-emission of an atomizer is ∼0.2; the probability of recombination of an atom is ∼0.2

  16. Ceramic Membrane Enabling Technology for Improved IGCC Efficiency

    Energy Technology Data Exchange (ETDEWEB)

    John Sirman; Bart vanHassel

    2005-06-01

    This final report summarizes work accomplished in the program from October 1, 1999 through December 31,2004. While many of the key technical objectives for this program were achieved, after a thorough economic and OTM (Oxygen Transport Membrane) reliability analysis were completed, a decision was made to terminate the project prior to construction of a second pilot reactor. In the program, oxygen with purity greater than 99% was produced in both single tube tests and multi-tube pilot plant tests for over 1000 hours. This demonstrated the technical viability of using ceramic OTM devices for producing oxygen from a high pressure air stream. The oxygen fluxes that were achieved in single tube tests exceeded the original target flux for commercial operation. However, extended testing showed that the mean time to failure of the ceramics was insufficient to enable a commercially viable system. In addition, manufacturing and material strength constraints led to size limitations of the OTM tubes that could be tested. This has a severe impact on the cost of both the ceramic devices, but also the cost of assembling the OTM tubes in a large reactor. As such and combined with significant progress in cost reduction of large cryogenic oxygen separation devices, an economic gain that justifies continued development could not be derived.

  17. Investigations and calculations toward increasing the efficiency of muon catalyzed fusion

    International Nuclear Information System (INIS)

    Monkhorst, H.J.

    1989-11-01

    A brief summary of results during this report period is given. Some of the topics investigated includes: (1) calculations of sticking fractions and d-t fusion from dtμ(JV) states, (2) ddμ sticking fractions, (3) the reactivation coefficient in d-t fusion, (4) fusion rates for all XYμ(JV)(JV=0,1), (5) nuclear effects on energy shifts and fusion rates for (J=O) states of dtμ, (6) and some comments on cold fusion

  18. Gβγ directly modulates vesicle fusion by competing with synaptotagmin for binding to neuronal SNARE proteins embedded in membranes.

    Science.gov (United States)

    Zurawski, Zack; Page, Brian; Chicka, Michael C; Brindley, Rebecca L; Wells, Christopher A; Preininger, Anita M; Hyde, Karren; Gilbert, James A; Cruz-Rodriguez, Osvaldo; Currie, Kevin P M; Chapman, Edwin R; Alford, Simon; Hamm, Heidi E

    2017-07-21

    G i/o -coupled G protein-coupled receptors can inhibit neurotransmitter release at synapses via multiple mechanisms. In addition to Gβγ-mediated modulation of voltage-gated calcium channels (VGCC), inhibition can also be mediated through the direct interaction of Gβγ subunits with the soluble N -ethylmaleimide attachment protein receptor (SNARE) complex of the vesicle fusion apparatus. Binding studies with soluble SNARE complexes have shown that Gβγ binds to both ternary SNARE complexes, t-SNARE heterodimers, and monomeric SNAREs, competing with synaptotagmin 1(syt1) for binding sites on t-SNARE. However, in secretory cells, Gβγ, SNAREs, and synaptotagmin interact in the lipid environment of a vesicle at the plasma membrane. To approximate this environment, we show that fluorescently labeled Gβγ interacts specifically with lipid-embedded t-SNAREs consisting of full-length syntaxin 1 and SNAP-25B at the membrane, as measured by fluorescence polarization. Fluorescently labeled syt1 undergoes competition with Gβγ for SNARE-binding sites in lipid environments. Mutant Gβγ subunits that were previously shown to be more efficacious at inhibiting Ca 2+ -triggered exocytotic release than wild-type Gβγ were also shown to bind SNAREs at a higher affinity than wild type in a lipid environment. These mutant Gβγ subunits were unable to inhibit VGCC currents. Specific peptides corresponding to regions on Gβ and Gγ shown to be important for the interaction disrupt the interaction in a concentration-dependent manner. In in vitro fusion assays using full-length t- and v-SNAREs embedded in liposomes, Gβγ inhibited Ca 2+ /synaptotagmin-dependent fusion. Together, these studies demonstrate the importance of these regions for the Gβγ-SNARE interaction and show that the target of Gβγ, downstream of VGCC, is the membrane-embedded SNARE complex. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  19. Energy efficient neural stimulation: coupling circuit design and membrane biophysics.

    Directory of Open Access Journals (Sweden)

    Thomas J Foutz

    Full Text Available The delivery of therapeutic levels of electrical current to neural tissue is a well-established treatment for numerous indications such as Parkinson's disease and chronic pain. While the neuromodulation medical device industry has experienced steady clinical growth over the last two decades, much of the core technology underlying implanted pulse generators remain unchanged. In this study we propose some new methods for achieving increased energy-efficiency during neural stimulation. The first method exploits the biophysical features of excitable tissue through the use of a centered-triangular stimulation waveform. Neural activation with this waveform is achieved with a statistically significant reduction in energy compared to traditional rectangular waveforms. The second method demonstrates energy savings that could be achieved by advanced circuitry design. We show that the traditional practice of using a fixed compliance voltage for constant-current stimulation results in substantial energy loss. A portion of this energy can be recuperated by adjusting the compliance voltage to real-time requirements. Lastly, we demonstrate the potential impact of axon fiber diameter on defining the energy-optimal pulse-width for stimulation. When designing implantable pulse generators for energy efficiency, we propose that the future combination of a variable compliance system, a centered-triangular stimulus waveform, and an axon diameter specific stimulation pulse-width has great potential to reduce energy consumption and prolong battery life in neuromodulation devices.

  20. Energy efficient neural stimulation: coupling circuit design and membrane biophysics.

    Science.gov (United States)

    Foutz, Thomas J; Ackermann, D Michael; Kilgore, Kevin L; McIntyre, Cameron C

    2012-01-01

    The delivery of therapeutic levels of electrical current to neural tissue is a well-established treatment for numerous indications such as Parkinson's disease and chronic pain. While the neuromodulation medical device industry has experienced steady clinical growth over the last two decades, much of the core technology underlying implanted pulse generators remain unchanged. In this study we propose some new methods for achieving increased energy-efficiency during neural stimulation. The first method exploits the biophysical features of excitable tissue through the use of a centered-triangular stimulation waveform. Neural activation with this waveform is achieved with a statistically significant reduction in energy compared to traditional rectangular waveforms. The second method demonstrates energy savings that could be achieved by advanced circuitry design. We show that the traditional practice of using a fixed compliance voltage for constant-current stimulation results in substantial energy loss. A portion of this energy can be recuperated by adjusting the compliance voltage to real-time requirements. Lastly, we demonstrate the potential impact of axon fiber diameter on defining the energy-optimal pulse-width for stimulation. When designing implantable pulse generators for energy efficiency, we propose that the future combination of a variable compliance system, a centered-triangular stimulus waveform, and an axon diameter specific stimulation pulse-width has great potential to reduce energy consumption and prolong battery life in neuromodulation devices.

  1. The role of the N-terminal segment of CCR5 in HIV-1 Env-mediated membrane fusion and the mechanism of virus adaptation to CCR5 lacking this segment

    Directory of Open Access Journals (Sweden)

    Kabat David

    2007-08-01

    Full Text Available Abstract Background HIV-1 envelope glycoprotein (Env induces membrane fusion as a result of sequential binding to CD4 and chemokine receptors (CCR5 or CXCR4. The critical determinants of CCR5 coreceptor function are the N-terminal domain (Nt and the second extracellular loop. However, mutations in gp120 adapt HIV-1 to grow on cells expressing the N-terminally truncated CCR5(Δ18 (Platt et al., J. Virol. 2005, 79: 4357–68. Results We have functionally characterized the adapted Env (designated Env(NYP using a quantitative cell-cell fusion assay. The rate of fusion with target cells expressing wild-type CCR5 and the resistance to fusion inhibitors was virtually identical for wild-type Env and Env(NYP, implying that the coreceptor affinity had not increased as a result of adaptation. In contrast, Env(NYP-induced fusion with cells expressing CCR5(Δ18 occurred at a slower rate and was extremely sensitive to the CCR5 binding inhibitor, Sch-C. Resistance to Sch-C drastically increased after pre-incubation of Env(NYP- and CCR5(Δ18-expressing cells at a temperature that was not permissive to fusion. This indicates that ternary Env(NYP-CD4-CCR5(Δ18 complexes accumulate at sub-threshold temperature and that low-affinity interactions with the truncated coreceptor are sufficient for triggering conformational changes in the gp41 of Env(NYP but not in wild-type Env. We also demonstrated that the ability of CCR5(Δ18 to support fusion and infection mediated by wild-type Env can be partially reconstituted in the presence of a synthetic sulfated peptide corresponding to the CCR5 Nt. Pre-incubation of wild-type Env- and CCR5(Δ18-expressing cells with the sulfated peptide at sub-threshold temperature markedly increased the efficiency of fusion. Conclusion We propose that, upon binding the Nt region of CCR5, wild-type Env acquires the ability to productively engage the extracellular loop(s of CCR5 – an event that triggers gp41 refolding and membrane merger

  2. Refining the Mechanisms of Heniparvirus-Mediated Membrane Fusion Through Mutagenesis of Hendra virus Envelope Glycoproteins

    Science.gov (United States)

    2007-09-06

    glycoprotein single point mutants…..67 Figure 9: Effects of multiple point mutations on fusion activity of HeV F……….……..69 Figure 10: Effects of multiple ...relapsing encephalitis (28). How or whether this latter manifestation of disease is at all analogous to Subacute Sclerosing Panencephalitis (SSPE), a...total DNA per T-25cm2 flask overnight followed by infection with wild-type vaccinia virus (strain WR) at a multiplicity of infection (MOI) of 10. At

  3. A cytochrome c fusion protein domain for convenient detection, quantification, and enhanced production of membrane proteins in Escherichia coli--expression and characterization of cytochrome-tagged Complex I subunits.

    Science.gov (United States)

    Gustavsson, Tobias; Trane, Maria; Moparthi, Vamsi K; Miklovyte, Egle; Moparthi, Lavanya; Górecki, Kamil; Leiding, Thom; Arsköld, Sindra Peterson; Hägerhäll, Cecilia

    2010-08-01

    Overproduction of membrane proteins can be a cumbersome task, particularly if high yields are desirable. NADH:quinone oxidoreductase (Complex I) contains several very large membrane-spanning protein subunits that hitherto have been impossible to express individually in any appreciable amounts in Escherichia coli. The polypeptides contain no prosthetic groups and are poorly antigenic, making optimization of protein production a challenging task. In this work, the C-terminal ends of the Complex I subunits NuoH, NuoL, NuoM, and NuoN from E. coli Complex I and the bona fide antiporters MrpA and MrpD were genetically fused to the cytochrome c domain of Bacillus subtilis cytochrome c(550). Compared with other available fusion-protein tagging systems, the cytochrome c has several advantages. The heme is covalently bound, renders the proteins visible by optical spectroscopy, and can be used to monitor, quantify, and determine the orientation of the polypeptides in a plethora of experiments. For the antiporter-like subunits NuoL, NuoM, and NuoN and the real antiporters MrpA and MrpD, unprecedented amounts of holo-cytochrome fusion proteins could be obtained in E. coli. The NuoHcyt polypeptide was also efficiently produced, but heme insertion was less effective in this construct. The cytochrome c(550) domain in all the fusion proteins exhibited normal spectra and redox properties, with an E(m) of about +170 mV. The MrpA and MrpD antiporters remained functional after being fused to the cytochrome c-tag. Finally, a his-tag could be added to the cytochrome domain, without any perturbations to the cytochrome properties, allowing efficient purification of the overexpressed fusion proteins.

  4. Fusion of liposomes with the plasma membrane of epithelial cells: Fate of incorporated lipids as followed by freeze fracture and autoradiography of plastic sections

    NARCIS (Netherlands)

    Knoll, G.; Burger, K.N.J.; Bron, R.; van Meer, G.; Verkleij, A.J.

    1988-01-01

    The fusion of liposomes with the plasma membrane of influenza virus-infected monolayers of an epithelial cell line, Madin-Darby canine kidney cells (van Meer et al., 1985. Biochemistry, 24: 3593-3602), has been analyzed by morphological techniques. The distribution of liposomal lipids over the

  5. A high throughput Cre–lox activated viral membrane fusion assay identifies pharmacological inhibitors of HIV entry

    International Nuclear Information System (INIS)

    Esposito, Anthony M.; Cheung, Pamela; Swartz, Talia H.; Li, Hongru; Tsibane, Tshidi; Durham, Natasha D.; Basler, Christopher F.; Felsenfeld, Dan P.; Chen, Benjamin K.

    2016-01-01

    Enveloped virus entry occurs when viral and cellular membranes fuse releasing particle contents into the target cell. Human immunodeficiency virus (HIV) entry occurs by cell-free virus or virus transferred between infected and uninfected cells through structures called virological synapses. We developed a high-throughput cell-based assay to identify small molecule inhibitors of cell-free or virological synapse-mediated entry. An HIV clone carrying Cre recombinase as a Gag-internal gene fusion releases active Cre into cells upon viral entry activating a recombinatorial gene switch changing dsRed to GFP-expression. A screen of a 1998 known-biological profile small molecule library identified pharmacological HIV entry inhibitors that block both cell-free and cell-to-cell infection. Many top hits were noted as HIV inhibitors in prior studies, but not previously recognized as entry antagonists. Modest therapeutic indices for simvastatin and nigericin were observed in confirmatory HIV infection assays. This robust assay is adaptable to study HIV and heterologous viral pseudotypes. - Highlights: • Cre recombinase viral fusion assay screens cell-free or cell–cell entry inhibitors. • This Gag-iCre based assay is specific for the entry step of HIV replication. • Screened a library of known pharmacologic compounds for HIV fusion antagonists. • Many top hits were previously noted as HIV inhibitors, but here are classified as entry antagonists. Many top hits were previously noted as HIV inhibitors, but not as entry antagonists. • The assay is compatible with pseudotyping with HIV and heterologous viruses.

  6. A high throughput Cre–lox activated viral membrane fusion assay identifies pharmacological inhibitors of HIV entry

    Energy Technology Data Exchange (ETDEWEB)

    Esposito, Anthony M. [Division of Infectious Diseases, Department of Medicine, Icahn School of Medicine at Mount Sinai, Immunology Institute, New York, NY (United States); Cheung, Pamela [Integrated Screening Core, Icahn School of Medicine at Mount Sinai, New York, NY (United States); Swartz, Talia H.; Li, Hongru [Division of Infectious Diseases, Department of Medicine, Icahn School of Medicine at Mount Sinai, Immunology Institute, New York, NY (United States); Tsibane, Tshidi [Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, NY (United States); Durham, Natasha D. [Division of Infectious Diseases, Department of Medicine, Icahn School of Medicine at Mount Sinai, Immunology Institute, New York, NY (United States); Basler, Christopher F. [Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, NY (United States); Felsenfeld, Dan P. [Integrated Screening Core, Icahn School of Medicine at Mount Sinai, New York, NY (United States); Chen, Benjamin K., E-mail: benjamin.chen@mssm.edu [Division of Infectious Diseases, Department of Medicine, Icahn School of Medicine at Mount Sinai, Immunology Institute, New York, NY (United States)

    2016-03-15

    Enveloped virus entry occurs when viral and cellular membranes fuse releasing particle contents into the target cell. Human immunodeficiency virus (HIV) entry occurs by cell-free virus or virus transferred between infected and uninfected cells through structures called virological synapses. We developed a high-throughput cell-based assay to identify small molecule inhibitors of cell-free or virological synapse-mediated entry. An HIV clone carrying Cre recombinase as a Gag-internal gene fusion releases active Cre into cells upon viral entry activating a recombinatorial gene switch changing dsRed to GFP-expression. A screen of a 1998 known-biological profile small molecule library identified pharmacological HIV entry inhibitors that block both cell-free and cell-to-cell infection. Many top hits were noted as HIV inhibitors in prior studies, but not previously recognized as entry antagonists. Modest therapeutic indices for simvastatin and nigericin were observed in confirmatory HIV infection assays. This robust assay is adaptable to study HIV and heterologous viral pseudotypes. - Highlights: • Cre recombinase viral fusion assay screens cell-free or cell–cell entry inhibitors. • This Gag-iCre based assay is specific for the entry step of HIV replication. • Screened a library of known pharmacologic compounds for HIV fusion antagonists. • Many top hits were previously noted as HIV inhibitors, but here are classified as entry antagonists. Many top hits were previously noted as HIV inhibitors, but not as entry antagonists. • The assay is compatible with pseudotyping with HIV and heterologous viruses.

  7. 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

    The growing scarcity of fresh water is a major political and economic challenge in the 21st century. Compared to thermal-based distillation technique of water production, pressure driven membrane-based water purification process, such as ultrafiltration (UF), nanofiltration (NF) and reverse osmosis (RO), can offer more energy-efficient and environmentally friendly solution to clean water production. Potential applications also include removal of hazardous chemicals (i.e., arsenic, pesticides, organics) from water. Although those membrane-separation technologies have been used to produce drinking water from seawater (desalination) and non-traditional water (i.e., municipal wastewater and brackish groundwater) over the last decades, they still have problems in order to be applied in large-scale operations. Currently, a major huddle of membrane-based water purification technology for large-scale commercialization is membrane fouling and its resulting increases in pressure and energy cost of filtration process. Membrane cleaning methods, which can restore the membrane properties to some degree, usually cause irreversible damage to the membranes. Considering that electricity for creating of pressure constitutes a majority of cost (~50%) in membrane-based water purification process, the development of new nano-porous membranes that are more resistant to degradation and less subject to fouling is highly desired. Styrene-ethylene/butylene-styrene (SEBS) block copolymer is one of the best known block copolymers that induces well defined morphologies. Due to the polarity difference of aromatic styrene unit and saturated ethylene/butylene unit, these two polymer chains self-assemble each other and form different phase-separated morphologies depending on the ratios of two polymer chain lengths. Because the surface of SEBS is hydrophobic which easily causes fouling of membrane, incorporation of ionic group (e,g, sulfonate) to the polymer is necessary to reduces fouling

  8. Efficiency of Polymer Electrolyte Membrane Fuel Cell Stack

    Directory of Open Access Journals (Sweden)

    Hans Bosma

    2011-08-01

    Full Text Available This paper applies a feedforward control of optimal oxygen excess ratio that maximize net power (improve efficiency of a NedStack P8.0-64 PEM fuel cell stack (FCS system. Net powers profile as a function of oxygen excess ratio for some points of operation are analyzed by using FCS model. The relationships between stack current and the corresponding control input voltage that gives an optimal oxygen excess ratio are used to design a feedforward control scheme. The results of this scheme are compared to the results of a feedforward control using a constant oxygen excess ratio. Simulation results show that optimal oxygen excess ratio improves fuel cell performance compared to the results of constant oxygen excess ratio. The same procedures are performed experimentally for the FCS system. The behaviour of the net power of the fuel cell stack with respect to the variation of oxygen excess ratio is analyzed to obtain optimal values. Data of stack current and the corresponding voltage input to the compressor that gives optimal values of oxygen excess ratio are used to develop a feedforward control. Feedforward control based on constant and optimal oxygen excess ratio control, are implemented in the NedStack P8.0-64 PEM fuel cell stack system by using LabVIEW. Implementation results shows that optimal oxygen excess ratio control improves the fuel cell performance compared to the constant oxygen excess ratio control.

  9. Solid-state NMR spectroscopy of the HIV gp41 membrane fusion protein supports intermolecular antiparallel β sheet fusion peptide structure in the final six-helix bundle state.

    Science.gov (United States)

    Sackett, Kelly; Nethercott, Matthew J; Zheng, Zhaoxiong; Weliky, David P

    2014-03-06

    The HIV gp41 protein catalyzes fusion between viral and target cell membranes. Although the ~20-residue N-terminal fusion peptide (FP) region is critical for fusion, the structure of this region is not well characterized in large gp41 constructs that model the gp41 state at different times during fusion. This paper describes solid-state NMR (SSNMR) studies of FP structure in a membrane-associated construct (FP-Hairpin), which likely models the final fusion state thought to be thermostable trimers with six-helix bundle structure in the region C-terminal of the FP. The SSNMR data show that there are populations of FP-Hairpin with either α helical or β sheet FP conformation. For the β sheet population, measurements of intermolecular (13)C-(13)C proximities in the FP are consistent with a significant fraction of intermolecular antiparallel β sheet FP structure with adjacent strand crossing near L7 and F8. There appears to be negligible in-register parallel structure. These findings support assembly of membrane-associated gp41 trimers through interleaving of N-terminal FPs from different trimers. Similar SSNMR data are obtained for FP-Hairpin and a construct containing the 70 N-terminal residues of gp41 (N70), which is a model for part of the putative pre-hairpin intermediate state of gp41. FP assembly may therefore occur at an early fusion stage. On a more fundamental level, similar SSNMR data are obtained for FP-Hairpin and a construct containing the 34 N-terminal gp41 residues (FP34) and support the hypothesis that the FP is an autonomous folding domain. Copyright © 2013 Elsevier Ltd. All rights reserved.

  10. Manipulating the membrane penetration mechanism of helical polypeptides via aromatic modification for efficient gene delivery.

    Science.gov (United States)

    Zheng, Nan; Song, Ziyuan; Yang, Jiandong; Liu, Yang; Li, Fangfang; Cheng, Jianjun; Yin, Lichen

    2017-08-01

    The delivery performance of non-viral gene vectors is greatly related to their intracellular kinetics. Cationic helical polypeptides with potent membrane penetration properties and gene transfection efficiencies have been recently developed by us. However, they suffer from severe drawbacks in terms of their membrane penetration mechanisms that mainly include endocytosis and pore formation. The endocytosis mechanism leads to endosomal entrapment of gene cargos, while the charge- and helicity-induced pore formation causes appreciable cytotoxicity at high concentrations. With the attempt to overcome such critical challenges, we incorporated aromatic motifs into the design of helical polypeptides to enhance their membrane activities and more importantly, to manipulate their membrane penetration mechanisms. The aromatically modified polypeptides exhibited higher cellular internalization level than the unmodified analogue by up to 2.5 folds. Such improvement is possibly because aromatic domains promoted the polypeptides to penetrate cell membranes via direct transduction, a non-endocytosis and non-pore formation mechanism. As such, gene cargos were more efficiently delivered into cells by bypassing endocytosis and subsequently avoiding endosomal entrapment, and the material toxicity associated with excessive pore formation was also reduced. The top-performing aromatic polypeptide containing naphthyl side chains at the incorporated content of 20mol% revealed notably higher transfection efficiencies than commercial reagents in melanoma cells in vitro (by 11.7 folds) and in vivo (by 9.1 folds), and thus found potential utilities toward topical gene delivery for cancer therapy. Cationic helical polypeptides, as efficient gene delivery materials, suffer from severe drawbacks in terms of their membrane penetration mechanisms. The main cell penetration mechanisms involved are endocytosis and pore formation. However, the endocytosis mechanism has the limitation of endosomal

  11. Reversible acid-induced inactivation of the membrane fusion protein of Semliki Forest virus

    NARCIS (Netherlands)

    Waarts, BL; Smit, JM; Aneke, OJC; McInerney, GM; Liljestrom, P; Bittman, R; Wilschut, J

    Previously, it has been shown that the exposure of Semliki Forest virus (SFV) to a mildly acidic environment induces a rapid and complete loss of the ability of the virus to bind and fuse to target membranes added subsequently. In the present study, incubation of SFV at low pH followed by a specific

  12. Huntingtin is required for ER-to-Golgi transport and for secretory vesicle fusion at the plasma membrane

    Directory of Open Access Journals (Sweden)

    Hemma Brandstaetter

    2014-12-01

    Full Text Available Huntingtin is a large membrane-associated scaffolding protein that associates with endocytic and exocytic vesicles and modulates their trafficking along cytoskeletal tracks. Although the progression of Huntington’s disease is linked to toxic accumulation of mutant huntingtin protein, loss of wild-type huntingtin function might also contribute to neuronal cell death, but its precise function is not well understood. Therefore, we investigated the molecular role of huntingtin in exocytosis and observed that huntingtin knockdown in HeLa cells causes a delay in endoplasmic reticulum (ER-to-Golgi transport and a reduction in the number of cargo vesicles leaving the trans-Golgi network. In addition, we found that huntingtin is required for secretory vesicle fusion at the plasma membrane. Similar defects in the early exocytic pathway were observed in primary fibroblasts from homozygous Htt140Q/140Q knock-in mice, which have the expansion inserted into the mouse huntingtin gene so lack wild-type huntingtin expression. Interestingly, heterozygous fibroblasts from a Huntington’s disease patient with a 180Q expansion displayed no obvious defects in the early secretory pathway. Thus, our results highlight the requirement for wild-type huntingtin at distinct steps along the secretory pathway.

  13. Fusion Machinery

    DEFF Research Database (Denmark)

    Sørensen, Jakob Balslev; Milosevic, Ira

    2015-01-01

    SNARE proteins constitute the minimal machinery needed for membrane fusion. SNAREs operate by forming a complex, which pulls the lipid bilayers into close contact and provides the mechanical force needed for lipid bilayer fusion. At the chemical synapse, SNARE-complex formation between...... the vesicular SNARE VAMP2/synaptobrevin-2 and the target (plasma membrane) SNAREs SNAP25 and syntaxin-1 results in fusion and release of neurotransmitter, synchronized to the electrical activity of the cell by calcium influx and binding to synaptotagmin. Formation of the SNARE complex is tightly regulated...... and appears to start with syntaxin-1 bound to an SM (Sec1/Munc18-like) protein. Proteins of the Munc13-family are responsible for opening up syntaxin and allowing sequential binding of SNAP-25 and VAMP2/synaptobrevin-2. N- to C-terminal “zippering” of the SNARE domains leads to membrane fusion...

  14. A High Performance Platform Based on cDNA Display for Efficient Synthesis of Protein Fusions and Accelerated Directed Evolution.

    Science.gov (United States)

    Naimuddin, Mohammed; Kubo, Tai

    2016-02-08

    We describe a high performance platform based on cDNA display technology by developing a new modified puromycin linker-oligonucleotide. The linker consists of four major characteristics: a "ligation site" for hybridization and ligation of mRNA by T4 RNA ligase, a "puromycin arm" for covalent linkage of the protein, a "polyadenosine site" for a longer puromycin arm and purification of protein fusions (optional) using oligo-dT matrices, and a "reverse transcription site" for the formation of stable cDNA protein fusions whose cDNA is covalently linked to its encoded protein. The linker was synthesized by a novel branching strategy and provided >8-fold higher yield than previous linkers. This linker enables rapid and highly efficient ligation of mRNA (>90%) and synthesis of protein fusions (∼ 50-95%) in various cell-free expression systems. Overall, this new cDNA display method provides 10-200 fold higher end-usage fusions than previous methods and benefits higher diversity libraries crucial for directed protein/peptide evolution. With the increased efficiency, this system was able to reduce the time for one selection cycle to cDNA display method. A three-finger protein library was evolved to isolate superior nanomolar range binding candidates for vascular endothelial growth factor. This method is expected to provide a beneficial impact to accelerated drug discovery and proteome analysis.

  15. Reassessment of the Lineage Fusion Hypothesis for the Origin of Double Membrane Bacteria

    OpenAIRE

    Swithers, Kristen S.; Fournier, Gregory P.; Green, Anna G.; Gogarten, J. Peter; Lapierre, Pascal

    2011-01-01

    In 2009, James Lake introduced a new hypothesis in which reticulate phylogeny reconstruction is used to elucidate the origin of gram-negative bacteria (Nature 460: 967-971). The presented data supported the gram-negative bacteria originating from an ancient endosymbiosis between the Actinobacteria and Clostridia. His conclusion was based on a presence-absence analysis of protein families that divided all prokaryotes into five groups: Actinobacteria, Double Membrane bacteria (DM), Clostridia, ...

  16. Exploring large-scale phenomena in composite membranes through an efficient implicit-solvent model

    Science.gov (United States)

    Laradji, Mohamed; Kumar, P. B. Sunil; Spangler, Eric J.

    2016-07-01

    Several microscopic and mesoscale models have been introduced in the past to investigate various phenomena in lipid membranes. Most of these models account for the solvent explicitly. Since in a typical molecular dynamics simulation, the majority of particles belong to the solvent, much of the computational effort in these simulations is devoted for calculating forces between solvent particles. To overcome this problem, several implicit-solvent mesoscale models for lipid membranes have been proposed during the last few years. In the present article, we review an efficient coarse-grained implicit-solvent model we introduced earlier for studies of lipid membranes. In this model, lipid molecules are coarse-grained into short semi-flexible chains of beads with soft interactions. Through molecular dynamics simulations, the model is used to investigate the thermal, structural and elastic properties of lipid membranes. We will also review here few studies, based on this model, of the phase behavior of nanoscale liposomes, cytoskeleton-induced blebbing in lipid membranes, as well as nanoparticles wrapping and endocytosis by tensionless lipid membranes. Topical Review article submitted to the Journal of Physics D: Applied Physics, May 9, 2016

  17. Exploring large-scale phenomena in composite membranes through an efficient implicit-solvent model

    International Nuclear Information System (INIS)

    Laradji, Mohamed; Sunil Kumar, P B; Spangler, Eric J

    2016-01-01

    Several microscopic and mesoscale models have been introduced in the past to investigate various phenomena in lipid membranes. Most of these models account for the solvent explicitly. Since in a typical molecular dynamics simulation, the majority of particles belong to the solvent, much of the computational effort in these simulations is devoted for calculating forces between solvent particles. To overcome this problem, several implicit-solvent mesoscale models for lipid membranes have been proposed during the last few years. In the present article, we review an efficient coarse-grained implicit-solvent model we introduced earlier for studies of lipid membranes. In this model, lipid molecules are coarse-grained into short semi-flexible chains of beads with soft interactions. Through molecular dynamics simulations, the model is used to investigate the thermal, structural and elastic properties of lipid membranes. We will also review here few studies, based on this model, of the phase behavior of nanoscale liposomes, cytoskeleton-induced blebbing in lipid membranes, as well as nanoparticles wrapping and endocytosis by tensionless lipid membranes. (topical review)

  18. Mutations in the Fusion Protein of Measles Virus That Confer Resistance to the Membrane Fusion Inhibitors Carbobenzoxy-d-Phe-l-Phe-Gly and 4-Nitro-2-Phenylacetyl Amino-Benzamide.

    Science.gov (United States)

    Ha, Michael N; Delpeut, Sébastien; Noyce, Ryan S; Sisson, Gary; Black, Karen M; Lin, Liang-Tzung; Bilimoria, Darius; Plemper, Richard K; Privé, Gilbert G; Richardson, Christopher D

    2017-12-01

    The inhibitors carbobenzoxy (Z)-d-Phe-l-Phe-Gly (fusion inhibitor peptide [FIP]) and 4-nitro-2-phenylacetyl amino-benzamide (AS-48) have similar efficacies in blocking membrane fusion and syncytium formation mediated by measles virus (MeV). Other homologues, such as Z-d-Phe, are less effective but may act through the same mechanism. In an attempt to map the site of action of these inhibitors, we generated mutant viruses that were resistant to the inhibitory effects of Z-d-Phe-l-Phe-Gly. These 10 mutations were localized to the heptad repeat B (HRB) region of the fusion protein, and no changes were observed in the viral hemagglutinin, which is the receptor attachment protein. Mutations were validated in a luciferase-based membrane fusion assay, using transfected fusion and hemagglutinin expression plasmids or with syncytium-based assays in Vero, Vero-SLAM, and Vero-Nectin 4 cell lines. The changes I452T, D458N, D458G/V459A, N462K, N462H, G464E, and I483R conferred resistance to both FIP and AS-48 without compromising membrane fusion. The inhibitors did not block hemagglutinin protein-mediated binding to the target cell. Edmonston vaccine/laboratory and IC323 wild-type strains were equally affected by the inhibitors. Escape mutations were mapped upon a three-dimensional (3D) structure modeled from the published crystal structure of parainfluenzavirus 5 fusion protein. The most effective mutations were situated in a region located near the base of the globular head and its junction with the alpha-helical stalk of the prefusion protein. We hypothesize that the fusion inhibitors could interfere with the structural changes that occur between the prefusion and postfusion conformations of the fusion protein. IMPORTANCE Due to lapses in vaccination worldwide that have caused localized outbreaks, measles virus (MeV) has regained importance as a pathogen. Antiviral agents against measles virus are not commercially available but could be useful in conjunction with Me

  19. Energy Efficient Aeration in a Single Low Pressure Hollow Sheet Membrane Filtration Module

    DEFF Research Database (Denmark)

    Bentzen, Thomas Ruby; Ratkovich, Nicolas Rios; Rasmussen, Michael R.

    2011-01-01

    The main drawback of membrane bioreactors (MBR) systems is the fouling of the membrane, which is decreased and/or prevented through gas sparging. However, this practice is based on rules of thumb or a trial-and-error approaches which are tedious, very time-consuming, do not necessarily provide...... optimal fouling control and they are not energy efficient. Therefore, dedicated experiments are needed to fully understand the hydrodynamics of it. A hollow sheet (HS) MBR was studied. Experimental velocity measurements were made using micro-propellers and compared to CFD results. A good agreement between...

  20. Efficient protein immobilization on polyethersolfone electrospun nanofibrous membrane via covalent binding for biosensing applications

    Energy Technology Data Exchange (ETDEWEB)

    Mahmoudifard, Matin [Institute for Nanoscience and Nanotechnology, Sharif University of Technology, Tehran (Iran, Islamic Republic of); Soudi, Sara [Stem Cell Biology Department, Stem Cell Technology Research Center, Tehran (Iran, Islamic Republic of); Department of Immunology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran (Iran, Islamic Republic of); Soleimani, Masoud [Department of Hematology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran (Iran, Islamic Republic of); Hosseinzadeh, Simzar [Nanotechnology and Tissue Engineering Department, Stem Cell Technology Research Center, Tehran (Iran, Islamic Republic of); School of Advanced Medical Technologies, Tehran University of Medical Sciences, Tehran (Iran, Islamic Republic of); Esmaeili, Elaheh [Institute for Nanoscience and Nanotechnology, Sharif University of Technology, Tehran (Iran, Islamic Republic of); Department of Hematology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran (Iran, Islamic Republic of); Vossoughi, Manouchehr, E-mail: vosoughi@sharif.edu [Institute for Nanoscience and Nanotechnology, Sharif University of Technology, Tehran (Iran, Islamic Republic of); Chemical and Petroleum Engineering Department, Sharif University of Technology, Tehran (Iran, Islamic Republic of)

    2016-01-01

    In this paper we introduce novel strategy for antibody immobilization using high surface area electrospun nanofibrous membrane based on ethyl-3-(3-dimethylaminopropyl)-carbodiimide/N-hydroxysuccinimide (EDC/NHS) coupling chemistry. To present the high performance of proposed biosensors, anti-staphylococcus enterotoxin B (anti-SEB) was used as a model to demonstrate the utility of our proposed system. Polymer solution of polyethersolfone was used to fabricate fine nanofibrous membrane. Moreover, industrial polyvinylidene fluoride membrane and conventional microtiter plate were also used to compare the efficiency of antibody immobilization. Scanning electron microscopy images were taken to study the morphology of the membranes. The surface activation of nanofibrous membrane was done with the help of O{sub 2} plasma. PES nanofibrous membrane with carboxyl functional groups for covalent attachment of antibodies were treated by EDC/NHS coupling agent. The quantity of antibody immobilization was measured by enzyme-linked immuno sorbent assay (ELISA) method. Attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR) spectroscopy was performed to confirm the covalent immobilization of antibody on membrane. Atomic force microscopy, scanning electron microscopy and invert fluorescence microscopy were used to analyze the antibody distribution pattern on solid surfaces. Results show that oxygen plasma treatment effectively increased the amount of antibody immobilization through EDC/NHS coupling chemistry. It was found that the use of nanofibrous membrane causes the improved detection signal of ELISA based biosensors in comparison to the standard assay carried out in the 96-well microtiter plate. This method has the potential to improve the ELISA-based biosensor and we believe that this technique can be used in various biosensing methods. - Highlights: • Introduction of novel strategy for antibody immobilization using high surface area electrospun

  1. A single amino acid substitution modulates low-pH-triggered membrane fusion of GP64 protein in Autographa californica and Bombyx mori nucleopolyhedroviruses

    International Nuclear Information System (INIS)

    Katou, Yasuhiro; Yamada, Hayato; Ikeda, Motoko; Kobayashi, Michihiro

    2010-01-01

    We have previously shown that budded viruses of Bombyx mori nucleopolyhedrovirus (BmNPV) enter the cell cytoplasm but do not migrate into the nuclei of non-permissive Sf9 cells that support a high titer of Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) multiplication. Here we show, using the syncytium formation assay, that low-pH-triggered membrane fusion of BmNPV GP64 protein (Bm-GP64) is significantly lower than that of AcMNPV GP64 protein (Ac-GP64). Mutational analyses of GP64 proteins revealed that a single amino acid substitution between Ac-GP64 H155 and Bm-GP64 Y153 can have significant positive or negative effects on membrane fusion activity. Studies using bacmid-based GP64 recombinant AcMNPV harboring point-mutated ac-gp64 and bm-gp64 genes showed that Ac-GP64 H155Y and Bm-GP64 Y153H substitutions decreased and increased, respectively, the multiplication and cell-to-cell spread of progeny viruses. These results indicate that Ac-GP64 H155 facilitates the low-pH-triggered membrane fusion reaction between virus envelopes and endosomal membranes.

  2. Efficient treatment of aniline containing wastewater in bipolar membrane microbial electrolysis cell-Fenton system.

    Science.gov (United States)

    Li, Xiaohu; Jin, Xiangdan; Zhao, Nannan; Angelidaki, Irini; Zhang, Yifeng

    2017-08-01

    Aniline-containing wastewater can cause significant environmental problems and threaten the humans's life. However, rapid degradation of aniline with cost-efficient methods remains a challenge. In this work, a novel microbial electrolysis cell with bipolar membrane was integrated with Fenton reaction (MEC-Fenton) for efficient treatment of real wastewater containing a high concentration (4460 ± 52 mg L -1 ) of aniline. In this system, H 2 O 2 was in situ electro-synthesized from O 2 reduction on the graphite cathode and was simultaneously used as source of OH for the oxidation of aniline wastewater under an acidic condition maintained by the bipolar membrane. The aniline was effectively degraded following first-order kinetics at a rate constant of 0.0166 h -1 under an applied voltage of 0.5 V. Meanwhile, a total organic carbon (TOC) removal efficiency of 93.1 ± 1.2% was obtained, revealing efficient mineralization of aniline. The applicability of bipolar membrane MEC-Fenton system was successfully demonstrated with actual aniline wastewater. Moreover, energy balance showed that the system could be a promising technology for removal of biorefractory organic pollutants from wastewaters. Copyright © 2017 Elsevier Ltd. All rights reserved.

  3. Advanced, Energy-Efficient Hybrid Membrane System for Industrial Water Reuse

    Energy Technology Data Exchange (ETDEWEB)

    Toy, Lora [RTI International, Research Triangle Park, NC (United States); Choi, Young Chul [RTI International, Research Triangle Park, NC (United States); Hendren, Zachary [RTI International, Research Triangle Park, NC (United States); Kim, Gyu Dong [RTI International, Research Triangle Park, NC (United States)

    2017-03-31

    In the U.S. manufacturing sector, current industrial water use practices are energy-intensive and utilize and discharge high volumes of waters, rendering them not sustainable especially in light of the growing scarcity of suitable water supplies. To help address this problem, the goal of this project was to develop an advanced, cost-effective, hybrid membrane-based water treatment system that can improve the energy efficiency of industrial wastewater treatment while allowing at least 50% water reuse efficiency. This hybrid process would combine emerging Forward Osmosis (FO) and Membrane Distillation (MD) technology components into an integrated FO-MD system that can beneficially utilize low-grade waste heat (i.e., T < 450 °F) in industrial facilities to produce distilled-quality product water for reuse. In this project, laboratory-, bench-, and pilot-scale experiments on the hybrid FO-MD system were conducted for industrial wastewater treatment. It was demonstrated at laboratory, bench, and pilot scales that FO-MD membrane technology can concentrate brine to very high total dissolved solids (TDS) levels (>200,000 ppm) that are at least 2.5 times higher than the TDS level to which RO can achieve. In laboratory testing, currently available FO and MD membranes were tested to select for high-performing membranes with high salt rejection and high water flux. Multiple FO membrane/draw-salt solution combinations that gave high water flux with higher than 98% salt rejection were also identified. Reverse draw-salt fluxes were observed to be much lower for divalent salts than for monovalent salts. MD membranes were identified that had 99.9+% salt rejection and water flux as high as 50-90 L/(m2·h) for flat-sheet membranes and >20 L/(m2·h) for hollow fibers. In bench-scale testing, a single unit of commercially available FO and MD membrane modules were evaluated for continuous, integrated operation. Using the laboratory- and bench-scale test data

  4. Inorganic Nanoparticles/Metal Organic Framework Hybrid Membrane Reactors for Efficient Photocatalytic Conversion of CO2.

    Science.gov (United States)

    Maina, James W; Schütz, Jürg A; Grundy, Luke; Des Ligneris, Elise; Yi, Zhifeng; Kong, Lingxue; Pozo-Gonzalo, Cristina; Ionescu, Mihail; Dumée, Ludovic F

    2017-10-11

    Photocatalytic conversion of carbon dioxide (CO 2 ) to useful products has potential to address the adverse environmental impact of global warming. However, most photocatalysts used to date exhibit limited catalytic performance, due to poor CO 2 adsorption capacity, inability to efficiently generate photoexcited electrons, and/or poor transfer of the photogenerated electrons to CO 2 molecules adsorbed on the catalyst surface. The integration of inorganic semiconductor nanoparticles across metal organic framework (MOF) materials has potential to yield new hybrid materials, combining the high CO 2 adsorption capacity of MOF and the ability of the semiconductor nanoparticles to generate photoexcited electrons. Herein, controlled encapsulation of TiO 2 and Cu-TiO 2 nanoparticles within zeolitic imidazolate framework (ZIF-8) membranes was successfully accomplished, using rapid thermal deposition (RTD), and their photocatalytic efficiency toward CO 2 conversion was investigated under UV irradiation. Methanol and carbon monoxide (CO) were found to be the only products of the CO 2 reduction, with yields strongly dependent upon the content and composition of the dopant semiconductor particles. CuTiO 2 nanoparticle doped membranes exhibited the best photocatalytic performance, with 7 μg of the semiconductor nanoparticle enhancing CO yield of the pristine ZIF-8 membrane by 233%, and methanol yield by 70%. This work opens new routes for the fabrication of hybrid membranes containing inorganic nanoparticles and MOFs, with potential application not only in catalysis but also in electrochemical, separation, and sensing applications.

