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1

AN IMPROVED CELL FRACTIONATION PROCEDURE FOR THE PREPARATION OF RAT LIVER MEMBRANE-BOUND RIBOSOMES  

PubMed Central

A cell fractionation procedure is described which allows the preparation from rat liver of a rough microsome population containing almost 50% of the membrane-bound ribosomes of the tissue. The fraction is not contaminated with free ribosomes or smooth microsomes, and, by various other criteria, is suitable for studies of ribosome-membrane interaction.

Adelman, M. R.; Blobel, Gunter; Sabatini, David D.

1973-01-01

2

Mechanical Forces Used for Cell Fractionation Can Create Hybrid Membrane Vesicles  

PubMed Central

The ability to understand the inner works of the cell requires methods for separation of intracellular membrane-enclosed compartments. Disruption of the plasma membrane (PM) by mechanical forces to investigate the content of the cell is common practice. Whether vesicles or membranes of different sources can fuse as a result is unclear. If such contamination occurs, conclusions based on these techniques should consider these. Utilizing an endoplasmic reticulum (ER) membrane marker and a PM marker, we were able to detect the source of membranes following the breakup of cells using flow cytometry and immuno Electron Microscopy (immuno EM). Fractionation processes produced a small fraction of new membrane entities from two distinctively different origins generated during the initial disruption steps in a temperature independent manner, stressing that defining organelles or intrinsic fusion events based on such procedures and markers are valid when exceeding the small number of vesciles fused during the fractionation process.

Salomon, Izhar; Janssen, Hans; Neefjes, Jacques

2010-01-01

3

FURTHER CHARACTERIZATION OF PARTICULATE FRACTIONS FROM LYSED CELL ENVELOPES OF HALOBACTERIUM HALOBIUM AND ISOLATION OF GAS VACUOLE MEMBRANES  

Microsoft Academic Search

Lysates of cell envelopes from Halobacterium halobium have been separated into four fractions. A soluble, colorless fraction (I) containing protein, hexosamines, and no lipid is apparently derived from the cell wall. A red fraction (II), containing approximately 40 per cent lipid, 60 per cent protein, and a small amount of hexosamines consists of cell membrane disag- gregated into fragments of

WALTHER STOECKENIUS; WOLF H. KUNAU

1968-01-01

4

Fraction reduction in membrane systems.  

PubMed

Fraction reduction is a basic computation for rational numbers. P system is a new computing model, while the current methods for fraction reductions are not available in these systems. In this paper, we propose a method of fraction reduction and discuss how to carry it out in cell-like P systems with the membrane structure and the rules with priority designed. During the application of fraction reduction rules, synchronization is guaranteed by arranging some special objects in these rules. Our work contributes to performing the rational computation in P systems since the rational operands can be given in the form of fraction. PMID:24772037

Guo, Ping; Zhang, Hong; Chen, Haizhu; Liu, Ran

2014-01-01

5

Fractional order models of viscoelasticity as an alternative in the analysis of red blood cell (RBC) membrane mechanics  

PubMed Central

New lumped-element models of red blood cell mechanics can be constructed using fractional order generalizations of springs and dashpots. Such ‘spring-pots’ exhibit a fractional order viscoelastic behavior that captures a wide spectrum of experimental results through power-law expressions in both the time and frequency domains. The system dynamics is fully described by linear fractional order differential equations derived from first order stress–strain relationships using the tools of fractional calculus. Changes in the composition or structure of the membrane are conveniently expressed in the fractional order of the model system. This approach provides a concise way to describe and quantify the biomechanical behavior of membranes, cells and tissues.

Craiem, Damian; Magin, Richard L

2011-01-01

6

Terpenes in sponge cell membranes: Cell separation and membrane fractionation studies with the tropical marine sponge Amphimedon sp  

Microsoft Academic Search

The differing sponge and symbiotic microbial cell types in the tropical marine spongeAmphimedon sp. were fractionated according to density, investigated by electron microscopy, and analyzed by high-performance liquid\\u000a chromatography and nuclear magnetic resonance for the presence of the terpene metabolite diisocyanoadociane (1) and ?5,7-sterols (2–7). A sample of whole sponge was dissected into superficial ectosome and deeper choanosome. The superficial

Mary J. Garsona; Janice E. Thompsonb; Rundi M. Larsen; Christopher N. Battershill; Peter T. Murphy; Patricia R. Bergquist

1992-01-01

7

Calcium translocation across the hen oviduct shell gland: subcellular fractionation of the epithelial cells and calcium transport studies with isolated membrane populations  

SciTech Connect

An attempt was made to develop methods for studying calcium transport in vitro using isolated membrane populations from the epithelium of the hen shell gland. First, subcellular components of the epithelial cells were fractionated by isopycnic centrifugation on sucrose density gradients and subsequent countercurrent distribution in aqueous polymer two-phase systems. Four membrane populations were identified based on the assignment of traditional enzyme markers; three contained primarily either endoplasmic reticulum, Golgi or basolateral cell membranes. Apical cell membranes could not be unequivocally assigned to the fourth membrane population. Microautoradiography of radioiodinated shell gland sections indicated a preferential, but not absolute, specificity of /sup 125/I for the epithelial cell apical membrane. Next, calcium transport by these isolated membrane fractions was investigated, calcium accumulation was dependent on time, temperature, ATP and the absence of the calcium ionophore A23187, strongly suggesting uptake into intact membrane vesicles.