  5. Cell cycle-regulated membrane binding of NuMA contributes to efficient anaphase chromosome separation.

    Science.gov (United States)

    Zheng, Zhen; Wan, Qingwen; Meixiong, Gerry; Du, Quansheng

    2014-03-01

    Accurate and efficient separation of sister chromatids during anaphase is critical for faithful cell division. It has been proposed that cortical dynein-generated pulling forces on astral microtubules contribute to anaphase spindle elongation and chromosome separation. In mammalian cells, however, definitive evidence for the involvement of cortical dynein in chromosome separation is missing. It is believed that dynein is recruited and anchored at the cell cortex during mitosis by the α subunit of heterotrimeric G protein (Gα)/mammalian homologue of Drosophila Partner of Inscuteable/nuclear mitotic apparatus (NuMA) ternary complex. Here we uncover a Gα/LGN-independent lipid- and membrane-binding domain at the C-terminus of NuMA. We show that the membrane binding of NuMA is cell cycle regulated-it is inhibited during prophase and metaphase by cyclin-dependent kinase 1 (CDK1)-mediated phosphorylation and only occurs after anaphase onset when CDK1 activity is down-regulated. Further studies indicate that cell cycle-regulated membrane association of NuMA underlies anaphase-specific enhancement of cortical NuMA and dynein. By replacing endogenous NuMA with membrane-binding-deficient NuMA, we can specifically reduce the cortical accumulation of NuMA and dynein during anaphase and demonstrate that cortical NuMA and dynein contribute to efficient chromosome separation in mammalian cells.

  6. Fusion neutronics

    CERN Document Server

    Wu, Yican

    2017-01-01

    This book provides a systematic and comprehensive introduction to fusion neutronics, covering all key topics from the fundamental theories and methodologies, as well as a wide range of fusion system designs and experiments. It is the first-ever book focusing on the subject of fusion neutronics research. Compared with other nuclear devices such as fission reactors and accelerators, fusion systems are normally characterized by their complex geometry and nuclear physics, which entail new challenges for neutronics such as complicated modeling, deep penetration, low simulation efficiency, multi-physics coupling, etc. The book focuses on the neutronics characteristics of fusion systems and introduces a series of theories and methodologies that were developed to address the challenges of fusion neutronics, and which have since been widely applied all over the world. Further, it introduces readers to neutronics design’s unique principles and procedures, experimental methodologies and technologies for fusion systems...

  7. Palmitoylation of SARS-CoV S protein is necessary for partitioning into detergent-resistant membranes and cell-cell fusion but not interaction with M protein

    International Nuclear Information System (INIS)

    McBride, Corrin E.; Machamer, Carolyn E.

    2010-01-01

    Coronaviruses are enveloped RNA viruses that generally cause mild disease in humans. However, the recently emerged coronavirus that caused severe acute respiratory syndrome (SARS-CoV) is the most pathogenic human coronavirus discovered to date. The SARS-CoV spike (S) protein mediates virus entry by binding cellular receptors and inducing fusion between the viral envelope and the host cell membrane. Coronavirus S proteins are palmitoylated, which may affect function. Here, we created a non-palmitoylated SARS-CoV S protein by mutating all nine cytoplasmic cysteine residues. Palmitoylation of SARS-CoV S was required for partitioning into detergent-resistant membranes and for cell-cell fusion. Surprisingly, however, palmitoylation of S was not required for interaction with SARS-CoV M protein. This contrasts with the requirement for palmitoylation of mouse hepatitis virus S protein for interaction with M protein and may point to important differences in assembly and infectivity of these two coronaviruses.

  8. Establishment of insect cell lines expressing green fluorescent protein on cell surface based on AcMNPV GP64 membrane fusion characteristic.

    Science.gov (United States)

    Qi, Ben-Xiang; Chen, Ying-Jian; Su, Rui; Li, Yi-Fei; Zheng, Gui-Ling; Li, Chang-You

    2017-10-01

    Displaying a protein on the surface of cells has been provided a very successful strategy to function research of exogenous proteins. Based on the membrane fusion characteristic of Autographa californica multiple nucleopolyhedrovirus envelope protein GP64, we amplified and cloned N-terminal signal peptide and C-terminal transmembrane domain as well as cytoplasmic tail domain of gp64 gene into vector pIZ/V5-His with multi-cloning sites to construct the cell surface expression vector pIZ/V5-gp64. To verify that the vector can be used to express proteins on the membrane of insect cells, a recombinant plasmid pIZ/V5-gp64-GFP was constructed by introducing the PCR amplified green fluorescent protein (GFP) gene and transfected into insect cell lines Sf9 and H5. The transected cells were screened with zeocin and cell cloning. PCR verification results showed that the GFP gene was successfully integrated into these cells. Green fluorescence in Sf9-GFP and H5-GFP cells was observed by using confocal laser scanning microscopy and immunofluorescence detection indicated that GFP protein was located on the cell membrane. Western blot results showed that a fusion protein GP64-GFP of about 40 kDa was expressed on the membrane of Sf9-GFP and H5-GFP cells. The expression system constructed in this paper can be used for localization and continuous expression of exogenous proteins on insect cell membrane.

  9. Unusual Self-Assembly of the Recombinant Chlamydia trachomatis Major Outer Membrane Protein-Based Fusion Antigen CTH522 Into Protein Nanoparticles

    DEFF Research Database (Denmark)

    Rose, Fabrice; Karlsen, Kasper; Jensen, Pernille

    2018-01-01

    but is a challenging vaccine candidate by being an integral membrane protein, and the immunogenicity depends on a correctly folded structure. We investigated the biophysical properties of the recombinant MOMP-based fusion antigen CTH522, which is tested in early human clinical trials. It consists of a truncated......Sexually transmitted Chlamydia trachomatis (Ct) infects more than 100 million people annually, and untreated chlamydia infections can cause severe complications. Therefore, there is an urgent need for a chlamydia vaccine. The Ct major outer membrane protein (MOMP) is highly immunogenic...

  10. Hexameric assembly of membrane fusion protein YknX of the sporulation delaying efflux pump from Bacillus amyloliquefaciens.

    Science.gov (United States)

    Xu, Yongbin; Jo, Inseong; Wang, Lulu; Chen, Jinli; Fan, Shengdi; Dong, Yuesheng; Quan, Chunshan; Ha, Nam-Chul

    2017-11-04

    Membrane fusion proteins (MFPs) play an essential role in the action of the drug efflux pumps and protein secretion systems in bacteria. The sporulation delaying protein (SDP) efflux pump YknWXYZ has been identified in diverse Bacillus species. The MFP YknX requires the ATP-binding cassette (ABC) transporter YknYZ and the Yip1 family protein YknW to form a functional complex. To date, the crystal structure, molecular function and mechanism of action of YknX remain unknown. In this study, to characterize the structural and biochemical roles of YknX in the functional assembly of YknWXYZ from B. amyloliquefaciens, we successfully obtained crystals of the YknX protein that diffracted X-rays to a resolution of 4.4 Å. We calculated an experimentally phased map using single-wavelength anomalous diffraction (SAD), revealing that YknX forms a hexameric assembly similar to that of MacA from Gram-negative bacteria. The hexameric assembly of YknX exhibited a funnel-like structure with a central channel and a conical mouth. Functional studies in vitro suggest that YknX can bind directly to peptidoglycan. Our study provides an improved understanding of the assembly of the YknWXYZ efflux pump and the role of YknX in the complex. Copyright © 2017 Elsevier Inc. All rights reserved.

  11. Mitochondrial matrix delivery using MITO-Porter, a liposome-based carrier that specifies fusion with mitochondrial membranes

    Energy Technology Data Exchange (ETDEWEB)

    Yasuzaki, Yukari; Yamada, Yuma [Laboratory for Molecular Design of Pharmaceutics, Faculty of Pharmaceutical Sciences, Hokkaido University, Kita-12, Nishi-6, Kita-ku, Sapporo 060-0812 (Japan); Harashima, Hideyoshi, E-mail: harasima@pharm.hokudai.ac.jp [Laboratory for Molecular Design of Pharmaceutics, Faculty of Pharmaceutical Sciences, Hokkaido University, Kita-12, Nishi-6, Kita-ku, Sapporo 060-0812 (Japan)

    2010-06-25

    Mitochondria are the principal producers of energy in cells of higher organisms. It was recently reported that mutations and defects in mitochondrial DNA (mtDNA) are associated with various mitochondrial diseases including a variety of neurodegenerative and neuromuscular diseases. Therefore, an effective mitochondrial gene therapy and diagnosis would be expected to have great medical benefits. To achieve this, therapeutic agents need to be delivered into the innermost mitochondrial space (mitochondrial matrix), which contains the mtDNA pool. We previously reported on the development of MITO-Porter, a liposome-based carrier that introduces macromolecular cargos into mitochondria via membrane fusion. In this study, we provide a demonstration of mitochondrial matrix delivery and the visualization of mitochondrial genes (mtDNA) in living cells using the MITO-Porter. We first prepared MITO-Porter containing encapsulated propidium iodide (PI), a fluorescent dye used to stain nucleic acids to detect mtDNA. We then confirmed the emission of red-fluorescence from PI by conjugation with mtDNA, when the carriers were incubated in the presence of isolated rat liver mitochondria. Finally, intracellular observation by confocal laser scanning microscopy clearly verified that the MITO-Porter delivered PI to the mitochondrial matrix.

  12. Mitochondrial matrix delivery using MITO-Porter, a liposome-based carrier that specifies fusion with mitochondrial membranes

    International Nuclear Information System (INIS)

    Yasuzaki, Yukari; Yamada, Yuma; Harashima, Hideyoshi

    2010-01-01

    Mitochondria are the principal producers of energy in cells of higher organisms. It was recently reported that mutations and defects in mitochondrial DNA (mtDNA) are associated with various mitochondrial diseases including a variety of neurodegenerative and neuromuscular diseases. Therefore, an effective mitochondrial gene therapy and diagnosis would be expected to have great medical benefits. To achieve this, therapeutic agents need to be delivered into the innermost mitochondrial space (mitochondrial matrix), which contains the mtDNA pool. We previously reported on the development of MITO-Porter, a liposome-based carrier that introduces macromolecular cargos into mitochondria via membrane fusion. In this study, we provide a demonstration of mitochondrial matrix delivery and the visualization of mitochondrial genes (mtDNA) in living cells using the MITO-Porter. We first prepared MITO-Porter containing encapsulated propidium iodide (PI), a fluorescent dye used to stain nucleic acids to detect mtDNA. We then confirmed the emission of red-fluorescence from PI by conjugation with mtDNA, when the carriers were incubated in the presence of isolated rat liver mitochondria. Finally, intracellular observation by confocal laser scanning microscopy clearly verified that the MITO-Porter delivered PI to the mitochondrial matrix.

  13. Catalytic partial oxidation coupled with membrane purification to improve resource and energy efficiency in syngas production.

    Science.gov (United States)

    Iaquaniello, G; Salladini, A; Palo, E; Centi, G

    2015-02-01

    Catalytic partial oxidation coupled with membrane purification is a new process scheme to improve resource and energy efficiency in a well-established and large scale-process like syngas production. Experimentation in a semi industrial-scale unit (20 Nm(3)  h(-1) production) shows that a novel syngas production scheme based on a pre-reforming stage followed by a membrane for hydrogen separation, a catalytic partial oxidation step, and a further step of syngas purification by membrane allows the oxygen-to-carbon ratio to be decreased while maintaining levels of feed conversion. For a total feed conversion of 40 %, for example, the integrated novel architecture reduces oxygen consumption by over 50 %, with thus a corresponding improvement in resource efficiency and an improved energy efficiency and economics, these factors largely depending on the air separation stage used to produce pure oxygen. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  14. High-Efficiency Artificial Photosynthesis Using a Novel Alkaline Membrane Cell

    Science.gov (United States)

    Narayan, Sri; Haines, Brennan; Blosiu, Julian; Marzwell, Neville

    2009-01-01

    A new cell designed to mimic the photosynthetic processes of plants to convert carbon dioxide into carbonaceous products and oxygen at high efficiency, has an improved configuration using a polymer membrane electrolyte and an alkaline medium. This increases efficiency of the artificial photosynthetic process, achieves high conversion rates, permits the use of inexpensive catalysts, and widens the range of products generated by this type of process. The alkaline membrane electrolyte allows for the continuous generation of sodium formate without the need for any additional separation system. The electrolyte type, pH, electrocatalyst type, and cell voltage were found to have a strong effect on the efficiency of conversion of carbon dioxide to formate. Indium electrodes were found to have higher conversion efficiency compared to lead. Bicarbonate electrolyte offers higher conversion efficiency and higher rates than water solutions saturated with carbon dioxide. pH values between 8 and 9 lead to the maximum values of efficiency. The operating cell voltage of 2.5 V, or higher, ensures conversion of the carbon dioxide to formate, although the hydrogen evolution reaction begins to compete strongly with the formate production reaction at higher cell voltages. Formate is produced at indium and lead electrodes at a conversion efficiency of 48 mg of CO2/kilojoule of energy input. This efficiency is about eight times that of natural photosynthesis in green plants. The electrochemical method of artificial photosynthesis is a promising approach for the conversion, separation and sequestration of carbon dioxide for confined environments as in space habitats, and also for carbon dioxide management in the terrestrial context. The heart of the reactor is a membrane cell fabricated from an alkaline polymer electrolyte membrane and catalyst- coated electrodes. This cell is assembled and held in compression in gold-plated hardware. The cathode side of the cell is supplied with carbon

  15. Herpes simplex virus glycoproteins gB and gH function in fusion between the virion envelope and the outer nuclear membrane.

    Science.gov (United States)

    Farnsworth, Aaron; Wisner, Todd W; Webb, Michael; Roller, Richard; Cohen, Gary; Eisenberg, Roselyn; Johnson, David C

    2007-06-12

    Herpesviruses must traverse the nuclear envelope to gain access to the cytoplasm and, ultimately, to exit cells. It is believed that herpesvirus nucleocapsids enter the perinuclear space by budding through the inner nuclear membrane (NM). To reach the cytoplasm these enveloped particles must fuse with the outer NM and the unenveloped capsids then acquire a second envelope in the trans-Golgi network. Little is known about the process by which herpesviruses virions fuse with the outer NM. Here we show that a herpes simplex virus (HSV) mutant lacking both the two putative fusion glycoproteins gB and gH failed to cross the nuclear envelope. Enveloped virions accumulated in the perinuclear space or in membrane vesicles that bulged into the nucleoplasm (herniations). By contrast, mutants lacking just gB or gH showed only minor or no defects in nuclear egress. We concluded that either HSV gB or gH can promote fusion between the virion envelope and the outer NM. It is noteworthy that fusion associated with HSV entry requires the cooperative action of both gB and gH, suggesting that the two types of fusion (egress versus entry) are dissimilar processes.

  16. Vesicle Fusion Mediated by Solanesol-Anchored DNA.

    Science.gov (United States)

    Flavier, Kristina M; Boxer, Steven G

    2017-09-19

    Fusion between two lipid bilayers is one of the central processes in cell biology, playing a key role in endocytosis, exocytosis, and vesicle transport. We have previously developed a model system that uses the hybridization of complementary DNA strands to model the formation of the SNARE four-helix bundle that mediates synaptic vesicle fusion and used it to study vesicle fusion to a tethered lipid bilayer. Using single vesicle assays, 70% of observed fusion events in the DNA-lipid system are arrested at the hemifusion stage, whereas only 5% eventually go to full fusion. This may be because the diglycerol ether that anchors the DNA in the membrane spans only half the bilayer: upon hemifusion and mixing of the outer leaflets, the DNA-lipid is free to diffuse into the target membrane and away from the vesicle. Here, we test the hypothesis that the length of the membrane anchor may impact the outcome by comparing single leaflet-spanning DNA-lipid mediated vesicle fusion with fusion mediated by DNA anchored by solanesol, a C45 isoprenoid of sufficient length to span the bilayer. When the solanesol anchor was present on the incoming vesicles, target membrane, or both, ∼2-3 times as much full fusion was observed as in the DNA-lipid mediated system, as measured by lipid mixing or content transfer. These results indicate that a transmembrane anchor increases the efficiency of full fusion. Copyright © 2017 Biophysical Society. Published by Elsevier Inc. All rights reserved.

  17. Lipophilic indocarbocyanine conjugates for efficient incorporation of enzymes, antibodies and small molecules into biological membranes.

    Science.gov (United States)

    Smith, Weston J; Tran, Huy; Griffin, James I; Jones, Jessica; Vu, Vivian P; Nilewski, Lizanne; Gianneschi, Nathan; Simberg, Dmitri

    2018-04-01

    Decoration of cell membranes with biomolecules, targeting ligands and imaging agents is an emerging strategy to improve functionality of cell-based therapies. Compared to covalent chemistry or genetic expression on the cell surface, lipid painting (i.e., incorporation of lipid-conjugated molecules into the cell bilayer) is a fast, non-damaging and less expensive approach. Previous studies demonstrated excellent incorporation and retention of distearyl indocarbocyanine dye DiI in membranes of cells in vitro and in vivo. In order to exploit the membrane stability of DiI, we synthesized an amino-DiI derivative, to which we subsequently conjugated an antibody (cetuximab), an enzyme (superoxide dismutase), and a small molecule (DyLight 800). Red blood cells have long been used as drug delivery vehicles so they were utilized as a model to study the incorporation of DiI conjugates in the plasma membrane. All the DiI constructs demonstrated fast and efficient ex vivo incorporation in the membrane of mouse RBCs, resulting in millions of exogenous molecules per RBC. Following an intravenous injection into mice, the molecules were detected on circulating RBCs for several days. DiI anchored molecules showed longer residence time in blood and significantly higher area under the curve (AUC) compared to free non-conjugated molecules. Thus, cetuximab, SOD and DyLight painted on RBC showed 5.5-fold, 6.5-fold and 78-fold increase in the AUC, respectively, compared to the non-modified molecules. Lipophilic indocarbocyanine anchors are a promising technology for incorporation of biomolecules and small molecules into biological membranes for in vivo applications. Copyright © 2018 Elsevier Ltd. All rights reserved.

  18. TRAIL death receptor 4 signaling via lysosome fusion and membrane raft clustering in coronary arterial endothelial cells: evidence from ASM knockout mice.

    Science.gov (United States)

    Li, Xiang; Han, Wei-Qing; Boini, Krishna M; Xia, Min; Zhang, Yang; Li, Pin-Lan

    2013-01-01

    Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and its receptor, death receptor 4 (DR4), have been implicated in the development of endothelial dysfunction and atherosclerosis. However, the signaling mechanism mediating DR4 activation leading to endothelial injury remains unclear. We recently demonstrated that ceramide production via hydrolysis of membrane sphingomyelin by acid sphingomyelinase (ASM) results in membrane raft (MR) clustering and the formation of important redox signaling platforms, which play a crucial role in amplifying redox signaling in endothelial cells leading to endothelial dysfunction. The present study aims to investigate whether TRAIL triggers MR clustering via lysosome fusion and ASM activation, thereby conducting transmembrane redox signaling and changing endothelial function. Using confocal microscopy, we found that TRAIL induced MR clustering and co-localized with DR4 in coronary arterial endothelial cells (CAECs) isolated from wild-type (Smpd1 (+/+)) mice. Furthermore, TRAIL triggered ASM translocation, ceramide production, and NADPH oxidase aggregation in MR clusters in Smpd1 ( +/+ ) CAECs, whereas these observations were not found in Smpd1 (-/-) CAECs. Moreover, ASM deficiency reduced TRAIL-induced O(2) (-[Symbol: see text]) production in CAECs and abolished TRAIL-induced impairment on endothelium-dependent vasodilation in small resistance arteries. By measuring fluorescence resonance energy transfer, we found that Lamp-1 (lysosome membrane marker protein) and ganglioside G(M1) (MR marker) were trafficking together in Smpd1 (+/+) CAECs, which was absent in Smpd1 (-/-) CAECs. Consistently, fluorescence imaging of living cells with specific lysosome probes demonstrated that TRAIL-induced lysosome fusion with membrane was also absent in Smpd1 (-/-) CAECs. Taken together, these results suggest that ASM is essential for TRAIL-induced lysosomal trafficking, membrane fusion and formation of MR redox signaling platforms

  19. Sulfated polysaccharide, curdlan sulfate, efficiently prevents entry/fusion and restricts antibody-dependent enhancement of dengue virus infection in vitro: a possible candidate for clinical application.

    Directory of Open Access Journals (Sweden)

    Koji Ichiyama

    Full Text Available Curdlan sulfate (CRDS, a sulfated 1→3-β-D glucan, previously shown to be a potent HIV entry inhibitor, is characterized in this study as a potent inhibitor of the Dengue virus (DENV. CRDS was identified by in silico blind docking studies to exhibit binding potential to the envelope (E protein of the DENV. CRDS was shown to inhibit the DENV replication very efficiently in different cells in vitro. Minimal effective concentration of CRDS was as low as 0.1 µg/mL in LLC-MK2 cells, and toxicity was observed only at concentrations over 10 mg/mL. CRDS can also inhibit DENV-1, 3, and 4 efficiently. CRDS did not inhibit the replication of DENV subgenomic replicon. Time of addition experiments demonstrated that the compound not only inhibited viral infection at the host cell binding step, but also at an early post-attachment step of entry (membrane fusion. The direct binding of CRDS to DENV was suggested by an evident reduction in the viral titers after interaction of the virus with CRDS following an ultrafiltration device separation, as well as after virus adsorption to an alkyl CRDS-coated membrane filter. The electron microscopic features also showed that CRDS interacted directly with the viral envelope, and caused changes to the viral surface. CRDS also potently inhibited DENV infection in DC-SIGN expressing cells as well as the antibody-dependent enhancement of DENV-2 infection. Based on these data, a probable binding model of CRDS to DENV E protein was constructed by a flexible receptor and ligand docking study. The binding site of CRDS was predicted to be at the interface between domains II and III of E protein dimer, which is unique to this compound, and is apparently different from the β-OG binding site. Since CRDS has already been tested in humans without serious side effects, its clinical application can be considered.

  20. Graphene Oxide Membranes with Heterogeneous Nanodomains for Efficient CO2 Separations.

    Science.gov (United States)

    Wang, Shaofei; Xie, Yu; He, Guangwei; Xin, Qingping; Zhang, Jinhui; Yang, Leixin; Li, Yifan; Wu, Hong; Zhang, Yuzhong; Guiver, Michael D; Jiang, Zhongyi

    2017-11-06

    Achieving high membrane performance in terms of gas permeance and carbon dioxide selectivity is an important target in carbon capture. Aiming to manipulate the channel affinity towards CO 2 to implement efficient separations, gas separation membranes containing CO 2 -philic and non-CO 2 -philic nanodomains in the interlayer channels of graphene oxide (GO) were formed by intercalating poly(ethylene glycol) diamines (PEGDA). PEGDA reacts with epoxy groups on the GO surface, constructing CO 2 -philic nanodomains and rendering a high sorption capacity, whereas unreacted GO surfaces give non-CO 2 -philic nanodomains, rendering low-friction diffusion. Owing to the orderly stacking of nanochannels through cross-linking and the heterogeneous nanodomains with moderate CO 2 affinity, a GO-PEGDA500 membrane exhibits a high CO 2 permeance of 175.5 GPU and a CO 2 /CH 4 selectivity of 69.5, which is the highest performance reported for dry-state GO-stacking membranes. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  1. Glass microporous fiber/nanoporous polytetrafluoroethene composite membranes for high efficient phosphoric acid fuel cell

    International Nuclear Information System (INIS)

    Lu, Chia-Lien; Lee, Wei-Jia; Tseng, Fan-Gang; Chang, Cheng-Ping

    2014-01-01

    This paper reports a high efficient phosphoric acid fuel cell by employing a micro/nano composite proton exchange membrane incorporating glass microfiber (GMF) sealed by polytetrafluoroethylene (PTFE) nano-porous film. This multilayer membrane not only possesses both thermal and chemical stability at phosphoric acid fuel cell working temperature at 150∼220°C but also is cost effective. As a result, the inclusion of the high porosity and proton conductivity from glass microfiber and the prevention of phosphoric acid leakage from PTEF nano film can be achieved at the same time.The composite membrane maximum proton conductivity achieves 0.71 S/cm at 150 °C from AC impedance analysis, much higher than common phosphoric acid porous membranes For single cell test, The GMF fuel cell provides a 63.6mW/cm2 power density at 200mA/cm2 current density while GMF plus methanol treated PTFE (GMF+mPTFE) provides 59.2mW/cm2 power density at 160mA/cm2 current density for hydrogen and oxygen supply at 150 °C. When we change the electrodes that are more suited for phosphoric acid fuel cell, the GMF+mPTFE single cell gets higher performance which achieve 296mW/cm2 power density at 900mA/cm2 current density for hydrogen and oxygen supply at 150 °C

  2. Boron removal efficiency from Red Sea water using different SWRO/BWRO membranes

    KAUST Repository

    Rahmawati, Karina

    2012-12-01

    Seawater reverse osmosis (SWRO) desalination process provides high quality of fresh water. However, due to some operational constraints mainly scaling control some trace contaminant removal, such as acceptable boron concentration, cannot be achieved in a single pass SWRO system. The objective of this study was to investigate the efficiency of five difference reverse osmosis (RO) membranes (seawater SW and brackish water BW) provided by different manufacturers for boron removal. RO experiments using pretreated real Red Sea water were conducted in parallel to compare membrane performance under the same operating conditions. As expected, results showed that boron rejection increased as the feed pH increased. This was caused by dissociation of boric acid to negatively charged borate ions and more negatively charged membrane surface at elevated pH which enhanced boron rejection. Single pass RO system, with and without elevating the pH, may not be sufficient for two reasons. First, boron concentration in permeate does not fulfill local regulations (<0.5ppm). Second, severe scaling occurs due to operation in alkaline condition, since Ca 2+ and Mg 2+ concentrations are still high to cause salts precipitation. Techno-economical study was performed to select the best configuration and membrane giving the highest performance in terms of boron and TDS rejections and energy consumption. © 2012 Elsevier B.V.

  3. Efficient synthesis of tension modulation in strings and membranes based on energy estimation.

    Science.gov (United States)

    Avanzini, Federico; Marogna, Riccardo; Bank, Balázs

    2012-01-01

    String and membrane vibrations cannot be considered as linear above a certain amplitude due to the variation in string or membrane tension. A relevant special case is when the tension is spatially constant and varies in time only in dependence of the overall string length or membrane surface. The most apparent perceptual effect of this tension modulation phenomenon is the exponential decay of pitch in time. Pitch glides due to tension modulation are an important timbral characteristic of several musical instruments, including the electric guitar and tom-tom drum, and many ethnic instruments. This paper presents a unified formulation to the tension modulation problem for one-dimensional (1-D) (string) and two-dimensional (2-D) (membrane) cases. In addition, it shows that the short-time average of the tension variation, which is responsible for pitch glides, is approximately proportional to the system energy. This proportionality allows the efficient physics-based sound synthesis of pitch glides. The proposed models require only slightly more computational resources than linear models as opposed to earlier tension-modulated models of higher complexity. © 2012 Acoustical Society of America.

  4. Phase-Separated Liposomes Enhance the Efficiency of Macromolecular Delivery to the Cellular Cytoplasm.

    Science.gov (United States)

    Imam, Zachary I; Kenyon, Laura E; Ashby, Grant; Nagib, Fatema; Mendicino, Morgan; Zhao, Chi; Gadok, Avinash K; Stachowiak, Jeanne C

    2017-10-01

    From viruses to organelles, fusion of biological membranes is used by diverse biological systems to deliver macromolecules across membrane barriers. Membrane fusion is also a potentially efficient mechanism for the delivery of macromolecular therapeutics to the cellular cytoplasm. However, a key shortcoming of existing fusogenic liposomal systems is that they are inefficient, requiring a high concentration of fusion-promoting lipids in order to cross cellular membrane barriers. Toward addressing this limitation, our experiments explore the extent to which membrane fusion can be amplified by using the process of lipid membrane phase separation to concentrate fusion-promoting lipids within distinct regions of the membrane surface. We used confocal fluorescence microscopy to investigate the integration of fusion-promoting lipids into a ternary lipid membrane system that separated into liquid-ordered and liquid-disordered membrane phases. Additionally, we quantified the impact of membrane phase separation on the efficiency with which liposomes transferred lipids and encapsulated macromolecules to cells, using a combination of confocal fluorescence imaging and flow cytometry. Here we report that concentrating fusion-promoting lipids within phase-separated lipid domains on the surfaces of liposomes significantly increases the efficiency of liposome fusion with model membranes and cells. In particular, membrane phase separation enhanced the delivery of lipids and model macromolecules to the cytoplasm of tumor cells by at least 4-fold in comparison to homogenous liposomes. Our findings demonstrate that phase separation can enhance membrane fusion by locally concentrating fusion-promoting lipids on the surface of liposomes. This work represents the first application of lipid membrane phase separation in the design of biomaterials-based delivery systems. Additionally, these results lay the ground work for developing fusogenic liposomes that are triggered by physical and

  5. Efficient and reusable polyamide-56 nanofiber/nets membrane with bimodal structures for air filtration.

    Science.gov (United States)

    Liu, Bowen; Zhang, Shichao; Wang, Xueli; Yu, Jianyong; Ding, Bin

    2015-11-01

    Nanofibrous media that both possess high airborne particle interception efficiency and robust air permeability would have broad technological implications for areas ranging from individual protection and industrial security to environmental governance; however, creating such filtration media has proved extremely challenging. Here we report a strategy to construct the bio-based polyamide-56 nanofiber/nets (PA-56 NFN) membranes with bimodal structures for effective air filtration via one-step electrospinning/netting. The PA-56 membranes are composed of completely covered two-dimensional (2D) ultrathin (∼20 nm) nanonets which are optimized by facilely regulating the solution concentration, and the bonded scaffold fibers constructed cavity structures which are synchronously created by using the CH3COOH inspiration. With integrated properties of small aperture, high porosity, and bonded scaffold, the resulting PA-56 NFN membranes exhibit high filtration efficiency of 99.995%, low pressure drop of 111 Pa, combined with large dust holding capacity of 49 g/m(2) and dust-cleaning regeneration ability, for filtrating ultrafine airborne particles in the most safe manner involving sieving principle and surface filtration. The successful synthesis of PA-56 NFN medium would not only make it a promising candidate for air filtration, but also provide new insights into the design and development of nanonet-based bimodal structures for various applications. Copyright © 2015 Elsevier Inc. All rights reserved.

  6. pH Optimum of Hemagglutinin-Mediated Membrane Fusion Determines Sensitivity of Influenza A Viruses to the Interferon-Induced Antiviral State and IFITMs.

    Science.gov (United States)

    Gerlach, Thomas; Hensen, Luca; Matrosovich, Tatyana; Bergmann, Janina; Winkler, Michael; Peteranderl, Christin; Klenk, Hans-Dieter; Weber, Friedemann; Herold, Susanne; Pöhlmann, Stefan; Matrosovich, Mikhail

    2017-06-01

    The replication and pathogenicity of influenza A viruses (IAVs) critically depend on their ability to tolerate the antiviral interferon (IFN) response. To determine a potential role for the IAV hemagglutinin (HA) in viral sensitivity to IFN, we studied the restriction of IAV infection in IFN-β-treated human epithelial cells by using 2:6 recombinant IAVs that shared six gene segments of A/Puerto Rico/8/1934 virus (PR8) and contained HAs and neuraminidases of representative avian, human, and zoonotic H5N1 and H7N9 viruses. In A549 and Calu-3 cells, viruses displaying a higher pH optimum of HA-mediated membrane fusion, H5N1-PR8 and H7N9-PR8, were less sensitive to the IFN-induced antiviral state than their counterparts with HAs from duck and human viruses, which fused at a lower pH. The association between a high pH optimum of fusion and reduced IFN sensitivity was confirmed by using HA point mutants of A/Hong Kong/1/1968-PR8 that differed solely by their fusion properties. Furthermore, similar effects of the viral fusion pH on IFN sensitivity were observed in experiments with (i) primary human type II alveolar epithelial cells and differentiated cultures of human airway epithelial cells, (ii) nonrecombinant zoonotic and pandemic IAVs, and (iii) preparations of IFN-α and IFN-λ1. A higher pH of membrane fusion and reduced sensitivity to IFN correlated with lower restriction of the viruses in MDCK cells stably expressing the IFN-inducible transmembrane proteins IFITM2 and IFITM3, which are known to inhibit viral fusion. Our results reveal that the pH optimum of HA-driven membrane fusion of IAVs is a determinant of their sensitivity to IFN and IFITM proteins. IMPORTANCE The IFN system constitutes an important innate defense against viral infection. Substantial information is available on how IAVs avoid detection by sensors of the IFN system and disable IFN signaling pathways. Much less is known about the ability of IAVs to tolerate the antiviral activity of IFN

  7. Experimental evaluation of enthalpy efficiency and gas-phase contaminant transfer in an enthalpy recovery unit with polymer membrane foils

    DEFF Research Database (Denmark)

    Nie, Jinzhe; Yang, Jianrong; Fang, Lei

    2015-01-01

    Experimental studies were conducted in a laboratory setting to investigate the enthalpy efficiency and gas-phase contaminant transfer in a polymer membrane enthalpy recovery unit. One commercially available polymer membrane enthalpy recovery unit was used as a reference unit. Simulated indoor air...

  8. Novel antimony doped tin oxide/carbon aerogel as efficient electrocatalytic filtration membrane

    Directory of Open Access Journals (Sweden)

    Zhimeng Liu

    2016-05-01

    Full Text Available A facile method was developed to prepare antimony doped tin oxide (Sb-SnO2/carbon aerogel (CA for use as an electrocatalytic filtration membrane. The preparation process included synthesis of a precursor sol, impregnation, and thermal decomposition. The Sb-SnO2, which was tetragonal in phase with an average crystallite size of 10.8 nm, was uniformly distributed on the CA surface and firmly attached via carbon-oxygen-tin chemical bonds. Preliminary filtration tests indicated that the Sb-SnO2/CA membrane had a high rate of total organic carbon removal for aqueous tetracycline owing to its high current efficiency and electrode stability.

  9. Flagellar membrane fusion and protein exchange in trypanosomes; a new form of cell-cell communication? [version 1; referees: 2 approved

    Directory of Open Access Journals (Sweden)

    Simon Imhof

    2016-04-01

    Full Text Available Diverse structures facilitate direct exchange of proteins between cells, including plasmadesmata in plants and tunnelling nanotubes in bacteria and higher eukaryotes.  Here we describe a new mechanism of protein transfer, flagellar membrane fusion, in the unicellular parasite Trypanosoma brucei. When fluorescently tagged trypanosomes were co-cultured, a small proportion of double-positive cells were observed. The formation of double-positive cells was dependent on the presence of extracellular calcium and was enhanced by placing cells in medium supplemented with fresh bovine serum. Time-lapse microscopy revealed that double-positive cells arose by bidirectional protein exchange in the absence of nuclear transfer.  Furthermore, super-resolution microscopy showed that this process occurred in ≤1 minute, the limit of temporal resolution in these experiments. Both cytoplasmic and membrane proteins could be transferred provided they gained access to the flagellum. Intriguingly, a component of the RNAi machinery (Argonaute was able to move between cells, raising the possibility that small interfering RNAs are transported as cargo. Transmission electron microscopy showed that shared flagella contained two axonemes and two paraflagellar rods bounded by a single membrane. In some cases flagellar fusion was partial and interactions between cells were transient. In other cases fusion occurred along the entire length of the flagellum, was stable for several hours and might be irreversible. Fusion did not appear to be deleterious for cell function: paired cells were motile and could give rise to progeny while fused. The motile flagella of unicellular organisms are related to the sensory cilia of higher eukaryotes, raising the possibility that protein transfer between cells via cilia or flagella occurs more widely in nature.

  10. Membrane-assisted vapor stripping: energy efficient hybrid distillation-vapor permeation process for alcohol-water separation

    Science.gov (United States)

    BACKGROUND: Energy efficient alternatives to distillation for alcohol recovery from dilute solution are needed to improve biofuel sustainability. A process integrating steam stripping with a vapor compression step and a vapor permeation membrane separation step is proposed. The...

  11. Evidence for a conserved inhibitory binding mode between the membrane fusion assembly factors Munc18 and syntaxin in animals.

    Science.gov (United States)

    Morey, Czuee; Kienle, C Nickias; Klöpper, Tobias H; Burkhardt, Pawel; Fasshauer, Dirk

    2017-12-15

    The membrane fusion necessary for vesicle trafficking is driven by the assembly of heterologous SNARE proteins orchestrated by the binding of Sec1/Munc18 (SM) proteins to specific syntaxin SNARE proteins. However, the precise mode of interaction between SM proteins and SNAREs is debated, as contrasting binding modes have been found for different members of the SM protein family, including the three vertebrate Munc18 isoforms. While different binding modes could be necessary, given their roles in different secretory processes in different tissues, the structural similarity of the three isoforms makes this divergence perplexing. Although the neuronal isoform Munc18a is well-established to bind tightly to both the closed conformation and the N-peptide of syntaxin 1a, thereby inhibiting SNARE complex formation, Munc18b and -c, which have a more widespread distribution, are reported to mainly interact with the N-peptide of their partnering syntaxins and are thought to instead promote SNARE complex formation. We have reinvestigated the interaction between Munc18c and syntaxin 4 (Syx4). Using isothermal titration calorimetry, we found that Munc18c, like Munc18a, binds to both the closed conformation and the N-peptide of Syx4. Furthermore, using a novel kinetic approach, we found that Munc18c, like Munc18a, slows down SNARE complex formation through high-affinity binding to syntaxin. This strongly suggests that secretory Munc18s in general control the accessibility of the bound syntaxin, probably preparing it for SNARE complex assembly. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  12. Efficient hydrogen isotopologues separation through a tunable potential barrier: The case of a C2N membrane

    OpenAIRE

    Qu, Yuanyuan; Li, Feng; Zhao, Mingwen

    2017-01-01

    Isotopes separation through quantum sieving effect of membranes is quite promising for industrial applications. For the light hydrogen isotopologues (eg. H2, D2), the confinement of potential wells in porous membranes to isotopologues was commonly regarded to be crucial for highly efficient separation ability. Here, we demonstrate from first-principles that a potential barrier is also favorable for efficient hydrogen isotopologues separation. Taking an already-synthesized two-dimensional carb...

  13. Rab3A is a new interacting partner of synaptotagmin I and may modulate synaptic membrane fusion through a competitive mechanism

    Energy Technology Data Exchange (ETDEWEB)

    Xie, Chunliang [Key Laboratory of Protein Chemistry and Developmental Biology of Ministry of Education, College of Life Sciences, Hunan Normal University, Changsha 410081 (China); Institute of Bast Fiber Crops, Chinese Academy of Agricultural Sciences, Changsha 410205 (China); Li, Jianglin; Guo, Tianyao; Yan, Yizhong; Tang, Cheng; Wang, Ying; Chen, Ping [Key Laboratory of Protein Chemistry and Developmental Biology of Ministry of Education, College of Life Sciences, Hunan Normal University, Changsha 410081 (China); Wang, Xianchun, E-mail: wang_xianchun@263.net [Key Laboratory of Protein Chemistry and Developmental Biology of Ministry of Education, College of Life Sciences, Hunan Normal University, Changsha 410081 (China); Liang, Songping, E-mail: liangsp@hunnu.edu.cn [Key Laboratory of Protein Chemistry and Developmental Biology of Ministry of Education, College of Life Sciences, Hunan Normal University, Changsha 410081 (China)

    2014-02-21

    Highlights: • Rab3A has been found to be a novel interacting protein of synaptotagmin I. • Rab3A binds to synaptotagmin I in a Ca{sup 2+}-independent manner. • KKKK motif in C2B domain of synaptotagmin I is a key site for Rab3A binding. • Rab3A competitively inhibits the binding of C2B in synaptotagmin I to syntaxin 1B. • Rab3A may regulate synaptic membrane fusion and exocytosis in a competitive manner. - Abstract: Rab3 and synaptotagmin have been reported to be the key proteins that have opposite actions but cooperatively play critical regulatory roles in selecting and limiting the number of vesicles released at central synapses. However, the exact mechanism has not been fully understood. In this study, Rab3A and synaptotagmin I, the most abundant isoforms of Rab3 and synaptotagmin, respectively, in brain were for the first time demonstrated to directly interact with each other in a Ca{sup 2+}-independent manner, and the KKKK motif in the C2B domain of synaptotagmin I was a key site for the Rab3A binding, which was further confirmed by the competitive inhibition of inositol hexakisphosphate. Further studies demonstrated that Rab3A competitively affected the synaptotagmin I interaction with syntaxin 1B that was involved in membrane fusion during the synaptic vesicle exocytosis. These data indicate that Rab3A is a new synaptotagmin I interacting partner and may participate in the regulation of synaptic membrane fusion and thus the vesicle exocytosis by competitively modulating the interaction of synaptotagmin with syntaxin of the t-SNARE complex in presynaptic membranes.

  14. Modeling and operation optimization of a proton exchange membrane fuel cell system for maximum efficiency

    International Nuclear Information System (INIS)

    Han, In-Su; Park, Sang-Kyun; Chung, Chang-Bock

    2016-01-01

    Highlights: • A proton exchange membrane fuel cell system is operationally optimized. • A constrained optimization problem is formulated to maximize fuel cell efficiency. • Empirical and semi-empirical models for most system components are developed. • Sensitivity analysis is performed to elucidate the effects of major operating variables. • The optimization results are verified by comparison with actual operation data. - Abstract: This paper presents an operation optimization method and demonstrates its application to a proton exchange membrane fuel cell system. A constrained optimization problem was formulated to maximize the efficiency of a fuel cell system by incorporating practical models derived from actual operations of the system. Empirical and semi-empirical models for most of the system components were developed based on artificial neural networks and semi-empirical equations. Prior to system optimizations, the developed models were validated by comparing simulation results with the measured ones. Moreover, sensitivity analyses were performed to elucidate the effects of major operating variables on the system efficiency under practical operating constraints. Then, the optimal operating conditions were sought at various system power loads. The optimization results revealed that the efficiency gaps between the worst and best operation conditions of the system could reach 1.2–5.5% depending on the power output range. To verify the optimization results, the optimal operating conditions were applied to the fuel cell system, and the measured results were compared with the expected optimal values. The discrepancies between the measured and expected values were found to be trivial, indicating that the proposed operation optimization method was quite successful for a substantial increase in the efficiency of the fuel cell system.

  15. The fusion of biology, computer science, and engineering: towards efficient and successful synthetic biology.

    Science.gov (United States)

    Linshiz, Gregory; Goldberg, Alex; Konry, Tania; Hillson, Nathan J

    2012-01-01

    Synthetic biology is a nascent field that emerged in earnest only around the turn of the millennium. It aims to engineer new biological systems and impart new biological functionality, often through genetic modifications. The design and construction of new biological systems is a complex, multistep process, requiring multidisciplinary collaborative efforts from "fusion" scientists who have formal training in computer science or engineering, as well as hands-on biological expertise. The public has high expectations for synthetic biology and eagerly anticipates the development of solutions to the major challenges facing humanity. This article discusses laboratory practices and the conduct of research in synthetic biology. It argues that the fusion science approach, which integrates biology with computer science and engineering best practices, including standardization, process optimization, computer-aided design and laboratory automation, miniaturization, and systematic management, will increase the predictability and reproducibility of experiments and lead to breakthroughs in the construction of new biological systems. The article also discusses several successful fusion projects, including the development of software tools for DNA construction design automation, recursive DNA construction, and the development of integrated microfluidics systems.

  16. Inner-membrane proteins PMI/TMEM11 regulate mitochondrial morphogenesis independently of the DRP1/MFN fission/fusion pathways.

    Science.gov (United States)

    Rival, Thomas; Macchi, Marc; Arnauné-Pelloquin, Laetitia; Poidevin, Mickael; Maillet, Frédéric; Richard, Fabrice; Fatmi, Ahmed; Belenguer, Pascale; Royet, Julien

    2011-03-01

    Mitochondria are highly dynamic organelles that can change in number and morphology during cell cycle, development or in response to extracellular stimuli. These morphological dynamics are controlled by a tight balance between two antagonistic pathways that promote fusion and fission. Genetic approaches have identified a cohort of conserved proteins that form the core of mitochondrial remodelling machineries. Mitofusins (MFNs) and OPA1 proteins are dynamin-related GTPases that are required for outer- and inner-mitochondrial membrane fusion respectively whereas dynamin-related protein 1 (DRP1) is the master regulator of mitochondrial fission. We demonstrate here that the Drosophila PMI gene and its human orthologue TMEM11 encode mitochondrial inner-membrane proteins that regulate mitochondrial morphogenesis. PMI-mutant cells contain a highly condensed mitochondrial network, suggesting that PMI has either a pro-fission or an anti-fusion function. Surprisingly, however, epistatic experiments indicate that PMI shapes the mitochondria through a mechanism that is independent of drp1 and mfn. This shows that mitochondrial networks can be shaped in higher eukaryotes by at least two separate pathways: one PMI-dependent and one DRP1/MFN-dependent.

  17. Gallium nitride electrodes for membrane-based electrochemical biosensors.

    Science.gov (United States)

    Schubert, T; Steinhoff, G; von Ribbeck, H-G; Stutzmannn, M; Eickhoff, M; Tanaka, M

    2009-10-01

    We report on the deposition of planar lipid bilayers (supported membranes) on gallium nitride (GaN) electrodes for potential applications as membrane-based biosensors. The kinetics of the lipid membrane formation upon vesicle fusion were monitored by simultaneous measurements of resistance and capacitance of the membrane using AC impedance spectroscopy in the frequency range between 50 mHz and 50 kHz. We could identify a two-step process of membrane spreading and self-healing. Despite its relatively low resistance, the membrane can be modeled by a parallel combination of an ideal resistor and capacitor, indicating that the membrane efficiently blocks the diffusion of ions.

  18. Zipping into fusion

    NARCIS (Netherlands)

    Zheng, Tingting

    2014-01-01

    Fusion of lipid bilayers in cells facilitates the active transport of chemicals. Non-viral membrane fusion is regulated by a cascade of proteins as the process is highly regulated both in space and time. In eukaryotic cells, the so-called SNARE protein complex is at the heart of fusion. However,

  19. SIPHORE: Conceptual Study of a High Efficiency Neutral Beam Injector Based on Photo-detachment for Future Fusion Reactors

    Science.gov (United States)

    Simonin, A.; Christin, L.; de Esch, H.; Garibaldi, P.; Grand, C.; Villecroze, F.; Blondel, C.; Delsart, C.; Drag, C.; Vandevraye, M.; Brillet, A.; Chaibi, W.

    2011-09-01

    An innovative high efficiency neutral beam injector concept for future fusion reactors is under investigation (simulation and R&D) between several laboratories in France, the goal being to perform a feasibility study for the neutralization of intense high energy (1 MeV) negative ion (NI) beams by photo-detachment. The objective of the proposed project is to put together the expertise of three leading groups in negative ion quantum physics, high power stabilized lasers and neutral beam injectors to perform studies of a new injector concept called SIPHORE (SIngle gap PHOto-neutralizer energy REcovery injector), based on the photo-detachment of negative ions and energy recovery of unneutralised ions; the main feature of SIPHORE being the relevance for the future Fusion reactors (DEMO), where high injector efficiency (up to 70-80%), technological simplicity and cost reduction are key issues to be addressed. The paper presents the on-going developments and simulations around this project, such as, a new concept of ion source which would fit with this injector topology and which could solve the remaining uniformity issue of the large size ion source, and, finally, the presentation of the R&D program in the laboratories (LAC, ARTEMIS) around the photo-neutralization for Siphore.

  20. Efficiency of a Traveling Wave Direct Energy Converter with High-Density Beam for Applications to Aneutronic Fusion Experiments

    Science.gov (United States)

    Tarditi, Alfonso

    2012-03-01

    Due to the appeal of aneutronic fusion, a variety of reactor concepts have been proposed in past. In most cases, to achieve a positive net power balance these reactor concepts rely on a significant re-circulation of the energy produced to maintain a non-equilibrium configuration (unlike ignited plasmas). The availability of a direct conversion process with high efficiency is then critical for determining the feasibility of a reactor (particularly when the ``almost true aneutronic'' reaction like p-^11B is considered). A Traveling Wave Direct Energy Converter (TWDEC, [1]) is considered for the energy conversion of a high-density beam formed by the fusion products (MeV-range α-particles). As in [2], a PIC code is utilized for a realistic beam model. The study is focused on the possibility of obtaining high-efficiency coupling between a modulated high-density ``bunched'' beam, accounting also for a neutralizing electron environment, and the TWDEC electrode collector structure.[4pt] [1] Momota et al. (1999) Fus. Tech., 35, 60[0pt] [2] Y.Yasaka et al. (2009), Nucl. Fus., 49, 075009

  1. An efficient formulation for linear and geometric non-linear membrane elements

    Directory of Open Access Journals (Sweden)

    Mohammad Rezaiee-Pajand

    Full Text Available Utilizing the straingradient notation process and the free formulation, an efficient way of constructing membrane elements will be proposed. This strategy can be utilized for linear and geometric non-linear problems. In the suggested formulation, the optimization constraints of insensitivity to distortion, rotational invariance and not having parasitic shear error are employed. In addition, the equilibrium equations will be established based on some constraints among the strain states. The authors' technique can easily separate the rigid body motions, and those belong to deformational motions. In this article, a novel triangular element, named SST10, is formulated. This element will be used in several plane problems having irregular mesh and complicated geometry with linear and geometrically nonlinear behavior. The numerical outcomes clearly demonstrate the efficiency of the new formulation.

  2. Pervaporation membrane bioreactor with permeate fractional condensation and mechanical vapor compression for energy efficient ethanol production

    International Nuclear Information System (INIS)

    Fan, Senqing; Xiao, Zeyi; Li, Minghai; Li, Sizhong

    2016-01-01

    Graphical abstract: Pervaporation membrane bioreactor with permeate partial condensation and mechanical vapor compression is developed for an energy efficient ethanol production. - Highlights: • PVMBR-MVC for energy efficient ethanol production. • Process separation factor of 20–44 for ethanol achieved by fractional condensation. • Energy production of 20.25 MJ and hourly energy production of 56.25 kJ/h achieved. • Over 50% of energy saved in PVMBR-MVC compared with PVMBR-LTC. • Integrated heat pump with COP of 7–9 for the energy recovery of the permeate. - Abstract: Improved process separation factor and heat integration are two key issues to increase the energy efficiency of ethanol production in a pervaporation membrane bioreactor (PVMBR). A PVMBR with permeate fractional condensation and mechanical vapor compression was developed for energy efficient ethanol production. A condensation model based on the mass balance and thermodynamic equilibrium in the partial vacuum condenser was developed for predicting the purification performance of the permeate vapor. Three runs of ethanol fermentation-pervaporation experiment were carried out and ethanol concentration of higher than 50 wt% could be achieved in the final condensate, with the separation factor of the process for ethanol increased to 20. Ethanol production could be enhanced in the bioreactor and 17.1 MJ of the energy could be produced in per liter of fermentation broth, owing to 27.0 MJ/kg heating value of the recovered ethanol. Compared with the traditional pervaporation process with low temperature condensation for ethanol production, 50% of the energy would be saved in the process. The energy consumption would be further reduced, if the available energy of the permeate vapor was utilized by integrating the mechanical vapor compression heat pump.

  3. Dimerization Efficiency of Canine Distemper Virus Matrix Protein Regulates Membrane-Budding Activity.

    Science.gov (United States)

    Bringolf, Fanny; Herren, Michael; Wyss, Marianne; Vidondo, Beatriz; Langedijk, Johannes P; Zurbriggen, Andreas; Plattet, Philippe

    2017-08-15

    Paramyxoviruses rely on the matrix (M) protein to orchestrate viral assembly and budding at the plasma membrane. Although the mechanistic details remain largely unknown, structural data suggested that M dimers and/or higher-order oligomers may facilitate membrane budding. To gain functional insights, we employed a structure-guided mutagenesis approach to investigate the role of canine distemper virus (CDV) M protein self-assembly in membrane-budding activity. Three six-alanine-block (6A-block) mutants with mutations located at strategic oligomeric positions were initially designed. While the first one includes residues potentially residing at the protomer-protomer interface, the other two display amino acids located within two distal surface-exposed α-helices proposed to be involved in dimer-dimer contacts. We further focused on the core of the dimeric interface by mutating asparagine 138 (N138) to several nonconservative amino acids. Cellular localization combined with dimerization and coimmunopurification assays, performed under various denaturing conditions, revealed that all 6A-block mutants were impaired in self-assembly and cell periphery accumulation. These phenotypes correlated with deficiencies in relocating CDV nucleocapsid proteins to the cell periphery and in virus-like particle (VLP) production. Conversely, all M-N138 mutants remained capable of self-assembly, though to various extents, which correlated with proper accumulation and redistribution of nucleocapsid proteins at the plasma membrane. However, membrane deformation and VLP assays indicated that the M-N138 variants exhibiting the most reduced dimerization propensity were also defective in triggering membrane remodeling and budding, despite proper plasma membrane accumulation. Overall, our data provide mechanistic evidence that the efficiency of CDV M dimerization/oligomerization governs both cell periphery localization and membrane-budding activity. IMPORTANCE Despite the availability of

  4. Induction of heterosubtypic cross-protection against influenza by a whole inactivated virus vaccine: the role of viral membrane fusion activity.

    Directory of Open Access Journals (Sweden)

    Natalija Budimir

    Full Text Available BACKGROUND: The inability of seasonal influenza vaccines to effectively protect against infection with antigenically drifted viruses or newly emerging pandemic viruses underlines the need for development of cross-reactive influenza vaccines that induce immunity against a variety of virus subtypes. Therefore, potential cross-protective vaccines, e.g., whole inactivated virus (WIV vaccine, that can target conserved internal antigens such as the nucleoprotein (NP and/or matrix protein (M1 need to be explored. METHODOLOGY/PRINCIPAL FINDINGS: In the current study we show that a WIV vaccine, through induction of cross-protective cytotoxic T lymphocytes (CTLs, protects mice from heterosubtypic infection. This protection was abrogated after depletion of CD8+ cells in vaccinated mice, indicating that CTLs were the primary mediators of protection. Previously, we have shown that different procedures used for virus inactivation influence optimal activation of CTLs by WIV, most likely by affecting the membrane fusion properties of the virus. Specifically, inactivation with formalin (FA severely compromises fusion activity of the virus, while inactivation with β-propiolactone (BPL preserves fusion activity. Here, we demonstrate that vaccination of mice with BPL-inactivated H5N1 WIV vaccine induces solid protection from lethal heterosubtypic H1N1 challenge. By contrast, vaccination with FA-inactivated WIV, while preventing death after lethal challenge, failed to protect against development of disease and severe body weight loss. Vaccination with BPL-inactivated WIV, compared to FA-inactivated WIV, induced higher levels of specific CD8+ T cells in blood, spleen and lungs, and a higher production of granzyme B in the lungs upon H1N1 virus challenge. CONCLUSION/SIGNIFICANCE: The results underline the potential use of WIV as a cross-protective influenza vaccine candidate. However, careful choice of the virus inactivation procedure is important to retain membrane

  5. Induction of heterosubtypic cross-protection against influenza by a whole inactivated virus vaccine: the role of viral membrane fusion activity.

    Science.gov (United States)

    Budimir, Natalija; Huckriede, Anke; Meijerhof, Tjarko; Boon, Louis; Gostick, Emma; Price, David A; Wilschut, Jan; de Haan, Aalzen

    2012-01-01

    The inability of seasonal influenza vaccines to effectively protect against infection with antigenically drifted viruses or newly emerging pandemic viruses underlines the need for development of cross-reactive influenza vaccines that induce immunity against a variety of virus subtypes. Therefore, potential cross-protective vaccines, e.g., whole inactivated virus (WIV) vaccine, that can target conserved internal antigens such as the nucleoprotein (NP) and/or matrix protein (M1) need to be explored. In the current study we show that a WIV vaccine, through induction of cross-protective cytotoxic T lymphocytes (CTLs), protects mice from heterosubtypic infection. This protection was abrogated after depletion of CD8+ cells in vaccinated mice, indicating that CTLs were the primary mediators of protection. Previously, we have shown that different procedures used for virus inactivation influence optimal activation of CTLs by WIV, most likely by affecting the membrane fusion properties of the virus. Specifically, inactivation with formalin (FA) severely compromises fusion activity of the virus, while inactivation with β-propiolactone (BPL) preserves fusion activity. Here, we demonstrate that vaccination of mice with BPL-inactivated H5N1 WIV vaccine induces solid protection from lethal heterosubtypic H1N1 challenge. By contrast, vaccination with FA-inactivated WIV, while preventing death after lethal challenge, failed to protect against development of disease and severe body weight loss. Vaccination with BPL-inactivated WIV, compared to FA-inactivated WIV, induced higher levels of specific CD8+ T cells in blood, spleen and lungs, and a higher production of granzyme B in the lungs upon H1N1 virus challenge. The results underline the potential use of WIV as a cross-protective influenza vaccine candidate. However, careful choice of the virus inactivation procedure is important to retain membrane fusion activity and full immunogenicity of the vaccine.

  6. Vertically Aligned Graphene Sheets Membrane for Highly Efficient Solar Thermal Generation of Clean Water.

    Science.gov (United States)

    Zhang, Panpan; Li, Jing; Lv, Lingxiao; Zhao, Yang; Qu, Liangti

    2017-05-23

    Efficient utilization of solar energy for clean water is an attractive, renewable, and environment friendly way to solve the long-standing water crisis. For this task, we prepared the long-range vertically aligned graphene sheets membrane (VA-GSM) as the highly efficient solar thermal converter for generation of clean water. The VA-GSM was prepared by the antifreeze-assisted freezing technique we developed, which possessed the run-through channels facilitating the water transport, high light absorption capacity for excellent photothermal transduction, and the extraordinary stability in rigorous conditions. As a result, VA-GSM has achieved average water evaporation rates of 1.62 and 6.25 kg m -2 h -1 under 1 and 4 sun illumination with a superb solar thermal conversion efficiency of up to 86.5% and 94.2%, respectively, better than that of most carbon materials reported previously, which can efficiently produce the clean water from seawater, common wastewater, and even concentrated acid and/or alkali solutions.

  7. Resource-efficient fusion with pre-compensated transmissions for cooperative spectrum sensing.

    Science.gov (United States)

    Guimarães, Dayan Adionel; Aquino, Guilherme Pedro; Cattaneo, Marco E G V

    2015-05-08

    Recently, a novel fusion scheme for cooperative spectrum sensing was proposed for saving resources in the control channel. Secondary users (SUs) simultaneously report their decisions using binary modulations with the same carrier frequencies. The transmitted symbols add incoherently at the fusion centre (FC), leading to a larger set of symbols in which a subset is associated with the presence of the primary user (PU) signal, and another subset is associated with the absence of such a signal. The decision criterion applied for discriminating these subsets works under the assumption that the channel gains are known at the FC. In this paper, we propose a new simultaneous transmission and decision scheme in which the task of channel estimation is shifted from the FC to the SUs, without the need for feeding-back of the estimates to the FC. The estimates are used at the SUs to pre-compensate for the reporting channel phase rotations and to partially compensate for the channel gains. This partial compensation is the result of signal clipping for peak-to-average power ratio (PAPR) control. We show, analytically and with simulations, that this new scheme can produce large performance improvements, yet reduces the implementation complexity when compared with the original one.

  8. Resource-Efficient Fusion with Pre-Compensated Transmissions for Cooperative Spectrum Sensing

    Directory of Open Access Journals (Sweden)

    Dayan Adionel Guimarães

    2015-05-01

    Full Text Available Recently, a novel fusion scheme for cooperative spectrum sensing was proposed for saving resources in the control channel. Secondary users (SUs simultaneously report their decisions using binary modulations with the same carrier frequencies. The transmitted symbols add incoherently at the fusion centre (FC, leading to a larger set of symbols in which a subset is associated with the presence of the primary user (PU signal, and another subset is associated with the absence of such a signal. The decision criterion applied for discriminating these subsets works under the assumption that the channel gains are known at the FC. In this paper, we propose a new simultaneous transmission and decision scheme in which the task of channel estimation is shifted from the FC to the SUs, without the need for feeding-back of the estimates to the FC. The estimates are used at the SUs to pre-compensate for the reporting channel phase rotations and to partially compensate for the channel gains. This partial compensation is the result of signal clipping for peak-to-average power ratio (PAPR control. We show, analytically and with simulations, that this new scheme can produce large performance improvements, yet reduces the implementation complexity when compared with the original one.

  9. Cell fusion and nuclear fusion in plants.

    Science.gov (United States)

    Maruyama, Daisuke; Ohtsu, Mina; Higashiyama, Tetsuya

    2016-12-01

    Eukaryotic cells are surrounded by a plasma membrane and have a large nucleus containing the genomic DNA, which is enclosed by a nuclear envelope consisting of the outer and inner nuclear membranes. Although these membranes maintain the identity of cells, they sometimes fuse to each other, such as to produce a zygote during sexual reproduction or to give rise to other characteristically polyploid tissues. Recent studies have demonstrated that the mechanisms of plasma membrane or nuclear membrane fusion in plants are shared to some extent with those of yeasts and animals, despite the unique features of plant cells including thick cell walls and intercellular connections. Here, we summarize the key factors in the fusion of these membranes during plant reproduction, and also focus on "non-gametic cell fusion," which was thought to be rare in plant tissue, in which each cell is separated by a cell wall. Copyright © 2016 Elsevier Ltd. All rights reserved.

  10. Distinct Requirements for HIV-Cell Fusion and HIV-mediated Cell-Cell Fusion*

    Science.gov (United States)

    Kondo, Naoyuki; Marin, Mariana; Kim, Jeong Hwa; Desai, Tanay M.; Melikyan, Gregory B.

    2015-01-01

    Whether HIV-1 enters cells by fusing with the plasma membrane or with endosomes is a subject of active debate. The ability of HIV-1 to mediate fusion between adjacent cells, a process referred to as “fusion-from-without” (FFWO), shows that this virus can fuse with the plasma membrane. To compare FFWO occurring at the cell surface with HIV-cell fusion through a conventional entry route, we designed an experimental approach that enabled the measurements of both processes in the same sample. The following key differences were observed. First, a very small fraction of viruses fusing with target cells participated in FFWO. Second, whereas HIV-1 fusion with adherent cells was insensitive to actin inhibitors, post-CD4/coreceptor binding steps during FFWO were abrogated. A partial dependence of HIV-cell fusion on actin remodeling was observed in CD4+ T cells, but this effect appeared to be due to the actin dependence of virus uptake. Third, deletion of the cytoplasmic tail of HIV-1 gp41 dramatically enhanced the ability of the virus to promote FFWO, while having a modest effect on virus-cell fusion. Distinct efficiencies and actin dependences of FFWO versus HIV-cell fusion are consistent with the notion that, except for a minor fraction of particles that mediate fusion between the plasma membranes of adjacent cells, HIV-1 enters through an endocytic pathway. We surmise, however, that cell-cell contacts enabling HIV-1 fusion with the plasma membrane could be favored at the sites of high density of target cells, such as lymph nodes. PMID:25589785

  11. Unusual Self-Assembly of the Recombinant Chlamydia trachomatis Major Outer Membrane Protein-Based Fusion Antigen CTH522 Into Protein Nanoparticles

    DEFF Research Database (Denmark)

    Rose, Fabrice; Karlsen, Kasper; Jensen, Pernille

    2018-01-01

    but is a challenging vaccine candidate by being an integral membrane protein, and the immunogenicity depends on a correctly folded structure. We investigated the biophysical properties of the recombinant MOMP-based fusion antigen CTH522, which is tested in early human clinical trials. It consists of a truncated......Sexually transmitted Chlamydia trachomatis (Ct) infects more than 100 million people annually, and untreated chlamydia infections can cause severe complications. Therefore, there is an urgent need for a chlamydia vaccine. The Ct major outer membrane protein (MOMP) is highly immunogenic......-defined secondary structural elements, and no thermal transitions were measurable. Chemical unfolding resulted monomers that upon removal of the denaturant self-assembled into higher order structures, comparable to the structure of the native protein. The conformation of CTH522 in nanoparticles is thus not entirely...

  12. Efficient source for the production of ultradense deuterium D(-1) for laser-induced fusion (ICF)

    International Nuclear Information System (INIS)

    Andersson, Patrik U.; Loenn, Benny; Holmlid, Leif

    2011-01-01

    A novel source which simplifies the study of ultradense deuterium D(-1) is now described. This means one step further toward deuterium fusion energy production. The source uses internal gas feed and D(-1) can now be studied without time-of-flight spectral overlap from the related dense phase D(1). The main aim here is to understand the material production parameters, and thus a relatively weak laser with focused intensity ≤10 12 W cm -2 is employed for analyzing the D(-1) material. The properties of the D(-1) material at the source are studied as a function of laser focus position outside the emitter, deuterium gas feed, laser pulse repetition frequency and laser power, and temperature of the source. These parameters influence the D(-1) cluster size, the ionization mode, and the laser fragmentation patterns.

  13. Efficient source for the production of ultradense deuterium D(-1) for laser-induced fusion (ICF)

    Science.gov (United States)

    Andersson, Patrik U.; Lönn, Benny; Holmlid, Leif

    2011-01-01

    A novel source which simplifies the study of ultradense deuterium D(-1) is now described. This means one step further toward deuterium fusion energy production. The source uses internal gas feed and D(-1) can now be studied without time-of-flight spectral overlap from the related dense phase D(1). The main aim here is to understand the material production parameters, and thus a relatively weak laser with focused intensity ≤1012 W cm-2 is employed for analyzing the D(-1) material. The properties of the D(-1) material at the source are studied as a function of laser focus position outside the emitter, deuterium gas feed, laser pulse repetition frequency and laser power, and temperature of the source. These parameters influence the D(-1) cluster size, the ionization mode, and the laser fragmentation patterns.

  14. Efficient Error Detection in Soft Data Fusion for Cooperative Spectrum Sensing

    KAUST Repository

    Saqib Bhatti, Dost Muhammad

    2018-03-18

    The primary objective of cooperative spectrum sensing (CSS) is to determine whether a particular spectrum is occupied by a licensed user or not, so that unlicensed users called secondary users (SUs) can utilize that spectrum, if it is not occupied. For CSS, all SUs report their sensing information through reporting channel to the central base station called fusion center (FC). During transmission, some of the SUs are subjected to fading and shadowing, due to which the overall performance of CSS is degraded. We have proposed an algorithm which uses error detection technique on sensing measurement of all SUs. Each SU is required to re-transmit the sensing data to the FC, if error is detected on it. Our proposed algorithm combines the sensing measurement of limited number of SUs. Using Proposed algorithm, we have achieved the improved probability of detection (PD) and throughput. The simulation results compare the proposed algorithm with conventional scheme.

  15. Reconstituted Fusion Pore

    OpenAIRE

    Jeremic, Aleksandar; Kelly, Marie; Cho, Sang-Joon; Stromer, Marvin H.; Jena, Bhanu P.

    2003-01-01

    Fusion pores or porosomes are basket-like structures at the cell plasma membrane, at the base of which, membrane-bound secretory vesicles dock and fuse to release vesicular contents. Earlier studies using atomic force microscopy (AFM) demonstrated the presence of fusion pores at the cell plasma membrane in a number of live secretory cells, revealing their morphology and dynamics at nm resolution and in real time. ImmunoAFM studies demonstrated the release of vesicular contents through the por...

  16. Highly efficient integration of the viral portal proteins from different types of phages into planar bilayers for the black lipid membrane analysis.

    Science.gov (United States)

    Jing, Peng; Paraiso, Hallel; Burris, Benjamin

    2016-02-01

    The planar lipid bilayer technology is a technique that yields incredibly useful structural function information about a single channel protein. It is also currently actively utilized as a powerful platform using biological protein nanopores for the development of single-molecule nanopore sensing technology, as well as ultrafast DNA sequencing technology. The portal protein, GP10, from the bacteriophage Φ29 was the first phage portal protein shown to be successfully inserted into planar bilayer membranes, thereby it may inspire more researchers to apply the techniques to portal proteins from the other bacteriophages. However, the technology is far from perfect since the insertion of the channel proteins into planar bilayer membranes is not only technically difficult but also time-consuming. For the fusion of phage portal proteins, vesicles are typically needed to be reconstituted with the portal proteins to form proteoliposomes. However, most of the phage portal proteins have low solubility, and may self-aggregate during the preparation of the proteoliposomes. Furthermore, the fusion of the formed proteoliposomes is sporadic, unpredictable and varied from person to person. Due to the lack of experimental consistency between labs, the results from different methodologies reported for generating fusible proteoliposomes are highly variable. In this research, we propose a new method for the preparation of the fusible proteoliposomes containing portal proteins from bacteriophages, to circumvent the problems aforementioned. Compared to the conventional methods, this method was able to avoid the protein aggregation issues during the vesicle preparation by eliminating the need for detergents and the subsequent time-consuming step for detergent removal. The proteoliposomes prepared by the method were shown to be more efficiently and rapidly inserted into planar bilayer membranes bathed in different conducting buffer solutions including those with nonelectrolytes such as

  17. Protein fusion tags for efficient expression and purification of recombinant proteins in the periplasmic space of E. coli.

    Science.gov (United States)

    Malik, Ajamaluddin

    2016-06-01

    Disulfide bonds occurred in majority of secreted protein. Formation of correct disulfide bonds are must for achieving native conformation, solubility and activity. Production of recombinant proteins containing disulfide bond for therapeutic, diagnostic and various other purposes is a challenging task of research. Production of such proteins in the reducing cytosolic compartment of E. coli usually ends up in inclusion bodies formation. Refolding of inclusion bodies can be difficult, time and labor consuming and uneconomical. Translocation of these proteins into the oxidative periplasmic compartment provides correct environment to undergo proper disulfide bonds formation and thus achieving native conformation. However, not all proteins can be efficiently translocated to the periplasm with the help of bacterial signal peptides. Therefore, fusion to a small well-folded and stable periplasmic protein is more promising for periplasmic production of disulfide bonded proteins. In the past decades, several full-length proteins or domains were used for enhancing translocation and solubility. Here, protein fusion tags that significantly increase the yields of target proteins in the periplasmic space are reviewed.

  18. Superhydrophilic graphene oxide@electrospun cellulose nanofiber hybrid membrane for high-efficiency oil/water separation.

    Science.gov (United States)

    Ao, Chenghong; Yuan, Wei; Zhao, Jiangqi; He, Xu; Zhang, Xiaofang; Li, Qingye; Xia, Tian; Zhang, Wei; Lu, Canhui

    2017-11-01

    Inspired from fishscales, membranes with special surface wettability have been applied widely for the treatment of oily waste water. Herein, a novel superhydrophilic graphene oxide (GO)@electrospun cellulose nanofiber (CNF) membrane was successfully fabricated. This membrane exhibited a high separation efficiency, excellent antifouling properties, as well as a high flux for the gravity-driven oil/water separation. Moreover, the GO@CNF membrane was capable to effectively separate oil/water mixtures in a broad pH range or with a high concentration of salt, suggesting that this membrane was quite promising for future real-world practice in oil spill cleanup and oily wastewater treatment. Copyright © 2017 Elsevier Ltd. All rights reserved.

  19. Improvement of proton exchange membrane fuel cell overall efficiency by integrating heat-to-electricity conversion

    Science.gov (United States)

    Xie, Chungang; Wang, Shuxin; Zhang, Lianhong; Hu, S. Jack

    Proton exchange membrane fuel cells (PEMFCs) have shown to be well suited for distributed power generation due to their excellent performance. However, a PEMFC produces a considerable amount of heat in the process of electrochemical reaction. It is desirable to use thermal energy for electricity generation in addition to heating applications. Based on the operating characteristics of a PEMFC, an advanced thermal energy conversion system using "ocean thermal energy conversion" (OTEC) technology is applied to exploit the thermal energy of the PEMFC for electricity generation. Through this combination of technology, this unique PEMFC power plant not only achieves the combined heat and power efficiency, but also adequately utilizes heat to generate more valuable electricity. Exergy analysis illustrates the improvement of overall efficiency and energy flow distribution in the power plant. Analytical results show that the overall efficiency of the PEMFC is increased by 0.4-2.3% due to the thermal energy conversion (TEC) system. It is also evident that the PEMFC should operate within the optimal load range by balancing the design parameters of the PEMFC and of the TEC system.

  20. Parallel artificial liquid membrane extraction as an efficient tool for removal of phospholipids from human plasma.

    Science.gov (United States)

    Ask, Kristine Skoglund; Bardakci, Turgay; Parmer, Marthe Petrine; Halvorsen, Trine Grønhaug; Øiestad, Elisabeth Leere; Pedersen-Bjergaard, Stig; Gjelstad, Astrid

    2016-09-10

    Generic Parallel Artificial Liquid Membrane Extraction (PALME) methods for non-polar basic and non-polar acidic drugs from human plasma were investigated with respect to phospholipid removal. In both cases, extractions in 96-well format were performed from plasma (125μL), through 4μL organic solvent used as supported liquid membranes (SLMs), and into 50μL aqueous acceptor solutions. The acceptor solutions were subsequently analysed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) using in-source fragmentation and monitoring the m/z 184→184 transition for investigation of phosphatidylcholines (PC), sphingomyelins (SM), and lysophosphatidylcholines (Lyso-PC). In both generic methods, no phospholipids were detected in the acceptor solutions. Thus, PALME appeared to be highly efficient for phospholipid removal. To further support this, qualitative (post-column infusion) and quantitative matrix effects were investigated with fluoxetine, fluvoxamine, and quetiapine as model analytes. No signs of matrix effects were observed. Finally, PALME was evaluated for the aforementioned drug substances, and data were in accordance with European Medicines Agency (EMA) guidelines. Copyright © 2016 Elsevier B.V. All rights reserved.

  1. Life cycle cost reduction through high efficiency membrane based air intake filters; Reduzierung der Lebensdauerzykluskosten durch hocheffiziente Zuluftfilter auf Membranbasis

    Energy Technology Data Exchange (ETDEWEB)

    Krah, Helmut [W.L. Gore and Associates GmbH, Putzbrunn (Germany)

    2011-07-01

    The use of highly efficient, membrane-based air intake filters means that massive savings can be made in the operation of gas turbines: on the one hand, a higher degree of efficiency can be achieved, which leads to lower fuel consumption and better turbine performance, and on the other, maintenance costs can be reduced thanks to the avoidance of erosion and corrosion. EPA (Efficient Particulate Air) filters based on fibreglass have the disadvantage that they exhibit a relatively high differential pressure, and they can frequently only be used by converting the filter house. This is where the tremendous advantage of membrane-based EPA filters comes in. Its core, a micro-porous PTFE membrane with excellent air permeability. (orig.)

  2. The use of polybenzimidazole membranes in vanadium redox flow batteries leading to increased coulombic efficiency and cycling performance

    International Nuclear Information System (INIS)

    Zhou, X.L.; Zhao, T.S.; An, L.; Wei, L.; Zhang, C.

    2015-01-01

    An issue with conventional vanadium redox flow batteries (VRFB) with Nafion membranes is the crossover of vanadium ions, resulting in low coulombic efficiency and rapid decay in capacity. In this work, a VRFB with a polybenzimidazole (PBI) membrane is tested and compared with the Nafion system. Results show that the PBI-based VRFB exhibits a substantially higher coulombic efficiency of up to 99% at current densities ranging from 20 mA cm −2 to 80 mA cm −2 . More importantly, it is demonstrated that the PBI-based VRFB has a capacity decay rate of as low as 0.3% per cycle, which is four times lower than that of the Nafion system (1.3% per cycle). The improved coulombic efficiency and cycling performance are attributed to the low crossover of vanadium ions through the PBI membrane

  3. Pandemic H1N1 influenza A directly induces a robust and acute inflammatory gene signature in primary human bronchial epithelial cells downstream of membrane fusion

    Energy Technology Data Exchange (ETDEWEB)

    Paquette, Stéphane G. [Division of Experimental Therapeutics, Toronto General Hospital Research Institute, University Health Network, Toronto, Ontario (Canada); Institute of Medical Science, Faculty of Medicine, University of Toronto, Toronto, Ontario (Canada); Banner, David [Division of Experimental Therapeutics, Toronto General Hospital Research Institute, University Health Network, Toronto, Ontario (Canada); Chi, Le Thi Bao [Department of Microbiology, Hue University of Medicine and Pharmacy, Thua Thien Hue (Viet Nam); Carlo Urbani Centre, Hue University of Medicine and Pharmacy, Thua Thien Hue (Viet Nam); Leon, Alberto J. [Division of Experimental Therapeutics, Toronto General Hospital Research Institute, University Health Network, Toronto, Ontario (Canada); International Institute of Infection and Immunity, Shantou University Medical College, Shantou, Guangdong (China); Xu, Luoling; Ran, Longsi [Division of Experimental Therapeutics, Toronto General Hospital Research Institute, University Health Network, Toronto, Ontario (Canada); Huang, Stephen S.H. [Division of Experimental Therapeutics, Toronto General Hospital Research Institute, University Health Network, Toronto, Ontario (Canada); Department of Immunology, Faculty of Medicine, University of Toronto, Toronto, Ontario (Canada); Farooqui, Amber [Division of Experimental Therapeutics, Toronto General Hospital Research Institute, University Health Network, Toronto, Ontario (Canada); International Institute of Infection and Immunity, Shantou University Medical College, Shantou, Guangdong (China); and others

    2014-01-05

    Pandemic H1N1 influenza A (H1N1pdm) elicits stronger pulmonary inflammation than previously circulating seasonal H1N1 influenza A (sH1N1), yet mechanisms of inflammatory activation in respiratory epithelial cells during H1N1pdm infection are unclear. We investigated host responses to H1N1pdm/sH1N1 infection and virus entry mechanisms in primary human bronchial epithelial cells in vitro. H1N1pdm infection rapidly initiated a robust inflammatory gene signature (3 h post-infection) not elicited by sH1N1 infection. Protein secretion inhibition had no effect on gene induction. Infection with membrane fusion deficient H1N1pdm failed to induce robust inflammatory gene expression which was rescued with restoration of fusion ability, suggesting H1N1pdm directly triggered the inflammatory signature downstream of membrane fusion. Investigation of intra-virion components revealed H1N1pdm viral RNA (vRNA) triggered a stronger inflammatory phenotype than sH1N1 vRNA. Thus, our study is first to report H1N1pdm induces greater inflammatory gene expression than sH1N1 in vitro due to direct virus–epithelial cell interaction. - Highlights: • We investigated H1N1pdm/sH1N1 infection in primary epithelial cells. • H1N1pdm directly initiated a robust inflammatory gene signature, sH1N1 did not. • H1N1pdm viral RNA triggered a stronger response than sH1N1. • H1N1pdm induces greater response due to direct virus–cell interaction. • These results have potential to impact vaccine and therapeutic development.

  4. Pandemic H1N1 influenza A directly induces a robust and acute inflammatory gene signature in primary human bronchial epithelial cells downstream of membrane fusion

    International Nuclear Information System (INIS)

    Paquette, Stéphane G.; Banner, David; Chi, Le Thi Bao; Leon, Alberto J.; Xu, Luoling; Ran, Longsi; Huang, Stephen S.H.; Farooqui, Amber

    2014-01-01

    Pandemic H1N1 influenza A (H1N1pdm) elicits stronger pulmonary inflammation than previously circulating seasonal H1N1 influenza A (sH1N1), yet mechanisms of inflammatory activation in respiratory epithelial cells during H1N1pdm infection are unclear. We investigated host responses to H1N1pdm/sH1N1 infection and virus entry mechanisms in primary human bronchial epithelial cells in vitro. H1N1pdm infection rapidly initiated a robust inflammatory gene signature (3 h post-infection) not elicited by sH1N1 infection. Protein secretion inhibition had no effect on gene induction. Infection with membrane fusion deficient H1N1pdm failed to induce robust inflammatory gene expression which was rescued with restoration of fusion ability, suggesting H1N1pdm directly triggered the inflammatory signature downstream of membrane fusion. Investigation of intra-virion components revealed H1N1pdm viral RNA (vRNA) triggered a stronger inflammatory phenotype than sH1N1 vRNA. Thus, our study is first to report H1N1pdm induces greater inflammatory gene expression than sH1N1 in vitro due to direct virus–epithelial cell interaction. - Highlights: • We investigated H1N1pdm/sH1N1 infection in primary epithelial cells. • H1N1pdm directly initiated a robust inflammatory gene signature, sH1N1 did not. • H1N1pdm viral RNA triggered a stronger response than sH1N1. • H1N1pdm induces greater response due to direct virus–cell interaction. • These results have potential to impact vaccine and therapeutic development

  5. Measurements of tokamak fusion test reactor D-T radiation shielding efficiency

    Energy Technology Data Exchange (ETDEWEB)

    Kugel, H.W. [Princeton Univ., NJ (United States). Plasma Physics Lab.; Ascione, G. [Princeton Univ., NJ (United States). Plasma Physics Lab.; Elwood, S. [Princeton Univ., NJ (United States). Plasma Physics Lab.; Gilbert, J. [Princeton Univ., NJ (United States). Plasma Physics Lab.; Ku, L.P. [Princeton Univ., NJ (United States). Plasma Physics Lab.; Levine, J. [Princeton Univ., NJ (United States). Plasma Physics Lab.; Rule, K. [Princeton Univ., NJ (United States). Plasma Physics Lab.; Azziz, N. [USDOE Environmental Measurements Lab., New York (United States); Goldhagen, P. [USDOE Environmental Measurements Lab., New York (United States); Hajnal, F. [USDOE Environmental Measurements Lab., New York (United States); Shebell, P. [USDOE Environmental Measurements Lab., New York (United States)

    1995-03-01

    Measurements of neutron and gamma dose-equivalents were performed in the test cell, at the outer test cell wall, in nearby work areas, and out to the nearest property lines at a distance of 180m. Argon ionization chambers, moderated {sup 3}He proportional counters, and fission chamber detectors were used to obtain measurements of neutron and gamma dose-equivalents per D-T neutron during individual tokamak fusion test reactor (TFTR) discharges. These measured neutron and gamma D-T dose-equivalents per TFTR neutron characterize the effects of local variations in material density resulting from the complex asymmetric site geometry. The measured dose-equivalents per TFTR D-T neutron and the cumulative neutron production were used to determine that the planned annual TFTR neutron production of 1 x 10{sup 21} D-T neutrons is consistent with the design objective of limiting the total dose-equivalent at the property line, from all radiation sources and pathways, to less than 10mrem per year. (orig.).

  6. Review of fusion synfuels

    International Nuclear Information System (INIS)

    Fillo, J.A.

    1980-01-01

    Thermonuclear fusion offers an inexhaustible source of energy for the production of hydrogen from water. Depending on design, electric generation efficiencies of approx. 40 to 60% and hydrogen production efficiencies by high-temperature electrolysis of approx. 50 to 65% are projected for fusion reactors using high-temperatures blankets. Fusion/coal symbiotic systems appear economically promising for the first generation of commercial fusion synfuels plants. Coal production requirements and the environmental effects of large-scale coal usage would be greatly reduced by a fusion/coal system. In the long term, there could be a gradual transition to an inexhaustible energy system based solely on fusion

  7. Highly-efficient forward osmosis membrane tailored by magnetically responsive graphene oxide/Fe3O4 nanohybrid

    Science.gov (United States)

    Rastgar, Masoud; Shakeri, Alireza; Bozorg, Ali; Salehi, Hasan; Saadattalab, Vahid

    2018-05-01

    Emerging forward osmosis (FO) process as a potentially more energy efficient method has recently gained remarkable attention. Herein, considering the unique features of graphene oxide (GO), a new facile method has been proposed to magnetically modify GO within the polyamide active layer to obtain highly efficient osmotically driven membranes. While exposed to magnetic field, thin film nanocomposite membranes modified by GO/Fe3O4 nanohybrids (TFN-MMGO/Fe3O4) were synthesized by in-situ interfacial polymerization of the prepared monomer solution and organic trimesoyl chloride. Water permeability, salt rejection, and fouling tendency of the modified membranes were then evaluated and compared with both pristine thin film composite (TFC) membrane and the ones modified by GO/Fe3O4 nanohybrides in the absence of magnetic field (TFN-GO/Fe3O4). According to the experimental results, when compared to the TFC and TFN-GO/Fe3O4 membranes, respectively, 117.4% and 63.2% water flux enhancements were achieved in TFN-MMGO/Fe3O4 membrane with optimal GO/Fe3O4 nanohybrid concentration of 100 ppm. In spite of such improvements in water flux, little compromise in reverse salt leakages were observed in the TFN-MMGO/Fe3O4 membranes compared to the TFC one. As well, the TFN-MMGO/Fe3O4 and TFN-GO/Fe3O4 membranes revealed higher fouling resistances than the TFC membrane due to their distinguished manipulated surface characteristics.

  8. The golgin GMAP-210 is required for efficient membrane trafficking in the early secretory pathway.

    Science.gov (United States)

    Roboti, Peristera; Sato, Keisuke; Lowe, Martin

    2015-04-15

    Golgins are coiled-coil proteins that participate in membrane-tethering events at the Golgi complex. Golgin-mediated tethering is thought to be important for vesicular trafficking and Golgi organization. However, the degree to which individual golgins contribute to these processes is poorly defined, and it has been proposed that golgins act in a largely redundant manner. Previous studies on the golgin GMAP-210 (also known as TRIP11), which is mutated in the rare skeletal disorder achondrogenesis type 1A, have yielded conflicting results regarding its involvement in trafficking. Here, we re-investigated the trafficking role of GMAP-210, and found that it is indeed required for efficient trafficking in the secretory pathway. GMAP-210 acts at both the endoplasmic reticulum (ER)-to-Golgi intermediate compartment (ERGIC) and Golgi complex during anterograde trafficking, and is also required for retrograde trafficking to the ER. Using co-depletion experiments, we also found that GMAP-210 acts in a partially redundant manner with the golgin GM130 to ensure efficient anterograde cargo delivery to the cis-Golgi. In summary, our results indicate a role for GMAP-210 in several trafficking steps at the ER-Golgi interface, some of which are partially redundant with another golgin, namely GM130 (also known as GOLGA2). © 2015. Published by The Company of Biologists Ltd.

  9. Efficient ethanol recovery from yeast fermentation broth with integrated distillation-membrane process

    Science.gov (United States)

    A hybrid process integrating vapor stripping with vapor compression and vapor permeation membrane separation, termed Membrane Assisted Vapor Stripping (MAVS), was evaluated for recovery and dehydration of ethanol from aqueous solution as an alternative to conventional distillatio...

  10. An efficient analytical model for baffled, multi-celled membrane-type acoustic metamaterial panels

    Science.gov (United States)

    Langfeldt, F.; Gleine, W.; von Estorff, O.