Brown, T.A.

1987-01-01

8

RIBONUCLEIC ACID IN A "MEMBRANE" FRACTION OF ESCHERICHIA COLI AND ITS RELATION TO CELL-WALL SYNTHESIS  

PubMed Central

Suit, Joan C. (The University of Texas M. D. Anderson Hospital and Tumor Institute, Houston). Ribonucleic acid in a “membranefraction of Escherichia coli and its relation to cell-wall synthesis. J. Bacteriol. 84:1061–1070. 1962.—A small amount of ribonucleic acid (RNA) was found in a “membranefraction prepared from osmotically sensitized Escherichia coli. It exhibited an elevated metabolic activity in that it attained the highest specific activity of any RNA in subcellular fractions of logarithmic-phase cells or spheroplasts prepared from logarithmic-phase cells which had been allowed to incorporate P32 briefly. The metabolic activity of this RNA, in terms of P32 incorporation, was found to be independent of cell-wall synthesis in the diaminopimelic acid (DAP)-less mutant, E. coli W 173-25, but was inhibited by penicillin in both this strain and in E. coli B. The latter effect is considered to be a result of other complex inhibitions of cellular metabolism by the antibiotic. The development of sensitivity to osmotic shock, capability of recovery, and synthesis of macromolecules in penicillin-treated and DAP-starved cultures, under these conditions, is described.

Suit, Joan C.

1962-01-01

9

Toxic Membrane Fractions from Mycoplasma fermentans1  

PubMed Central

A recent isolate of Mycoplasma fermentans (strain K10, from human leukemic bone marrow) induced a lethal toxicity syndrome in mice. High doses of both viable and inactivated cells were toxic when injected intraperitoneally. Whole lysates and membranes from osmotically shocked cells killed mice, but cytoplasm did not. When membranes were dissolved in detergents and reaggregated by dialysis in the presence of Mg2+, the lipid-protein complex thus formed was toxic. Lipids extracted from membranes with chloroform-methanol did not kill mice. Protein-rich fractions (obtained by reaggregation plus acetone washes or ammonium sulfate precipitation of dissolved membranes) were also not toxic. No qualitative differences in proteins from three toxic isolates and three nontoxic laboratory strains of M. fermentans were detectable by polyacrylamide gel electrophoresis. The toxic factor contained in reaggregated membranes was heat-stable but sensitive to Pronase, trypsin, and lipase. Images

Gabridge, Michael G.; Murphy, William H.

1971-01-01

10

Microscale isoelectric fractionation using photopolymerized membranes.  

PubMed

In this work, we introduce microscale isoelectric fractionation (?IF) for isolation and enrichment of molecular species at any desired location in a microfluidic chip. Narrow pH-specific polyacrylamide membranes are photopatterned in situ for customizable device fabrication; multiple membranes of precise pH are easily incorporated throughout existing channel layouts. Samples are electrophoretically driven across the membranes such that charged species, for example, proteins and peptides, are rapidly discretized into fractions based on their isoelectric points (pI) without the use of carrier ampholytes. This format makes fractions easy to compartmentalize and access for integrated preparative or analytical operations on-chip. We present and discuss the key design considerations and trade-offs associated with proper system operation and optimal run conditions. Efficient and reproducible fractionation of model fluorescent pI markers and proteins is achieved using single membrane fractionators at pH 6.5 and 5.3 from both buffer and Escherichia coli cell lysate sample conditions. Effective fractionation is also shown using a serial 3-membrane fractionator tailored for isolating analytes-of-interest from high abundance components of serum. We further demonstrate that proteins focused in pH specific bins can be rapidly and efficiently transferred to another location in the same chip without unwanted dilution or dispersive effects. ?IF provides a rapid and versatile option for integrated sample prep or multidimensional analysis, and addresses the glaring proteomic need to isolate trace analytes from high-abundance species in minute volumes of complex samples. PMID:21417312

Sommer, Greg J; Mai, Junyu; Singh, Anup K; Hatch, Anson V

2011-04-15

11

Antimicrobial properties of milkfat globule membrane fractions.  