    2018-03-01

    A new analytical model for the oblique incidence sound transmission loss prediction of baffled panels with multiple subwavelength sized membrane-type acoustic metamaterial (MAM) unit cells is proposed. The model employs a novel approach via the concept of the effective surface mass density and approximates the unit cell vibrations in the form of piston-like displacements. This yields a coupled system of linear equations that can be solved efficiently using well-known solution procedures. A comparison with results from finite element model simulations for both normal and diffuse field incidence shows that the analytical model delivers accurate results as long as the edge length of the MAM unit cells is smaller than half the acoustic wavelength. The computation times for the analytical calculations are 100 times smaller than for the numerical simulations. In addition to that, the effect of flexible MAM unit cell edges compared to the fixed edges assumed in the analytical model is studied numerically. It is shown that the compliance of the edges has only a small impact on the transmission loss of the panel, except at very low frequencies in the stiffness-controlled regime. The proposed analytical model is applied to investigate the effect of variations of the membrane prestress, added mass, and mass eccentricity on the diffuse transmission loss of a MAM panel with 120 unit cells. Unlike most previous investigations of MAMs, these results provide a better understanding of the acoustic performance of MAMs under more realistic conditions. For example, it is shown that by varying these parameters deliberately in a checkerboard pattern, a new anti-resonance with large transmission loss values can be introduced. A random variation of these parameters, on the other hand, is shown to have only little influence on the diffuse transmission loss, as long as the standard deviation is not too large. For very large random variations, it is shown that the peak transmission loss

  11. A Simple, Cost-Efficient Method to Separate Microalgal Lipids from Wet Biomass Using Surface Energy-Modified Membranes.

    Science.gov (United States)

    Kwak, Moo Jin; Yoo, Youngmin; Lee, Han Sol; Kim, Jiyeon; Yang, Ji-Won; Han, Jong-In; Im, Sung Gap; Kwon, Jong-Hee

    2016-01-13

    For the efficient separation of lipid extracted from microalgae cells, a novel membrane was devised by introducing a functional polymer coating onto a membrane surface by means of an initiated chemical vapor deposition (iCVD) process. To this end, a steel-use-stainless (SUS) membrane was modified in a way that its surface energy was systemically modified. The surface modification by conformal coating of functional polymer film allowed for selective separation of oil-water mixture, by harnessing the tuned interfacial energy between each liquid phase and the membrane surface. The surface-modified membrane, when used with chloroform-based solvent, exhibited superb permeate flux, breakthrough pressure, and also separation yield: it allowed separation of 95.5 ± 1.2% of converted lipid (FAME) in the chloroform phase from the water/MeOH phase with microalgal debris. This result clearly supported that the membrane-based lipid separation is indeed facilitated by way of membrane being functionalized, enabling us to simplify the whole downstream process of microalgae-derived biodiesel production.

  12. Simulation and Experimental Study on the Efficiency of Traveling Wave Direct Energy Conversion for Application to Aneutronic Fusion Reactions

    Science.gov (United States)

    Tarditi, Alfonso; Chap, Andrew; Miley, George; Scott, John

    2013-10-01

    A study based on both Particle-in-cell (PIC) simulation and experiments is being developed to study the physics of the Traveling Wave Direct Energy Converter (TWDEC,) with the perspective of application to aneutronic fusion reaction products and space propulsion. The PIC model is investigating in detail the key TWDEC physics process by simulating the time-dependent transfer of energy from the ion beam to an electric load connected to ring-type electrodes in cylindrical symmetry. An experimental effort is in progress on a TWDEC test article at NASA, Johnson Space Center with the purpose of studying the conditions for improving the efficiency of the direct energy conversion process. Using a scaled-down ion energy source, the experiment is primarily focused on the effect of the (bunched) beam density on the efficiency and on the optimization of the electrode design. The simulation model is guiding the development of the experimental configuration and will provide details of the beam dynamics for direct comparison with experimental diagnostics. Work supported by NASA, Johnson Space Center.

  13. Assembly of a methodology for determination of membrane efficiency in preserved shales

    International Nuclear Information System (INIS)

    Villabona Camacho, Jhoao; Orozco Orozco, Sergio; Caldero Carrillo, Zuly; Saavedra, Netor F

    2009-01-01

    Determination of Membrane Efficiency (ME) is a very useful tool in the study of the chemical component of well bore stability since it is a variable input in chemical-elastic models (Lomba, Chenevert and Sharma, 2000). This article presents a novel methodology for the determination of ME using the Electrochemical Potential Test (EPT) in shale rocks. This method is based on the development of correlations with Ionic Selectivity (IS) values in presence of NaCl, KCl and CaCl 2 at diverse solution concentrations. The correlation, not reported previously in the literature, depends on the type of salt used. The EPT is a generic test easily applied to any rock type from any well or basin. It is simpler and quicker than other tests used for the ME determination, like the Pressure Transmission Test (PTT). Correlations between ME and IS are applicable to any type of argillaceous rock. Samples of unperturbed plugs with diverse properties belonging to different Colombian formations were used. The results obtained with the application of the proposed methodology indicate that it is possible to obtain IS and ME values through EPT in any type of argillaceous rock by applying the developed correlations.

  14. Biochemical factors affecting the quantum efficiency of hydrogen production by membranes of green photosynthetic bacteria

    Energy Technology Data Exchange (ETDEWEB)

    Bernstein, J.D.; Olson, J.M.

    1981-01-01

    Photohydrogen production, 200-700 ..mu..mol H/sub 2/ h/sup -1/ (mg bacteriochlorophyll a)/sup -1/ has been obtained in a system containing unit membrane vesicles (Complex I) from the green photosynthetic bacterium Chlorobium limicola f. thiosulfatophilum, ascorbate, N,N,N',N'-tetramethyl-p-phenylenediamine, dithioerythritol, an oxygen scavenging mixture, either methyl viologen (MV) or clostridial ferredoxin (CPS Fd) as electron carrier, and either CPS hydrogenase or platinum asbestos as catalyst. All components are necessary for maximum activity, and spinach Fd cannot be substituted for CPS Fd. Higher rates of photohydrogen production are obtained using MV or CPS Fd with hydrogenase than with MV and Pt asbestos. The highest quantum efficiencies (7-10% at 0.2-0.9 mW absorbed light and over 20% at lower light) were obtained with CPS Fd, hydrogenase and non-saturating 812 nm light. With saturating white light, however, rates of photohydrogen production varied relatively little among the various combinations of electron carrier and catalyst tested. The reaction rate is unaffected by 0.03% Triton X-100, and is insensitive to treatment with antimycin a or m-chloro-carbonyl cyanide phenylhydrazone.This indicates that neither electron flow through an endogenous cyclic chain, nor maintenance of a proton gradient are involved in this process.

  15. High-efficiency targets for high-gain inertial confinement fusion

    International Nuclear Information System (INIS)

    Gardner, J.H.; Bodner, S.E.

    1986-01-01

    Rocket efficiencies as high as 15% are possible using short wavelength lasers and moderately high aspect ratio pellet designs. These designs are made possible by two recent breakthroughs in physics constraints. First is the development of the induced spatial incoherence (ISI) technique, which allows uniform illumination of the pellet and relaxes the constraint of thermal smoothing, permitting the use of short wavelength laser light. Second is the discovery that the Rayleigh--Taylor growth rate is considerably reduced at short laser wavelengths. By taking advantage of the reduced constraints imposed by nonuniform laser illumination and Rayleigh--Taylor instability, pellets using (1)/(4) μm laser light and initial aspect ratios of about 10 (with in flight aspect ratios of about 150--200) may produce energy gains as high as 200--250

  16. Efficient DNP NMR of Membrane Proteins: Sample Preparation Protocols, Sensitivity, and Radical Location

    Science.gov (United States)

    Liao, Shu Y.; Lee, Myungwoon; Wang, Tuo; Sergeyev, Ivan V.; Hong, Mei

    2016-01-01

    Although dynamic nuclear polarization (DNP) has dramatically enhanced solid-state NMR spectral sensitivities of many synthetic materials and some biological macromolecules, recent studies of membrane-protein DNP using exogenously doped paramagnetic radicals as polarizing agents have reported varied and sometimes surprisingly limited enhancement factors. This motivated us to carry out a systematic evaluation of sample preparation protocols for optimizing the sensitivity of DNP NMR spectra of membrane-bound peptides and proteins at cryogenic temperatures of ~110 K. We show that mixing the radical with the membrane by direct titration instead of centrifugation gives a significant boost to DNP enhancement. We quantify the relative sensitivity enhancement between AMUPol and TOTAPOL, two commonly used radicals, and between deuterated and protonated lipid membranes. AMUPol shows ~4 fold higher sensitivity enhancement than TOTAPOL, while deuterated lipid membrane does not give net higher sensitivity for the membrane peptides than protonated membrane. Overall, a ~100 fold enhancement between the microwave-on and microwave-off spectra can be achieved on lipid-rich membranes containing conformationally disordered peptides, and absolute sensitivity gains of 105–160 can be obtained between low-temperature DNP spectra and high-temperature non-DNP spectra. We also measured the paramagnetic relaxation enhancement of lipid signals by TOTAPOL and AMUPol, to determine the depths of these two radicals in the lipid bilayer. Our data indicate a bimodal distribution of both radicals, a surface-bound fraction and a membrane-bound fraction where the nitroxides lie at ~10 Å from the membrane surface. TOTAPOL appears to have a higher membrane-embedded fraction than AMUPol. These results should be useful for membrane-protein solid-state NMR studies under DNP conditions and provide insights into how biradicals interact with phospholipid membranes. PMID:26873390

  17. Herpesvirus gB-induced fusion between the virion envelope and outer nuclear membrane during virus egress is regulated by the viral US3 kinase.

    Science.gov (United States)

    Wisner, Todd W; Wright, Catherine C; Kato, Akihisa; Kawaguchi, Yasushi; Mou, Fan; Baines, Joel D; Roller, Richard J; Johnson, David C

    2009-04-01

    Herpesvirus capsids collect along the inner surface of the nuclear envelope and bud into the perinuclear space. Enveloped virions then fuse with the outer nuclear membrane (NM). We previously showed that herpes simplex virus (HSV) glycoproteins gB and gH act in a redundant fashion to promote fusion between the virion envelope and the outer NM. HSV mutants lacking both gB and gH accumulate enveloped virions in herniations, vesicles that bulge into the nucleoplasm. Earlier studies had shown that HSV mutants lacking the viral serine/threonine kinase US3 also accumulate herniations. Here, we demonstrate that HSV gB is phosphorylated in a US3-dependent manner in HSV-infected cells, especially in a crude nuclear fraction. Moreover, US3 directly phosphorylated the gB cytoplasmic (CT) domain in in vitro assays. Deletion of gB in the context of a US3-null virus did not add substantially to defects in nuclear egress. The majority of the US3-dependent phosphorylation of gB involved the CT domain and amino acid T887, a residue present in a motif similar to that recognized by US3 in other proteins. HSV recombinants lacking gH and expressing either gB substitution mutation T887A or a gB truncated at residue 886 displayed substantial defects in nuclear egress. We concluded that phosphorylation of the gB CT domain is important for gB-mediated fusion with the outer NM. This suggested a model in which the US3 kinase is incorporated into the tegument layer (between the capsid and envelope) in HSV virions present in the perinuclear space. By this packaging, US3 might be brought close to the gB CT tail, leading to phosphorylation and triggering fusion between the virion envelope and the outer NM.

  18. Efficient hydrogen isotopologues separation through a tunable potential barrier: The case of a C2N membrane.

    Science.gov (United States)

    Qu, Yuanyuan; Li, Feng; Zhao, Mingwen

    2017-05-03

    Isotopes separation through quantum sieving effect of membranes is quite promising for industrial applications. For the light hydrogen isotopologues (eg. H 2 , D 2 ), the confinement of potential wells in porous membranes to isotopologues was commonly regarded to be crucial for highly efficient separation ability. Here, we demonstrate from first-principles that a potential barrier is also favorable for efficient hydrogen isotopologues separation. Taking an already-synthesized two-dimensional carbon nitride (C 2 N-h2D) as an example, we predict that the competition between quantum tunneling and zero-point-energy (ZPE) effects regulated by the tensile strain leads to high selectivity and permeance. Both kinetic quantum sieving and equilibrium quantum sieving effects are considered. The quantum effects revealed in this work offer a prospective strategy for highly efficient hydrogen isotopologues separation.

  19. Efficient cellular solid-state NMR of membrane proteins by targeted protein labeling

    International Nuclear Information System (INIS)

    Baker, Lindsay A.; Daniëls, Mark; Cruijsen, Elwin A. W. van der; Folkers, Gert E.; Baldus, Marc

    2015-01-01

    Solid-state NMR spectroscopy (ssNMR) has made significant progress towards the study of membrane proteins in their native cellular membranes. However, reduced spectroscopic sensitivity and high background signal levels can complicate these experiments. Here, we describe a method for ssNMR to specifically label a single protein by repressing endogenous protein expression with rifampicin. Our results demonstrate that treatment of E. coli with rifampicin during induction of recombinant membrane protein expression reduces background signals for different expression levels and improves sensitivity in cellular membrane samples. Further, the method reduces the amount of time and resources needed to produce membrane protein samples, enabling new strategies for studying challenging membrane proteins by ssNMR

  20. Graphene oxide doped ionic liquid ultrathin composite membranes for efficient CO2 capture

    KAUST Repository

    Karunakaran, Madhavan

    2016-11-28

    Advanced membrane systems with high flux and sufficient selectivity are required for industrial gas separation processes. In order to achieve high flux and high selectivity, the membrane material should be as thin as possible and it should have selective sieving channels and long term stability. This could be achieved by designing a three component material consisting of a blend of an ionic liquid and graphene oxide covered by a highly permeable low selective polymeric coating. By using a simple dip coating technique, we prepared high flux and CO selective ultrathin graphene oxide (GO)/ionic liquid membranes on a porous ultrafiltration support. The ultrathin composite membranes derived from GO/ionic liquid complex displays remarkable combinations of permeability (CO flux: 37 GPU) and selectivity (CO/N selectivity: 130) that surpass the upper bound of ionic liquid membranes for CO/N separation. Moreover, the membranes were stable when tested for 120 hours.

  1. Highly efficient forward osmosis based on porous membranes--applications and implications.

    Science.gov (United States)

    Qi, Saren; Li, Ye; Zhao, Yang; Li, Weiyi; Tang, Chuyang Y

    2015-04-07

    For the first time, forward osmosis (FO) was performed using a porous membrane with an ultrafiltration (UF)-like rejection layer and its feasibility for high performance FO filtration was demonstrated. Compared to traditional FO membranes with dense rejection layers, the UF-like FO membrane was 2 orders of magnitude more permeable. This gave rise to respectable FO water flux even at ultralow osmotic driving force, for example, 7.6 L/m(2).h at an osmotic pressure of merely 0.11 bar (achieved by using a 0.1% poly(sodium 4-styrene-sulfonate) draw solution). The membrane was applied to oil/water separation, and a highly stable FO water flux was achieved. The adoption of porous FO membranes opens a door to many new opportunities, with potential applications ranging from wastewater treatment, valuable product recovery, and biomedical applications. The potential applications and implications of porous FO membranes are addressed in this paper.

  2. A rapid and efficient newly established method to detect COL1A1–PDGFB gene fusion in dermatofibrosarcoma protuberans

    International Nuclear Information System (INIS)

    Yokoyama, Yoko; Shimizu, Akira; Okada, Etsuko; Ishikawa, Osamu; Motegi, Sei-ichiro

    2012-01-01

    Highlights: ► We developed new method to rapidly identify COL1A1–PDGFB fusion in DFSP. ► New PCR method using a single primer pair detected COL1A1–PDGFB fusion in DFSP. ► This is the first report of DFSP with a novel COL1A1 breakpoint in exon 5. -- Abstract: The detection of fusion transcripts of the collagen type 1α1 (COL1A1) and platelet-derived growth factor-BB (PDGFB) genes by genetic analysis has recognized as a reliable and valuable molecular tool for the diagnosis of dermatofibrosarcoma protuberans (DFSP). To detect the COL1A1–PDGFB fusion, almost previous reports performed reverse transcription polymerase chain reaction (RT-PCR) using multiplex forward primers from COL1A1. However, it has possible technical difficulties with respect to the handling of multiple primers and reagents in the procedure. The objective of this study is to establish a rapid, easy, and efficient one-step method of PCR using only a single primer pair to detect the fusion transcripts of the COL1A1 and PDGFB in DFSP. To validate new method, we compared the results of RT-PCR in five patients of DFSP between the previous method using multiplex primers and our established one-step RT-PCR using a single primer pair. In all cases of DFSP, the COL1A1–PDGFB fusion was detected by both previous method and newly established one-step PCR. Importantly, we detected a novel COL1A1 breakpoint in exon 5. The newly developed method is valuable to rapidly identify COL1A1–PDGFB fusion transcripts in DFSP.

  3. Solid-state nuclear magnetic resonance measurements of HIV fusion peptide 13CO to lipid 31P proximities support similar partially inserted membrane locations of the α helical and β sheet peptide structures.

    Science.gov (United States)

    Gabrys, Charles M; Qiang, Wei; Sun, Yan; Xie, Li; Schmick, Scott D; Weliky, David P

    2013-10-03

    Fusion of the human immunodeficiency virus (HIV) membrane and the host cell membrane is an initial step of infection of the host cell. Fusion is catalyzed by gp41, which is an integral membrane protein of HIV. The fusion peptide (FP) is the ∼25 N-terminal residues of gp41 and is a domain of gp41 that plays a key role in fusion catalysis likely through interaction with the host cell membrane. Much of our understanding of the FP domain has been accomplished with studies of "HFP", i.e., a ∼25-residue peptide composed of the FP sequence but lacking the rest of gp41. HFP catalyzes fusion between membrane vesicles and serves as a model system to understand fusion catalysis. HFP binds to membranes and the membrane location of HFP is likely a significant determinant of fusion catalysis perhaps because the consequent membrane perturbation reduces the fusion activation energy. In the present study, many HFPs were synthesized and differed in the residue position that was (13)CO backbone labeled. Samples were then prepared that each contained a singly (13)CO labeled HFP incorporated into membranes that lacked cholesterol. HFP had distinct molecular populations with either α helical or oligomeric β sheet structure. Proximity between the HFP (13)CO nuclei and (31)P nuclei in the membrane headgroups was probed by solid-state NMR (SSNMR) rotational-echo double-resonance (REDOR) measurements. For many samples, there were distinct (13)CO shifts for the α helical and β sheet structures so that the proximities to (31)P nuclei could be determined for each structure. Data from several differently labeled HFPs were then incorporated into a membrane location model for the particular structure. In addition to the (13)CO labeled residue position, the HFPs also differed in sequence and/or chemical structure. "HFPmn" was a linear peptide that contained the 23 N-terminal residues of gp41. "HFPmn_V2E" contained the V2E mutation that for HIV leads to greatly reduced extent of fusion and

  4. Solid-State Nuclear Magnetic Resonance Measurements of HIV Fusion Peptide 13CO to Lipid 31P Proximities Support Similar Partially Inserted Membrane Locations of the α Helical and β Sheet Peptide Structures

    Science.gov (United States)

    Gabrys, Charles M.; Qiang, Wei; Sun, Yan; Xie, Li; Schmick, Scott D.; Weliky, David P.

    2013-10-01

    Fusion of the human immunodeficiency virus (HIV) membrane and the host cell membrane is an initial step of infection of the host cell. Fusion is catalyzed by gp41, which is an integral membrane protein of HIV. The fusion peptide (FP) is the -25 N-terminal residues of gp41 and is a domain of gp41 that plays a key role in fusion catalysis likely through interaction with the host cell membrane. Much of our understanding of the FP domain has been accomplished with studies of -HFP-, i.e., a -25-residue peptide composed of the FP sequence but lacking the rest of gp41. HFP catalyzes fusion between membrane vesicles and serves as a model system to understand fusion catalysis. HFP binds to membranes and the membrane location of HFP is likely a significant determinant of fusion catalysis perhaps because the consequent membrane perturbation reduces the fusion activation energy. In the present study, many HFPs were synthesized and differed in the residue position that was 13CO backbone labeled. Samples were then prepared that each contained a singly 13CO labeled HFP incorporated into membranes that lacked cholesterol. HFP had distinct molecular populations with either α helical or oligomeric - sheet structure. Proximity between the HFP 13CO nuclei and 31P nuclei in the membrane headgroups was probed by solid-state NMR (SSNMR) rotational-echo double-resonance (REDOR) measurements. For many samples, there were distinct 13CO shifts for the α helical and - sheet structures so that the proximities to 31P nuclei could be determined for each structure. Data from several differently labeled HFPs were then incorporated into a membrane location model for the particular structure. In addition to the 13CO labeled residue position, the HFPs also differed in sequence and/or chemical structure. -HFPmn- was a linear peptide that contained the 23 N-terminal residues of gp41. -HFPmn_V2E- contained the V2E mutation that for HIV leads to greatly reduced extent of fusion and infection. The

  5. High-performance nanocomposite membranes realized by efficient molecular sieving with CuBDC nanosheets.

    Science.gov (United States)

    Yang, Yanqin; Goh, Kunli; Wang, Rong; Bae, Tae-Hyun

    2017-04-11

    Two-dimensional (2-D) CuBDC nanosheets (ns-CuBDC) with high-aspect-ratios were deliberately paired with polymers possessing high free volumes to fabricate high performance gas separation membranes. Owing to the molecular sieving effect of the filler, a small ns-CuBDC loading (2-4 wt%) could significantly improve the CO 2 /CH 4 selectivities of membranes, resulting in performances that surpass the upper bound limit for polymer membranes.

  6. Annexin A4 and A6 induce membrane curvature and constriction during cell membrane repair

    DEFF Research Database (Denmark)

    Boye, Theresa Louise; Maeda, Kenji; Pezeshkian, Weria

    2017-01-01

    that annexin A4 binds to artificial membranes and generates curvature force initiated from free edges, whereas annexin A6 induces constriction force. In cells, plasma membrane injury and Ca2+ influx recruit annexin A4 to the vicinity of membrane wound edges where its homo-trimerization leads to membrane......Efficient cell membrane repair mechanisms are essential for maintaining membrane integrity and thus for cell life. Here we show that the Ca2+- and phospholipid-binding proteins annexin A4 and A6 are involved in plasma membrane repair and needed for rapid closure of micron-size holes. We demonstrate...... curvature near the edges. We propose that curvature force is utilized together with annexin A6-mediated constriction force to pull the wound edges together for eventual fusion. We show that annexin A4 can counteract various plasma membrane disruptions including holes of several micrometers indicating...

  7. Survival of the fittest: positive selection of CD4+ T cells expressing a membrane-bound fusion inhibitor following HIV-1 infection.

    Science.gov (United States)

    Kimpel, Janine; Braun, Stephen E; Qiu, Gang; Wong, Fay Eng; Conolle, Michelle; Schmitz, Jörn E; Brendel, Christian; Humeau, Laurent M; Dropulic, Boro; Rossi, John J; Berger, Annemarie; von Laer, Dorothee; Johnson, R Paul

    2010-08-23

    Although a variety of genetic strategies have been developed to inhibit HIV replication, few direct comparisons of the efficacy of these inhibitors have been carried out. Moreover, most studies have not examined whether genetic inhibitors are able to induce a survival advantage that results in an expansion of genetically-modified cells following HIV infection. We evaluated the efficacy of three leading genetic strategies to inhibit HIV replication: 1) an HIV-1 tat/rev-specific small hairpin (sh) RNA; 2) an RNA antisense gene specific for the HIV-1 envelope; and 3) a viral entry inhibitor, maC46. In stably transduced cell lines selected such that >95% of cells expressed the genetic inhibitor, the RNA antisense envelope and viral entry inhibitor maC46 provided the strongest inhibition of HIV-1 replication. However, when mixed populations of transduced and untransduced cells were challenged with HIV-1, the maC46 fusion inhibitor resulted in highly efficient positive selection of transduced cells, an effect that was evident even in mixed populations containing as few as 1% maC46-expressing cells. The selective advantage of the maC46 fusion inhibitor was also observed in HIV-1-infected cultures of primary T lymphocytes as well as in HIV-1-infected humanized mice. These results demonstrate robust inhibition of HIV replication with the fusion inhibitor maC46 and the antisense Env inhibitor, and importantly, a survival advantage of cells expressing the maC46 fusion inhibitor both in vitro and in vivo. Evaluation of the ability of genetic inhibitors of HIV-1 replication to confer a survival advantage on genetically-modified cells provides unique information not provided by standard techniques that may be important in the in vivo efficacy of these genes.

  8. Sequence motif upstream of the Hendra virus fusion protein cleavage site is not sufficient to promote efficient proteolytic processing

    International Nuclear Information System (INIS)

    Craft, Willie Warren; Dutch, Rebecca Ellis

    2005-01-01

    The Hendra virus fusion (HeV F) protein is synthesized as a precursor, F 0 , and proteolytically cleaved into the mature F 1 and F 2 heterodimer, following an HDLVDGVK 109 motif. This cleavage event is required for fusogenic activity. To determine the amino acid requirements for processing of the HeV F protein, we constructed multiple mutants. Individual and simultaneous alanine substitutions of the eight residues immediately upstream of the cleavage site did not eliminate processing. A chimeric SV5 F protein in which the furin site was substituted for the VDGVK 109 motif of the HeV F protein was not processed but was expressed on the cell surface. Another chimeric SV5 F protein containing the HDLVDGVK 109 motif of the HeV F protein underwent partial cleavage. These data indicate that the upstream region can play a role in protease recognition, but is neither absolutely required nor sufficient for efficient processing of the HeV F protein

  9. Calculation of the absolute detection efficiency of a moderated 235U neutron detector on the tokamak fusion test reactor

    Science.gov (United States)

    Ku, L. P.; Hendel, H. W.; Liew, S. L.

    1989-08-01

    Neutron transport simulations have been carried out to calculate the absolute detection efficiency of a moderated 235U neutron detector which is used on the TFTR as part of the primary fission detector diagnostic system for measuring fusion power yields. Transport simulations provide a means by which the effects of variations in various shielding and geometrical parameters can be explored. These effects are difficult to study in calibration experiments. The calculational model, benchmarked against measurements, can be used to complement future detector calibrations, when the high level of radioactivity resulting from machine operation may severely restrict access to the tokamak. We present a coupled forward-adjoint algorithm, employing both the deterministic and Monte Carlo sampling methods, to model the neutron transport in the complex tokamak and detector geometries. Sensitivities of the detector response to the major and minor radii, and angular anisotropy of the neutron emission are discussed. A semiempirical model based on matching the calculational results with a small set of experiments produces good agreement (± 15%) for a wide range of source energies and geometries.

  10. Study of conversion efficiencies for laser fusion rocket like a VISTA in the laser plasma experiments and PIC simulation

    International Nuclear Information System (INIS)

    Zakharov, Yu. P.; Vchivkov, K. V.; Melekhov, A. V.; Posukh, V. G.; Boyarintsev, E. L.; Shaikhislamov, I. F.; Nakashima, V. G.

    2007-01-01

    Φ at axial distance 4.2 cm (in a plane of one field-coil with N t = 15 of turns, acting there as one-half of pick-up coil) gives us expected ideal current value J m (A) = ΔΦ/L ≅ 2E0(J) * N t / L(H) * B(G) * R t (cm) * (1+ α 2 ) 3 /2 ≅ 45 A. But for finer grid 0.75 mm, when a cavity radius was decreased by more pronounced effect of flute-type instability (observed in MHD experiment also), the calculated flux-changing ΔΦ was smaller in a good agreement with the measured current 33 A in MHD. Thus we can use this simplified expression for J m to estimate the efficiency η e ∝ J m 2 /LE 0 , but due to effects of real plasma we should use only one-half of its ideal value, denoted as * η e . First results of VISTA-KI numerical simulation confirm that under the same assumptions we can use such J m -estimation of η e in a quasi-dipole field also (at least for ae≤0.1) and therefore we can rewrite it into general form [7] as * η e (%) ≅ (5/10)ae /α 3 from which we can conclude that for parameters of VISTA its efficiency * η e ≤ 2% could be very low. So the main goal of VISTA-KI experiment and its further numerical simulations will be verification of proposed η e (ae,α) relation under conditions of η p ≥ 50% (and ε b < 1), to optimize original VISTA-scheme [4]. References: 1. Nagamine Y. and Nakashima H., Fusion Technology, 35 (1999) 62. 2. Vchivkov K., Nakashima H., Zakharov Yu.P., et al., Jpn. J. Appl. Phys., 42 (2003) 6590. 3. Zakharov Yu.P., et al., in Proc. 4th Symp. 'Current Trends in Intern. Fusion Research' (Wash., DC, 2001) 31. 4. Orth, C.D., LLNL Report VISTA, University of California: UCRL-LR-110500 (2003) 143 p. 5. Zakharov Yu.P., et al. Trans. Fusion Science and Technology, 41 (2005) 187. 6. Zakharov Yu.P., et al., Plasma Physics Reports, 32 (2006) 183. 7. Zakharov Yu. P., In 'Inertial Fusion Sciences and Applications: State of the Art 2003' (American Nuclear Society, 2004) 1106

  11. Cross-flow-assembled ultrathin and robust graphene oxide membranes for efficient molecule separation

    Science.gov (United States)

    Ying, Yulong; Ying, Wen; Guo, Yi; Peng, Xinsheng

    2018-04-01

    A graphene oxide (GO) membrane is promising for molecule separation. However, it is still a big challenge to achieve highly stable pristine GO membranes, especially in water. In this work, an ultrathin and robust GO membrane is assembled via the cross-flow method. The as-prepared 12 nm thick GO membrane (GOCF membrane) presents high stability with water permeance of 1505 ± 65 litres per hour per square meter per bar (LHM bar-1) and Evans Blue (EB) rejection of 98.7 ± 0.4%, 21-fold enhancement in water permeance compared with that of a pristine GO membrane (50-70 LHM bar-1) and 100 times higher than that of commercial ultrafiltration membranes (15 LHM.bar-1, GE2540F30, MWCO 1000, GE Co., Ltd) with similar rejection. Attributed to the surface cross-flow, the GO nanosheets will be refolded, crumpled, or wrinkled, resulting in a very strong inter-locking structure among the GO membrane, which significantly enhances the stability and facilitates their separation performance. This cross-flow assembling technique is also easily extended to assemble GO membranes onto other various backing filter supports. Based on the Donnan effect and size sieving mechanism, selective membrane separation of dyes with a similar molecular structure from their mixture (such as Rhodamine B (RhB) and Rose Bengal, and RhB and EB) are achieved with a selectivity of 133 ± 10 and 227 ± 15, respectively. Assembly of this ultrathin GO membrane with high stability and separation performance, via a simple cross-flow method, shows great potential for water purification.

  12. Functional fluorescent protein insertions in herpes simplex virus gB report on gB conformation before and after execution of membrane fusion.

    Directory of Open Access Journals (Sweden)

    John R Gallagher

    2014-09-01

    Full Text Available Entry of herpes simplex virus (HSV into a target cell requires complex interactions and conformational changes by viral glycoproteins gD, gH/gL, and gB. During viral entry, gB transitions from a prefusion to a postfusion conformation, driving fusion of the viral envelope with the host cell membrane. While the structure of postfusion gB is known, the prefusion conformation of gB remains elusive. As the prefusion conformation of gB is a critical target for neutralizing antibodies, we set out to describe its structure by making genetic insertions of fluorescent proteins (FP throughout the gB ectodomain. We created gB constructs with FP insertions in each of the three globular domains of gB. Among 21 FP insertion constructs, we found 8 that allowed gB to remain membrane fusion competent. Due to the size of an FP, regions in gB that tolerate FP insertion must be solvent exposed. Two FP insertion mutants were cell-surface expressed but non-functional, while FP insertions located in the crown were not surface expressed. This is the first report of placing a fluorescent protein insertion within a structural domain of a functional viral fusion protein, and our results are consistent with a model of prefusion HSV gB constructed from the prefusion VSV G crystal structure. Additionally, we found that functional FP insertions from two different structural domains could be combined to create a functional form of gB labeled with both CFP and YFP. FRET was measured with this construct, and we found that when co-expressed with gH/gL, the FRET signal from gB was significantly different from the construct containing CFP alone, as well as gB found in syncytia, indicating that this construct and others of similar design are likely to be powerful tools to monitor the conformation of gB in any model system accessible to light microscopy.

  13. J-TEXT WebScope: An efficient data access and visualization system for long pulse fusion experiment

    Energy Technology Data Exchange (ETDEWEB)

    Zheng, Wei, E-mail: zhenghaku@gmail.com [State Key Laboratory of Advanced Electromagnetic Engineering and Technology in Huazhong University of Science and Technology, Wuhan 430074 (China); School of Electrical and Electronic Engineering in Huazhong University of Science and Technology, Wuhan 430074 (China); Wan, Kuanhong; Chen, Zhi; Hu, Feiran; Liu, Qiang [State Key Laboratory of Advanced Electromagnetic Engineering and Technology in Huazhong University of Science and Technology, Wuhan 430074 (China); School of Electrical and Electronic Engineering in Huazhong University of Science and Technology, Wuhan 430074 (China)

    2016-11-15

    Highlights: • No matter how large the data is, the response time is always less than 500 milliseconds. • It is intelligent and just gives you the data you want. • It can be accessed directly over the Internet without installing special client software if you already have a browser. • Adopt scale and segment technology to organize data. • To support a new database for the WebScope is quite easy. • With the configuration stored in user’s profile, you have your own portable WebScope. - Abstract: Fusion research is an international collaboration work. To enable researchers across the world to visualize and analyze the experiment data, a web based data access and visualization tool is quite important [1]. Now, a new WebScope based on RIA (Rich Internet Application) is designed and implemented to meet these requirements. On the browser side, a fluent and intuitive interface is provided for researchers at J-TEXT laboratory and collaborators from all over the world to view experiment data and related metadata. The fusion experiments will feature long pulse and high sampling rate in the future. The data access and visualization system in this work has adopted segment and scale concept. Large data samples are re-sampled in different scales and then split into segments for instant response. It allows users to view extremely large data on the web browser efficiently, without worrying about the limitation on the size of the data. The HTML5 and JavaScript based web front-end can provide intuitive and fluent user experience. On the server side, a RESTful (Representational State Transfer) web API, which is based on ASP.NET MVC (Model View Controller), allows users to access the data and its metadata through HTTP (HyperText Transfer Protocol). An interface to the database has been designed to decouple the data access and visualization system from the data storage. It can be applied upon any data storage system like MDSplus or JTEXTDB, and this system is very easy to

  14. J-TEXT WebScope: An efficient data access and visualization system for long pulse fusion experiment

    International Nuclear Information System (INIS)

    Zheng, Wei; Wan, Kuanhong; Chen, Zhi; Hu, Feiran; Liu, Qiang

    2016-01-01

    Highlights: • No matter how large the data is, the response time is always less than 500 milliseconds. • It is intelligent and just gives you the data you want. • It can be accessed directly over the Internet without installing special client software if you already have a browser. • Adopt scale and segment technology to organize data. • To support a new database for the WebScope is quite easy. • With the configuration stored in user’s profile, you have your own portable WebScope. - Abstract: Fusion research is an international collaboration work. To enable researchers across the world to visualize and analyze the experiment data, a web based data access and visualization tool is quite important [1]. Now, a new WebScope based on RIA (Rich Internet Application) is designed and implemented to meet these requirements. On the browser side, a fluent and intuitive interface is provided for researchers at J-TEXT laboratory and collaborators from all over the world to view experiment data and related metadata. The fusion experiments will feature long pulse and high sampling rate in the future. The data access and visualization system in this work has adopted segment and scale concept. Large data samples are re-sampled in different scales and then split into segments for instant response. It allows users to view extremely large data on the web browser efficiently, without worrying about the limitation on the size of the data. The HTML5 and JavaScript based web front-end can provide intuitive and fluent user experience. On the server side, a RESTful (Representational State Transfer) web API, which is based on ASP.NET MVC (Model View Controller), allows users to access the data and its metadata through HTTP (HyperText Transfer Protocol). An interface to the database has been designed to decouple the data access and visualization system from the data storage. It can be applied upon any data storage system like MDSplus or JTEXTDB, and this system is very easy to

  15. Macrophage entry mediated by HIV Envs from brain and lymphoid tissues is determined by the capacity to use low CD4 levels and overall efficiency of fusion

    International Nuclear Information System (INIS)

    Thomas, Elaine R.; Dunfee, Rebecca L.; Stanton, Jennifer; Bogdan, Derek; Taylor, Joann; Kunstman, Kevin; Bell, Jeanne E.; Wolinsky, Steven M.; Gabuzda, Dana

    2007-01-01

    HIV infects macrophages and microglia in the central nervous system (CNS), which express lower levels of CD4 than CD4+ T cells in peripheral blood. To investigate mechanisms of HIV neurotropism, full-length env genes were cloned from autopsy brain and lymphoid tissues from 4 AIDS patients with HIV-associated dementia (HAD). Characterization of 55 functional Env clones demonstrated that Envs with reduced dependence on CD4 for fusion and viral entry are more frequent in brain compared to lymphoid tissue. Envs that mediated efficient entry into macrophages were frequent in brain but were also present in lymphoid tissue. For most Envs, entry into macrophages correlated with overall fusion activity at all levels of CD4 and CCR5. gp160 nucleotide sequences were compartmentalized in brain versus lymphoid tissue within each patient. Proline at position 308 in the V3 loop of gp120 was associated with brain compartmentalization in 3 patients, but mutagenesis studies suggested that P308 alone does not contribute to reduced CD4 dependence or macrophage-tropism. These results suggest that HIV adaptation to replicate in the CNS selects for Envs with reduced CD4 dependence and increased fusion activity. Macrophage-tropic Envs are frequent in brain but are also present in lymphoid tissues of AIDS patients with HAD, and entry into macrophages in the CNS and other tissues is dependent on the ability to use low receptor levels and overall efficiency of fusion

  16. Efficient gas-liquid contact using microfluidic membrane devices with staggered herringbone mixers.

    Science.gov (United States)

    Femmer, Tim; Eggersdorfer, Max L; Kuehne, Alexander J C; Wessling, Matthias

    2015-08-07

    We describe a novel membrane based gas-liquid-contacting device with increased mass transport and reduced pressure loss by combining a membrane with a staggered herringbone static mixer. Herringbone structures are imposed on the microfluidic channel geometry via soft lithography, acting as mixers which introduce secondary flows at the membrane interface. Such flows include Dean vortices and Taylor flows generating effective mixing while improving mass transport and preventing concentration polarization in microfluidic channels. Furthermore, our static herringbone mixer membranes effectively reduce pressure losses leading to devices with enhanced transfer properties for microfluidic gas-liquid contact. We investigate the red blood cell distribution to tailor our devices towards miniaturised extracorporeal membrane oxygenation and improved comfort of patients with lung insufficiencies.

  17. Lamellipodia and Membrane Blebs Drive Efficient Electrotactic Migration of Rat Walker Carcinosarcoma Cells WC 256.

    Science.gov (United States)

    Sroka, Jolanta; Krecioch, Izabela; Zimolag, Eliza; Lasota, Slawomir; Rak, Monika; Kedracka-Krok, Sylwia; Borowicz, Pawel; Gajek, Marta; Madeja, Zbigniew

    2016-01-01

    The endogenous electric field (EF) may provide an important signal for directional cell migration during wound healing, embryonic development and cancer metastasis but the mechanism of cell electrotaxis is poorly understood. Additionally, there is no research addressing the question on the difference in electrotactic motility of cells representing various strategies of cell movement-specifically blebbing vs. lamellipodial migration. In the current study we constructed a unique experimental model which allowed for the investigation of electrotactic movement of cells of the same origin but representing different modes of cell migration: weakly adherent, spontaneously blebbing (BC) and lamellipodia forming (LC) WC256 cells. We report that both BC and LC sublines show robust cathodal migration in a physiological EF (1-3 V/cm). The directionality of cell movement was completely reversible upon reversing the field polarity. However, the full reversal of cell direction after the change of EF polarity was much faster in the case of BC (10 minutes) than LC cells (30 minutes). We also investigated the distinct requirements for Rac, Cdc42 and Rho pathways and intracellular Ca2+ in electrotaxis of WC256 sublines forming different types of cell protrusions. It was found that Rac1 is required for directional movement of LC to a much greater extent than for BC, but Cdc42 and RhoA are more crucial for BC than for LC cells. The inhibition of ROCK did not affect electrotaxis of LC in contrast to BC cells. The results also showed that intracellular Ca2+ is essential only for the electrotactic reaction of BC cells. Moreover, inhibition of MLCK and myosin II did not affect the electrotaxis of LC in contrast to BC cells. In conclusion, our results revealed that both lamellipodia and membrane blebs can efficiently drive electrotactic migration of WC 256 carcinosarcoma cells, however directional migration is mediated by different signalling pathways.

  18. Lamellipodia and Membrane Blebs Drive Efficient Electrotactic Migration of Rat Walker Carcinosarcoma Cells WC 256.

    Directory of Open Access Journals (Sweden)

    Jolanta Sroka

    Full Text Available The endogenous electric field (EF may provide an important signal for directional cell migration during wound healing, embryonic development and cancer metastasis but the mechanism of cell electrotaxis is poorly understood. Additionally, there is no research addressing the question on the difference in electrotactic motility of cells representing various strategies of cell movement-specifically blebbing vs. lamellipodial migration. In the current study we constructed a unique experimental model which allowed for the investigation of electrotactic movement of cells of the same origin but representing different modes of cell migration: weakly adherent, spontaneously blebbing (BC and lamellipodia forming (LC WC256 cells. We report that both BC and LC sublines show robust cathodal migration in a physiological EF (1-3 V/cm. The directionality of cell movement was completely reversible upon reversing the field polarity. However, the full reversal of cell direction after the change of EF polarity was much faster in the case of BC (10 minutes than LC cells (30 minutes. We also investigated the distinct requirements for Rac, Cdc42 and Rho pathways and intracellular Ca2+ in electrotaxis of WC256 sublines forming different types of cell protrusions. It was found that Rac1 is required for directional movement of LC to a much greater extent than for BC, but Cdc42 and RhoA are more crucial for BC than for LC cells. The inhibition of ROCK did not affect electrotaxis of LC in contrast to BC cells. The results also showed that intracellular Ca2+ is essential only for the electrotactic reaction of BC cells. Moreover, inhibition of MLCK and myosin II did not affect the electrotaxis of LC in contrast to BC cells. In conclusion, our results revealed that both lamellipodia and membrane blebs can efficiently drive electrotactic migration of WC 256 carcinosarcoma cells, however directional migration is mediated by different signalling pathways.

  19. Thin, High-Flux, Self-Standing, Graphene Oxide Membranes for Efficient Hydrogen Separation from Gas Mixtures.

    Science.gov (United States)

    Bouša, Daniel; Friess, Karel; Pilnáček, Kryštof; Vopička, Ondřej; Lanč, Marek; Fónod, Kristián; Pumera, Martin; Sedmidubský, David; Luxa, Jan; Sofer, Zdeněk

    2017-08-22

    The preparation and gas-separation performance of self-standing, high-flux, graphene oxide (GO) membranes is reported. Defect-free, 15-20 μm thick, mechanically stable, unsupported GO membranes exhibited outstanding gas-separation performance towards H 2 /CO 2 that far exceeded the corresponding 2008 Robeson upper bound. Remarkable separation efficiency of GO membranes for H 2 and bulky C 3 or C 4 hydrocarbons was achieved with high flux and good selectivity at the same time. On the contrary, N 2 and CH 4 molecules, with larger kinetic diameter and simultaneously lower molecular weight, relative to that of CO 2 , remained far from the corresponding H 2 /N 2 or H 2 /CH 4 upper bounds. Pore size distribution analysis revealed that the most abundant pores in GO material were those with an effective pore diameter of 4 nm; therefore, gas transport is not exclusively governed by size sieving and/or Knudsen diffusion, but in the case of CO 2 was supplemented by specific interactions through 1) hydrogen bonding with carboxyl or hydroxyl functional groups and 2) the quadrupole moment. The self-standing GO membranes presented herein demonstrate a promising route towards the large-scale fabrication of high-flux, hydrogen-selective gas membranes intended for the separation of H 2 /CO 2 or H 2 /alkanes. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  20. Efficient replacement of plasma membrane outer leaflet phospholipids and sphingolipids in cells with exogenous lipids.

    Science.gov (United States)

    Li, Guangtao; Kim, JiHyun; Huang, Zhen; St Clair, Johnna R; Brown, Deborah A; London, Erwin

    2016-12-06

    Our understanding of membranes and membrane lipid function has lagged far behind that of nucleic acids and proteins, largely because it is difficult to manipulate cellular membrane lipid composition. To help solve this problem, we show that methyl-α-cyclodextrin (MαCD)-catalyzed lipid exchange can be used to maximally replace the sphingolipids and phospholipids in the outer leaflet of the plasma membrane of living mammalian cells with exogenous lipids, including unnatural lipids. In addition, lipid exchange experiments revealed that 70-80% of cell sphingomyelin resided in the plasma membrane outer leaflet; the asymmetry of metabolically active cells was similar to that previously defined for erythrocytes, as judged by outer leaflet lipid composition; and plasma membrane outer leaflet phosphatidylcholine had a significantly lower level of unsaturation than phosphatidylcholine in the remainder of the cell. The data also provided a rough estimate for the total cellular lipids residing in the plasma membrane (about half). In addition to such lipidomics applications, the exchange method should have wide potential for investigations of lipid function and modification of cellular behavior by modification of lipids.

  1. Improvement on performance and efficiency of direct methanol fuel cells using hydrocarbon-based membrane electrode assembly

    International Nuclear Information System (INIS)

    Kim, Joon-Hee; Yang, Min-Jee; Park, Jun-Young

    2014-01-01

    Highlights: • Faradaic efficiency and water transfer coefficient (WTC) of DMFC MEAs are calculated based on mass balance measurements. • Faradaic efficiency of the HC-based MEAs is generally improved over the Nafion-based MEAs. • Nafion-based MEAs show a WTC of 3, whereas the HC-based MEAs show a very low WTC of -2. • Low WTC of the HC-based MEAs indicates the back-diffusion of water from the cathode to the anode. • Performance of HC-based MEAs is improved as the fuel stoichiometry increases, maintaining high Faradaic efficiency. - Abstract: In order to improve the energy efficiency (fuel efficiency and electrical power) of direct methanol fuel cells (DMFCs), the hydrocarbon (HC) membrane-based membrane electrode assemblies (MEAs) are investigated under various operating conditions. The MEAs are then compared with the conventional Nafion-based MEA in terms of their efficiency and performance. The Faradaic efficiency and water transfer coefficient (WTC) are calculated based on mass balance measurements. The Faradaic efficiency of the HC-based MEAs is improved over the Nafion-based MEAs since methanol crossover decreased. The performance of HC-based MEAs shows strong dependency on the anode stoichiometry at high current densities probably because of the limited mass transport of fuel, which is not observed for the Nafion-based MEAs. The Nafion-based MEAs show a WTC of 3, whereas the HC-based MEAs show a very low WTC of −2, indicating the back-diffusion of water from the cathode to the anode. This may have limited mass transport by interrupting proton conduction at high current densities. The performance of HC-based MEAs at high current densities is improved as the fuel stoichiometry increases; High Faradaic efficiency is maintained by decreasing the cathode stoichiometry

  2. Porous Poly(Ionic Liquid) Membranes as Efficient and Recyclable Absorbents for Heavy Metal Ions.

    Science.gov (United States)

    Ren, Yongyuan; Zhang, Jiandong; Guo, Jiangna; Chen, Fei; Yan, Feng

    2017-07-01

    Heavy metal ion pollution has become a serious environmental problem. Herein, this study reports the synthesis of poly(ionic liquid) (PIL) membranes via in situ photo-crosslinking of vinyl imidazole with both hydrophilic and hydrophobic ionic liquid monomers. The resultant amphiphilic polymer membranes are porous and exhibit high absorption capacity of metal ions (including Hg 2+ , Pb 2+ , Cu 2+ , Cd 2+ , and Zn 2+ ) in both high (1000 mg L -1 ) and low (10 mg L -1 ) concentration metal ion solutions. These metal ionic absorption membranes are easily regenerated in acid solution and can be reused without significant decreases of absorption capacity after many cycles. These PIL membranes may have potential applications as eco-friendly and safe heavy metal ion removal materials. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  3. Efficient ethanol recovery from fermentation broths with integrated distillation-membrane process

    Science.gov (United States)

    The energy demand of distillation-molecular sieve systems for ethanol recovery/dehydration can be significant, particularly for dilute solutions. An alternative process integrating vapor stripping (like a beer still) with vapor compression and a vapor permeation membrane separati...

  4. Efficiency-optimized CO2 separation in IGCC power plants by water-gas shift membrane reactors

    International Nuclear Information System (INIS)

    Schiebahn, Sebastian Thomas

    2013-01-01

    The conversion of solid fuels such as coal and biomass into syngas in the integrated gasification combined cycle (IGCC) process is carried out at elevated pressure. Since, from a thermodynamic point of view, this is a crucial prerequisite for an efficient CO 2 separation step, IGCC has great potential for incorporating CO 2 separation with a low energy consumption. However, studies predict efficiency penalties in the range of 6-11 %-points depending on the respective gasification process utilized, thus revealing that the thermodynamic potential is not fully exploited. In this thesis, a specially adapted IGCC power plant concept for the optimized implementation of gas separation membranes was developed and investigated in order to evaluate the extent to which the auxiliary boundary conditions can be advantageously designed. To create a standard of comparison, a reference IGCC power plant as well as a Selexol-based CO 2 scrubbing process were designed and simulated, resulting in an overall efficiency reduction from 48.0 % to 38.4 %. This corresponds to an increase of 25 % in coal consumption. The analysis of the simulation results revealed that, besides the auxiliary demand of Selexol scrubbing and CO 2 compression subsequent to the low pressure regeneration of the solvent, the main contributor to the loss is the water-gas shift reaction. To reduce this high efficiency penalty, an integration concept was developed to optimize the use of the gas permeation membrane, with parameters better adapted to its special characteristics and mode of operation. The design process resulted in the use of an H 2 -selective membrane, which was combined with the water-gas shift reaction to create the water-gas shift membrane reactor (WGS-MR), and which was swept with recirculated flue gas at elevated pressure in countercurrent 4-End mode. In addition, the ''membrane steam recuperator'' was introduced as a new process unit and integrated to enhance the steam utilization within the

  5. Peptide-Mediated Liposome Fusion: The Effect of Anchor Positioning

    Directory of Open Access Journals (Sweden)

    Niek S. A. Crone

    2018-01-01

    Full Text Available A minimal model system for membrane fusion, comprising two complementary peptides dubbed “E” and “K” joined to a cholesterol anchor via a polyethyleneglycol spacer, has previously been developed in our group. This system promotes the fusion of large unilamellar vesicles and facilitates liposome-cell fusion both in vitro and in vivo. Whilst several aspects of the system have previously been investigated to provide an insight as to how fusion is facilitated, anchor positioning has not yet been considered. In this study, the effects of placing the anchor at either the N-terminus or in the center of the peptide are investigated using a combination of circular dichroism spectroscopy, dynamic light scattering, and fluorescence assays. It was discovered that anchoring the “K” peptide in the center of the sequence had no effect on its structure, its ability to interact with membranes, or its ability to promote fusion, whereas anchoring the ‘E’ peptide in the middle of the sequence dramatically decreases fusion efficiency. We postulate that anchoring the ‘E’ peptide in the middle of the sequence disrupts its ability to form homodimers with peptides on the same membrane, leading to aggregation and content leakage.

  6. Optimization of membrane stack configuration for efficient hydrogen production in microbial reverse-electrodialysis electrolysis cells coupled with thermolytic solutions

    KAUST Repository

    Luo, Xi

    2013-07-01

    Waste heat can be captured as electrical energy to drive hydrogen evolution in microbial reverse-electrodialysis electrolysis cells (MRECs) by using thermolytic solutions such as ammonium bicarbonate. To determine the optimal membrane stack configuration for efficient hydrogen production in MRECs using ammonium bicarbonate solutions, different numbers of cell pairs and stack arrangements were tested. The optimum number of cell pairs was determined to be five based on MREC performance and a desire to minimize capital costs. The stack arrangement was altered by placing an extra low concentration chamber adjacent to anode chamber to reduce ammonia crossover. This additional chamber decreased ammonia nitrogen losses into anolyte by 60%, increased the coulombic efficiency to 83%, and improved the hydrogen yield to a maximum of 3.5mol H2/mol acetate, with an overall energy efficiency of 27%. These results improve the MREC process, making it a more efficient method for renewable hydrogen gas production. © 2013 Elsevier Ltd.

  7. An efficient strategy for small-scale screening and production of archaeal membrane transport proteins in Escherichia coli.

    Science.gov (United States)

    Ma, Pikyee; Varela, Filipa; Magoch, Malgorzata; Silva, Ana Rita; Rosário, Ana Lúcia; Brito, José; Oliveira, Tânia Filipa; Nogly, Przemyslaw; Pessanha, Miguel; Stelter, Meike; Kletzin, Arnulf; Henderson, Peter J F; Archer, Margarida

    2013-01-01

    Membrane proteins play a key role in many fundamental cellular processes such as transport of nutrients, sensing of environmental signals and energy transduction, and account for over 50% of all known drug targets. Despite their importance, structural and functional characterisation of membrane proteins still remains a challenge, partially due to the difficulties in recombinant expression and purification. Therefore the need for development of efficient methods for heterologous production is essential. Fifteen integral membrane transport proteins from Archaea were selected as test targets, chosen to represent two superfamilies widespread in all organisms known as the Major Facilitator Superfamily (MFS) and the 5-Helix Inverted Repeat Transporter superfamily (5HIRT). These proteins typically have eleven to twelve predicted transmembrane helices and are putative transporters for sugar, metabolite, nucleobase, vitamin or neurotransmitter. They include a wide range of examples from the following families: Metabolite-H(+)-symporter; Sugar Porter; Nucleobase-Cation-Symporter-1; Nucleobase-Cation-Symporter-2; and neurotransmitter-sodium-symporter. Overproduction of transporters was evaluated with three vectors (pTTQ18, pET52b, pWarf) and two Escherichia coli strains (BL21 Star and C43 (DE3)). Thirteen transporter genes were successfully expressed; only two did not express in any of the tested vector-strain combinations. Initial trials showed that seven transporters could be purified and six of these yielded quantities of ≥ 0.4 mg per litre suitable for functional and structural studies. Size-exclusion chromatography confirmed that two purified transporters were almost homogeneous while four others were shown to be non-aggregating, indicating that they are ready for up-scale production and crystallisation trials. Here, we describe an efficient strategy for heterologous production of membrane transport proteins in E. coli. Small-volume cultures (10 mL) produced sufficient

  8. Mutation of the dengue virus type 2 envelope protein heparan sulfate binding sites or the domain III lateral ridge blocks replication in Vero cells prior to membrane fusion

    International Nuclear Information System (INIS)

    Roehrig, John T.; Butrapet, Siritorn; Liss, Nathan M.; Bennett, Susan L.; Luy, Betty E.; Childers, Thomas; Boroughs, Karen L.; Stovall, Janae L.; Calvert, Amanda E.; Blair, Carol D.; Huang, Claire Y.-H.

    2013-01-01

    Using an infectious cDNA clone we engineered seven mutations in the putative heparan sulfate- and receptor-binding motifs of the envelope protein of dengue virus serotype 2, strain 16681. Four mutant viruses, KK122/123EE, E202K, G304K, and KKK305/307/310EEE, were recovered following transfection of C6/36 cells. A fifth mutant, KK291/295EE, was recovered from C6/36 cells with a compensatory E295V mutation. All mutants grew in and mediated fusion of virus-infected C6/36 cells, but three of the mutants, KK122/123EE, E202K, G304K, did not grow in Vero cells without further modification. Two Vero cell lethal mutants, KK291/295EV and KKK307/307/310EEE, failed to replicate in DC-SIGN-transformed Raji cells and did not react with monoclonal antibodies known to block DENV attachment to Vero cells. Additionally, both mutants were unable to initiate negative-strand vRNA synthesis in Vero cells by 72 h post-infection, suggesting that the replication block occurred prior to virus-mediated membrane fusion. - Highlights: • Heparan sulfate- and receptor-binding motifs of DENV2 envelope protein were mutated. • Four mutant viruses were isolated—all could fuse C6/36 cells. • Two of these mutants were lethal in Vero cells without further modification. • Lethal mutations were KK291/295EV and KKK305/307/310EEE. • Cell attachment was implicated as the replication block for both mutants

  9. Mutation of the dengue virus type 2 envelope protein heparan sulfate binding sites or the domain III lateral ridge blocks replication in Vero cells prior to membrane fusion

    Energy Technology Data Exchange (ETDEWEB)

    Roehrig, John T., E-mail: jtr1@cdc.gov [Division of Vector-Borne Diseases, Centers for Disease Control and Prevention, Fort Collins, CO 80521 (United States); Butrapet, Siritorn; Liss, Nathan M. [Division of Vector-Borne Diseases, Centers for Disease Control and Prevention, Fort Collins, CO 80521 (United States); Bennett, Susan L. [Arthropod-borne and Infectious Diseases Laboratory, Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, CO 80523 (United States); Luy, Betty E.; Childers, Thomas; Boroughs, Karen L.; Stovall, Janae L.; Calvert, Amanda E. [Division of Vector-Borne Diseases, Centers for Disease Control and Prevention, Fort Collins, CO 80521 (United States); Blair, Carol D. [Arthropod-borne and Infectious Diseases Laboratory, Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, CO 80523 (United States); Huang, Claire Y.-H. [Division of Vector-Borne Diseases, Centers for Disease Control and Prevention, Fort Collins, CO 80521 (United States)

    2013-07-05

    Using an infectious cDNA clone we engineered seven mutations in the putative heparan sulfate- and receptor-binding motifs of the envelope protein of dengue virus serotype 2, strain 16681. Four mutant viruses, KK122/123EE, E202K, G304K, and KKK305/307/310EEE, were recovered following transfection of C6/36 cells. A fifth mutant, KK291/295EE, was recovered from C6/36 cells with a compensatory E295V mutation. All mutants grew in and mediated fusion of virus-infected C6/36 cells, but three of the mutants, KK122/123EE, E202K, G304K, did not grow in Vero cells without further modification. Two Vero cell lethal mutants, KK291/295EV and KKK307/307/310EEE, failed to replicate in DC-SIGN-transformed Raji cells and did not react with monoclonal antibodies known to block DENV attachment to Vero cells. Additionally, both mutants were unable to initiate negative-strand vRNA synthesis in Vero cells by 72 h post-infection, suggesting that the replication block occurred prior to virus-mediated membrane fusion. - Highlights: • Heparan sulfate- and receptor-binding motifs of DENV2 envelope protein were mutated. • Four mutant viruses were isolated—all could fuse C6/36 cells. • Two of these mutants were lethal in Vero cells without further modification. • Lethal mutations were KK291/295EV and KKK305/307/310EEE. • Cell attachment was implicated as the replication block for both mutants.

  10. Acid-induced movements in the glycoprotein shell of an alphavirus turn the spikes into membrane fusion mode

    OpenAIRE

    Haag, Lars; Garoff, Henrik; Xing, Li; Hammar, Lena; Kan, Sin-Tau; Cheng, R.Holland

    2002-01-01

    In the icosahedral (T = 4) Semliki Forest virus, the envelope protomers, i.e. E1–E2 heterodimers, make one-to-one interactions with capsid proteins below the viral lipid bilayer, transverse the membrane and form an external glycoprotein shell with projections. The shell is organized by protomer domains interacting as hexamers and pentamers around shell openings at icosahedral 2- and 5-fold axes, respectively, and the projections by other domains associating as trimers at 3- and quasi 3-fold a...

  11. Self-supported fibrous porous aromatic membranes for efficient CO2/N2 separations.

    Science.gov (United States)

    Meng, Lingbo; Zou, Xiaoqin; Guo, Shukun; Ma, Heping; Zhao, Yongnan; Zhu, Guangshan

    2015-07-22

    In this paper, we describe a new synthesis protocol for the preparation of self-supported hollow fiber membranes composed of porous aromatic framework PAF-56P and PSF. PAF-56P was facilely prepared by the cross-coupling reaction of triangle-shaped cyanuric chloride and linear p-terophenyl monomers. The prepared PAF-56P material possesses an extended conjugated network, the structure of which is confirmed by nuclear magnetic resonance and infrared characterizations, as well as a permanent porosity with a BET surface area of 553.4 m(2) g(-1) and a pore size of 1.2 nm. PAF-56P was subsequently integrated with PSF matrix into PAF-56P/PSF asymmetric hollow fiber membranes via the dry jet-wet quench method employing PAF-56P/PSF suspensions. Scanning electron microscopy studies show that PAF-56P particles are embedded in the PSF matrix to form continuous membranes. Fabricated PAF-56P/PSF membranes were further exploited for CO2 capture, which was exemplified by gas separations of CO2/N2 mixtures. The PAF-56P/PSF membranes show a high selectivity of CO2 over N2 with a separation factor of 38.9 due to the abundant nitrogen groups in the PAF-56P framework. A preferred permeance for CO2 in the binary CO2/N2 gas mixture is obtained in the range of 93-141 GPU due to the large CO2 adsorption capacity and a large pore size of PAF-56P. Additionally, PAF-56P/PSF membranes exhibit excellent thermal and mechanical stabilities, which were examined by thermal analysis and gas separation tests with the dependencies of temperatures and pressures. The merits of high selectivity for CO2, good stability, and easy scale up make PAF-56P/PSF hollow fiber membranes of great interest for the industrial separations of CO2 from the gas exhausts.

  12. Out-coupling membrane for large-size organic light-emitting panels with high efficiency and improved uniformity

    Energy Technology Data Exchange (ETDEWEB)

    Ding, Lei, E-mail: dinglei@sust.edu.cn [College of Electrical and Information Engineering, Shaanxi University of Science and Technology, Xi’an, Shaanxi 710021 (China); Wang, Lu-Wei [College of Electrical and Information Engineering, Shaanxi University of Science and Technology, Xi’an, Shaanxi 710021 (China); Zhou, Lei, E-mail: zhzhlei@gmail.com [Faculty of Mathematics and Physics, Huaiyin Institute of Technology, Huai' an 223003 (China); Zhang, Fang-hui [College of Electrical and Information Engineering, Shaanxi University of Science and Technology, Xi’an, Shaanxi 710021 (China)

    2016-12-15

    Highlights: • An out-coupling membrane embedded with a scattering film of SiO{sub 2} spheres and polyethylene terephthalate (PET) plastic was successfully developed for 150 × 150 mm{sup 2} OLEDs. • Remarkable enhancement in efficiency was achieved from the OLEDs with out- coupling membrane. • The uniformity of large-size GOLED lighting panel is remarkably improved. - Abstract: An out-coupling membrane embedded with a scattering film of SiO{sub 2} spheres and polyethylene terephthalate (PET) plastic was successfully developed for 150 × 150 mm{sup 2} green OLEDs. Comparing with a reference OLED panel, an approximately 1-fold enhancement in the forward emission was obtained with an out-coupling membrane adhered to the surface of the external glass substrate of the panel. Moreover, it was verified that the emission color at different viewing angles can be stabilized without apparent spectral distortion. Particularly, the uniformity of the large-area OLEDs was greatly improved. Theoretical calculation clarified that the improved performance of the lighting panels is primarily attributed to the effect of particle scattering.

  13. Integrated membrane and microbial fuel cell technologies for enabling energy-efficient effluent Re-use in power plants.

    Science.gov (United States)

    Shrestha, Namita; Chilkoor, Govinda; Xia, Lichao; Alvarado, Catalina; Kilduff, James E; Keating, John J; Belfort, Georges; Gadhamshetty, Venkataramana

    2017-06-15

    Municipal wastewater is an attractive alternative to freshwater sources to meet the cooling water needs of thermal power plants. Here we offer an energy-efficient integrated microbial fuel cell (MFC)/ultrafiltration (UF) process to purify primary clarifier effluent from a municipal wastewater treatment plant for use as cooling water. The microbial fuel cell was shown to significantly reduce chemical oxygen demand (COD) in the primary settled wastewater effluent upstream of the UF module, while eliminating the energy demand required to deliver dissolved oxygen in conventional aerobic treatment. We investigated surface modification of the UF membranes to control fouling. Two promising hydrophilic monomers were identified in a high-throughput search: zwitterion (2-(Methacryloyloxy)-ethyl-dimethyl-(3-sulfopropyl ammoniumhydroxide, abbreviated BET SO 3 - ), and amine (2-(Methacryloyloxy) ethyl trimethylammonium chloride, abbreviated N(CH 3 ) 3 + ). Monomers were grafted using UV-induced polymerization on commercial poly (ether sulfone) membranes. Filtration of MFC effluent by membranes modified with BET SO 3 - and N(CH 3 ) 3 + exhibited a lower rate of resistance increase and lower energy consumption than the commercially available membrane. The MFC/UF process produced high quality cooling water that meets the Electrical Power Research Institute (EPRI) recommendations for COD, a suite of metals (Fe, Al, Cu, Zn, Si, Mn, S, Ca and Mg), and offered extremely low corrosion rates (<0.05 mm/yr). A series of AC and DC diagnostic tests were used to evaluate the MFC performance. Copyright © 2017 Elsevier Ltd. All rights reserved.

  14. More Efficient Media Design for Enhanced Biofouling Control in a Membrane Bioreactor: Quorum Quenching Bacteria Entrapping Hollow Cylinder.

    Science.gov (United States)

    Lee, Sang H; Lee, Seonki; Lee, Kibaek; Nahm, Chang H; Kwon, Hyeokpil; Oh, Hyun-Suk; Won, Young-June; Choo, Kwang-Ho; Lee, Chung-Hak; Park, Pyung-Kyu

    2016-08-16

    Recently, membrane bioreactors (MBRs) with quorum quenching (QQ) bacteria entrapping beads have been reported as a new paradigm in biofouling control because, unlike conventional post-biofilm control methods, bacterial QQ can inhibit biofilm formation through its combined effects of physical scouring of the membrane and inhibition of quorum sensing (QS). In this study, using a special reporter strain (Escherichia coli JB525), the interaction between QS signal molecules and quorum quenching bacteria entrapping beads (QQ-beads) was elucidated through visualization of the QS signal molecules within a QQ-bead using a fluorescence microscope. As a result, under the conditions considered in this study, the surface area of QQ-media was likely to be a dominant parameter in enhancing QQ activity over total mass of entrapped QQ bacteria because QQ bacteria located near the core of a QQ-bead were unable to display their QQ activities. On the basis of this information, a more efficient QQ-medium, a QQ hollow cylinder (QQ-HC), was designed and prepared. In batch experiments, QQ-HCs showed greater QQ activity than QQ-beads as a result of their higher surface area and enhanced physical washing effect because of their larger impact area against the membrane surface. Furthermore, it was shown that such advantages of QQ-HCs resulted in more effective mitigation of membrane fouling than from QQ-beads in lab-scale continuous MBRs.

  15. Transparent exopolymer particles (TEP) removal efficiency by a combination of coagulation and ultrafiltration to minimize SWRO membrane fouling

    KAUST Repository

    Li, Sheng

    2016-07-02

    This study investigated the impact of coagulation on the transformation between colloidal and particulate transparent exopolymer particles (TEP) in seawater; and the effectiveness of a combined pretreatment consisting of coagulation and UF on minimizing TEP fouling of seawater reverse osmosis (SWRO) membranes. Coagulation with ferric chloride at pH 5 substantially transformed colloidal TEP (0.1–0.4) into particulate TEP (>0.4) leading to a better membrane fouling control. Both 50 and 100 kDa molecular weight cut-off (MWCO) UF membranes removed most of particulate and colloidal TEP without the assistance of coagulation, but coagulation is still necessary for better UF fouling control. The improvement of combined SWRO pretreatment with coagulation and 50 kDa UF membranes was not that much significant compared to UF pretreatment with 50 KDa alone. Therefore, the minimal coagulant dosage for seawater containing TEP should be based on the UF fouling control requirements rather than removal efficiency. © 2016 Elsevier Ltd

  16. Efficiency of RO/NF membranes at the removal of veterinary antibiotics.

    Science.gov (United States)

    Dolar, D; Vuković, A; Ašperger, D; Košutić, K

    2012-01-01

    The production of pharmaceuticals has increased rapidly during the last several decades as they have been used for the health of both humans and animals. Routes of environmental exposure include the release of treated wastewater, the land disposal of livestock manures and municipal biosolids (i.e. sewage sludge), as well as the use of medicated aquaculture feed. This study deals with application of reverse osmosis (RO) and nanofiltration (NF) membranes for removing of antibiotic residues (sulfamethoxazole, trimethoprim, ciprofloxacin, dexamethasone and febantel) and their mixture. According to the results obtained in this work the use of RO (LFC-1 and XLE) and the tight NF (NF90) membranes are recommended to achieve a high level of retention (>95%) of all selected veterinary antibiotics (VAs). Nanofiltration NF270, NF and HL membranes showed a lower rejection of individual components, but much higher in a mixture solution, due to the synergistic effect.

  17. Avian sarcoma and leukosis virus-receptor interactions: From classical genetics to novel insights into virus-cell membrane fusion

    International Nuclear Information System (INIS)

    Barnard, R.J.O.; Elleder, D.; Young, J.A.T.

    2006-01-01

    For over 40 years, avian sarcoma and leukosis virus (ASLV)-receptor interactions have been employed as a useful model system to study the mechanism of retroviral entry into cells. Pioneering studies on this system focused upon the genetic basis of the differential susceptibilities of different lines of chickens to infection by distinct subgroups of ASLV. These studies led to the definition of three distinct autosomal recessive genes that were predicted to encode cellular receptors for different viral subgroups. They also led to the concept of viral interference, i.e. the mechanism by which infection by one virus can render cells resistant to reinfection by other viruses that use the same cellular receptor. Here, we review the contributions that analyses of the ASLV-receptor system have made in unraveling the mechanisms of retroviral entry into cells and focus on key findings such as identification and characterization of the ASLV receptor genes and the subsequent elucidation of an unprecedented mechanism of virus-cell fusion. Since many of the initial findings on this system were published in the early volumes of Virology, this subject is especially well suited to this special anniversary issue of the journal

  18. Parallel artificial liquid membrane extraction as an efficient tool for removal of phospholipids from human plasma

    DEFF Research Database (Denmark)

    Ask, Kristine Skoglund; Bardakci, Turgay; Parmer, Marthe Petrine

    2016-01-01

    Generic Parallel Artificial Liquid Membrane Extraction (PALME) methods for non-polar basic and non-polar acidic drugs from human plasma were investigated with respect to phospholipid removal. In both cases, extractions in 96-well format were performed from plasma (125μL), through 4μL organic...... solvent used as supported liquid membranes (SLMs), and into 50μL aqueous acceptor solutions. The acceptor solutions were subsequently analysed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) using in-source fragmentation and monitoring the m/z 184→184 transition for investigation...

  19. Fusion of the Dhfr/Mtx and IR/MAR gene amplification methods produces a rapid and efficient method for stable recombinant protein production.

    Science.gov (United States)

    Noguchi, Chiemi; Araki, Yoshio; Miki, Daisuke; Shimizu, Noriaki

    2012-01-01

    Amplification of the dihydrofolate reductase gene (Dhfr) by methotrexate (Mtx) exposure is commonly used for recombinant protein expression in Chinese hamster ovary (CHO) cells. However, this method is both time- and labor-intensive, and the high-producing cells that are generated are frequently unstable in culture. Another gene amplification method is based on using a plasmid bearing a mammalian replication initiation region (IR) and a matrix attachment region (MAR), which result in the spontaneous initiation of gene amplification in transfected cells. The IR/MAR and Dhfr/Mtx methods of gene amplification are based on entirely different principles. In this study, we combine these two methods to yield a novel method, termed the IR/MAR-Dhfr fusion method, which was used to express three proteins, the Fc receptor, GFP, and recombinant antibody. The fusion method resulted in a dramatic increase in expression of all three proteins in two CHO sub-lines, DXB-11, and DG44. The IR/MAR-Dhfr fusion amplified the genes rapidly and efficiently, and produced larger amounts of antibody than the Dhfr/Mtx or IR/MAR methods alone. While the amplified structure produced by the Dhfr/Mtx method was highly unstable, and the antibody production rate rapidly decreased with the culture time of the cells, the IR/MAR-Dhfr fusion method resulted in stable amplification and generated clonal cells that produced large amounts of antibody protein over a long period of time. In summary, the novel IR/MAR-Dhfr fusion method enables isolation of stable cells that produce larger amounts of a target recombinant protein more rapidly and easily than either the Dhfr/Mtx or IR/MAR methods alone.

  20. Experimental demonstration of the reverse flow catalytic membrane reactor concept for energy efficient syngas production. Part 1: Influence of operating conditions

    NARCIS (Netherlands)

    Smit, J.; Bekink, G.J.; van Sint Annaland, M.; Kuipers, J.A.M.

    2007-01-01

    In this contribution the technical feasibility of the reverse flow catalytic membrane reactor (RFCMR) concept with porous membranes for energy efficient syngas production is investigated. In earlier work an experimental proof of principle was already provided [Smit, J., Bekink, G.J., van Sint

  1. Efficiency of Polymeric Membrane Graphene Oxide-TiO2 for Removal of Azo Dye

    Directory of Open Access Journals (Sweden)

    Elahe Dadvar

    2017-01-01

    Full Text Available Achieving the desired standard of drinking water quality has been one of the concerns across water treatment plants in the developing countries. Processes such as grid chamber, coagulation, sedimentation, clarification, filtration, and disinfection are typically used in water purification plants. Among these methods, unit filtration which employs polymers is one of the new technologies. There have been many studies about the use of semiconductive TiO2 with graphene oxide (GO on the base of different polymeric membranes for the removal of azo dyes, especially methylene blue (MB. Polymeric GO-TiO2 membranes have high photocatalytic, antifouling property and permeate the flux removal of organic pollutants. The aim of this study was to investigate the characteristics of different polymeric membranes such as anionic perfluorinated polymer (Nafion, cellulose acetate, polycarbonate (PC, polysulfone fluoride (PSF, and polyvinylidene fluoride (PVDF. The result of this study showed that the GO-TiO2 membrane can be used in the field of water treatment and will be used for the removal of polycyclic aromatic hydrocarbons (PAHs from wastewater.

  2. A trivalent Apx-fusion protein delivered by E. coli outer membrane vesicles induce protection against Actinobacillus pleuropneumoniae of serotype 1 and 7 challenge in a murine model.

    Science.gov (United States)

    Xu, Kui; Zhao, Qin; Wen, Xintian; Wu, Rui; Wen, Yiping; Huang, Xiaobo; Huang, Yong; Yan, Qigui; Han, Xinfeng; Ma, Xiaoping; Chang, Yung-Fu; Cao, Sanjie

    2018-01-01

    Actinobacillus pleuropneumoniae (APP) causes serious economic losses in the swine industry, and is the etiologic agent of porcine pleuropneumonia. In this study we have engineered a trivalent Apx fusion protein enclosed in outer membrane vesicles (Apxr-OMV) and studied its immunoprotective efficacy against APP serotypes 1 and 7 challenge in mice. The results showed that the IgG levels in the Apxr-OMVs immune group were significantly higher than those of the negative control (P < 0.05). Up-regulation of both Th1 (IFN-γ, IL-2) and Th2 (IL-4) cytokines were detected in splenocytes of Apxr-OMVs immune group. The survival rates 87.5% and 62.5% were observed against APP strain 1516 of serotype 7 and APP strain 2701 of serotype 1 in the groups of Apxr-OMVs immune group, respectively. Histopathological lesions of the pulmonary structure alveoli were found to be minimal in APX-OMV group challenged with APP serotypes 1 and 7. These results strongly indicated that engineered OMVs could effectively induce specific humoral or cellular immune responses. Moreover, Apxr-OMVs used as novel vaccine provides cross-protective immunity against different serotype 1 and 7 of APP infection in a mouse model. In contrast, the OMV-empty and PBS as negative controls or inactivated strain of APP-2701 and APP-1516 as positive controls for the animal study cannot provide protection or cross-protection.

  3. MacA, a periplasmic membrane fusion protein of the macrolide transporter MacAB-TolC, binds lipopolysaccharide core specifically and with high affinity.

    Science.gov (United States)

    Lu, Shuo; Zgurskaya, Helen I

    2013-11-01

    The Escherichia coli MacAB-TolC transporter has been implicated in efflux of macrolide antibiotics and secretion of enterotoxin STII. In this study, we found that purified MacA, a periplasmic membrane fusion protein, contains one tightly bound rough core lipopolysaccharide (R-LPS) molecule per MacA molecule. R-LPS was bound specifically to MacA protein with affinity exceeding that of polymyxin B. Sequence analyses showed that MacA contains two high-density clusters of positively charged amino acid residues located in the cytoplasmic N-terminal domain and the periplasmic C-terminal domain. Substitutions in the C-terminal cluster reducing the positive-charge density completely abolished binding of R-LPS. At the same time, these substitutions significantly reduced the functionality of MacA in the protection of E. coli against macrolides in vivo and in the in vitro MacB ATPase stimulation assays. Taken together, our results suggest that R-LPS or a similar glycolipid is a physiological substrate of MacAB-TolC.

  4. Kar5p is required for multiple functions in both inner and outer nuclear envelope fusion in Saccharomyces cerevisiae.

    Science.gov (United States)

    Rogers, Jason V; Rose, Mark D

    2014-12-02

    During mating in the budding yeast Saccharomyces cerevisiae, two haploid nuclei fuse via two sequential membrane fusion steps. SNAREs (i.e., soluble N-ethylmaleimide-sensitive factor attachment protein receptors) and Prm3p mediate outer nuclear membrane fusion, but the inner membrane fusogen remains unknown. Kar5p is a highly conserved transmembrane protein that localizes adjacent to the spindle pole body (SPB), mediates nuclear envelope fusion, and recruits Prm3p adjacent to the SPB. To separate Kar5p's functions, we tested localization, Prm3p recruitment, and nuclear fusion efficiency in various kar5 mutants. All domains and the conserved cysteine residues were essential for nuclear fusion. Several kar5 mutant proteins localized properly but did not mediate Prm3p recruitment; other kar5 mutant proteins localized and recruited Prm3p but were nevertheless defective for nuclear fusion, demonstrating additional functions beyond Prm3p recruitment. We identified one Kar5p domain required for SPB localization, which is dependent on the half-bridge protein Mps3p. Electron microscopy revealed a kar5 mutant that arrests with expanded nuclear envelope bridges, suggesting that Kar5p is required after outer nuclear envelope fusion. Finally, a split-GFP assay demonstrated that Kar5p localizes to both the inner and outer nuclear envelope. These insights suggest a mechanism by which Kar5p mediates inner nuclear membrane fusion. Copyright © 2015 Rogers and Rose.

  5. Self-floating carbon nanotube membrane on macroporous silica substrate for highly efficient solar-driven interfacial water evaporation

    KAUST Repository

    Wang, Yuchao

    2016-01-22

    Given the emerging energy and water challenges facing the mankind, solar-driven water evaporation has been gaining renewed research attention from both academia and industry as an energy efficient means of wastewater treatment and clean water production. In this project, a bi-layered material, consisting of a top self-floating hydrophobic CNT membrane and a bottom hydrophilic macroporous silica substrate, was rationally designed and fabricated for highly energy-efficient solar driven water evaporation based on the concept of interfacial heating. The top thin CNT membrane with excellent light adsorption capability, acted as photothermal component, which harvested and converted almost the entire incident light to heat for exclusively heating of interfacial water. On the other hand, the macroporous silica substrate provided multi-functions toward further improvement of operation stability and water evaporation performance of the material, including water pumping, mechanical support and heat barriers. The silica substrate was conducive in forming the rough surface structures of the CNT top layers during vacuum filtration and thus indirectly contributed to high light adsorption by the top CNT layers. With optimized thicknesses of the CNT top layer and silica substrate, a solar thermal conversion efficiency of 82 % was achieved in this study. The bi-layered material also showed great performance toward water evaporation from seawater and contaminated water, realizing the separation of water from pollutants, and indicating its application versatility.

  6. Efficiency of membrane technology, activated charcoal, and a micelle-clay complex for removal of the acidic pharmaceutical mefenamic acid.

    Science.gov (United States)

    Khalaf, Samer; Al-Rimawi, Fuad; Khamis, Mustafa; Nir, Shlomo; Bufo, Sabino A; Scrano, Laura; Mecca, Gennaro; Karaman, Rafik

    2013-01-01

    The efficiency of sequential advanced membrane technology wastewater treatment plant towards removal of a widely used non-steroid anti-inflammatory drug (NSAID) mefenamic acid was investigated. The sequential system included activated sludge, ultrafiltration by hollow fibre membranes with 100 kDa cutoff, and spiral wound membranes with 20 kDa cutoff, activated carbon and a reverse osmosis (RO) unit. The performance of the integrated plant showed complete removal of mefenamic acid from spiked wastewater samples. The activated carbon column was the most effective component in removing mefenamic acid with a removal efficiency of 97.2%. Stability study of mefenamic acid in pure water and Al-Quds activated sludge revealed that the anti-inflammatory drug was resistant to degradation in both environments. Batch adsorption of mefenamic acid by activated charcoal and a composite micelle (otadecyltrimethylammonium (ODTMA)-clay (montmorillonite) was determined at 25.0°C. Langmuir isotherm was found to fit the data with Qmax of 90.9 mg g(-1) and 100.0 mg g(-1) for activated carbon and micelle-clay complex, respectively. Filtration experiment by micelle-clay columns mixed with sand in the mg L(-1) range revealed complete removal of the drug with much larger capacity than activated carbon column. The combined results demonstrated that an integration of a micelle-clay column in the plant system has a good potential to improve the removal efficiency of the plant towards NSAID drugs such as mefenamic acid.

  7. The chicken embryo as an efficient model to test the function of muscle fusion genes in amniotes.

    Directory of Open Access Journals (Sweden)

    Daniel Sieiro

    Full Text Available The fusion of myoblasts into multinucleated myotubes is a crucial step of muscle growth during development and of muscle repair in the adult. While multiple genes were shown to play a role in this process, a vertebrate model where novel candidates can be tested and analyzed at high throughput and relative ease has been lacking. Here, we show that the early chicken embryo is a fast and robust model in which functional testing of muscle fusion candidate genes can be performed. We have used known modulators of muscle fusion, Rac1 and Cdc42, along with the in vivo electroporation of integrated, inducible vectors, to show that the chicken embryo is a suitable model in which their function can be tested and quantified. In addition to nuclei content, specific characteristics of the experimental model allow a fine characterization of additional morphological features that are nearly impossible to assess in other model organisms. This study should establish the chicken embryo as a cheap, reliable and powerful model in which novel vertebrate muscle fusion candidates can be evaluated.

  8. The golgin GMAP-210 is required for efficient membrane trafficking in the early secretory pathway

    OpenAIRE

    Roboti, Peristera; Sato, Keisuke; Lowe, Martin

    2015-01-01

    Golgins are coiled-coil proteins that participate in membrane-tethering events at the Golgi complex. Golgin-mediated tethering is thought to be important for vesicular trafficking and Golgi organization. However, the degree to which individual golgins contribute to these processes is poorly defined, and it has been proposed that golgins act in a largely redundant manner. Previous studies on the golgin GMAP-210 (also known as TRIP11), which is mutated in the rare skeletal disorder achondrogene...

  9. Delta-Opioid receptors exhibit high efficiency when activating trimeric G proteins in membrane domains

    Czech Academy of Sciences Publication Activity Database

    Bouřová, Lenka; Koštrnová, Alexandra; Hejnová, Lucie; Moravcová, Zuzana; Moon, H. E.; Novotný, Jiří; Milligan, G.; Svoboda, Petr

    2003-01-01

    Roč. 85, č. 1 (2003), s. 34-49 ISSN 0022-3042 R&D Projects: GA MŠk LN00A026; GA ČR GA309/01/0255 Grant - others:Welcome Trust(GB) xx Institutional research plan: CEZ:AV0Z5011922 Keywords : membrane domains * GTPase activity * G protein coupling Subject RIV: CE - Biochemistry Impact factor: 4.825, year: 2003

  10. Applications of MMPBSA to Membrane Proteins I: Efficient Numerical Solutions of Periodic Poisson-Boltzmann Equation.

    Science.gov (United States)

    Botello-Smith, Wesley M; Luo, Ray

    2015-10-26

    Continuum solvent models have been widely used in biomolecular modeling applications. Recently much attention has been given to inclusion of implicit membranes into existing continuum Poisson-Boltzmann solvent models to extend their applications to membrane systems. Inclusion of an implicit membrane complicates numerical solutions of the underlining Poisson-Boltzmann equation due to the dielectric inhomogeneity on the boundary surfaces of a computation grid. This can be alleviated by the use of the periodic boundary condition, a common practice in electrostatic computations in particle simulations. The conjugate gradient and successive over-relaxation methods are relatively straightforward to be adapted to periodic calculations, but their convergence rates are quite low, limiting their applications to free energy simulations that require a large number of conformations to be processed. To accelerate convergence, the Incomplete Cholesky preconditioning and the geometric multigrid methods have been extended to incorporate periodicity for biomolecular applications. Impressive convergence behaviors were found as in the previous applications of these numerical methods to tested biomolecules and MMPBSA calculations.

  11. Mixed matrix membranes with fast and selective transport pathways for efficient CO2 separation

    Science.gov (United States)

    Hou, Jinpeng; Li, Xueqin; Guo, Ruili; Zhang, Jianshu; Wang, Zhongming

    2018-03-01

    To improve CO2 separation performance, porous carbon nanosheets (PCNs) were used as a filler into a Pebax MH 1657 (Pebax) matrix, fabricating mixed matrix membranes (MMMs). The PCNs exhibited a preferential horizontal orientation within the Pebax matrix because of the extremely large 2D plane and nanoscale thickness of the matrix. Therefore, the micropores of the PCNs provided fast CO2 transport pathways, which led to increased CO2 permeability. The reduced pore size of the PCNs was a consequence of the overlapping of PCNs and the polymer chains penetrating into the pores of the PCNs. The reduction in the pore size of the PCNs improved the CO2/gas selectivity. As a result, the CO2 permeability and CO2/CH4 selectivity of the Pebax membrane with 10 wt% PCNs-loading (Pebax-PCNs-10) were 520 barrer and 51, respectively, for CO2/CH4 mixed-gas. The CO2 permeability and CO2/N2 selectivity of the Pebax-PCNs-10 membrane were 614 barrer and 61, respectively, for CO2/N2 mixed-gas.