PubMed

Milkfat globule membranes (MFGMs) were prepared from bovine cream according to standard procedures. These membranes and peptide hydrolysates, which were generated by proteolysis with immobilized digestive enzymes, were screened for antibacterial activity against Escherichia coli O157:H7, Listeria monocytogenes, Salmonella enterica Typhimurium, Pseudomonas fluorescens, Bacillus cereus, Lactobacillus acidophilus, and Lactobacillus gasseri. Assays were first performed on beef heart infusion (BHI) plates spotted with test protein-peptide fractions and then seeded with lawns of indicator cells to monitor the zone of growth inhibition. Under these experimental conditions, MFGMs were most active against Salmonella Typhimurium and P. fluorescens. However, antibacterial activity was not seen after plating on Luria-Bertani (LB) medium. We determined that the antimicrobial effects observed on BHI plates were due to the generation of H2O2 by xanthine oxidase, a major protein constituent of the MFGMs, as a result of purine catalysis. This substrate is present in BHI but lacking in LB medium. Evaluation of purified xanthine oxidase alone resulted in analogous data trends. The growth of probiotic Lactobacillus strains were affected only marginally when grown on lactobacilli deMan Rogosa Sharpe plates, suggesting the decreased sensitivity of these bacteria to H2O2. In this study, several MFGM hydrolysates exhibited variable antibacterial activity against test food pathogens on agar plates prepared with M9 minimal media, and this variation was not attributable to xanthine oxidase enzymatic activity. The probiotic microorganisms were mostly resilient to these antibacterial fractions. Bovine MFGM fractions may represent an excellent resource material from which to generate native, naturally occurring biodefensive proteins and/or peptides. PMID:18236672

Clare, Debra A; Zheng, Zuoxing; Hassan, Hosni M; Swaisgood, Harold E; Catignani, George L

2008-01-01

12

Quantitative proteomics of fractionated membrane and lumen exosome proteins from isogenic metastatic and nonmetastatic bladder cancer cells reveal differential expression of EMT factors.  

PubMed

Cancer cells secrete soluble factors and various extracellular vesicles, including exosomes, into their tissue microenvironment. The secretion of exosomes is speculated to facilitate local invasion and metastatic spread. Here, we used an in vivo metastasis model of human bladder carcinoma cell line T24 without metastatic capacity and its two isogenic derivate cell lines SLT4 and FL3, which form metastases in the lungs and liver of mice, respectively. Cultivation in CLAD1000 bioreactors rather than conventional culture flasks resulted in a 13- to 16-fold increased exosome yield and facilitated quantitative proteomics of fractionated exosomes. Exosomes from T24, SLT4, and FL3 cells were partitioned into membrane and luminal fractions and changes in protein abundance related to the gain of metastatic capacity were identified by quantitative iTRAQ proteomics. We identified several proteins linked to epithelial-mesenchymal transition, including increased abundance of vimentin and hepatoma-derived growth factor in the membrane, and casein kinase II ? and annexin A2 in the lumen of exosomes, respectively, from metastatic cells. The change in exosome protein abundance correlated little, although significant for FL3 versus T24, with changes in cellular mRNA expression. Our proteomic approach may help identification of proteins in the membrane and lumen of exosomes potentially involved in the metastatic process. PMID:24376083

Jeppesen, Dennis Kjølhede; Nawrocki, Arkadiusz; Jensen, Steffen Grann; Thorsen, Kasper; Whitehead, Bradley; Howard, Kenneth A; Dyrskjøt, Lars; Ørntoft, Torben Falck; Larsen, Martin R; Ostenfeld, Marie Stampe

2014-03-01

13

Composite fuel cell membranes  

DOEpatents

A bilayer or trilayer composite ion exchange membrane is described suitable for use in a fuel cell. The composite membrane has a high equivalent weight thick layer in order to provide sufficient strength and low equivalent weight surface layers for improved electrical performance in a fuel cell. In use, the composite membrane is provided with electrode surface layers. The composite membrane can be composed of a sulfonic fluoropolymer in both core and surface layers.

Plowman, K.R.; Rehg, T.J.; Davis, L.W.; Carl, W.P.; Cisar, A.J.; Eastland, C.S.

1997-08-05

14

Composite fuel cell membranes  

DOEpatents

A bilayer or trilayer composite ion exchange membrane suitable for use in a fuel cell. The composite membrane has a high equivalent weight thick layer in order to provide sufficient strength and low equivalent weight surface layers for improved electrical performance in a fuel cell. In use, the composite membrane is provided with electrode surface layers. The composite membrane can be composed of a sulfonic fluoropolymer in both core and surface layers.

Plowman, Keith R. (Lake Jackson, TX) [Lake Jackson, TX; Rehg, Timothy J. (Lake Jackson, TX) [Lake Jackson, TX; Davis, Larry W. (West Columbia, TX) [West Columbia, TX; Carl, William P. (Marble Falls, TX) [Marble Falls, TX; Cisar, Alan J. (Cypress, TX) [Cypress, TX; Eastland, Charles S. (West Columbia, TX) [West Columbia, TX

1997-01-01

15

Plant cell membranes  

SciTech Connect

The contents of this book are: Cells, Protoplasts, Vacuoles and Liposomes; Tonoplasts; Nuclei, Endolplasmic Reticulum, and Plasma Membrane; Peroxisomes; Plastids; Teneral Physical and Biochemical Methods; and Mitochondira.

Packer, L.; Douce, R.

1987-01-01

16

Fractionation and analysis of mitochondria with polycarbonate membrane filters.  