  12. A facile method to prepare dual-functional membrane for efficient oil removal and in situ reversible mercury ions adsorption from wastewater

    Science.gov (United States)

    Zhang, Qingdong; Liu, Na; Cao, Yingze; Zhang, Weifeng; Wei, Yen; Feng, Lin; Jiang, Lei

    2018-03-01

    In this work, a novel thiol covered polyamide (nylon 66) microfiltration membrane was fabricated by combining mussel-inspired chemistry and coupling reaction, which owns excellent dual-function that can simultaneously remove oil from water efficiently and adsorb the mercury ions contained in the wastewater reversibly. Such membrane exhibited high oil/water separation efficiency, outstanding mercury adsorption ability, and good stability. Moreover, it can be regenerated in nitric acid solution, and maintain its good adsorption performance. The as-prepared membrane showed great potentials for water purification to reduce the heavy metal ion pollution and complicated industrial oily wastewater and living wastewater.

  13. Lysosomal fusion and SNARE function are impaired by cholesterol accumulation in lysosomal storage disorders.

    Science.gov (United States)

    Fraldi, Alessandro; Annunziata, Fabio; Lombardi, Alessia; Kaiser, Hermann-Josef; Medina, Diego Luis; Spampanato, Carmine; Fedele, Anthony Olind; Polishchuk, Roman; Sorrentino, Nicolina Cristina; Simons, Kai; Ballabio, Andrea

    2010-11-03

    The function of lysosomes relies on the ability of the lysosomal membrane to fuse with several target membranes in the cell. It is known that in lysosomal storage disorders (LSDs), lysosomal accumulation of several types of substrates is associated with lysosomal dysfunction and impairment of endocytic membrane traffic. By analysing cells from two severe neurodegenerative LSDs, we observed that cholesterol abnormally accumulates in the endolysosomal membrane of LSD cells, thereby reducing the ability of lysosomes to efficiently fuse with endocytic and autophagic vesicles. Furthermore, we discovered that soluble N-ethylmaleimide-sensitive factor attachment protein (SNAP) receptors (SNAREs), which are key components of the cellular membrane fusion machinery are aberrantly sequestered in cholesterol-enriched regions of LSD endolysosomal membranes. This abnormal spatial organization locks SNAREs in complexes and impairs their sorting and recycling. Importantly, reducing membrane cholesterol levels in LSD cells restores normal SNARE function and efficient lysosomal fusion. Our results support a model by which cholesterol abnormalities determine lysosomal dysfunction and endocytic traffic jam in LSDs by impairing the membrane fusion machinery, thus suggesting new therapeutic targets for the treatment of these disorders.

  14. Cell fusions in mammals

    DEFF Research Database (Denmark)

    Larsson, Lars-Inge; Bjerregaard, Bolette; Talts, Jan Fredrik

    2008-01-01

    Cell fusions are important to fertilization, placentation, development of skeletal muscle and bone, calcium homeostasis and the immune defense system. Additionally, cell fusions participate in tissue repair and may be important to cancer development and progression. A large number of factors appear...... to regulate cell fusions, including receptors and ligands, membrane domain organizing proteins, proteases, signaling molecules and fusogenic proteins forming alpha-helical bundles that bring membranes close together. The syncytin family of proteins represent true fusogens and the founding member, syncytin-1......, has been documented to be involved in fusions between placental trophoblasts, between cancer cells and between cancer cells and host ells. We review the literature with emphasis on the syncytin family and propose that syncytins may represent universal fusogens in primates and rodents, which work...

  15. Efficient expression of stable recombinant human insulin-like growth factor-1 fusion with human serum albumin in Chinese hamster ovary cells.

    Science.gov (United States)

    Wan, Aini; Xu, Dongsheng; Liu, Kedong; Peng, Lin; Cai, Yanfei; Chen, Yun; He, Yang; Yang, Jianfeng; Jin, Jian; Li, Huazhong

    2017-08-09

    Insulin-like growth factor-1 (IGF-1) plays a crucial role in cell development, differentiation, and metabolism, and has been a potential therapeutic agent for many diseases. Chinese hamster ovary (CHO) cells are widely used for production of recombinant therapeutic proteins, but the expression level of IGF-1 in CHO cells is very low (1,500 µg/L) and the half-life of IGF-1 in blood circulation is only 4.5 min according to previous studies. Therefore, IGF-1 was fused to long-circulating serum protein human serum albumin (HSA) and expressed in CHO cells. After 8-day fed-batch culture, the expression level of HSA-IGF-1 reached 100 mg/L. The fusion protein HSA-IGF-1 was purified with a recovery of 35% using a two-step chromatographic procedure. According to bioactivity assay, the purified HSA-IGF-1 could stimulate the proliferation of NIH3T3 cells in a dose-dependent fashion and promote the cell-cycle progression. Besides this, HSA-IGF-1 could bind to IGF-1 receptor on cell membrane and activate the intracellular PI3K/AKT signaling pathway. Our study suggested that HSA fusion technology carried out in CHO cells not only provided bioactivity in HSA-IGF-1 for further research but also offered a beneficial strategy to produce other similar cytokines in CHO cells.

  16. Modeling efficiency and water balance in PEM fuel cell systems with liquid fuel processing and hydrogen membranes

    Science.gov (United States)

    Pearlman, Joshua B.; Bhargav, Atul; Shields, Eric B.; Jackson, Gregory S.; Hearn, Patrick L.

    Integrating PEM fuel cells effectively with liquid hydrocarbon reforming requires careful system analysis to assess trade-offs associated with H 2 production, purification, and overall water balance. To this end, a model of a PEM fuel cell system integrated with an autothermal reformer for liquid hydrocarbon fuels (modeled as C 12H 23) and with H 2 purification in a water-gas-shift/membrane reactor is developed to do iterative calculations for mass, species, and energy balances at a component and system level. The model evaluates system efficiency with parasitic loads (from compressors, pumps, and cooling fans), system water balance, and component operating temperatures/pressures. Model results for a 5-kW fuel cell generator show that with state-of-the-art PEM fuel cell polarization curves, thermal efficiencies >30% can be achieved when power densities are low enough for operating voltages >0.72 V per cell. Efficiency can be increased by operating the reformer at steam-to-carbon ratios as high as constraints related to stable reactor temperatures allow. Decreasing ambient temperature improves system water balance and increases efficiency through parasitic load reduction. The baseline configuration studied herein sustained water balance for ambient temperatures ≤35 °C at full power and ≤44 °C at half power with efficiencies approaching ∼27 and ∼30%, respectively.

  17. An efficient strategy for small-scale screening and production of archaeal membrane transport proteins in Escherichia coli.

    Directory of Open Access Journals (Sweden)

    Pikyee Ma

    Full Text Available Membrane proteins play a key role in many fundamental cellular processes such as transport of nutrients, sensing of environmental signals and energy transduction, and account for over 50% of all known drug targets. Despite their importance, structural and functional characterisation of membrane proteins still remains a challenge, partially due to the difficulties in recombinant expression and purification. Therefore the need for development of efficient methods for heterologous production is essential.Fifteen integral membrane transport proteins from Archaea were selected as test targets, chosen to represent two superfamilies widespread in all organisms known as the Major Facilitator Superfamily (MFS and the 5-Helix Inverted Repeat Transporter superfamily (5HIRT. These proteins typically have eleven to twelve predicted transmembrane helices and are putative transporters for sugar, metabolite, nucleobase, vitamin or neurotransmitter. They include a wide range of examples from the following families: Metabolite-H(+-symporter; Sugar Porter; Nucleobase-Cation-Symporter-1; Nucleobase-Cation-Symporter-2; and neurotransmitter-sodium-symporter. Overproduction of transporters was evaluated with three vectors (pTTQ18, pET52b, pWarf and two Escherichia coli strains (BL21 Star and C43 (DE3. Thirteen transporter genes were successfully expressed; only two did not express in any of the tested vector-strain combinations. Initial trials showed that seven transporters could be purified and six of these yielded quantities of ≥ 0.4 mg per litre suitable for functional and structural studies. Size-exclusion chromatography confirmed that two purified transporters were almost homogeneous while four others were shown to be non-aggregating, indicating that they are ready for up-scale production and crystallisation trials.Here, we describe an efficient strategy for heterologous production of membrane transport proteins in E. coli. Small-volume cultures (10 mL produced

  18. Perfect focusing fusion system

    International Nuclear Information System (INIS)

    Miyamoto, G.; Takeda, T.; Iwata, G.; Mori, S.; Inoue, K.; Tanaka, M.

    1994-01-01

    We propose new perfect focusing (perfo) fusion systems in which ion- or atom-beam is used as a target for the fusion reaction, and ions (perfo particles) of different species moving in the perfo field collide with them. The 'efficiency' defined as the ratio of the fusion energy output to the radiation loss is ∼300, ∼20, and ∼4 for the T+D, 3 He+D, and 11 B+H reactions, respectively. (author)

  19. Enhanced selectivity in mixed matrix membranes for CO2 capture through efficient dispersion of amine-functionalized MOF nanoparticles

    Science.gov (United States)

    Ghalei, Behnam; Sakurai, Kento; Kinoshita, Yosuke; Wakimoto, Kazuki; Isfahani, Ali Pournaghshband; Song, Qilei; Doitomi, Kazuki; Furukawa, Shuhei; Hirao, Hajime; Kusuda, Hiromu; Kitagawa, Susumu; Sivaniah, Easan

    2017-07-01

    Mixed matrix membranes (MMMs) for gas separation applications have enhanced selectivity when compared with the pure polymer matrix, but are commonly reported with low intrinsic permeability, which has major cost implications for implementation of membrane technologies in large-scale carbon capture projects. High-permeability polymers rarely generate sufficient selectivity for energy-efficient CO2 capture. Here we report substantial selectivity enhancements within high-permeability polymers as a result of the efficient dispersion of amine-functionalized, nanosized metal-organic framework (MOF) additives. The enhancement effects under optimal mixing conditions occur with minimal loss in overall permeability. Nanosizing of the MOF enhances its dispersion within the polymer matrix to minimize non-selective microvoid formation around the particles. Amination of such MOFs increases their interaction with thepolymer matrix, resulting in a measured rigidification and enhanced selectivity of the overall composite. The optimal MOF MMM performance was verified in three different polymer systems, and also over pressure and temperature ranges suitable for carbon capture.

  20. A high efficient micro-proton exchange membrane fuel cell by integrating micro-nano synergical structures

    Science.gov (United States)

    Peng, Hsien-Chih; Wang, Chung-Nan; Yeh, Tsung-Kuang; Su, Yu-Chuan; Pan, Chin; Tseng, Fan-Gang

    2013-03-01

    A silicon-based micro-proton exchange membrane fuel cell (μ-PEMFC) which carries out an outstanding cell performance and high efficient catalyst utilization ratio is proposed in this paper by the employment of three micro-nano synergical techniques. Firstly, a novel design incorporating multi-walled carbon nano-tubes (MWCNTs) on radial shaped micro-channel walls to form micro/nano reaction chambers which significantly enhance the reaction surface areas for the performance improvement. In the second, the dispensing of ionomer onto the three-phase zones in the reaction chambers is controlled by different rotation speed and the optimized distribution can therefore be achieved for the improvement of catalyst utilization. In the third, the cell interfacial strength is greatly enhanced by the employment of micro-interlocks for maintaining excellent interface between Nafion membrane and reaction chambers while minimizing the system size. A μ-PEMFC is successfully fabricated by integrating aforementioned synergical micro/nano structures altogether. The best cell efficiency is 26 mW cm-2 which corresponds to a catalyst utilization ratio of 38.2 W g-pt-1, and can be considered among the best performed micro-fuel cells.

  1. High-efficient acetate production from carbon dioxide using a bioanode microbial electrosynthesis system with bipolar membrane.

    Science.gov (United States)

    Xiang, Yinbo; Liu, Guangli; Zhang, Renduo; Lu, Yaobin; Luo, Haiping

    2017-06-01

    The aim of this study was to develop an efficient bioanode microbial electrosynthesis system (MES) to convert carbon dioxide into acetate using bioenergy from the wastewater. The bioanode MESs were constructed using proton exchange membrane (PEM) and bipolar membrane (BPM) as separator, respectively, and operated under different voltages (i.e., 0.8, 1.0, 1.2, and 1.4V). Since BPM could dissociate H 2 O into H + and OH - in situ to buffer the pH change in the chambers, the BPM-MES achieved 238% improvement in cathodic acetate production rate, 45% increase in anodic substrate removal efficiency, and more than five times enhancement in current output, as compared to the PEM-MES. The biomass on the surface of anode and cathode, and the relative abundance of Acetobacterium in the cathode of BPM-MES was higher than that in PEM-MES. Bioanode MES with BPM should be a useful microbial electrosynthesis strategy for acetate production using bioenergy from wastewater treatment. Copyright © 2017 Elsevier Ltd. All rights reserved.

  2. Measurement of the efficiency of cell membrane electroporation using pulsed ac fields

    International Nuclear Information System (INIS)

    Chen, C; Robinson, M P; Evans, J A; Smye, S W; O'Toole, P

    2008-01-01

    Electroporation is a long-established technique used to deliver molecules to cells. Most in vivo electroporation protocols entail applying square-wave, or monotonically-decreasing pulses but relatively few have explored the use of pulsed ac fields. This study measures the efficiency of electroporation in human kidney embryonal cells, using pulsed ac electric fields of peak amplitude 30-200 kV m -1 . The results indicate that optimum electroporation efficiencies of up to 70% can be achieved using pulses at frequencies of 20-160 kHz. Increasing the field strength results in higher electroporation efficiency, but also increases cell kill. This study confirms that efficient electroporation may be achieved using pulsed ac fields. This finding raises the possibility of a wider range of clinical and laboratory applications based on ac technology and avoiding the use of invasive needle electrodes

  3. Energy efficient recovery and dehydration of ethanol from fermentation broths by Membrane Assisted Vapor Stripping technology

    Science.gov (United States)

    Distillation combined with molecular sieve dehydration is the current state of the art for fuel grade ethanol production from fermentation broths. To improve the sustainability of bioethanol production, energy efficient separation alternatives are needed, particularly for lower ...

  4. Surface Density of the Hendra G Protein Modulates Hendra F Protein-Promoted Membrane Fusion: Role for Hendra G Protein Trafficking and Degradation

    OpenAIRE

    Whitman, Shannon D.; Dutch, Rebecca Ellis

    2007-01-01

    Hendra virus, like most paramyxoviruses, requires both a fusion (F) and attachment (G) protein for promotion of cell-cell fusion. Recent studies determined that Hendra F is proteolytically processed by the cellular protease cathepsin L after endocytosis. This unique cathepsin L processing results in a small percentage of Hendra F on the cell surface. To determine how the surface densities of the two Hendra glycoproteins affect fusion promotion, we performed experiments that varied the levels ...

  5. Polymer-Derived Membranes for Large Scale Energy-Efficient Separations

    Science.gov (United States)

    Koros, William

    2013-03-01

    A significant fraction of global energy is consumed to meet separation and purification needs of society, since existing processes are based primarily on energy intensive operations such as distillation. In fact, movement to alternative raw material sources tends to increase this consumption, since separation needs are more difficult to meet in such cases. Energy intensity and carbon dioxide emissions associated with many large scale separations can be reduced by a full order of magnitude by substituting membrane processes for traditional thermally-driven separation approaches. This presentation will provide a framework illustrating how such a strategy can be applied. An advanced manufacturing perspective relying upon polymer-derived materials is stressed within this framework.

  6. Membrane processes for alcohol-water separation: Improving the energy efficiency of biofuel production

    Science.gov (United States)

    The economics and environmental impact of producing fuels and chemicals biologically can be a strong function of the efficiency with which the fermentation products are removed from the biological media. Due to growth inhibition by some fermentation products, including ethanol an...

  7. Efficient treatment of aniline containing wastewater in bipolar membrane microbial electrolysis cell-Fenton system

    DEFF Research Database (Denmark)

    Li, Xiaohu; Jin, Xiangdan; Zhao, Nannan

    2017-01-01

    reaction (MEC-Fenton) for efficient treatment of real wastewater containing a high concentration (4460 ± 52 mg L−1) of aniline. In this system, H2O2 was in situ electro-synthesized from O2 reduction on the graphite cathode and was simultaneously used as source of radical dotOH for the oxidation of aniline...

  8. Spinal fusion

    Science.gov (United States)

    ... Herniated disk - fusion; Spinal stenosis - fusion; Laminectomy - fusion Patient Instructions Bathroom safety - adults Preventing falls Preventing falls - what to ask your doctor Spine surgery - discharge Surgical wound care - open Images Scoliosis Spinal ...

  9. Specific and efficient targeting of cyanobacterial bicarbonate transporters to the inner envelope membrane of chloroplasts in Arabidopsis

    Directory of Open Access Journals (Sweden)

    Susumu eUehara

    2016-02-01

    Full Text Available Installation of cyanobacterial bicarbonate transporters to the inner envelope membrane (IEM of chloroplasts in C3 plants has been thought to improve photosynthetic performance. However, the method to deliver cyanobacterial bicarbonate transporters to the chloroplast IEM remains to be established. In this study, we provide evidence that the cyanobacterial bicarbonate transporters, BicA and SbtA, can be specifically installed into the chloroplast IEM using the chloroplast IEM targeting signal in conjunction with the transit peptide. We fused the transit peptide and the mature portion of Cor413im1, whose targeting mechanism to the IEM has been characterized in detail, to either BicA or SbtA isolated from Synechocystis sp. PCC6803. Among the seven chimeric constructs tested, we confirmed that four chimeric bicarbonate transporters, designated as BicAI, BicAII, SbtAII, and SbtAIII, were expressed in Arabidopsis. Furthermore, these chimeric transporters were specifically targeted to the chloroplast IEM. They were also resistant to alkaline extraction but can be solubilized by Triton X-100, indicating that they are integral membrane proteins in the chloroplast IEM. One of the transporters, BicA, could reside in the chloroplast IEM even after removal of the IEM targeting signal. Taken together, our results indicate that the addition of IEM targeting signal, as well as the transit peptide, to bicarbonate transporters allows us to efficiently target nuclear-encoded chimeric bicarbonate transporters to the chloroplast IEM.

  10. Aluminum Oxide Nanoparticles for Highly Efficient Asphaltene Separation from Crude Oil Using Ceramic Membrane Technology

    Directory of Open Access Journals (Sweden)

    Rezakazemi Mashallah

    2017-11-01

    Full Text Available The effects of aluminum oxide nanoparticles on the removal of asphaltenes from an Iranian crude oil (Soroush using a ceramic membrane with pore size of 0.2 µm were investigated. In order to achieve superior asphaltene separation by ultrafiltration, it is essential to make some changes for destabilizing asphaltene in crude oil. The asphaltene destabilization was done using crude oil contact with an acid containing dissolved metal ions. Metal oxide nanoparticles adsorbed asphaltene molecules and increased their molecular size. The nanoparticle of aluminum oxide was applied to alter precipitation and peptization properties of asphaltenes. Dynamic Light Scattering (DLS was used to measurement of the asphaltene molecular size dissolved in toluene. Raman spectroscopy and the Tuinstra equation were used to determine the aromatic sheet diameter (La via the integrated intensities of the G and D1 modes. This revealed that the asphaltene particles react with nano aluminum oxide and the average molecular size of asphaltene was raised from 512.754 to 2949.557 nm and La from 5.482 to 13.787. The obtained results showed that using nano aluminum oxides, asphaltene separation increased from 60–85 wt% to 90–97 wt% based on the asphaltene content of crude oil.

  11. Efficient Electrocatalytic Reduction of CO2by Nitrogen-Doped Nanoporous Carbon/Carbon Nanotube Membranes: A Step Towards the Electrochemical CO2Refinery.

    Science.gov (United States)

    Wang, Hong; Jia, Jia; Song, Pengfei; Wang, Qiang; Li, Debao; Min, Shixiong; Qian, Chenxi; Wang, Lu; Li, Young Feng; Ma, Chun; Wu, Tom; Yuan, Jiayin; Antonietti, Markus; Ozin, Geoffrey A

    2017-06-26

    Herein we introduce a straightforward, low cost, scalable, and technologically relevant method to manufacture an all-carbon, electroactive, nitrogen-doped nanoporous-carbon/carbon-nanotube composite membrane, dubbed "HNCM/CNT". The membrane is demonstrated to function as a binder-free, high-performance gas diffusion electrode for the electrocatalytic reduction of CO 2 to formate. The Faradaic efficiency (FE) for the production of formate is 81 %. Furthermore, the robust structural and electrochemical properties of the membrane endow it with excellent long-term stability. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  12. Cell fusion by ionizing radiation

    International Nuclear Information System (INIS)

    Khair, M.B.

    1993-08-01

    The relevance and importance of cell fusion are illustrated by the notion that current interest in this phenomenon is shared by scientists in quite varied disciplines. The diversity of cellular membrane fusion phenomena could provoke one to think that there must be a multitude of mechanisms that can account for such diversity. But, in general, the mechanism for the fusion reaction itself could be very similar in many, or even all, cases. Cell fusion can be induced by several factors such as virus Sendai, polyethylene glycol, electric current and ionizing radiation. This article provides the reader with short view of recent progress in research on cell fusion and gives some explanations about fusion mechanisms. This study shows for the first time, the results of the cell fusion induced by ionizing radiations that we have obtained in our researches and the work performed by other groups. (author). 44 refs

  13. ER-associated SNAREs and Sey1p mediate nuclear fusion at two distinct steps during yeast mating.

    Science.gov (United States)

    Rogers, Jason V; Arlow, Tim; Inkellis, Elizabeth R; Koo, Timothy S; Rose, Mark D

    2013-12-01

    During yeast mating, two haploid nuclei fuse membranes to form a single diploid nucleus. However, the known proteins required for nuclear fusion are unlikely to function as direct fusogens (i.e., they are unlikely to directly catalyze lipid bilayer fusion) based on their predicted structure and localization. Therefore we screened known fusogens from vesicle trafficking (soluble N-ethylmaleimide-sensitive factor attachment protein receptors [SNAREs]) and homotypic endoplasmic reticulum (ER) fusion (Sey1p) for additional roles in nuclear fusion. Here we demonstrate that the ER-localized SNAREs Sec20p, Ufe1p, Use1p, and Bos1p are required for efficient nuclear fusion. In contrast, Sey1p is required indirectly for nuclear fusion; sey1Δ zygotes accumulate ER at the zone of cell fusion, causing a block in nuclear congression. However, double mutants of Sey1p and Sec20p, Ufe1p, or Use1p, but not Bos1p, display extreme ER morphology defects, worse than either single mutant, suggesting that retrograde SNAREs fuse ER in the absence of Sey1p. Together these data demonstrate that SNAREs mediate nuclear fusion, ER fusion after cell fusion is necessary to complete nuclear congression, and there exists a SNARE-mediated, Sey1p-independent ER fusion pathway.

  14. A resilient and efficient CFD framework: Statistical learning tools for multi-fidelity and heterogeneous information fusion

    Science.gov (United States)

    Lee, Seungjoon; Kevrekidis, Ioannis G.; Karniadakis, George Em

    2017-09-01

    Exascale-level simulations require fault-resilient algorithms that are robust against repeated and expected software and/or hardware failures during computations, which may render the simulation results unsatisfactory. If each processor can share some global information about the simulation from a coarse, limited accuracy but relatively costless auxiliary simulator we can effectively fill-in the missing spatial data at the required times by a statistical learning technique - multi-level Gaussian process regression, on the fly; this has been demonstrated in previous work [1]. Based on the previous work, we also employ another (nonlinear) statistical learning technique, Diffusion Maps, that detects computational redundancy in time and hence accelerate the simulation by projective time integration, giving the overall computation a "patch dynamics" flavor. Furthermore, we are now able to perform information fusion with multi-fidelity and heterogeneous data (including stochastic data). Finally, we set the foundations of a new framework in CFD, called patch simulation, that combines information fusion techniques from, in principle, multiple fidelity and resolution simulations (and even experiments) with a new adaptive timestep refinement technique. We present two benchmark problems (the heat equation and the Navier-Stokes equations) to demonstrate the new capability that statistical learning tools can bring to traditional scientific computing algorithms. For each problem, we rely on heterogeneous and multi-fidelity data, either from a coarse simulation of the same equation or from a stochastic, particle-based, more "microscopic" simulation. We consider, as such "auxiliary" models, a Monte Carlo random walk for the heat equation and a dissipative particle dynamics (DPD) model for the Navier-Stokes equations. More broadly, in this paper we demonstrate the symbiotic and synergistic combination of statistical learning, domain decomposition, and scientific computing in

  15. Safety and Efficiency of Biomimetic Nanohydroxyapatite/Polyamide 66 Composite in Rabbits and Primary Use in Anterior Cervical Discectomy and Fusion

    Directory of Open Access Journals (Sweden)

    Hui Xu

    2014-01-01

    Full Text Available This study was conducted to validate the safety and efficiency of biomimetic nanohydroxyapatite/polyamide 66 (n-HA/PA66 composite in animal model (rabbit and report its application in anterior cervical discectomy and fusion (ACDF for 4, 12, and 24 weeks. N-HA/PA66 composite was implanted into one-side hind femur defects and the control defects were kept empty as blank controls. A combination of macroscopic and histomorphometric studies was performed up to 24 weeks postoperatively and compared with normal healing. 60 cervical spondylosis myelopathy and radiculopathy patients who were subjected to ACDF using n-HA/PA66 and PEEK cage were involved in this study with six-month minimum follow-up. Their radiographic (cage subsidence, fusion status, and segmental sagittal alignment (SSA and clinical (VAS and JOA scales data before surgery and at each follow-up were recorded and compared. Nanohydroxyapatite/polyamide 66 composite is safe and effective in animal experiment and ACDF.

  16. Investigating time-efficiency of forward masking paradigms for estimating basilar membrane input-output characteristics

    DEFF Research Database (Denmark)

    Fereczkowski, Michal; Jepsen, Morten Løve; Dau, Torsten

    2017-01-01

    appears to be the main drawback of the conventional TMC method. The results from the present study were consistent with the literature when estimating KP levels, but showed some limitations regarding the estimation of the CR values. Perspectives and limitations of both approaches are discussed.......-output (I/O) function have been proposed. However, such measures are very time consuming. The present study investigated possible modifications of the temporal masking curve (TMC) paradigm to improve time and measurement efficiency. In experiment 1, estimates of knee point (KP) and compression ratio (CR......It is well known that pure-tone audiometry does not sufficiently describe individual hearing loss (HL) and that additional measures beyond pure-tone sensitivity might improve the diagnostics of hearing deficits. Specifically, forward masking experiments to estimate basilarmembrane (BM) input...

  17. Highly efficient transition metal and nitrogen co-doped carbide-derived carbon electrocatalysts for anion exchange membrane fuel cells

    Science.gov (United States)

    Ratso, Sander; Kruusenberg, Ivar; Käärik, Maike; Kook, Mati; Puust, Laurits; Saar, Rando; Leis, Jaan; Tammeveski, Kaido

    2018-01-01

    The search for an efficient electrocatalyst for oxygen reduction reaction (ORR) to replace platinum in fuel cell cathode materials is one of the hottest topics in electrocatalysis. Among the many non-noble metal catalysts, metal/nitrogen/carbon composites made by pyrolysis of cheap materials are the most promising with control over the porosity and final structure of the catalyst a crucial point. In this work we show a method of producing a highly active ORR catalyst in alkaline media with a controllable porous structure using titanium carbide derived carbon as a base structure and dicyandiamide along with FeCl3 or CoCl2 as the dopants. The resulting transition metal-nitrogen co-doped carbide derived carbon (M/N/CDC) catalyst is highly efficient for ORR electrocatalysis with the activity in 0.1 M KOH approaching that of commercial 46.1 wt.% Pt/C. The catalyst materials are also investigated by scanning electron microscopy, Raman spectroscopy and X-ray photoelectron spectroscopy to characterise the changes in morphology and composition causing the raise in electrochemical activity. MEA performance of M/N/CDC cathode materials in H2/O2 alkaline membrane fuel cell is tested with the highest power density reached being 80 mW cm-2 compared to 90 mW cm-2 for Pt/C.

  18. High energy efficiency and high power density proton exchange membrane fuel cells: Electrode kinetics and mass transport

    Science.gov (United States)

    Srinivasan, Supramaniam; Velev, Omourtag A.; Parthasathy, Arvind; Manko, David J.; Appleby, A. John

    1991-01-01

    The development of proton exchange membrane (PEM) fuel cell power plants with high energy efficiencies and high power densities is gaining momentum because of the vital need of such high levels of performance for extraterrestrial (space, underwater) and terrestrial (power source for electric vehicles) applications. Since 1987, considerable progress has been made in achieving energy efficiencies of about 60 percent at a current density of 200 mA/sq cm and high power densities (greater than 1 W/sq cm) in PEM fuel cells with high (4 mg/sq cm) or low (0.4 mg/sq cm) platinum loadings in electrodes. The following areas are discussed: (1) methods to obtain these high levels of performance with low Pt loading electrodes - by proton conductor impregnation into electrodes, localization of Pt near front surface; (2) a novel microelectrode technique which yields electrode kinetic parameters for oxygen reduction and mass transport parameters; (3) demonstration of lack of water transport from anode to cathode; (4) modeling analysis of PEM fuel cell for comparison with experimental results and predicting further improvements in performance; and (5) recommendations of needed research and development for achieving the above goals.

  19. Microbial-based evaluation of anaerobic membrane bioreactors (AnMBRs) for the sustainable and efficient treatment of municipal wastewater

    KAUST Repository

    Harb, Moustapha

    2017-03-01

    Conventional activated sludge-based wastewater treatment is an energy and resource-intensive process. Historically it has been successful at producing safely treated wastewater effluents in the developed world, specifically in places that have the infrastructure and space to support its operation. However, with a growing need for safe and efficient wastewater treatment across the world in both urban and rural settings, a paradigm shift in waste treatment is proving to be necessary. The sustainability of the future of wastewater treatment, in a significant way, hinges on moving towards energy neutrality and wastewater effluent reuse. This potential for reuse is threatened by the recent emergence and study of contaminants that have not been previously taken into consideration, such as antibiotics and other organic micropollutants (OMPs), antibiotic resistance genes, and persistent pathogenic bacteria. This dissertation focuses on investigating the use of anaerobic membrane bioreactor (AnMBR) technology for the sustainable treatment of municipal-type wastewaters. Specifically, a microbial approach to understanding biofouling and methane recovery potential in anaerobic MBR systems has been employed to assess different reactor systems’ efficiency. This dissertation further compares AnMBRs to their more widely used aerobic counterparts. This comparison specifically focuses on the removal and biodegradation of OMPs and antibiotics in both anaerobic and aerobic MBRs, while also investigating their effect on the proliferation of antibiotic resistance genes. Due to rising interest in wastewater effluent reuse and the lack of a comprehensive understanding of MBR systems’ effects on pathogen proliferation, this dissertation also investigates the presence of pathogens in both aerobic and anaerobic MBR effluents by using molecularbased detection methods. The findings of this dissertation demonstrate that membrane-associated anaerobic digestion processes have significant

  20. Magnetic fusion reactor economics

    International Nuclear Information System (INIS)

    Krakowski, R.A.

    1995-01-01

    An almost primordial trend in the conversion and use of energy is an increased complexity and cost of conversion systems designed to utilize cheaper and more-abundant fuels; this trend is exemplified by the progression fossil fission → fusion. The present projections of the latter indicate that capital costs of the fusion ''burner'' far exceed any commensurate savings associated with the cheapest and most-abundant of fuels. These projections suggest competitive fusion power only if internal costs associate with the use of fossil or fission fuels emerge to make them either uneconomic, unacceptable, or both with respect to expensive fusion systems. This ''implementation-by-default'' plan for fusion is re-examined by identifying in general terms fusion power-plant embodiments that might compete favorably under conditions where internal costs (both economic and environmental) of fossil and/or fission are not as great as is needed to justify the contemporary vision for fusion power. Competitive fusion power in this context will require a significant broadening of an overly focused program to explore the physics and simbiotic technologies leading to more compact, simplified, and efficient plasma-confinement configurations that reside at the heart of an attractive fusion power plant

  1. Development and characterization of a Rift Valley fever virus cell-cell fusion assay using alphavirus replicon vectors

    International Nuclear Information System (INIS)

    Filone, Claire Marie; Heise, Mark; Doms, Robert W.; Bertolotti-Ciarlet, Andrea

    2006-01-01

    Rift Valley fever virus (RVFV), a member of the Phlebovirus genus in the Bunyaviridae family, is transmitted by mosquitoes and infects both humans and domestic animals, particularly cattle and sheep. Since primary RVFV strains must be handled in BSL-3+ or BSL-4 facilities, a RVFV cell-cell fusion assay will facilitate the investigation of RVFV glycoprotein function under BSL-2 conditions. As for other members of the Bunyaviridae family, RVFV glycoproteins are targeted to the Golgi, where the virus buds, and are not efficiently delivered to the cell surface. However, overexpression of RVFV glycoproteins using an alphavirus replicon vector resulted in the expression of the glycoproteins on the surface of multiple cell types. Brief treatment of RVFV glycoprotein expressing cells with mildly acidic media (pH 6.2 and below) resulted in rapid and efficient syncytia formation, which we quantified by β-galactosidase α-complementation. Fusion was observed with several cell types, suggesting that the receptor(s) for RVFV is widely expressed or that this acid-dependent virus does not require a specific receptor to mediate cell-cell fusion. Fusion occurred over a broad temperature range, as expected for a virus with both mosquito and mammalian hosts. In contrast to cell fusion mediated by the VSV-G glycoprotein, RVFV glycoprotein-dependent cell fusion could be prevented by treating target cells with trypsin, indicating that one or more proteins (or protein-associated carbohydrate) on the host cell surface are needed to support membrane fusion. The cell-cell fusion assay reported here will make it possible to study the membrane fusion activity of RVFV glycoproteins in a high-throughput format and to screen small molecule inhibitors for the ability to block virus-specific membrane fusion

  2. A plasma modified cellulose-chitosan porous membrane allows efficient DNA binding and provides antibacterial properties: A step towards developing a new DNA collecting card.

    Science.gov (United States)

    Chumwangwapee, Sasiwimon; Chingsungnoen, Artit; Siri, Sineenat

    2016-11-01

    In forensic DNA analyses, biological specimens are collected and stored for subsequent recovery and analysis of DNA. A cost-effective and efficient DNA recovery approach is therefore a need. This study aims to produce a plasma modified cellulose-chitosan membrane (pCE-CS) that efficiently binds and retains DNA as a potential DNA collecting card. The pCE-CS membrane was produced by a phase separation of ionic liquid dissolving CE and CS in water with subsequent surface-modification by a two-step exposure of argon plasma and nitrogen gas. Through plasma modification, the pCE-CS membrane demonstrated better DNA retention after a washing process and higher rate of DNA recovery as compared with the original CE-CS membrane and the commercial FTA card. In addition, the pCE-CS membrane exhibited anti-bacterial properties against both Escherichia coli and Staphylococcus aureus. The results of this work suggest a potential function of the pCE-CS membrane as a DNA collecting card with a high recovery rate of captured DNA. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  3. EMP Fusion

    OpenAIRE

    KUNTAY, Isık

    2010-01-01

    This paper introduces a novel fusion scheme, called EMP Fusion, which has the promise of achieving breakeven and realizing commercial fusion power. The method is based on harnessing the power of an electromagnetic pulse generated by the now well-developed flux compression technology. The electromagnetic pulse acts as a means of both heating up the plasma and confining the plasma, eliminating intermediate steps. The EMP Fusion device is simpler compared to other fusion devices and this reduces...

  4. Role of Conserved Histidine Residues in the Low-pH Dependence of the Semliki Forest Virus Fusion Protein▿

    Science.gov (United States)

    Qin, Zhao-ling; Zheng, Yan; Kielian, Margaret

    2009-01-01

    A wide variety of enveloped viruses infects cells by taking advantage of the low pH in the endocytic pathway to trigger virus-membrane fusion. For alphaviruses such as Semliki Forest virus (SFV), acidic pH initiates a series of conformational changes in the heterodimeric virus envelope proteins E1 and E2. Low pH dissociates the E2/E1 dimer, releasing the membrane fusion protein E1. E1 inserts into the target membrane and refolds to a trimeric hairpin conformation, thus driving the fusion reaction. The means by which E1 senses and responds to low pH is unclear, and protonation of conserved E1 histidine residues has been proposed as a possible mechanism. We tested the role of four conserved histidines by mutagenesis of the wild-type (wt) SFV infectious clone to create virus mutants with E1 H3A, H125A, H331A, and H331A/H333A mutations. The H125A, H331A, and H331A/H333A mutants had growth properties similar to those of wt SFV and showed modest change or no change in the pH dependence of virus-membrane fusion. By contrast, the E1 H3A mutation produced impaired virus growth and a markedly more acidic pH requirement for virus-membrane fusion. The dissociation of the H3A heterodimer and the membrane insertion of the mutant E1 protein were comparable to those of the wt in efficiency and pH dependence. However, the formation of the H3A homotrimer required a much lower pH and showed reduced efficiency. Together, these results and the location of H3 suggest that this residue acts to regulate the low-pH-dependent refolding of E1 during membrane fusion. PMID:19244325

  5. Role of conserved histidine residues in the low-pH dependence of the Semliki Forest virus fusion protein.

    Science.gov (United States)

    Qin, Zhao-Ling; Zheng, Yan; Kielian, Margaret

    2009-05-01

    A wide variety of enveloped viruses infects cells by taking advantage of the low pH in the endocytic pathway to trigger virus-membrane fusion. For alphaviruses such as Semliki Forest virus (SFV), acidic pH initiates a series of conformational changes in the heterodimeric virus envelope proteins E1 and E2. Low pH dissociates the E2/E1 dimer, releasing the membrane fusion protein E1. E1 inserts into the target membrane and refolds to a trimeric hairpin conformation, thus driving the fusion reaction. The means by which E1 senses and responds to low pH is unclear, and protonation of conserved E1 histidine residues has been proposed as a possible mechanism. We tested the role of four conserved histidines by mutagenesis of the wild-type (wt) SFV infectious clone to create virus mutants with E1 H3A, H125A, H331A, and H331A/H333A mutations. The H125A, H331A, and H331A/H333A mutants had growth properties similar to those of wt SFV and showed modest change or no change in the pH dependence of virus-membrane fusion. By contrast, the E1 H3A mutation produced impaired virus growth and a markedly more acidic pH requirement for virus-membrane fusion. The dissociation of the H3A heterodimer and the membrane insertion of the mutant E1 protein were comparable to those of the wt in efficiency and pH dependence. However, the formation of the H3A homotrimer required a much lower pH and showed reduced efficiency. Together, these results and the location of H3 suggest that this residue acts to regulate the low-pH-dependent refolding of E1 during membrane fusion.

  6. The influence of selected parameters on the efficiency and economic charactersistics of the oxy-type coal unit with a membrane-cryogenic oxygen separator

    Directory of Open Access Journals (Sweden)

    Kotowicz Janusz

    2016-03-01

    Full Text Available In this paper a 600 MW oxy-type coal unit with a pulverized bed boiler and a membrane-cryogenic oxygen separator and carbon capture installation was analyzed. A membrane-cryogenic oxygen separation installation consists of a membrane module and two cryogenic distillation columns. In this system oxygen is produced with the purity equal to 95%. Installation of carbon capture was based on the physical separation method and allows to reduce the CO2 emission by 90%. In this work the influence of the main parameter of the membrane process – the selectivity coefficient, on the efficiency of the coal unit was presented. The economic analysis with the use of the break-even point method was carried out. The economic calculations were realized in view of the break-even price of electricity depending on a coal unit availability.

  7. The influence of selected parameters on the efficiency and economic charactersistics of the oxy-type coal unit with a membrane-cryogenic oxygen separator

    Science.gov (United States)

    Kotowicz, Janusz; Berdowska, Sylwia

    2016-03-01

    In this paper a 600 MW oxy-type coal unit with a pulverized bed boiler and a membrane-cryogenic oxygen separator and carbon capture installation was analyzed. A membrane-cryogenic oxygen separation installation consists of a membrane module and two cryogenic distillation columns. In this system oxygen is produced with the purity equal to 95%. Installation of carbon capture was based on the physical separation method and allows to reduce the CO2 emission by 90%. In this work the influence of the main parameter of the membrane process - the selectivity coefficient, on the efficiency of the coal unit was presented. The economic analysis with the use of the break-even point method was carried out. The economic calculations were realized in view of the break-even price of electricity depending on a coal unit availability.

  8. Real-time and encryption efficiency improvements of simultaneous fusion, compression and encryption method based on chaotic generators

    Science.gov (United States)

    Jridi, Maher; Alfalou, Ayman

    2018-03-01

    In this paper, enhancement of an existing optical simultaneous fusion, compression and encryption (SFCE) scheme in terms of real-time requirements, bandwidth occupation and encryption robustness is proposed. We have used and approximate form of the DCT to decrease the computational resources. Then, a novel chaos-based encryption algorithm is introduced in order to achieve the confusion and diffusion effects. In the confusion phase, Henon map is used for row and column permutations, where the initial condition is related to the original image. Furthermore, the Skew Tent map is employed to generate another random matrix in order to carry out pixel scrambling. Finally, an adaptation of a classical diffusion process scheme is employed to strengthen security of the cryptosystem against statistical, differential, and chosen plaintext attacks. Analyses of key space, histogram, adjacent pixel correlation, sensitivity, and encryption speed of the encryption scheme are provided, and favorably compared to those of the existing crypto-compression system. The proposed method has been found to be digital/optical implementation-friendly which facilitates the integration of the crypto-compression system on a very broad range of scenarios.

  9. Enhancement of the power system efficiency using the hybrid-type harmonic filters for a KSTAR nuclear fusion experimental system

    International Nuclear Information System (INIS)

    Yoon, Dong-Hee; Lee, Hansang; Park, Byungju; Jang, Gilsoo

    2011-01-01

    Highlights: → The low power factor and power quality problems are occurred by the operation of the PF facility in KSTAR system. We model the power system of KSTAR system including the PF facility. We show a method of the filter insertion to improve the problem and conduct the simulations to verify our method. - Abstract: The KSTAR system, which includes a large amount of nonlinear load, is a relatively high reactive power consumption load which injects harmonic currents into the power system which could result in the possibility for a power system perturbation to occur in the transmission lines, affecting nearby customers. In order to maintain the power quality and power factor in the inner system of the KSTAR system and the adjacent distribution lines, the assessment of the effect of the DC power supply in the KSTAR system is required for appropriate countermeasures to be put in place. In this paper, on the basis of a preliminary inspection of the power system near a KSTAR system, the simulation of a compensating device is performed for the prevention of abnormal voltage variations caused by a large amount of reactive and nonlinear load. In addition, through the comparison of the pre- and post-application of compensation devices in the actual power system, it is verified that a stable operation of the KSTAR nuclear fusion experimental system can be achieved.