PubMed

Polycarbonate membrane filters were used to fractionate mitochondrial populations depending on their aggregation or association with other subcellular structures. Isolated rat liver mitochondria penetrated through filters which have pore sizes larger than 1 micron. In contrast, mitochondria which were induced to aggregate in vitro by incubation at low pH were retained by the filters and thus could be separated from the single or small aggregates of mitochondria. Use of this membrane filtration method to analyze release of mitochondria from isolated hepatocytes showed that treatment with digitonin at concentrations only sufficient to lyse the plasma membrane did not release mitochondria. Homogenization or sonication following digitonin treatment released 25-50% of the mitochondria, but only a small fraction was intact. A high yield of intact mitochondria was released from digitonin-treated cells by a brief treatment with a low concentration of the proteolytic enzyme nagarse. Thus, this membrane filtration method provides a simple and rapid approach to analyze the extent of mitochondrial aggregation and association with other subcellular structures. PMID:2757183

Bai, C; Slife, C W; Aw, T Y; Jones, D P

1989-05-15

17

Regulation of actin polymerization by membrane fraction of platelets.  

PubMed

We studied the interaction between the purified membrane fraction of human platelets and the polymerization of skeletal actin. The viscosity of actin was measured by the falling ball method. The fraction suppressed the polymerization of actin in the presence of 20 mM KCl and 0.4 mM EGTA. The addition of calcium ion or thrombin to the fraction did not cause suppression. A DNase I affinity column bound the membrane fraction in the presence of calcium ion. The frozen membrane fraction and the vesicles reconstituted with lipids from the platelet membrane enhanced the polymerization of actin. Trypsinized membrane fraction and the membrane fraction treated with phospolipase A2 enhanced the polymerization of actin, but membrane fraction treated with phospholipase C had no effect. The reconstituted membrane vesicles mentioned above lowered the critical concentration for actin polymerization. These findings suggested that the polymerization of intracellular actin is enhanced not only by the mobilization of calcium ion, but also by biochemical changes in the membrane lipids. PMID:3365264

Hashimoto, K; Tatsumi, N

1988-02-01

18

Gas phase fractionation method using porous ceramic membrane  

DOEpatents

Flaw-free porous ceramic membranes fabricated from metal sols and coated onto a porous support are advantageously used in gas phase fractionation methods. Mean pore diameters of less than 40 .ANG., preferably 5-20 .ANG. and most preferably about 15 .ANG., are permeable at lower pressures than existing membranes. Condensation of gases in small pores and non-Knudsen membrane transport mechanisms are employed to facilitate and increase membrane permeability and permselectivity.

Peterson, Reid A. (Madison, WI); Hill, Jr., Charles G. (Madison, WI); Anderson, Marc A. (Madison, WI)

1996-01-01

19

Hepatic Golgi fractions resolved into membrane and content subfractions  

PubMed Central

Golgi fractions isolated from rat liver homogenates have been resolved into membrane and content subfractions by treatment with 100 mM Na2CO3 pH 11.3. This procedure permitted extensive extraction of content proteins and lipoproteins, presumably because it caused an alteration of Golgi membranes that minimized the reformation of closed vesicles. The type and degree of contamination of the fractions was assessed by electron microscopy and biochemical assays. The membrane subfraction retained 15% of content proteins and lipids, and these could not be removed by various washing procedures. The content subfraction was contaminated by both membrane fragments and vesicles and accounted for 5 to 10% of the membrane enzyme activities of the original Golgi fraction. The lipid compositions of the subfractions was determined, and the phospholipids of both membrane and content were found to be uniformly labeled with [33P]phosphate administered in vivo.

1982-01-01

20

Membrane Cells for Brine Electrolysis.  

ERIC Educational Resources Information Center

Membrane cells were developed as alternatives to mercury and diaphragm cells for the electrolysis of brine. Compares the three types of cells, focusing on the advantages and disadvantages of membrane cells. (JN)

Tingle, M.

1982-01-01

21

Fractionation of olive mill wastewaters by membrane separation techniques.  

PubMed

This study aims to evaluate the potential of an integrated membrane system in the treatment of olive mill wastewaters (OMWs) to produce a purified fraction enriched in low molecular weight polyphenols, a concentrated fraction of organic substances and a water stream which can be reused in the extractive process of olive oil. In particular, a sequence of two ultrafiltration (UF) processes followed by a final nanofiltration (NF) step was investigated on laboratory scale operating in selected process parameters. The produced fractions were analyzed for their total content of polyphenols, total antioxidant activity (TAA), free low molecular weight polyphenols and total organic carbon (TOC). The performance of selected membranes in terms of productivity, fouling index and selectivity toward compounds of interest was also evaluated and discussed. An integrated membrane process was proposed to achieve high levels of purification of OMWs and a water fraction which can be discharged in aquatic systems or to be reused in the olive oil extraction process. PMID:23376489

Cassano, Alfredo; Conidi, Carmela; Giorno, Lidietta; Drioli, Enrico

2013-03-15

22

Cell Membranes Tutorial  

NSDL National Science Digital Library

New from The Biology Project of the University of Arizona, this online tutorial "introduces the dynamic complexes of proteins, carbohydrates, and lipids that comprise cell membranes," and relates how membranes "are important for regulating ion and molecular traffic flow between cells." Each section of this Web site takes the form of a multiple choice question. Answer the question correctly, and a brief explanation of each answer choice will be displayed. Answer the question incorrectly, and a short but helpful tutorial with colorful diagrams will help get you on the right track. This would be an valuable Web site for students wishing to test themselves on cell membrane structure and function, but would not be especially useful for those new to the subject.

2002-01-01

23

Enzymatic activities in cell fractions of mycoplasmalike organisms purified from aster yellows-infected plants.  

PubMed Central

Mycoplasmalike organisms (MLOs), purified from aster yellows-infected plants were osmotically lysed, and the membranes were separated from the cytoplasmic fraction through differential centrifugation. Electron microscopic examinations of sections of the purified MLOs and the isolated membranes showed pleomorphic bodies and unit membranous empty vesicles, respectively. Cell fractions were tested for NADH oxidase, NADPH oxidase, ATPase, RNase, DNase, and p-nitrophenyl phosphatase activity. NADH oxidase and ATPase were confined to the membrane fraction and NADPH oxidase to the cytoplasmic fraction of the MLOs. para-Nitrophenyl phosphatase, RNase, and DNase activities were detected in both membrane and cytoplasmic fractions, but p-nitrophenyl phosphatase and RNase appeared to be associated with membranes and DNase with the cytoplasmic fraction. Glucose-6-phosphate dehydrogenase was found in the cytoplasmic fraction of the MLO cells. Our findings on the distribution of enzymes in MLO cells and cell fractions are the first basic documentation on nonhelical, nonculturable microbes parasitic to plants. Images

Arora, Y K; Sinha, R C

1985-01-01

24

[Effect of different organic fraction on membrane flux declines].  

PubMed

Organic matter in the tap water was isolated into strongly hydrophobic acids, weakly hydrophobic acids, charged hydrophilic and neutral hydrophilic by DAX-8, XAD-4 and IRA-958 synthetic resins. Filtration tests using polyvinylidene fluoride (PVDF), polyethersulphone (PES) and cellulose acetate (CA) membranes were conducted to investigate the contribution of different organic fractions to membrane fouling. The results show that in filtration of raw water, flux declines with PES, PVDF and CA membrane are 67%, 59% and 19% of the initial flux, indicating that the more hydrophobic membrane resulted in more severe fouling. For the effect of different fractions on flux, flux decline with neutral hydrophilic is 41%-75% of the initial flux, whereas weakly hydrophobic acids is 6%-33%, suggesting that neutral hydrophilic has a great impact on filtration flux. Among three membranes tested, CA membrane shows the lowest flux decline compared with other membranes in spite of rejection of as high as 14.69% of neutral hydrophilic, suggesting that the extent of flux decline may not be associated with the total amount of NOM removed. The mechanism of fouling was discussed and found that the neutral hydrophilic fraction with greater than 3 x 10(4) of molecular weight caused a significant flux decline, through blocking the pore for the MF or UF having greater relative molecular mass cut-off (MWCO), but resulted in a little impact on flux with the UF having lower MWCO, through forming cake layer on the surface of membrane due to not entering the inside of pore. PMID:19402494

Zhou, Xian-Jiao; Dong, Bing-Zhi

2009-02-15

25

Protein fractionation using fast flow immobilized metal chelate affinity membranes.  

PubMed

A new group-specific affinity membrane using metal chelates as ligands and inorganic glass hollow fiber microfiltration membranes as support matrices is developed and tested. The study focused on developing the optimum activation and coupling procedures to bind the chelating agent (iminodiacetic acid, IDA) to the surface of the microporous glass hollow fiber membrane and testing the resultant affinity membrane. Starting with three different glass surfaces, five modification reactions were evaluated. All the modified "active surfaces" were first tested for their protein adsorptive properties in batch mode with suspended microporous glass grains using model proteins with known binding characteristics with Cu-IDA systems. The metal loading capacities of the surfaces exhibiting favorable fractionation were then measured by atomic absorption spectroscopy.The results were compared with the results obtained with a commercial material used in immobilized metal affinity column chromatography. The protein binding characteristics of the hollow fiber affinity membranes were also evaluated under conditions of convective flow. This was performed by flowing single solute protein solutions through the microporous membrane at different flow rates. These results were then used to estimate the optimum loading and elution times for the process. A mathematical model incorporating radial diffusion was solved using a finite difference discretization method. Comparison between model predictions and experimental results was performed for four different proteins at one flow rate. These results suggested that the kinetics of adsorption was concentration dependent. Finally, the hollow fiber affinity membranes were challenged with two component mixtures to test their ability to fractionate mixed protein solutions. Efficient separation and good purity were obtained.The results presented here represent the development of a new fast flow affinity membrane process-immobilized metal affinity membranes (IMAM). (c) 1994 John Wiley & Sons, Inc. PMID:18613307

Serafica, G C; Belfort, G; Pimbley, J

1994-01-01

26

The First Cell Membranes  

NASA Technical Reports Server (NTRS)

Organic compounds are synthesized in the interstellar medium and can be delivered to planetary surfaces such as the early Earth, where they mix with endogenous organic mixtures. Some of these compounds are amphiphilic, having polar and non-polar groups on the same molecule. Amphiphilic compounds spontaneously self-assembly into more complex structures such as bimolecular layers, which in turn form closed membranous vesicles. The first forms of cellular life required self-assembled membranes that were likely to be available on the prebiotic Earth. Laboratory simulations show that such vesicles readily encapsulate functional macromolecules, including nucleic acids and polymerases. A goal of future investigations is to fabricate artificial cells as models of the origin of life.