  10. Fusion of Enveloped Viruses in Endosomes

    Science.gov (United States)

    White, Judith M.; Whittaker, Gary R.

    2016-01-01

    Ari Helenius launched the field of enveloped virus fusion in endosomes with a seminal paper in the Journal of Cell Biology in 1980. In the intervening years a great deal has been learned about the structures and mechanisms of viral membrane fusion proteins as well as about the endosomes in which different enveloped viruses fuse and the endosomal cues that trigger fusion. We now recognize three classes of viral membrane fusion proteins based on structural criteria and four mechanisms of fusion triggering. After reviewing general features of viral membrane fusion proteins and viral fusion in endosomes, we delve into three characterized mechanisms for viral fusion triggering in endosomes: by low pH, by receptor binding plus low pH, and by receptor binding plus the action of a protease. We end with a discussion of viruses that may employ novel endosomal fusion triggering mechanisms. A key take home message is that enveloped viruses that enter cells by fusing in endosomes traverse the endocytic pathway until they reach an endosome that has all of the environmental conditions (pH, proteases, ions, intracellular receptors, and lipid composition) to (if needed) prime and (in all cases) trigger the fusion protein and to support membrane fusion. PMID:26935856

  11. Efficient Electrocatalytic Reduction of CO2 by Nitrogen-Doped Nanoporous Carbon/Carbon Nanotube Membranes - A Step Towards the Electrochemical CO2 Refinery

    KAUST Repository

    Wang, Hong

    2017-05-12

    The search for earth abundant, efficient and stable electrocatalysts that can enable the chemical reduction of CO2 to value-added chemicals and fuels at an industrially relevant scale, is a high priority for the development of a global network of renewable energy conversion and storage systems that can meaningfully impact greenhouse gas induced climate change. Here we introduce a straightforward, low cost, scalable and technologically relevant method to manufacture an all-carbon, electroactive, nitrogen-doped nanoporous carbon-carbon nanotube composite membrane. The membrane is demonstrated to function as a binder-free, high-performance electrode for the electrocatalytic reduction of CO2 to formate. The Faradaic efficiency for the production of formate is 81%. Furthermore, the robust structural and electrochemical properties of the membrane endow it with excellent long-term stability.

  12. Solid-state nuclear magnetic resonance (NMR) spectroscopy of human immunodeficiency virus gp41 protein that includes the fusion peptide: NMR detection of recombinant Fgp41 in inclusion bodies in whole bacterial cells and structural characterization of purified and membrane-associated Fgp41.

    Science.gov (United States)

    Vogel, Erica P; Curtis-Fisk, Jaime; Young, Kaitlin M; Weliky, David P

    2011-11-22

    Human immunodeficiency virus (HIV) infection of a host cell begins with fusion of the HIV and host cell membranes and is mediated by the gp41 protein, a single-pass integral membrane protein of HIV. The 175 N-terminal residues make up the ectodomain that lies outside the virus. This work describes the production and characterization of an ectodomain construct containing the 154 N-terminal gp41 residues, including the fusion peptide (FP) that binds to target cell membranes. The Fgp41 sequence was derived from one of the African clade A strains of HIV-1 that have been less studied than European/North American clade B strains. Fgp41 expression at a level of ~100 mg/L of culture was evidenced by an approach that included amino acid type (13)CO and (15)N labeling of recombinant protein and solid-state NMR (SSNMR) spectroscopy of lyophilized whole cells. The approach did not require any protein solubilization or purification and may be a general approach for detection of recombinant protein. The purified Fgp41 yield was ~5 mg/L of culture. SSNMR spectra of membrane-associated Fgp41 showed high helicity for the residues C-terminal of the FP. This was consistent with a "six-helix bundle" (SHB) structure that is the final gp41 state during membrane fusion. This observation and negligible Fgp41-induced vesicle fusion supported a function for SHB gp41 of membrane stabilization and fusion arrest. SSNMR spectra of residues in the membrane-associated FP provided evidence of a mixture of molecular populations with either helical or β-sheet FP conformation. These and earlier SSNMR data strongly support the existence of these populations in the SHB state of membrane-associated gp41. © 2011 American Chemical Society

  13. Loss-of-function of the ciliopathy protein Cc2d2a disorganizes the vesicle fusion machinery at the periciliary membrane and indirectly affects Rab8-trafficking in zebrafish photoreceptors.

    Science.gov (United States)

    Ojeda Naharros, Irene; Gesemann, Matthias; Mateos, José M; Barmettler, Gery; Forbes, Austin; Ziegler, Urs; Neuhauss, Stephan C F; Bachmann-Gagescu, Ruxandra

    2017-12-01

    Ciliopathies are human disorders caused by dysfunction of primary cilia, ubiquitous organelles involved in transduction of environmental signals such as light sensation in photoreceptors. Concentration of signal detection proteins such as opsins in the ciliary membrane is achieved by RabGTPase-regulated polarized vesicle trafficking and by a selective barrier at the ciliary base, the transition zone (TZ). Dysfunction of the TZ protein CC2D2A causes Joubert/Meckel syndromes in humans and loss of ciliary protein localization in animal models, including opsins in retinal photoreceptors. The link between the TZ and upstream vesicle trafficking has been little explored to date. Moreover, the role of the small GTPase Rab8 in opsin-carrier vesicle (OCV) trafficking has been recently questioned in a mouse model. Using correlative light and electron microscopy and live imaging in zebrafish photoreceptors, we provide the first live characterization of Rab8-mediated trafficking in photoreceptors in vivo. Our results support a possibly redundant role for both Rab8a/b paralogs in OCV trafficking, based on co-localization of Rab8 and opsins in vesicular structures, and joint movement of Rab8-tagged particles with opsin. We further investigate the role of the TZ protein Cc2d2a in Rab8-mediated trafficking using cc2d2a zebrafish mutants and identify a requirement for Cc2d2a in the latest step of OCV trafficking, namely vesicle fusion. Progressive accumulation of opsin-containing vesicles in the apical portion of photoreceptors lacking Cc2d2a is caused by disorganization of the vesicle fusion machinery at the periciliary membrane with mislocalization and loss of the t-SNAREs SNAP25 and Syntaxin3 and of the exocyst component Exoc4. We further observe secondary defects on upstream Rab8-trafficking with cytoplasmic accumulation of Rab8. Taken together, our results support participation of Rab8 in OCV trafficking and identify a novel role for the TZ protein Cc2d2a in fusion of incoming

  14. Analysis of the system efficiency of an intermediate temperature proton exchange membrane fuel cell at elevated temperature and relative humidity conditions

    International Nuclear Information System (INIS)

    Jeon, Seung Won; Cha, Dowon; Kim, Hyung Soon; Kim, Yongchan

    2016-01-01

    Highlights: • System efficiency of PEMFC is evaluated at elevated temperature and humidity. • Operating parameters are optimized using response surface methodology. • The optimal operating parameters are T = 90.6 °C, RH = 100.0%, and ζ = 2.07. • The power output and system efficiency are 1.28 W and 15.8% at the optimum. • The system efficiency can be effectively improved by increasing relative humidity. - Abstract: Humidification of the membrane is very important in a proton exchange membrane fuel cell (PEMFC), to maintain high ionic conductivity. At an elevated temperature, a large amount of thermal energy is required for humidification because of the exponentially increased saturation vapor pressure. In this study, the system efficiency of a PEMFC was evaluated by considering the heat required for preheating/humidification and compression work. Three-dimensional steady-state simulations were conducted using Fluent 14 to simulate the electrochemical reactions. The operating conditions were optimized using response surface methodology by considering both the fuel cell output and system efficiency. In addition, the effects of operating parameters such as the temperature, relative humidity, and stoichiometric ratio were investigated. The system efficiency can be improved more effectively by increasing relative humidity rather than increasing operating temperature because the ionic conductivity of the membrane was strongly influenced by the relative humidity.

  15. From biological membranes to biomimetic model membranes

    Directory of Open Access Journals (Sweden)

    Eeman, M.

    2010-01-01

    Full Text Available Biological membranes play an essential role in the cellular protection as well as in the control and the transport of nutrients. Many mechanisms such as molecular recognition, enzymatic catalysis, cellular adhesion and membrane fusion take place into the biological membranes. In 1972, Singer et al. provided a membrane model, called fluid mosaic model, in which each leaflet of the bilayer is formed by a homogeneous environment of lipids in a fluid state including globular assembling of proteins and glycoproteins. Since its conception in 1972, many developments were brought to this model in terms of composition and molecular organization. The main development of the fluid mosaic model was made by Simons et al. (1997 and Brown et al. (1997 who suggested that membrane lipids are organized into lateral microdomains (or lipid rafts with a specific composition and a molecular dynamic that are different to the composition and the dynamic of the surrounding liquid crystalline phase. The discovery of a phase separation in the plane of the membrane has induced an explosion in the research efforts related to the biology of cell membranes but also in the development of new technologies for the study of these biological systems. Due to the high complexity of biological membranes and in order to investigate the biological processes that occur on the membrane surface or within the membrane lipid bilayer, a large number of studies are performed using biomimicking model membranes. This paper aims at revisiting the fundamental properties of biological membranes in terms of membrane composition, membrane dynamic and molecular organization, as well as at describing the most common biomimicking models that are frequently used for investigating biological processes such as membrane fusion, membrane trafficking, pore formation as well as membrane interactions at a molecular level.

  16. Microfluidic device for high-yield pairing and fusion of stem cells with somatic cells

    Science.gov (United States)

    Gel, Murat; Hirano, Kunio; Oana, Hidehiro; Kotera, Hidetoshi; Tada, Takashi; Washizu, Masao

    2011-12-01

    Electro cell fusion has significant potential as a biotechnology tool with applications ranging from antibody production to cellular reprogramming. However due to low fusion efficiency of the conventional electro fusion methodology the true potential of the technique has not been reached. In this paper, we report a new method which takes cell fusion efficiency two orders magnitude higher than the conventional electro fusion method. The new method, based on one-toone pairing, fusion and selection of fused cells was developed using a microfabricated device. The device was composed of two microfluidic channels, a micro slit array and a petri dish integrated with electrodes. The electrodes positioned in each channel were used to generate electric field lines concentrating in the micro slits. Cells were introduced into channels and brought in to contact through the micro slit array using dielectrophoresis. The cells in contact were fused by applying a DC pulse to electrodes. As the electric field lines were concentrated at the micro slits the membrane potential was induced only at the vicinity of the micro slits, namely only at the cell-cell contact point. This mechanism assured the minimum damage to cells in the fusion as well as the ability to control the strength and location of induced membrane potential. We introduced mouse embryonic stem cells and mouse embryonic fibroblasts to the microfluidic channels and demonstrated high-yield fusion (> 80%). Post-fusion study showed the method can generate viable hybrids of stem cells and embryonic fibroblasts. Multinucleated hybrid cells adhering on the chip surface were routinely obtained by using this method and on-chip culturing.

  17. Effects of radiation transport on mass ablation rate and conversion efficiency in numerical simulations of inertial confinement fusion

    International Nuclear Information System (INIS)

    Gupta, N.K.

    2002-01-01

    The effects of radiation transport on hydrodynamic parameters of laser produced plasmas are studied. LTE and non-LTE atomic models are used to calculate multi group opacities and emissivities. Screened hydrogenic atom model is used to calculate the energy levels. The population densities of neutral to fully ionized ions are obtained by solving the steady state rate equations. Radiation transport is treated in multi-group diffusion or Sn method. A comparison is made between 1 and 100 group radiation transport and LTE and non-LTE models. For aluminium, multi group radiation transport leads to much higher mass ablation as compared to the 1 group and no radiation transport cases. This in turn leads to higher ablation pressures. However, for gold gray approximation gives higher mass ablation as compared to multi group simulations. LTE conversion efficiency of laser light into x-rays is more than the non-LTE estimates. For LTE as well as non-LTE cases, the one group approximation over-predicts the conversion efficiency Multi group non-LTE simulations predict that the conversion efficiency increases with laser intensity up to a maximum and then it decreases. (author)

  18. Optimizing E. coli-based membrane protein production using Lemo21(DE3) or pReX and GFP-Fusions

    NARCIS (Netherlands)

    Kuipers, Grietje; Peschke, Markus; Ismail, Nurzian Bernsel; Hjelm, Anna; Schlegel, Susan; Vikström, David; Luirink, Joen; de Gier, Jan Willem

    2017-01-01

    Optimizing the conditions for the production of membrane proteins in E. coli is usually a laborious and time-consuming process. Combining the Lemo21(DE3) strain or the pReX T7-based expression vector with membrane proteins C-terminally fused to Green Fluorescent Protein (GFP) greatly facilitates the

  19. MOF-mixed matrix membranes: Precise dispersion of MOF particles with better compatibility via a particle fusion approach for enhanced gas separation properties

    NARCIS (Netherlands)

    Shahid, S.; Nijmeijer, Dorothea C.; Nehache, Sabrina; Vankelecom, Ivo; Deratani, Andre; Quemener, Damien

    2015-01-01

    Mixed matrix membranes (MMMs) incorporating conventional fillers frequently suffer from insufficient adhesion between the polymer matrix and the fillers. This often results in the formation of non-selective voids at the filler/polymer interface, which decreases the performance of the membrane. A

  20. A Visual Attention Model Based Image Fusion

    OpenAIRE

    Rishabh Gupta; M.R.Vimala Devi; M. Devi

    2013-01-01

    To develop an efficient image fusion algorithm based on visual attention model for images with distinct objects. Image fusion is a process of combining complementary information from multiple images of the same scene into an image, so that the resultant image contains a more accurate description of the scene than any of the individual source images. The two basic fusion techniques are pixel level and region level fusion. Pixel level fusion deals with the operations on each and every pixel sep...

  1. A Fusion Protein of the p53 Transaction Domain and the p53-Binding Domain of the Oncoprotein MdmX as an Efficient System for High-Throughput Screening of MdmX Inhibitors.

    Science.gov (United States)

    Chen, Rong; Zhou, Jingjing; Qin, Lingyun; Chen, Yao; Huang, Yongqi; Liu, Huili; Su, Zhengding

    2017-06-27

    In nearly half of cancers, the anticancer activity of p53 protein is often impaired by the overexpressed oncoprotein Mdm2 and its homologue, MdmX, demanding efficient therapeutics to disrupt the aberrant p53-MdmX/Mdm2 interactions to restore the p53 activity. While many potent Mdm2-specific inhibitors have already undergone clinical investigations, searching for MdmX-specific inhibitors has become very attractive, requiring a more efficient screening strategy for evaluating potential scaffolds or leads. In this work, considering that the intrinsic fluorescence residue Trp23 in the p53 transaction domain (p53p) plays an important role in determining the p53-MdmX/Mdm2 interactions, we constructed a fusion protein to utilize this intrinsic fluorescence signal to monitor high-throughput screening of a compound library. The fusion protein was composed of the p53p followed by the N-terminal domain of MdmX (N-MdmX) through a flexible amino acid linker, while the whole fusion protein contained a sole intrinsic fluorescence probe. The fusion protein was then evaluated using fluorescence spectroscopy against model compounds. Our results revealed that the variation of the fluorescence signal was highly correlated with the concentration of the ligand within 65 μM. The fusion protein was further evaluated with respect to its feasibility for use in high-throughput screening using a model compound library, including controls. We found that the imidazo-indole scaffold was a bona fide scaffold for template-based design of MdmX inhibitors. Thus, the p53p-N-MdmX fusion protein we designed provides a convenient and efficient tool for high-throughput screening of new MdmX inhibitors. The strategy described in this work should be applicable for other protein targets to accelerate drug discovery.

  2. A Novel Simple and Efficient Procedure for the Pervaporation Transport Study of Binary Mixtures through Polymeric Membranes: Tested Systems Propanol Isomers — Water–Polyethylene Membrane.

    Czech Academy of Sciences Publication Activity Database

    Setničková, Kateřina; Petričkovič, Roman; Řezníčková Čermáková, Jiřina; Uchytil, Petr

    2016-01-01

    Roč. 58, JAN 2016 (2016), s. 49-56 ISSN 1876-1070 R&D Projects: GA ČR GA104/09/1165 Institutional support: RVO:67985858 Keywords : membrane separation * pervaporation * propanol/water mixture Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 4.217, year: 2016

  3. Immobilization of catalase on electrospun PVA/PA6-Cu(II) nanofibrous membrane for the development of efficient and reusable enzyme membrane reactor.

    Science.gov (United States)

    Feng, Quan; Zhao, Yong; Wei, Anfang; Li, Changlong; Wei, Qufu; Fong, Hao

    2014-09-02

    In this study, a mat/membrane consisting of overlaid PVA/PA6-Cu(II) composite nanofibers was prepared via the electrospinning technique followed by coordination/chelation with Cu(II) ions; an enzyme of catalase (CAT) was then immobilized onto the PVA/PA6-Cu(II) nanofibrous membrane. The amount of immobilized catalase reached a high value of 64 ± 4.6 mg/g, while the kinetic parameters (Vmax and Km) of enzyme were 3774 μmol/mg·min and 41.13 mM, respectively. Furthermore, the thermal stability and storage stability of immobilized catalase were improved significantly. Thereafter, a plug-flow type of immobilized enzyme membrane reactor (IEMR) was assembled from the PVA/PA6-Cu(II)-CAT membrane. With the increase of operational pressure from 0.02 to 0.2 MPa, the flux value of IEMR increased from 0.20 ± 0.02 to 0.76 ± 0.04 L/m(2)·min, whereas the conversion ratio of H2O2 decreased slightly from 92 ± 2.5% to 87 ± 2.1%. After 5 repeating cycles, the production capacity of IEMR was merely decreased from 0.144 ± 0.006 to 0.102 ± 0.004 mol/m(2)·min. These results indicated that the assembled IEMR possessed high productivity and excellent reusability, suggesting that the IEMR based on electrospun PVA/PA6-Cu(II) nanofibrous membrane might have great potential for various applications, particularly those related to environmental protection.

  4. Fusion power

    International Nuclear Information System (INIS)

    Hancox, R.

    1981-01-01

    The principles of fusion power, and its advantages and disadvantages, are outlined. Present research programmes and future plans directed towards the development of a fusion power reactor, are summarized. (U.K.)

  5. Fusion rings and fusion ideals

    DEFF Research Database (Denmark)

    Andersen, Troels Bak

    This dissertation investigates fusion rings, which are Grothendieck groups of rigid, monoidal, semisimple, abelian categories. Special interest is in rational fusion rings, i.e., fusion rings which admit a finite basis, for as commutative rings they may be presented as quotients of polynomial rings...... by the so-called fusion ideals. The fusion rings of Wess-Zumino-Witten models have been widely studied and are well understood in terms of precise combinatorial descriptions and explicit generating sets of the fusion ideals. They also appear in another, more general, setting via tilting modules for quantum...

  6. Fusion: introduction

    International Nuclear Information System (INIS)

    Decreton, M.

    2006-01-01

    The article gives an overview and introduction to the activities of SCK-CEN's research programme on fusion. The decision to construct the ITER international nuclear fusion experiment in Cadarache is highlighted. A summary of the Belgian contributions to fusion research is given with particular emphasis on studies of radiation effects on diagnostics systems, radiation effects on remote handling sensing systems, fusion waste management and socio-economic studies

  7. Fusion Canada

    International Nuclear Information System (INIS)

    1987-07-01

    This first issue of a quarterly newsletter announces the startup of the Tokamak de Varennes, describes Canada's national fusion program, and outlines the Canadian Fusion Fuels Technology Program. A map gives the location of the eleven principal fusion centres in Canada. (L.L.)

  8. Cost-Effective Method for Producing Self Supported Palladium Alloy Membranes for Use in Efficient Production of Coal Derived Hydrogen

    Energy Technology Data Exchange (ETDEWEB)

    K. Coulter

    2008-03-31

    Southwest Research Institute{reg_sign} (SwRI{reg_sign}) has utilized its expertise in large-area vacuum deposition methods to conduct research into the fabrication of dense, freestanding Pd-alloy membranes that are 3-5 microns thick and over 100 in{sup 2} in area. The membranes were deposited onto flexible and rigid supports that were subsequently removed and separated using novel techniques developed over the course of the project. Using these methods, the production of novel alloy compositions centered around the Pd-Cu system were developed with the objective of producing a thermally stable, nano-crystalline grain structure with the highest flux recorded as 242 SCFH/ft{sup 2} for a 2 {micro}m thick Pd{sub 53}Cu{sub 47} at 400 C and 20 psig feed pressure which when extrapolated is over twice the 2010 Department of Energy pure H{sub 2} flux target. Several membranes were made with the same permeability, but with different thicknesses and these membranes were highly selective. Researchers at the Colorado School of Mines supported the effort with extensive testing of experimental membranes as well as design and modeling of novel alloy composite structures. IdaTech provided commercial bench testing and analysis of SwRI-manufactured membranes. The completed deliverables for the project include test data on the performance of experimental membranes fabricated by vacuum deposition and several Pd-alloy membranes that were supplied to IdaTech for testing.

  9. Identification and characterization of a naturally occurring, efficiently cleaved, membrane-bound, clade A HIV-1 Env, suitable for immunogen design, with properties comparable to membrane-bound BG505.

    Science.gov (United States)

    Das, Supratik; Boliar, Saikat; Samal, Sweety; Ahmed, Shubbir; Shrivastava, Tripti; Shukla, Brihaspati N; Goswami, Sandeep; Bansal, Manish; Chakrabarti, Bimal K

    2017-10-01

    Efficient cleavage of HIV-1 Env gp160 into its constituent subunits correlates with selective binding to neutralizing antibodies and are the closest mimetic of native, functional Envs. This was first demonstrated with the clade B Env, JRFL. The correlation between efficient cleavage and selective binding to neutralizing antibodies is the guiding principle for immunogen design for HIV vaccine. We have recently reported that Envs 4-2.J41 (clade C) and JRCSF (clade B) are also efficiently cleaved and show similar properties. However, an efficiently cleaved, membrane-bound clade A Env suitable for genetic vaccination has not been directly demonstrated. Here we report that BG505 and a new clade A Env, QB726.70M.ENV.C4 (or A5) are efficiently cleaved on cell membrane. A5 shows desirable antigenic properties comparable with BG505 on cell surface. A5SOSIP in supernatant displays majority of bNAb binding epitopes. Thus, both BG505 and A5 Envs can be used in DNA prime-protein boost vaccination studies. Copyright © 2017 Elsevier Inc. All rights reserved.

  10. Membrane-controlled processes for the energy-efficient conversion of sludges to fuels and marketable chemicals

    Energy Technology Data Exchange (ETDEWEB)

    None

    1982-03-01

    Studies were carried out on the concentration of primary and secondary sludges by ultrafiltration, and the operation of a membrane-assisted anaerobic digester to treat these sludges. Auxiliary devices including water-spilling and membrane solvent extraction were tested for their feasibility in the ehhancement of digester operations and the recovery of valuable byproducts. It was shown that membrane-facilitated digestion can increase the rate of these processes by a factor of ten, together with a substantial decrease in the amount going to waste, and with the ultrafiltration permeate containing appreciable concentrations of valuable byproducts which could be concentrated and recovered using a combination of other membrane technologies. The utility of electrodialytic water-splitting and membrane solvent extraction was demonstrated. All of this was accomplished with a small three-liter bench-scale digester, the operation of which presented many problems because of its very small size and the difficulty in handling real sewage sludges.

  11. A Novel Energy-Efficient Multi-Sensor Fusion Wake-Up Control Strategy Based on a Biomimetic Infectious-Immune Mechanism for Target Tracking.

    Science.gov (United States)

    Zhou, Jie; Liang, Yan; Shen, Qiang; Feng, Xiaoxue; Pan, Quan

    2018-04-18

    A biomimetic distributed infection-immunity model (BDIIM), inspired by the immune mechanism of an infected organism, is proposed in order to achieve a high-efficiency wake-up control strategy based on multi-sensor fusion for target tracking. The resultant BDIIM consists of six sub-processes reflecting the infection-immunity mechanism: occurrence probabilities of direct-infection (DI) and cross-infection (CI), immunity/immune-deficiency of DI and CI, pathogen amount of DI and CI, immune cell production, immune memory, and pathogen accumulation under immunity state. Furthermore, a corresponding relationship between the BDIIM and sensor wake-up control is established to form the collaborative wake-up method. Finally, joint surveillance and target tracking are formulated in the simulation, in which we show that the energy cost and position tracking error are reduced to 50.8% and 78.9%, respectively. Effectiveness of the proposed BDIIM algorithm is shown, and this model is expected to have a significant role in guiding the performance improvement of multi-sensor networks.

  12. Efficient broadband simulation of fluid-structure coupling for membrane-type acoustic transducer arrays using the multi-level fast multipole algorithm

    Science.gov (United States)

    Shieh, Bernard; Sabra, Karim; Degertekin, F. Levent

    2016-01-01

    A boundary element model provides great flexibility for the simulation of membrane-type micromachined ultrasonic transducers (MUTs) in terms of membrane shape, actuating mechanism, and array layout. Acoustic crosstalk is accounted for through a mutual impedance matrix which captures the primary crosstalk mechanism of dispersive-guided modes generated at the fluid-solid interface. However, finding the solution to the fully-populated boundary element matrix equation using standard techniques requires computation time and memory usage which scales by the cube and by the square of the number of nodes, respectively, limiting simulation to a small number of membranes. We implement a solver with improved speed and efficiency through the application of a multi-level fast multipole algorithm (FMA). By approximating the fields of collections of nodes using multipole expansions of the free-space Green’s function, an FMA solver can enable the simulation of hundreds of thousands of nodes while incurring an approximation error that is controllable. Convergence is drastically improved using a problem-specific block-diagonal preconditioner. We demonstrate the solver’s capabilities by simulating a 32-element 7 MHz 1-D CMUT phased array with 2880 membranes. The array is simulated using 233,280 nodes for a very wide frequency band up to 50 MHz. For a simulation with 15,210 nodes, the FMA solver performed 10-times faster and used 32-times less memory than a standard solver based on LU decomposition We investigate the effects of mesh density and phasing on the predicted array response and find that it is necessary to use about 7 nodes over the width of the membrane to observe convergence of the solution–even below the first membrane resonance frequency–due to the influence of higher-order membrane modes. PMID:27824572

  13. ALKALI FUSION OF ROSETTA ZIRCON

    International Nuclear Information System (INIS)

    DAHER, A.

    2008-01-01

    The decomposition of Rosetta zircon by fusion with different types of alkalis has been investigated. These alkalis include sodium hydroxide, potassium hydroxide and eutectic mixture of both. The influences of the reaction temperature, zircon to alkalis ratio, fusion time and the stirring of the reactant on the fusion reaction have been evaluated. The obtained results favour the decomposition of zircon with the eutectic alkalis mixture by a decomposition efficiency of 96% obtained at 500 0 C after one hour

  14. Development of more efficient and cheaper MEA's for PEM fuel cells; Membrane-electrode-assembly

    Energy Technology Data Exchange (ETDEWEB)

    Yde Andersen, S. (IRD Fuel Cell A/S, Svendborg (Denmark)); Nilsson, M.S. (Danish Power System Aps, Charlottenlund (Denmark)); Siu, A.; Plackett, D. (Technical Univ. of Denmark. Risoe National Lab. for Sustainable Energy, Dansk Polymer Center, Roskilde (Denmark)); Li, Q. (Technical Univ. of Denmark, Dept. of Chemistry, Kgs. Lyngby (Denmark))

    2008-06-15

    The project covered 5 main areas: 1) polymer and membranes; 2) electrocatalysts; 3) gas diffusion electrodes; 4) MEAs; and 5) evaluation techniques. For the polymers, by purification of monomers and optimizing parameters, high molecular weight polybenzimidazoles have been synthesized in batches of 50 g with good reproducibility. Based on the polymer, two types of new membranes have been prepared. One is the cross-linked (covalently and acid-base) PBI blend membranes. The blend membranes were systematically characterized and show excellent properties such as very high acid doping levels, conductivity, mechanical strength and durability. The other type is composite membranes based on PBI and nanoclay. Using the modified nanoclay, good dispersion and transparent composite membranes have been achieved. For catalyst preparation, the carbon supports have been modified with thermal treatment. Improved corrosion resistance was achieved with little sacrificing of the catalytic activity. High Pt loading catalysts were prepared, based on which high performance gas diffusion electrodes were fabricated. The performance target of both cathode and anode was achieved, as evaluated by the PTFE half cell tests. New gas diffusion layer (GDL) materials have been developed and tested in different MEA configurations. Significant performance improvement has been achieved with also potential to reduce the cost. Techniques for applying micro porous layers and catalyst layers have been optimized, including tape casting, spraying, and catalyst-coated membrane (CCM). Using the developed membranes and gas diffusion electrodes, membrane-electrode assemblies (MEAs) were fabricated for both single cell and stack tests. Selection of sealing materials and design of integrated gaskets have been made for both low and high temperature MEAs. Parameters for hot-pressing such as temperature, pressure and duration were systematically studied. 44 MEAs with an active area of 256 cm{sup 2} have been prepared

  15. Fusion cost normalization

    International Nuclear Information System (INIS)

    Schulte, S.C.; Willke, T.L.

    1978-01-01

    The categorization and accounting methods described in this paper provide a common format that can be used to assess the economic character of magnetically confined fusion reactor design concepts. The format was developed with assistance from the fusion economics community, thus ensuring that the methods meet with the approval of potential users. The format will aid designers in the preparation of design concept cost estimates and also provide policy makers with a tool to assist in appraising which design concepts may be economically promising. Adherence to the format when evaluating prospective fusion reactor design concepts will result in the identification of the more promising concepts, thus enabling the fusion power alternatives with better economic potential to be quickly and efficiently developed

  16. Cold nuclear fusion device

    International Nuclear Information System (INIS)

    Ogino, Shinji.

    1991-01-01

    Selection of cathode material is a key to the attainment of cold nuclear fusion. However, there are only few reports on the cathode material at present and an effective development has been demanded. The device comprises an anode and a cathode and an electrolytic bath having metal salts dissolved therein and containing heavy water in a glass container. The anode is made of gold or platinum and the cathode is made of metals of V, Sr, Y, Nb, Hf or Ta, and a voltage of 3-25V is applied by way of a DC power source between them. The metal comprising V, Sr, Y, Nb, Hf or Ta absorbs deuterium formed by electrolysis of heavy water effectively to cause nuclear fusion reaction at substantially the same frequency and energy efficiency as palladium and titanium. Accordingly, a cold nuclear fusion device having high nuclear fusion generation frequency can be obtained. (N.H.)

  17. Vpr14-88-Apobec3G fusion protein is efficiently incorporated into Vif-positive HIV-1 particles and inhibits viral infection.

    Science.gov (United States)

    Ao, Zhujun; Yu, Zhe; Wang, Lina; Zheng, Yingfeng; Yao, Xiaojian

    2008-04-16

    APOBEC3G (A3G), a deoxycytidine deaminase, is a potent host antiviral factor that can restrict HIV-1 infection. During Vif-negative HIV-1 replication, A3G is incorporated into HIV-1 particles, induces mutations in reverse transcribed viral DNA and inhibits reverse transcription. However, HIV-1 Vif counteracts A3G's activities by inducing its degradation and by blocking its incorporation into HIV-1 particles. Thus, it is interesting to elucidate a mechanism that would allow A3G to escape the effects of Vif in order to rescue its potent antiviral activity and to provide a possible novel therapeutic strategy for treating HIV-1 infection. In this study, we generated an R88-A3G fusion protein by fusing A3G to a virion-targeting polypeptide (R14-88) derived from HIV-1 Vpr protein and compared its antiviral effects relative to those of HA-tagged native A3G (HA-A3G). Our study showed that transient expression of the R88-A3G fusion protein in both Vif(-) and Vif(+) HIV-1 producing cells drastically inhibited viral infection in HeLa-CD4-CCR5-cells, CD4(+) C8166 T cells and human primary PBMCs. Moreover, we established CD4(+) C8166 T cell lines that stably express either R88-A3G or HA-A3G by transduction with VSV-G-pseudotyped lentiviral vector that harbor expression cassettes for R88-A3G or HA-A3G, respectively, and tested their susceptibility to Vif(+) HIV-1 infection. Our results clearly reveal that expression of R88-A3G in transduced CD4(+) C8166 cells significantly blocked Vif(+) HIV-1 infection. In an attempt to understand the mechanism underlying the antiviral activity of R88-A3G, we demonstrated that R88-A3G was efficiently incorporated into viral particles in the presence of Vif. Moreover, PCR analysis revealed that R88-A3G significantly inhibited viral cDNA synthesis during the early stage of Vif(+) virus infection. Our results clearly indicate that R88 delivers A3G into Vif(+) HIV-1 particles and inhibits infectivity and spread of the virions among CD4(+) T cells

  18. Specific interaction of CXCR4 with CD4 and CD8α: Functional analysis of the CD4/CXCR4 interaction in the context of HIV-1 envelope glycoprotein-mediated membrane fusion

    International Nuclear Information System (INIS)

    Basmaciogullari, Stephane; Pacheco, Beatriz; Bour, Stephan; Sodroski, Joseph

    2006-01-01

    We investigated possible interactions between HIV-1 receptor (CD4) and the main coreceptors CXCR4 and CCR5. We found that CD4 and CXCR4 coexpressed in 293T cells form a complex that can be immunoprecipitated with antibodies directed against the extracellular domain of either protein. Mutagenesis revealed that the CD4/CXCR4 interaction maps to two previously uncharacterized basic motifs in the cytoplasmic domain of CD4. HIV-1 envelope glycoprotein-mediated membrane fusion was found to be independent of the ability of CD4 and CXCR4 to interact, whether fusion was studied in a virus-cell or a cell-cell model. However, this interaction might explain the adaptation of HIV-1 to CXCR4 as an alternative to CCR5. We found that CXCR4 also interacts with the cytoplasmic domain of CD8α in a way that is similar to the CD4/CXCR4 interaction. The CD4/CXCR4 and CD8α/CXCR4 interactions may thus be involved in cellular signaling pathways shared by the CD4 and CD8α molecules

  19. Efficient overproduction of membrane proteins in Lactococcus lactis requires the cell envelope stress sensor/regulator couple CesSR

    NARCIS (Netherlands)

    Pinto, Joao P C; Kuipers, Oscar P; Marreddy, Ravi K R; Poolman, Bert; Kok, Jan

    2011-01-01

    BACKGROUND: Membrane proteins comprise an important class of molecules whose study is largely frustrated by several intrinsic constraints, such as their hydrophobicity and added requirements for correct folding. Additionally, the complexity of the cellular mechanisms that are required to insert

  20. Alkaline Exchange Membrane (AEM) for High-Efficiency Fuel Cells, Electrolyzers and Regenerative Fuel Cell Systems Project

    Data.gov (United States)

    National Aeronautics and Space Administration — Develop an alkaline exchange membrane (AEM)for use as a polymer electrolyte in both fuel cell and electrolyzer systems.  The ultimate goal in AEM development is...

  1. Evaluating the efficiency of different microfiltration and ultrafiltration membranes used as pretreatment for Red Sea water reverse osmosis desalination

    KAUST Repository

    Almashharawi, Samir

    2013-01-01

    Conventional processes are widely used as pretreatment for reverse osmosis (RO) desalination technology since its development. However, these processes require a large footprint and have some limitation issues such as difficulty to maintain a consistent silt density index, coagulation control at low total suspended solids, and management of higher waste sludge. Recently, there has been a rapid growth in the use of low-pressure membranes as pretreatment for RO systems replacing the conventional processes. However, despite the numerous advantages of using this integrated membrane system mainly providing good and stable water quality to RO membranes, many issues have to be addressed. The primary limitation is membrane fouling which reduces the permeate flux; therefore, higher pumping intensity is required to maintain a consistent volume of product. This paper aims to optimize the permeation flux and cleaning frequency by providing high permeate quality. Different low-pressure polyethersulfone membranes with different pore sizes ranging from 0.1 lm to 50 kDa were tested. Eight different filtration configurations have been applied including the variation of coagulant doses aiming to control membrane fouling. Results showed that all the configurations with/without coagulation, provided permeate with excellent water quality which improves the stability of RO performance. However, more stable fluxes with less-energy consumption were achieved by using the 0.1 lm and 100 kDa membranes with 1 mg/L FeCl3 coagulation. The use of UF membranes, having tight pores, without coagulation also proved to be an excellent option for Red Sea water RO pretreatment. © 2013 Desalination Publications.

  2. COST-EFFECTIVE METHOD FOR PRODUCING SELF SUPPORTED PALLADIUM ALLOY MEMBRANES FOR USE IN EFFICIENT PRODUCTION OF COAL DERIVED HYDROGEN

    Energy Technology Data Exchange (ETDEWEB)

    J. Arps; K. Coulter

    2006-09-30

    In the past quarter, we have conducted additional characterization and permeation tests on different Pd alloy membranes including PdCuTa ternary alloy materials. We attempted to address some discrepancies between SwRI{reg_sign} and CSM relating to PdCu stoichiometry by preparing a range of PdCu membranes with compositions from {approx}58-65 at% Pd (bal. Cu). While some difficulties in cutting and sealing these thin membranes at CSM continue, some progress has been made in identifying improved membrane support materials. We have also completed an initial cost analysis for large-scale vacuum deposition and fabrication of thin Pd ally membranes and project that the process can meet DOE cost targets. Minimal progress was made in the past quarter relating to the testing of prototype membrane modules at Idatech. In the past quarter Idatech was acquired by a UK investment firm, which we believe may have impacted the ability of key technical personnel to devote sufficient time to support this effort. We are hopeful their work can be completed by the end of the calendar year.

  3. Tunable integration of absorption-membrane-adsorption for efficiently separating low boiling gas mixtures near normal temperature.

    Science.gov (United States)

    Liu, Huang; Pan, Yong; Liu, Bei; Sun, Changyu; Guo, Ping; Gao, Xueteng; Yang, Lanying; Ma, Qinglan; Chen, Guangjin

    2016-02-19

    Separation of low boiling gas mixtures is widely concerned in process industries. Now their separations heavily rely upon energy-intensive cryogenic processes. Here, we report a pseudo-absorption process for separating low boiling gas mixtures near normal temperature. In this process, absorption-membrane-adsorption is integrated by suspending suitable porous ZIF material in suitable solvent and forming selectively permeable liquid membrane around ZIF particles. Green solvents like water and glycol were used to form ZIF-8 slurry and tune the permeability of liquid membrane surrounding ZIF-8 particles. We found glycol molecules form tighter membrane while water molecules form looser membrane because of the hydrophobicity of ZIF-8. When using mixing solvents composed of glycol and water, the permeability of liquid membrane becomes tunable. It is shown that ZIF-8/water slurry always manifests remarkable higher separation selectivity than solid ZIF-8 and it could be tuned to further enhance the capture of light hydrocarbons by adding suitable quantity of glycol to water. Because of its lower viscosity and higher sorption/desorption rate, tunable ZIF-8/water-glycol slurry could be readily used as liquid absorbent to separate different kinds of low boiling gas mixtures by applying a multistage separation process in one traditional absorption tower, especially for the capture of light hydrocarbons.

  4. Tunable integration of absorption-membrane-adsorption for efficiently separating low boiling gas mixtures near normal temperature

    Science.gov (United States)

    Liu, Huang; Pan, Yong; Liu, Bei; Sun, Changyu; Guo, Ping; Gao, Xueteng; Yang, Lanying; Ma, Qinglan; Chen, Guangjin

    2016-01-01

    Separation of low boiling gas mixtures is widely concerned in process industries. Now their separations heavily rely upon energy-intensive cryogenic processes. Here, we report a pseudo-absorption process for separating low boiling gas mixtures near normal temperature. In this process, absorption-membrane-adsorption is integrated by suspending suitable porous ZIF material in suitable solvent and forming selectively permeable liquid membrane around ZIF particles. Green solvents like water and glycol were used to form ZIF-8 slurry and tune the permeability of liquid membrane surrounding ZIF-8 particles. We found glycol molecules form tighter membrane while water molecules form looser membrane because of the hydrophobicity of ZIF-8. When using mixing solvents composed of glycol and water, the permeability of liquid membrane becomes tunable. It is shown that ZIF-8/water slurry always manifests remarkable higher separation selectivity than solid ZIF-8 and it could be tuned to further enhance the capture of light hydrocarbons by adding suitable quantity of glycol to water. Because of its lower viscosity and higher sorption/desorption rate, tunable ZIF-8/water-glycol slurry could be readily used as liquid absorbent to separate different kinds of low boiling gas mixtures by applying a multistage separation process in one traditional absorption tower, especially for the capture of light hydrocarbons. PMID:26892255

  5. Sugarcane vinasse treatment by two-stage anaerobic membrane bioreactor: Effect of hydraulic retention time on changes in efficiency, biogas production and membrane fouling.

    Science.gov (United States)

    Santos, Fábio S; Ricci, Bárbara C; França Neta, Luzia S; Amaral, Míriam C S

    2017-12-01

    This research investigated the effect of hydraulic retention time (HRT) on two-stage anaerobic membrane bioreactor (2-SAnMBR) performance treating sugarcane vinasse. The experimental setup consisted of an upflow acidogenic reactor and a continuous stirred methanogenic reactor, fitted with submersed microfiltration hollow-fiber membranes. The results indicated excellent performance and robustness of 2-SAnMBR. The reduction in HRT of 5.3-3.1days did not cause loss of its performance. The 2-SAnMBR showed high capacity of removing organic matter (97%), producing biogas (6.3Nm 3 of CH 4 per m 3 of treated vinasse) and did not completely remove important nutrients to fertigation. Reducing the HRT, the average mass of soluble microbial products (SMP) and extracellular polymeric substances (EPS) per mass of mixed liquor volatile suspended solids (MLVSS) increased. Consequently, the transmembrane pressure (TPM) rate and fouling resistance rise. Despite the fouling effect, physical and chemical cleaning processes were able to recover operational permeability. Copyright © 2017 Elsevier Ltd. All rights reserved.

  6. Hybrid pressure retarded osmosis-membrane distillation system for power generation from low-grade heat: thermodynamic analysis and energy efficiency.

    Science.gov (United States)

    Lin, Shihong; Yip, Ngai Yin; Cath, Tzahi Y; Osuji, Chinedum O; Elimelech, Menachem

    2014-05-06

    We present a novel hybrid membrane system that operates as a heat engine capable of utilizing low-grade thermal energy, which is not readily recoverable with existing technologies. The closed-loop system combines membrane distillation (MD), which generates concentrated and pure water streams by thermal separation, and pressure retarded osmosis (PRO), which converts the energy of mixing to electricity by a hydro-turbine. The PRO-MD system was modeled by coupling the mass and energy flows between the thermal separation (MD) and power generation (PRO) stages for heat source temperatures ranging from 40 to 80 °C and working concentrations of 1.0, 2.0, and 4.0 mol/kg NaCl. The factors controlling the energy efficiency of the heat engine were evaluated for both limited and unlimited mass and heat transfer kinetics in the thermal separation stage. In both cases, the relative flow rate between the MD permeate (distillate) and feed streams is identified as an important operation parameter. There is an optimal relative flow rate that maximizes the overall energy efficiency of the PRO-MD system for given working temperatures and concentration. In the case of unlimited mass and heat transfer kinetics, the energy efficiency of the system can be analytically determined based on thermodynamics. Our assessment indicates that the hybrid PRO-MD system can theoretically achieve an energy efficiency of 9.8% (81.6% of the Carnot efficiency) with hot and cold working temperatures of 60 and 20 °C, respectively, and a working solution of 1.0 M NaCl. When mass and heat transfer kinetics are limited, conditions that more closely represent actual operations, the practical energy efficiency will be lower than the theoretically achievable efficiency. In such practical operations, utilizing a higher working concentration will yield greater energy efficiency. Overall, our study demonstrates the theoretical viability of the PRO-MD system and identifies the key factors for performance

  7. Fusion breeder

    International Nuclear Information System (INIS)

    Moir, R.W.