Vondrak, Richard R. (Technical Monitor); Demner, David; Dworkin, Jason P.; Sandford, Scott A.; Bernstein, Max P.; Allamandola, Louis J.

2002-01-01

27

The First Cell Membranes  

NASA Astrophysics Data System (ADS)

Organic compounds are synthesized in the interstellar medium and can be delivered to planetary surfaces such as the early Earth, where they mix with endogenous species. Some of these compounds are amphiphilic, having polar and nonpolar groups on the same molecule. Amphiphilic compounds spontaneously self-assemble into more complex structures such as bimolecular layers, which in turn form closed membranous vesicles. The first forms of cellular life required self-assembled membranes that were likely to have been produced from amphiphilic compounds on the prebiotic Earth. Laboratory simulations show that such vesicles readily encapsulate functional macromolecules, including nucleic acids and polymerases. The goal of future investigations will be to fabricate artificial cells as models of the origin of life.

Deamer, David; Dworkin, Jason P.; Sandford, Scott A.; Bernstein, Max P.; Allamandola, Louis J.

2002-12-01

28

Membrane rafts of the human red blood cell.  

PubMed

The cell type of election for the study of cell membranes, the mammalian non-nucleated erythrocyte, has been scarcely considered in the research of membrane rafts of the plasma membrane. However, detergent-resistant-membranes (DRM) were actually first described in human erythrocytes, as a fraction resisting solubilization by the nonionic detergent Triton X-100. These DRMs were insoluble entities of high density, easily pelleted by centrifugation, as opposed to the now accepted concept of lipid raft-like membrane fractions as material floating in low-density regions of sucrose gradients. The present article reviews the available literature on membrane rafts/DRMs in human erythrocytes from an historical point of view, describing the experiments that provided the solution to the above described discrepancy and suggesting possible avenue of research in the field of membrane rafts that, moving from the most studied model of living cell membrane, the erythrocyte's, could be relevant also for other cell types. PMID:24720522

Ciana, Annarita; Achilli, Cesare; Minetti, Giampaolo

2014-01-01

29

Origin of subdiffusion of water molecules on cell membrane surfaces  

NASA Astrophysics Data System (ADS)

Water molecules play an important role in providing unique environments for biological reactions on cell membranes. It is widely believed that water molecules form bridges that connect lipid molecules and stabilize cell membranes. Using all-atom molecular dynamics simulations, we show that translational and rotational diffusion of water molecules on lipid membrane surfaces exhibit subdiffusion and aging. Moreover, we provide evidence that both divergent mean trapping time (continuous-time random walk) and long-correlated noise (fractional Brownian motion) contribute to this subdiffusion. These results suggest that subdiffusion on cell membranes causes the water retardation, an enhancement of cell membrane stability, and a higher reaction efficiency.

Yamamoto, Eiji; Akimoto, Takuma; Yasui, Masato; Yasuoka, Kenji

2014-04-01

30

Blue Light-Reducible Cytochromes in Membrane Fractions from Neurospora crassa1  

PubMed Central

We have assayed absorbance changes generated by blue light in plasma membranes, endoplasmic reticulum, and mitochondrial membranes from Neurospora crassa. Light minus dark difference spectra, obtained anaerobically in the presence of ethylenediaminetetraacetate, indicated that b-type cytochromes could be photoreduced in all three membranes. In plasma membranes, a b-type cytochrome with a distinct difference spectrum was photoreducible without addition of exogenous flavin. Addition of riboflavin greatly stimulated the photoreduction of cytochromes in endoplasmic reticulum and mitochondrial membranes. In its spectral characteristics the cytochrome on the endoplasmic reticulum resembled cytochrome b5 or nitrate reductase, while the cytochrome in mitochondrial membranes had the same spectrum as cytochrome b of the mitochondrial respiratory chain. Cytochromes in the three membrane fractions reacted differently to blue light in the presence of various inhibitors. Potassium azide inhibited reduction of plasma membrane cytochrome b, with 50% inhibition at 1.0 millimolar. The same concentration of azide stimulated photoreduction of cytochromes in both endoplasmic reticulum and mitochondria. Although photoreduction of cytochromes in all three membranes was inhibited by salicylhydroxamic acid, cytochromes in plasma membranes were more sensitive to this inhibitor than those in endoplasmic reticulum and mitochondria. Cells grown to induce nitrate reductase activity showed an elevated amount of blue light-reducible cytochrome b in the endoplasmic reticulum.

Borgeson, Charlotte E.; Bowman, Barry J.

1985-01-01

31

Fractionation of membrane vesicles from coliphage M13-infected Escherichia coli.  

PubMed Central

Membrane vesicles were prepared by osmotic lysis of spheroplasts from M13-infected Escherichia coli. Reduced nicotinamide adenine dinucleotide (NADH) oxidase (reduced NAD: oxidoreductase, EC 1.6.99.3) and Mg2+-Ca2+-activated adenosine triphosphatase (ATP phosphohydrolase, EC 3.6.1.3), which are normally localized to the inner surface of the cytoplasmic membrane, were 50% acceesible to their polar substrates in these vesicles. The major coat protein of coliphage M13 is also bound to the cytoplasmic membrane (prior to phage assembly) but with its antigenic sites exposed to the exterior of the cell. Antibody to M13 coat protein was used to fractionate membrane vesicles. Neither agglutinated nor unagglutinated vesicles had altered NADH oxidase and adenosine triphosphatase specific activities. This is inconsistent with such vesicles being a mixture of correctly oriented and completely inverted membrane sacs and suggests that NADH oxidase, adenosine triphosphatase, M13 coat protein, or all three proteins rearrange during vesicle preparation.

Wickner, W

1976-01-01

32

Fuel cell with ionization membrane  

NASA Technical Reports Server (NTRS)

A fuel cell is disclosed comprising an ionization membrane having at least one area through which gas is passed, and which ionizes the gas passing therethrough, and a cathode for receiving the ions generated by the ionization membrane. The ionization membrane may include one or more openings in the membrane with electrodes that are located closer than a mean free path of molecules within the gas to be ionized. Methods of manufacture are also provided.

Hartley, Frank T. (Inventor)

2007-01-01

33

Thermoelasticity of red blood cell membrane.  

PubMed

The elastic properties of the human red blood cell membrane have been measured as functions of temperature. The area compressibility modulus and the elastic shear modulus, which together characterize the surface elastic behavior of the membrane, have been measured over the temperature range of 2-50 degrees C with micropipette aspiration of flaccid and osmotically swollen red cells. In addition, the fractional increase in membrane surface area from 2-50 degrees C has been measured to give a value for the thermal area expansivity. The value of the elastic shear modulus at 25 degrees C was measured to be 6.6 X 10(-3) dyne/cm. The change in the elastic shear modulus with temperature was -6 X 10(-5) dyne/cm degrees C. Fractional forces were shown to be only on the order of 10-15%. The area compressibility modulus at 25 degrees C was measured to be 450 dyne/cm. The change in the area compressibility modulus with temperature was -6 dyne/cm degrees C. The thermal area expansivity for red cell membrane was measured to be 1.2 X 10(-3)/degrees C. With this data and thermoelastic relations the heat of expansion is determined to be 110-200 ergs/cm2; the heat of extension is 2 X 10(-2) ergs/cm2 for unit extension of the red cell membrane. The heat of expansion is of the order anticipated for a lipid bilayer idealized as twice the behavior of a monolayer at an oil-water interface. The observation that the heat of extension is positive demonstrates that the entropy of the material increases with extension, and that the dominant mechanism of elastic energy storage is energetic. Assuming that the red cell membrane shear rigidity is associated with "spectrin," unit extension of the membrane increases the configurational entropy of spectrin by 500 cal/mol. PMID:262408

Waugh, R; Evans, E A

1979-04-01

34

Subcellular Fractionation Enhances Proteome Coverage of Pancreatic Duct Cells  

PubMed Central

Objectives Subcellular fractionation of whole cell lysates offers a means of simplifying protein mixtures, potentially permitting greater depth of proteomic analysis. Here we compare proteins identified from pancreatic duct cells (PaDC) following organelle enrichment to those identified from PaDC whole cell lysates to determine if the additional procedures of subcellular fractionation increases proteome coverage. Methods We used differential centrifugation to enrich for nuclear, mitochondrial, membrane, and cytosolic proteins. We then compared - via mass spectrometry-based analysis - the number of proteins identified from these four fractions with four biological replicates of PaDC whole cell lysates. Results We identified similar numbers of proteins among all samples investigated. In total, 1658 non-redundant proteins were identified in the replicate samples, while 2196 were identified in the subcellular fractionation samples, corresponding to a 30% increase. Additionally, we noted that each organelle fraction was in fact enriched with proteins specific to the targeted organelle. Conclusions Subcellular fractionation of PaDC resulted in greater proteome coverage compared to PaDC whole cell lysate analysis. Although more labor intensive and time consuming, subcellular fractionation provides greater proteome coverage, and enriches for compartmentalized sub-populations of proteins. Application of this subcellular fractionation strategy allows for a greater depth of proteomic analysis and thus a better understanding of the cellular mechanisms of pancreatic disease.

Paulo, Joao A.; Gaun, Aleksandr; Kadiyala, Vivek; Ghoulidi, Ali; Banks, Peter A.; Conwell, Darwin L.; Steen, Hanno

2013-01-01

35

Membrane Transport in Primitive Cells  

PubMed Central

Although model protocellular membranes consisting of monoacyl lipids are similar to membranes composed of contemporary diacyl lipids, they differ in at least one important aspect. Model protocellular membranes allow for the passage of polar solutes and thus can potentially support cell-to functions without the aid of transport machinery. The ability to transport polar molecules likely stems from increased lipid dynamics. Selectively permeable vesicle membranes composed of monoacyl lipids allow for many lifelike processes to emerge from a remarkably small set of molecules.