    1982-01-01

    The fusion breeder is a fusion reactor designed with special blankets to maximize the transmutation by 14 MeV neutrons of uranium-238 to plutonium or thorium to uranium-233 for use as a fuel for fission reactors. Breeding fissile fuels has not been a goal of the US fusion energy program. This paper suggests it is time for a policy change to make the fusion breeder a goal of the US fusion program and the US nuclear energy program. The purpose of this paper is to suggest this policy change be made and tell why it should be made, and to outline specific research and development goals so that the fusion breeder will be developed in time to meet fissile fuel needs

  8. Subcellular localization of a PhoE-LacZ fusion protein in E. coli by protease accessibility experiments reveals an inner-membrane-spanning form of the protein

    NARCIS (Netherlands)

    Tommassen, J.P.M.; Kroon, T. de

    1987-01-01

    Protease accessibility experiments were employed to localize a PhoE-LacZ hybrid protein, encompassing a large N-terminal fragment of the outer membrane PhoE protein of E. coli, fused to β-galactosidase, at the subcellular level. In previous studies, this protein was shown to co-fractionate with the

  9. Efficient Overproduction of Membrane Proteins in Lactococcus lactis Requires the Cell Envelope Stress Sensor/Regulator Couple CesSR

    Science.gov (United States)

    Pinto, Joao P. C.; Kuipers, Oscar P.; Marreddy, Ravi K. R.; Poolman, Bert; Kok, Jan

    2011-01-01

    Background Membrane proteins comprise an important class of molecules whose study is largely frustrated by several intrinsic constraints, such as their hydrophobicity and added requirements for correct folding. Additionally, the complexity of the cellular mechanisms that are required to insert membrane proteins functionally in the membrane and to monitor their folding state makes it difficult to foresee the yields at which one can obtain them or to predict which would be the optimal production host for a given protein. Methods and Findings We describe a rational design approach to improve the lactic acid bacterium Lactococcus lactis as a producer of membrane proteins. Our transcriptome data shows that the two-component system CesSR, which senses cell envelope stresses of different origins, is one of the major players when L. lactis is forced to overproduce the endogenous membrane protein BcaP, a branched-chain amino acid permease. Growth of the BcaP-producing L. lactis strain and its capability to produce membrane proteins are severely hampered when the CesSR system itself or particular members of the CesSR regulon are knocked out, notably the genes ftsH, oxaA2, llmg_2163 and rmaB. Overexpressing cesSR reduced the growth defect, thus directly improving the production yield of BcaP. Applying this rationale to eukaryotic proteins, some of which are notoriously more difficult to produce, such as the medically-important presenilin complex, we were able to significantly diminish the growth defect seen in the wild-type strain and improve the production yield of the presenilin variant PS1Δ9-H6 more than 4-fold. Conclusions The results shed light into a key, and perhaps central, membrane protein quality control mechanism in L. lactis. Modulating the expression of CesSR benefited the production yields of membrane proteins from different origins. These findings reinforce L. lactis as a legitimate alternative host for the production of membrane proteins. PMID:21818275

  10. Lactococcus lactis is an Efficient Expression System for Mammalian Membrane Proteins Involved in Liver Detoxification, CYP3A4, and MGST1.

    Science.gov (United States)

    Bakari, Sana; Lembrouk, Mehdi; Sourd, Laura; Ousalem, Fares; André, François; Orlowski, Stéphane; Delaforge, Marcel; Frelet-Barrand, Annie

    2016-04-01

    Despite the great importance of human membrane proteins involved in detoxification mechanisms, their wide use for biochemical approaches is still hampered by several technical difficulties considering eukaryotic protein expression in order to obtain the large amounts of protein required for functional and/or structural studies. Lactococcus lactis has emerged recently as an alternative heterologous expression system to Escherichia coli for proteins that are difficult to express. The aim of this work was to check its ability to express mammalian membrane proteins involved in liver detoxification, i.e., CYP3A4 and two isoforms of MGST1 (rat and human). Genes were cloned using two different strategies, i.e., classical or Gateway-compatible cloning, and we checked the possible influence of two affinity tags (6×-His-tag and Strep-tag II). Interestingly, all proteins could be successfully expressed in L. lactis at higher yields than those previously obtained for these proteins with classical expression systems (E. coli, Saccharomyces cerevisiae) or those of other eukaryotic membrane proteins expressed in L. lactis. In addition, rMGST1 was fairly active after expression in L. lactis. This study highlights L. lactis as an attractive system for efficient expression of mammalian detoxification membrane proteins at levels compatible with further functional and structural studies.

  11. Efficient Gene Delivery Mediated by a Helical Polypeptide: Controlling the Membrane Activity via Multivalency and Light-Assisted Photochemical Internalization (PCI).

    Science.gov (United States)

    Xu, Xin; Li, Yongjuan; Liang, Qiujun; Song, Ziyuan; Li, Fangfang; He, Hua; Wang, Jinhui; Zhu, Lipeng; Lin, Zhifeng; Yin, Lichen

    2018-01-10

    The development of robust and nontoxic membrane-penetrating materials is highly demanded for nonviral gene delivery. Herein, a photosensitizer (PS)-embedded, star-shaped helical polypeptide was developed, which combines the advantages of multivalency-enhanced intracellular DNA uptake and light-strengthened endosomal escape to enable highly efficient gene delivery with low toxicity. 5,10,15,20-Tetrakis-(4-aminophenyl) porphyrin as a selected PS initiated ring-opening polymerization of N-carboxyanhydride and yielded a star-shaped helical polypeptide after side-chain functionalization with guanidine groups. The star polypeptide afforded a notably higher transfection efficiency and lower cytotoxicity than those of its linear analogue. Light irradiation caused almost complete (∼90%) endosomal release of the DNA cargo via the photochemical internalization (PCI) mechanism and further led to a 6-8-fold increment of the transfection efficiency in HeLa, B16F10, and RAW 264.7 cells, outperforming commercial reagent 25k PEI by up to 3 orders of magnitude. Because the PS and DNA cargoes were compartmentalized distantly in the core and polypeptide layers, respectively, the generated reactive oxygen species caused minimal damage to DNA molecules to preserve their transfection potency. Such multivalency- and PCI-potentiated gene delivery efficiency was also demonstrated in vivo in melanoma-bearing mice. This study thus provides a promising strategy to overcome the multiple membrane barriers against nonviral gene delivery.

  12. Enhancement in biological response of Ag-nano composite polymer membranes using plasma treatment for fabrication of efficient bio materials

    Science.gov (United States)

    Agrawal, Narendra Kumar; Sharma, Tamanna Kumari; Chauhan, Manish; Agarwal, Ravi; Vijay, Y. K.; Swami, K. C.

    2016-05-01

    Biomaterials are nonviable material used in medical devices, intended to interact with biological systems, which are becoming necessary for the development of artificial material for biological systems such as artificial skin diaphragm, valves for heart and kidney, lenses for eye etc. Polymers having novel properties like antibacterial, antimicrobial, high adhesion, blood compatibility and wettability are most suitable for synthesis of biomaterial, but all of these properties does not exist in any natural or artificial polymeric material. Nano particles and plasma treatment can offer these properties to the polymers. Hence a new nano-biomaterial has been developed by modifying the surface and chemical properties of Ag nanocomposite polymer membranes (NCPM) by Argon ion plasma treatment. These membranes were characterized using different techniques for surface and chemical modifications occurred. Bacterial adhesion and wettability were also tested for these membranes, to show direct use of this new class of nano-biomaterial for biomedical applications.

  13. Economic potential of inertial fusion

    International Nuclear Information System (INIS)

    Nuckolls, J.H.

    1984-04-01

    Beyond the achievement of scientific feasibility, the key question for fusion energy is: does it have the economic potential to be significantly cheaper than fission and coal energy. If fusion has this high economic potential then there are compelling commercial and geopolitical incentives to accelerate the pace of the fusion program in the near term, and to install a global fusion energy system in the long term. Without this high economic potential, fusion's success depends on the failure of all alternatives, and there is no real incentive to accelerate the program. If my conjectures on the economic potential of inertial fusion are approximately correct, then inertial fusion energy's ultimate costs may be only half to two-thirds those of advanced fission and coal energy systems. Relative cost escalation is not assumed and could increase this advantage. Both magnetic and inertial approaches to fusion potentially have a two-fold economic advantage which derives from two fundamental properties: negligible fuel costs and high quality energy which makes possible more efficient generation of electricity. The wining approach to fusion may excel in three areas: electrical generating efficiency, minimum material costs, and adaptability to manufacture in automated factories. The winning approach must also rate highly in environmental potential, safety, availability factor, lifetime, small 0 and M costs, and no possibility of utility-disabling accidents

  14. Controlled fusion; La fusion controlee

    Energy Technology Data Exchange (ETDEWEB)

    Bobin, J.L

    2005-07-01

    During the last fifty years the researches on controlled thermonuclear fusion reached great performance in the magnetic confinement (tokamaks) as in the inertial confinement (lasers). But the state of the art is not in favor of the apparition of the fusion in the energy market before the second half of the 21 century. To explain this opinion the author presents the fusion reactions of light nuclei and the problems bound to the magnetic confinement. (A.L.B.)

  15. Fusion devices

    International Nuclear Information System (INIS)

    Fowler, T.K.

    1977-01-01

    Three types of thermonuclear fusion devices currently under development are reviewed for an electric utilities management audience. Overall design features of laser fusion, tokamak, and magnetic mirror type reactors are described and illustrated. Thrusts and trends in current research on these devices that promise to improve performance are briefly reviewed. Twenty photographs and drawings are included

  16. EPR Spectroscopy Targets Structural Changes in the E. coli Membrane Fusion CusB upon Cu(I) Binding.

    Science.gov (United States)

    Meir, Aviv; Abdelhai, Ahmad; Moskovitz, Yoni; Ruthstein, Sharon

    2017-06-20

    Bacterial cells have developed sophisticated systems to deal with the toxicity of metal ions. Escherichia coli CusCFBA is a complex efflux system, responsible for transferring Cu(I) and Ag(I) ions; this system, located in the periplasm, involves four proteins, CusA, CusB, CusC, and CusF. CusA, CusB, and CusC are connected to one another in an oligomerization ratio of 3:6:3 CusA/CusB/CusC to form the CusCBA periplasm membrane transporter. CusB is an adaptor protein that connects the two membrane proteins CusA (inner membrane) and CusC (outer membrane). CusF is a metallochaperone that transfers Cu(I) and Ag(I) to the CusCBA transporter from the periplasm. The crystal structures of CusB, CusC, CusF, and the CusBA complex have been resolved, shedding some light on the efflux mechanism underlying this intriguing system. However, since CusB is an adaptor protein, its role in operating this system is significant, and should be understood in detail. Here, we utilize EPR spectroscopy to target the conformational changes that take place in the full CusB protein upon binding Cu(I). We reveal that CusB is a dimer in solution, and that the orientation of one molecule with respect to the other molecule changes upon Cu(I) coordination, resulting in a more compact CusB structure. These structural and topological changes upon Cu(I) binding probably play the role of a switch for opening the channel and transferring metal ions from CusB to CusC and out of the cell. Copyright © 2017 Biophysical Society. Published by Elsevier Inc. All rights reserved.

  17. Membrane-Based Inverse Transition Cycling: An Improved Means for Purifying Plant-Derived Recombinant Protein-Elastin-Like Polypeptide Fusions

    Directory of Open Access Journals (Sweden)

    Udo Conrad

    2011-04-01

    Full Text Available Elastin-like peptide (ELP was fused to two different avian flu H5N1 antigens and expressed in transgenic tobacco plants. The presence of the ELP tag enhanced the accumulation of the heterologous proteins in the tobacco leaves. An effective membrane-based Inverse Transition Cycling was developed to recover the ELPylated antigens and antibodies from plant material. The functionality of both the ELPylated neuraminidase and an ELPylated nanobody was demonstrated.

  18. Quantitative analysis of lipid droplet fusion: inefficient steady state fusion but rapid stimulation by chemical fusogens.

    Directory of Open Access Journals (Sweden)

    Samantha Murphy

    2010-12-01

    Full Text Available Lipid droplets (LDs are dynamic cytoplasmic organelles containing neutral lipids and bounded by a phospholipid monolayer. Previous studies have suggested that LDs can undergo constitutive homotypic fusion, a process linked to the inhibitory effects of fatty acids on glucose transporter trafficking. Using strict quantitative criteria for LD fusion together with refined light microscopic methods and real-time analysis, we now show that LDs in diverse cell types show low constitutive fusogenic activity under normal growth conditions. To investigate the possible modulation of LD fusion, we screened for agents that can trigger fusion. A number of pharmacological agents caused homotypic fusion of lipid droplets in a variety of cell types. This provided a novel cell system to study rapid regulated fusion between homotypic phospholipid monolayers. LD fusion involved an initial step in which the two adjacent membranes became continuous (<10 s, followed by the slower merging (100 s of the neutral lipid cores to produce a single spherical LD. These fusion events were accompanied by changes to the LD surface organization. Measurements of LDs undergoing homotypic fusion showed that fused LDs maintained their initial volume, with a corresponding decrease in surface area suggesting rapid removal of membrane from the fused LD. This study provides estimates for the level of constitutive LD fusion in cells and questions the role of LD fusion in vivo. In addition, it highlights the extent of LD restructuring which occurs when homotypic LD fusion is triggered in a variety of cell types.

  19. Membrane fusion proteins of type I secretion system and tripartite efflux pumps share a binding motif for TolC in gram-negative bacteria.

    Directory of Open Access Journals (Sweden)

    Minho Lee

    Full Text Available The Hly translocator complex of Escherichia coli catalyzes type I secretion of the toxin hemolysin A (HlyA. In this complex, HlyB is an inner membrane ABC (ATP Binding Cassette-type transporter, TolC is an outer membrane channel protein, and HlyD is a periplasmic adaptor anchored in the inner membrane that bridges HlyB to TolC. This tripartite organization is reminiscent of that of drug efflux systems such as AcrA-AcrB-TolC and MacA-MacB-TolC of E. coli. We have previously shown the crucial role of conserved residues located at the hairpin tip region of AcrA and MacA adaptors during assembly of their cognate systems. In this study, we investigated the role of the putative tip region of HlyD using HlyD mutants with single amino acid substitutions at the conserved positions. In vivo and in vitro data show that all mutations abolished HlyD binding to TolC and resulted in the absence of HlyA secretion. Together, our results suggest that, similarly to AcrA and MacA, HlyD interacts with TolC in a tip-to-tip manner. A general model in which these conserved interactions induce opening of TolC during drug efflux and type I secretion is discussed.

  20. Membrane Fusion Proteins of Type I Secretion System and Tripartite Efflux Pumps Share a Binding Motif for TolC in Gram-Negative Bacteria

    Science.gov (United States)

    Yoon, Bo-Young; Song, Saemee; Lee, Kangseok; Ha, Nam-Chul

    2012-01-01

    The Hly translocator complex of Escherichia coli catalyzes type I secretion of the toxin hemolysin A (HlyA). In this complex, HlyB is an inner membrane ABC (ATP Binding Cassette)-type transporter, TolC is an outer membrane channel protein, and HlyD is a periplasmic adaptor anchored in the inner membrane that bridges HlyB to TolC. This tripartite organization is reminiscent of that of drug efflux systems such as AcrA-AcrB-TolC and MacA-MacB-TolC of E. coli. We have previously shown the crucial role of conserved residues located at the hairpin tip region of AcrA and MacA adaptors during assembly of their cognate systems. In this study, we investigated the role of the putative tip region of HlyD using HlyD mutants with single amino acid substitutions at the conserved positions. In vivo and in vitro data show that all mutations abolished HlyD binding to TolC and resulted in the absence of HlyA secretion. Together, our results suggest that, similarly to AcrA and MacA, HlyD interacts with TolC in a tip-to-tip manner. A general model in which these conserved interactions induce opening of TolC during drug efflux and type I secretion is discussed. PMID:22792337

  1. Poly(ethylene glycol) and poly(dimethyl siloxane): Combining their advantages into efficient CO2 gas separation membranes

    NARCIS (Netherlands)

    Reijerkerk, Sander; Knoef, Michel H.; Nijmeijer, Dorothea C.; Wessling, Matthias

    2010-01-01

    Polymer blending is a versatile tool to combine the beneficial properties of two or more components in one single material. Here, we present the preparation, thermal- and mass transport properties of a series of blend membranes made from the commercially available PEBAX® MH 1657 and a poly(ethylene

  2. Dominant factors controlling the efficiency of two-phase flow cleaning in spiral-wound membrane elements

    NARCIS (Netherlands)

    Wibisono, Y.; Ahmad, F.; Cornelissen, Emile; Cornelissen, E.R.; Kemperman, Antonius J.B.; Nijmeijer, Dorothea C.

    2015-01-01

    Two-phase flow cleaning has been successfully applied to control fouling in spiral wound membrane elements. This study focuses on its experimental optimization using a Taguchi Design of Experiment method (L-25 orthogonal arrays) to elucidate the influence of different factors and to reveal the

  3. The evolutionarily conserved protein PHOTOSYNTHESIS AFFECTED MUTANT71 is required for efficient manganese uptake at the thylakoid membrane in Arabidopsis

    DEFF Research Database (Denmark)

    Schneider, Anja; Steinberger, Iris; Herdean, Andrei

    2016-01-01

    lumen remains poorly understood. Here, we show that Arabidopsis thaliana PHOTOSYNTHESIS AFFECTED MUTANT71 (PAM71) is an integral thylakoid membrane protein involved in Mn2+ and Ca2+ homeostasis in chloroplasts. This protein is required for normal operation of the oxygen-evolving complex (as evidenced...

  4. Hydrogen Production in Fusion Reactors

    OpenAIRE

    S., Sudo; Y., Tomita; S., Yamaguchi; A., Iiyoshi; H., Momota; O., Motojima; M., Okamoto; M., Ohnishi; M., Onozuka; C., Uenosono

    1993-01-01

    As one of methods of innovative energy production in fusion reactors without having a conventional turbine-type generator, an efficient use of radiation produced in a fusion reactor with utilizing semiconductor and supplying clean fuel in a form of hydrogen gas are studied. Taking the candidates of reactors such as a toroidal system and an open system for application of the new concepts, the expected efficiency and a concept of plant system are investigated.

  5. Osteoclast Fusion

    DEFF Research Database (Denmark)

    Marie Julie Møller, Anaïs; Delaissé, Jean-Marie; Søe, Kent

    2017-01-01

    Investigations addressing the molecular keys of osteoclast fusion are primarily based on end-point analyses. No matter if investigations are performed in vivo or in vitro the impact of a given factor is predominantly analyzed by counting the number of multi-nucleated cells, the number of nuclei per...... multinucleated cell or TRAcP activity. But end-point analyses do not show how the fusion came about. This would not be a problem if fusion of osteoclasts was a random process and occurred by the same molecular mechanism from beginning to end. However, we and others have in the recent period published data...... suggesting that fusion partners may specifically select each other and that heterogeneity between the partners seems to play a role. Therefore, we set out to directly test the hypothesis that fusion factors have a heterogenic involvement at different stages of nuclearity. Therefore, we have analyzed...

  6. Fusion physics

    CERN Document Server

    Lackner, Karl; Tran, Minh Quang

    2012-01-01

    This publication is a comprehensive reference for graduate students and an invaluable guide for more experienced researchers. It provides an introduction to nuclear fusion and its status and prospects, and features specialized chapters written by leaders in the field, presenting the main research and development concepts in fusion physics. It starts with an introduction to the case for the development of fusion as an energy source. Magnetic and inertial confinement are addressed. Dedicated chapters focus on the physics of confinement, the equilibrium and stability of tokamaks, diagnostics, heating and current drive by neutral beam and radiofrequency waves, and plasma–wall interactions. While the tokamak is a leading concept for the realization of fusion, other concepts (helical confinement and, in a broader sense, other magnetic and inertial configurations) are also addressed in the book. At over 1100 pages, this publication provides an unparalleled resource for fusion physicists and engineers.

  7. Research and development of basic technologies for next-generation industry. Ultimate evaluation report on research and development of highly efficient polymeric separation membrane material; Jisedai sangyo kiban gijutsu kenkyu kaihatsu. Kokoritsu kobunshi bunrimaku zairyo saishu kenkyu kaihatsu hyoka

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1991-05-01

    For the enhancement of separation process efficiency and energy efficiency in the chemical industry, etc., basic technologies are developed involving high-performance separation membrane materials which are excellent in durability and usable in the field where separation by membranes has been impractical. The liquid mixtures subjected to separation are a neutral organic compound/water system, an acidic organic compound/water system, and a polar organic compound/water system; the gas mixtures subjected to separation are an oxygen/nitrogen system and a carbon monoxide/nitrogen system. After a 10-year/3-phase development endeavors, the initially intended goals are sufficiently achieved. Among those that have to be mentioned is the development of a nonaqueous separation membrane, a supported liquid membrane with amino acid optically active high performance separation capability, a high-precision evaluation unit for gas separation membrane characteristics, a selective permeation membrane with high-level oxygen and carbon monoxide carriers and reactivation technology, a high-performance ethanol separation membrane, a water/polar organic compound separation membrane, and a water/acetic acid separation membrane and stability providing technology. In particular, the water selective permeation membrane for a mixture of water and alcohol has already arrived at the stage of bench plant demonstration. (NEDO)

  8. Multispectral Image Enhancement Through Adaptive Wavelet Fusion

    Science.gov (United States)

    2016-09-14

    AFRL-AFOSR-UK-TR-2017-0005 Multispectral image enhancement through adaptive wavelet fusion Alexander Toet Nederlandse Organisatie voor Toegepast...image enhancement through adaptive wavelet fusion 5a.  CONTRACT NUMBER 5b.  GRANT NUMBER FA9550-15-1-0433 5c.  PROGRAM ELEMENT NUMBER 61102F 6. AUTHOR(S...efficient. 15. SUBJECT TERMS adaptive wavelet fusion, Multispectral image enhancement, Multispectral image fusion, multiband image interpolation

  9. Effect of specific amino acid substitutions in the putative fusion peptide of structural glycoprotein E2 on Classical Swine Fever Virus replication

    Energy Technology Data Exchange (ETDEWEB)

    Fernández-Sainz, I.J. [Plum Island Animal Disease Center, ARS, USDA (United States); Largo, E. [Biophysics Unit (CSIC-UPV/EHU), Department of Biochemistry and Molecular Biology, University of the Basque Country (UPV/EHU), P.O. Box 644, 48080 Bilbao (Spain); Gladue, D.P.; Fletcher, P. [Plum Island Animal Disease Center, ARS, USDA (United States); O’Donnell, V. [Plum Island Animal Disease Center, ARS, USDA (United States); Plum Island Animal Disease Center, DHS, Greenport, NY 11944 (United States); Holinka, L.G. [Plum Island Animal Disease Center, ARS, USDA (United States); Carey, L.B. [Department of Experimental and Health Sciences, Universitat Pompeu Fabra (UPF), E-08003 Barcelona (Spain); Lu, X. [Plum Island Animal Disease Center, DHS, Greenport, NY 11944 (United States); Nieva, J.L. [Biophysics Unit (CSIC-UPV/EHU), Department of Biochemistry and Molecular Biology, University of the Basque Country (UPV/EHU), P.O. Box 644, 48080 Bilbao (Spain); Borca, M.V., E-mail: manuel.borca@ars.usda.gov [Plum Island Animal Disease Center, ARS, USDA (United States)

    2014-05-15

    E2, along with E{sup rns} and E1, is an envelope glycoprotein of Classical Swine Fever Virus (CSFV). E2 is involved in several virus functions: cell attachment, host range susceptibility and virulence in natural hosts. Here we evaluate the role of a specific E2 region, {sup 818}CPIGWTGVIEC{sup 828}, containing a putative fusion peptide (FP) sequence. Reverse genetics utilizing a full-length infectious clone of the highly virulent CSFV strain Brescia (BICv) was used to evaluate how individual amino acid substitutions within this region of E2 may affect replication of BICv. A synthetic peptide representing the complete E2 FP amino acid sequence adopted a β-type extended conformation in membrane mimetics, penetrated into model membranes, and perturbed lipid bilayer integrity in vitro. Similar peptides harboring amino acid substitutions adopted comparable conformations but exhibited different membrane activities. Therefore, a preliminary characterization of the putative FP {sup 818}CPIGWTGVIEC{sup 828} indicates a membrane fusion activity and a critical role in virus replication. - Highlights: • A putative fusion peptide (FP) region in CSFV E2 protein was shown to be critical for virus growth. • Synthetic FPs were shown to efficiently penetrate into lipid membranes using an in vitro model. • Individual residues in the FP affecting virus replication were identified by reverse genetics. • The same FP residues are also responsible for mediating membrane fusion.

  10. Molecular dynamics simulations of lipid vesicle fusion in atomic detail

    NARCIS (Netherlands)

    Knecht, Volker; Marrink, Siewert-Jan

    The fusion of a membrane-bounded vesicle with a target membrane is a key step in intracellular trafficking, exocytosis, and drug delivery. Molecular dynamics simulations have been used to study the fusion of small unilamellar vesicles composed of a dipalmitoyl-phosphatidylcholine (DPPC)/palmitic

  11. Role of the synaptobrevin C terminus in fusion pore formation

    DEFF Research Database (Denmark)

    Ngatchou, Annita N; Kisler, Kassandra; Fang, Qinghua

    2010-01-01

    stimulation, the SNARE complex pulls the C terminus of sybII deeper into the vesicle membrane. We propose that this movement disrupts the vesicular membrane continuity leading to fusion pore formation. In contrast to current models, the experiments suggest that fusion pore formation begins with molecular......Neurotransmitter release is mediated by the SNARE proteins synaptobrevin II (sybII, also known as VAMP2), syntaxin, and SNAP-25, generating a force transfer to the membranes and inducing fusion pore formation. However, the molecular mechanism by which this force leads to opening of a fusion pore...

  12. Tension-induced vesicle fusion: pathways and pore dynamics

    DEFF Research Database (Denmark)

    Shillcock, Julian C.

    2008-01-01

    The dynamics of tension-induced fusion of two vesicles is studied using dissipative particle dynamics (DPD) simulations. The vesicle membranes use an improved DPD parameter set that results in their sustaining only a 10–30% relative area stretch before rupturing on the microsecond timescale...... fusion time on membrane tension implies that the fusion process is completed by overcoming two energy barriers with scales of 13kBT and 11kBT. The fusion pore radius as a function of time has also been extracted from the simulations, and provides a quantitative measure of the fusion dynamics which...

  13. In-tube electro-membrane extraction with a sub-microliter organic solvent consumption as an efficient technique for synthetic food dyes determination in foodstuff samples.

    Science.gov (United States)

    Bazregar, Mohammad; Rajabi, Maryam; Yamini, Yadollah; Asghari, Alireza; Abdossalami asl, Yousef

    2015-09-04

    A simple and efficient extraction technique with a sub-microliter organic solvent consumption termed as in-tube electro-membrane extraction (IEME) is introduced. This method is based upon the electro-kinetic migration of ionized compounds by the application of an electrical potential difference. For this purpose, a thin polypropylene (PP) sheet placed inside a tube acts as a support for the membrane solvent, and 30μL of an aqueous acceptor solution is separated by this solvent from 1.2mL of an aqueous donor solution. This method yielded high extraction recoveries (63-81%), and the consumption of the organic solvent used was only 0.5μL. By performing this method, the purification is high, and the utilization of the organic solvent, used as a mediator, is very simple and repeatable. The proposed method was evaluated by extraction of four synthetic food dyes (Amaranth, Ponceau 4R, Allura Red, and Carmoisine) as the model analytes. Optimization of variables affecting the method was carried out in order to achieve the best extraction efficiency. These variables were the type of membrane solvent, applied extraction voltage, extraction time, pH range, and concentration of salt added. Under the optimized conditions, IEME-HPLC-UV provided a good linearity in the range of 1.00-800ngmL(-1), low limits of detection (0.3-1ngmL(-1)), and good extraction repeatabilities (RSDs below 5.2%, n=5). It seems that this design is a proper one for the automation of the method. Also the consumption of the organic solvent in a sub-microliter scale, and its simplicity, high efficiency, and high purification can help one getting closer to the objectives of the green chemistry. Copyright © 2015 Elsevier B.V. All rights reserved.

  14. Time efficient methods for scanning a fluorescent membrane with a fluorescent microscopic imager for the quality assurance of food

    Science.gov (United States)

    Lerm, Steffen; Holder, Silvio; Schellhorn, Mathias; Brückner, Peter; Linß, Gerhard

    2013-05-01

    An important part of the quality assurance of meat is the estimation of germs in the meat exudes. The kind and the number of the germs in the meat affect the medical risk for the consumer of the meat. State-of-the-art analyses of meat are incubator test procedures. The main disadvantages of such incubator tests are the time consumption, the necessary equipment and the need of special skilled employees. These facts cause in high inspection cost. For this reason a new method for the quality assurance is necessary which combines low detection limits and less time consumption. One approach for such a new method is fluorescence microscopic imaging. The germs in the meat exude are caught in special membranes by body-antibody reactions. The germ typical signature could be enhanced with fluorescent chemical markers instead of reproduction of the germs. Each fluorescent marker connects with a free germ or run off the membrane. An image processing system is used to detect the number of fluorescent particles. Each fluorescent spot should be a marker which is connected with a germ. Caused by the small object sizes of germs, the image processing system needs a high optical magnification of the camera. However, this leads to a small field of view and a small depth of focus. For this reasons the whole area of the membrane has to be scanned in three dimensions. To minimize the time consumption, the optimal path has to be found. This optimization problem is influenced by features of the hardware and is presented in this paper. The traversing range in each direction, the step width, the velocity, the shape of the inspection volume and the field of view have influence on the optimal path to scan the membrane.

  15. Advanced fusion reactor

    Energy Technology Data Exchange (ETDEWEB)

    Tomita, Yukihiro [National Inst. for Fusion Science, Toki, Gifu (Japan)

    2003-04-01

    The main subjects on fusion research are now on D-T fueled fusion, mainly due to its high fusion reaction rate. However, many issues are still remained on the wall loading by the 14 MeV neutrons. In the case of D-D fueled fusion, the neutron wall loading is still remained, though the technology related to tritium breeding is not needed. The p-{sup 6}Li and p-{sup 11}B fueled fusions are not estimated to be the next generation candidate until the innovated plasma confinement technologies come in useful to achieve the high performance plasma parameters. The fusion reactor of D-{sup 3}He fuels has merits on the smaller neutron wall loading and tritium handling. However, there are difficulties on achieving the high temperature plasma more than 100 keV. Furthermore the high beta plasma is needed to decrease synchrotron radiation loss. In addition, the efficiency of the direct energy conversion from protons coming out from fusion reaction is one of the key parameters in keeping overall power balance. Therefore, open magnetic filed lines should surround the plasma column. In this paper, we outlined the design of the commercial base reactor (ARTEMIS) of 1 GW electric output power configured by D-{sup 3}He fueled FRC (Field Reversed Configuration). The ARTEMIS needs 64 kg of {sup 3}He per a year. On the other hand, 1 million tons of {sup 3}He is estimated to be in the moon. The {sup 3}He of about 10{sup 23} kg are to exist in gaseous planets such as Jupiter and Saturn. (Y. Tanaka)

  16. Efficiency measurement and uncertainty discussion of an electric engine powered by a "self-breathing" and "self-humidified" proton exchange membrane fuel cell.

    Science.gov (United States)

    Schiavetti, Pierluigi; Del Prete, Zaccaria

    2007-08-01

    The efficiency of an automotive engine based on a "self-breathing" and "self-humidified" proton exchange membrane fuel cell stack (PEM FC) connected to a dc brushless electrical motor was measured under variable power load conditions. Experiments have been carried out on a small scale 150 W engine model. After determining the fuel cell static polarization curve and the time response to power steps, the system was driven to copy on the test bench a "standard urban load cycle" and its instantaneous efficiencies were measured at an acquisition rate of 5 Hz. The integral system efficiency over the entire urban load cycle, comprising the losses of the unavoidable auxiliary components of the engine, was then calculated. The fuel cell stack was operated mainly in "partial" dead-end mode, with a periodic anode flow channel purging, and one test was carried out in "pure" dead-end mode, with no anode channel purging. An uncertainty analysis of the efficiencies was carried out, taking into account either type A and type B evaluation methods, strengthening the discussion about the outcomes obtained for a system based on this novel simplified FC type. For our small scale engine we measured over the standard urban cycle, on the basis of the H(2) high heating value (HHV), a tank-to-wheel integral efficiency of (18.2+/-0.8)%, when the fuel cell was operated with periodic flow channel purging, and of (21.5+/-1.3)% in complete dead-end operation mode.

  17. Deduced sequences of the membrane fusion and attachment proteins of canine distemper viruses isolated from dogs and wild animals in Korea.

    Science.gov (United States)

    Bae, Chae-Wun; Lee, Joong-Bok; Park, Seung-Yong; Song, Chang-Seon; Lee, Nak-Hyung; Seo, Kun-Ho; Kang, Young-Sun; Park, Choi-Kyu; Choi, In-Soo

    2013-08-01

    Canine distemper virus (CDV) causes highly contagious respiratory, gastrointestinal, and neurological diseases in wild and domestic animal species. Despite a broad vaccination campaign, the disease is still a serious problem worldwide. In this study, six field CDV strains were isolated from three dogs, two raccoon dogs, and one badger in Korea. The full sequence of the genes encoding fusion (F) and hemagglutinin (H) proteins were compared with those of other CDVs including field and vaccine strains. The phylogenetic analysis for the F and H genes indicated that the two CDV strains isolated from dogs were most closely related to Chinese strains in the Asia-1 genotype. Another four strains were closely related to Japanese strains in the Asia-2 genotype. The six currently isolated strains shared 90.2-92.1% and 88.2-91.8% identities with eight commercial vaccine strains in their nucleotide and amino acid sequences of the F protein, respectively. They also showed 90.1-91.4% and 87.8-90.7% identities with the same vaccine strains in their nucleotide and deduced amino acid sequences of the H protein, respectively. Different N-linked glycosylation sites were identified in the F and H genes of the six isolates from the prototype vaccine strain Onderstepoort. Collectively, these results demonstrate that at least two different CDV genotypes currently exist in Korea. The considerable genetic differences between the vaccine strains and wild-type isolates would be a major factor of the incomplete protection of dogs from CDV infections.

  18. Cell-Delivered Entry Inhibitors for HIV-1: CCR5 Downregulation and Blocking Virus/Membrane Fusion in Defending the Host Cell Population.

    Science.gov (United States)

    Symonds, Geoff; Bartlett, Jeffrey S; Kiem, Hans-Peter; Tsie, Marlene; Breton, Louis

    2016-12-01

    HIV-1 infection requires the presence of the CD4 receptor on the target cell surface and a coreceptor, predominantly CC-chemokine receptor 5 (CCR5). It has been shown that individuals who are homozygous for a defective CCR5 gene are protected from HIV-1 infection. A novel self-inactivating lentiviral vector LVsh5/C46 (Cal-1) has been engineered to block HIV-1 infection with two viral entry inhibitors, conferring resistance to HIV-1 infection from both CCR5 and CXCR4 tropic strains. Cal-1 encodes a short hairpin RNA (sh5) to downregulate CCR5 and C46, an HIV-1 fusion inhibitor. Gene therapy by Cal-1 is aimed at transducing CD4 + T cells and CD34 + hematopoietic stem/progenitor cells in an autologous transplant setting. Pre-clinical safety and efficacy studies in vitro and in vivo (humanized mouse model and nonhuman primates) have shown that Cal-1 is safe with no indication of any toxicity risk and acts to decrease viral load and increase CD4 counts. Two clinical trials are underway using Cal-1: a phase I/II study to assess safety and feasibility in an adult HIV-1-positive population not on antiretroviral therapy (ART); and a second Fred Hutchinson Investigator Initiated phase I study to assess safety and feasibility in adults with HIV-1-associated non-Hodgkin or Hodgkin lymphoma.

  19. Cold fusion

    International Nuclear Information System (INIS)

    Suh, Suk Yong; Sung, Ki Woong; Kang, Joo Sang; Lee, Jong Jik

    1995-02-01

    So called 'cold fusion phenomena' are not confirmed yet. Excess heat generation is very delicate one. Neutron generation is most reliable results, however, the records are erratic and the same results could not be repeated. So there is no reason to exclude the malfunction of testing instruments. The same arguments arise in recording 4 He, 3 He, 3 H, which are not rich in quantity basically. An experiment where plenty of 4 He were recorded is attached in appendix. The problem is that we are trying to search cold fusion which is permitted by nature or not. The famous tunneling effect in quantum mechanics will answer it, however, the most fusion rate is known to be negligible. The focus of this project is on the theme that how to increase that negligible fusion rate. 6 figs, 4 tabs, 1512 refs. (Author)

  20. Cold fusion

    Energy Technology Data Exchange (ETDEWEB)

    Suh, Suk Yong; Sung, Ki Woong; Kang, Joo Sang; Lee, Jong Jik [Korea Atomic Energy Research Institute, Taejon (Korea, Republic of)

    1995-02-01

    So called `cold fusion phenomena` are not confirmed yet. Excess heat generation is very delicate one. Neutron generation is most reliable results, however, the records are erratic and the same results could not be repeated. So there is no reason to exclude the malfunction of testing instruments. The same arguments arise in recording {sup 4}He, {sup 3}He, {sup 3}H, which are not rich in quantity basically. An experiment where plenty of {sup 4}He were recorded is attached in appendix. The problem is that we are trying to search cold fusion which is permitted by nature or not. The famous tunneling effect in quantum mechanics will answer it, however, the most fusion rate is known to be negligible. The focus of this project is on the theme that how to increase that negligible fusion rate. 6 figs, 4 tabs, 1512 refs. (Author).

  1. The nectin-1α transmembrane domain, but not the cytoplasmic tail, influences cell fusion induced by HSV-1 glycoproteins

    International Nuclear Information System (INIS)

    Subramanian, Ravi P.; Dunn, Jennifer E.; Geraghty, Robert J.

    2005-01-01

    Nectin-1 is a receptor for herpes simplex virus (HSV), a member of the immunoglobulin superfamily, and a cellular adhesion molecule. To study domains of nectin-1α involved in cell fusion, we measured the ability of nectin-1α/nectin-2α chimeras, nectin-1α/CD4 chimeras, and transmembrane domain and cytoplasmic tail mutants of nectin-1α to promote cell fusion induced by HSV-1 glycoproteins. Our results demonstrate that only chimeras and mutants containing the entire V-like domain and a link to the plasma membrane conferred cell-fusion activity. The transmembrane domain and cytoplasmic tail of nectin-1 were not required for any viral receptor or cell adhesion function tested. Cellular cytoplasmic factors that bind to the nectin-1α cytoplasmic tail, therefore, did not influence virus entry or cell fusion. Interestingly, the efficiency of cell fusion was reduced when membrane-spanning domains of nectin-1α and gD were replaced by glycosylphosphatidylinositol tethers, indicating that transmembrane domains may play a modulatory role in the gD/nectin-1α interaction in fusion

  2. Insulators for fusion applications

    International Nuclear Information System (INIS)

    1987-04-01

    Design studies for fusion devices and reactors have become more detailed in recent years and with this has come a better understanding of requirements and operating conditions for insulators in these machines. Ceramic and organic insulators are widely used for many components of fusion devices and reactors namely: radio frequency (RF) energy injection systems (BeO, Al 2 O 3 , Mg Al 2 O 4 , Si 3 N 4 ); electrical insulation for the torus structure (SiC, Al 2 O 3 , MgO, Mg Al 2 O 4 , Si 4 Al 2 O 2 N 6 , Si 3 N 4 , Y 2 O 3 ); lightly-shielded magnetic coils (MgO, MgAl 2 O 4 ); the toroidal field coil (epoxies, polyimides), neutron shield (B 4 C, TiH 2 ); high efficiency electrical generation; as well as the generation of very high temperatures for high efficiency hydrogen production processes (ZrO 2 and Al 2 O 3 - mat, graphite and carbon - felt). Timely development of insulators for fusion applications is clearly necessary. Those materials to be used in fusion machines should show high resistance to radiation damage and maintain their structural integrity. Now the need is urgent for a variety of radiation resistant materials, but much effort in these areas is required for insulators to be considered seriously by the design community. This document contains 14 papers from an IAEA meeting. It was the objective of this meeting to identify existing problems in analysing various situations of applications and requirements of electrical insulators and ceramics in fusion and to recommend strategies and different stages of implementation. This meeting was endorsed by the International Fusion Research Council

  3. The hallmarks of cell-cell fusion.

    Science.gov (United States)

    Hernández, Javier M; Podbilewicz, Benjamin

    2017-12-15

    Cell-cell fusion is essential for fertilization and organ development. Dedicated proteins known as fusogens are responsible for mediating membrane fusion. However, until recently, these proteins either remained unidentified or were poorly understood at the mechanistic level. Here, we review how fusogens surmount multiple energy barriers to mediate cell-cell fusion. We describe how early preparatory steps bring membranes to a distance of ∼10 nm, while fusogens act in the final approach between membranes. The mechanical force exerted by cell fusogens and the accompanying lipidic rearrangements constitute the hallmarks of cell-cell fusion. Finally, we discuss the relationship between viral and eukaryotic fusogens, highlight a classification scheme regrouping a superfamily of fusogens called Fusexins, and propose new questions and avenues of enquiry. © 2017. Published by The Company of Biologists Ltd.

  4. Synergistic efficiency of the desilication of brackish underground water in Saudi Arabia by coupling γ-radiation and Fenton process: Membrane scaling prevention in reverse osmosis process

    Science.gov (United States)

    Aljohani, Mohammed S.

    2017-12-01

    One of the main water resources in arid Saudi Arabia is underground water. However, this brackish water has high silica content which can cause a recalcitrant deposit on the membrane in the reverse osmosis units during its desalination. In this study, we examined the synergistic efficiency of the removal of silica from the Buwaib water sample, when combining two advanced oxidation processes, γ-irradiation and the Fenton process, using hydrogen peroxide and zero valent metal iron as source of Fe3+. This latter adsorbs effectively on silica and co-precipitate. The influence of absorbed dose, iron dosage and pH effect were investigated. This preliminary study showed that these attractive and effective hybrid processes are very efficient in removing silica.

  5. Fusion energy

    International Nuclear Information System (INIS)

    Anon.

    1979-01-01

    The efforts of the Chemical Technology Division in fusion energy include the areas of fuel handling, processing, and containment. Current studies are concerned largely with the development of vacuum pumps for fusion reactors and experiments and with development and evaluation of techniqu