Mansy, Sheref S.

2010-01-01

36

Cytoplasmic membrane fraction that promotes septation in an Escherichia coli lon mutant  

SciTech Connect

A particulate fraction derived from bacterial cells stimulates septation in irradiated Escherichia coli lon mutants when added to postirradiation plating media. It was established that the particles are derived from the cytoplasmic membrane and that they have been partially purified by sucrose density gradient centrifugation. These particles also contain the cytochrome-based respiratory activity of the cell. A variety of experiments established a correlation between the septation-promoting activity of the particles and their ability to remove oxygen from the postirradiation plating medium. It was suggested that the efficient removal of oxygen from the medium allowed the lon cells to repair radiation-induced damage to the septation mechanism.

Adler, H.I.; Carrasco, A.; Crow, W.; Gill, J.S.

1981-08-01

37

Theoretical analysis of membrane tension in moving cells.  

PubMed

Lateral tension in cell plasma membranes plays an essential role in regulation of a number of membrane-related intracellular processes and cell motion. Understanding the physical factors generating the lateral tension and quantitative determination of the tension distribution along the cell membrane is an emerging topic of cell biophysics. Although experimental data are accumulating on membrane tension values in several cell types, the tension distribution along the membranes of moving cells remains largely unexplored. Here we suggest and analyze a theoretical model predicting the tension distribution along the membrane of a cell crawling on a flat substrate. We consider the tension to be generated by the force of actin network polymerization against the membrane at the cell leading edge. The three major factors determining the tension distribution are the membrane interaction with anchors connecting the actin network to the lipid bilayer, the membrane interaction with cell adhesions, and the force developing at the rear boundary due to the detachment of the remaining cell adhesion from the substrate in the course of cell crawling. Our model recovers the experimentally measured values of the tension in fish keratocytes and their dependence on the number of adhesions. The model predicts, quantitatively, the tension distribution between the leading and rear membrane edges as a function of the area fractions of the anchors and the adhesions. PMID:24411240

Schweitzer, Yonatan; Lieber, Arnon D; Keren, Kinneret; Kozlov, Michael M

2014-01-01

38

In Vitro Enzymatic Reduction Kinetics of Mineral Oxides by Membrane Fractions from Shewanella oneidensis MR-1  

SciTech Connect

This study documents the first example of in vitro solid-phase mineral oxide reduction by enzyme-containing membrane fractions. Previous in vitro studies have only reported the reduction of aqueous ions. Total membrane (TM) fractions from iron-grown cultures of Shewanella oneidensis MR-1 were isolated and shown to catalyze the reduction of goethite, hematite, birnessite, and ramsdellite/pyrolusite using formate. In contrast, nicotinamide adenine dinucleotide (NADH) and succinate cannot function as electron donors. The significant implications of observations related to this cell-free system are: (i) both iron and manganese mineral oxides are reduced by the TM fraction, but aqueous U(VI) is not; (ii) TM fractions from anaerobically grown, but not aerobically grown, cells can reduce the mineral oxides; (iii) electron shuttles and iron chelators are not needed for this in vitro reduction, documenting conclusively that reduction can occur by direct contact with the mineral oxide; (iv) electron shuttles and EDTA stimulate the in vitro Fe(III) reduction, documenting that exogenous molecules can enhance rates of enzymatic mineral reduction; and (v) multiple membrane components are involved in solid-phase oxide reduction. The membrane fractions, consisting of liposomes of cytoplasmic and outer membrane segments, contain at least 100 proteins including the enzyme that oxidizes formate, formate dehydrogenase. Mineral oxide reduction was inhibited by the addition of detergent Triton X-100, which solubilizes membranes and their associated proteins, consistent with the involvement of multiple electron carriers that are disrupted by detergent addition. In contrast, formate dehydrogenase activity was not inhibited by Triton X-100. The addition of anthraquinone-2,6-disulfonate (AQDS) and menaquinone-4 was unable to restore activity; however, menadione (MD) restored 33% of the activity. The addition of AQDS and MD to reactions without added detergent increased the rate of goethite reduction. The Michaelis-Menten K{sub m} values of 71 {+-} 22 m{sup 2}/L for hematite and 50 {+-} 16 m{sup 2}/L for goethite were calculated as a function of surface area of the two insoluble minerals. V{sub max} was determined to be 123 {+-} 14 and 156 {+-} 13 nmol Fe(II)/min/mg of TM protein for hematite and goethite, respectively. These values are consistent with in vivo rates of reduction reported in the literature. These observations are consistent with our conclusion that the enzymatic reduction of mineral oxides is an effective probe that will allow elucidation of molecular chemistry of the membrane-mineral interface where electron transfer occurs.

Ruebush,S.; Icopini, G.; Brantley, S.; Tien, M.

2006-01-01

39

In vitro enzymatic reduction kinetics of mineral oxides by membrane fractions from Shewanella oneidensis MR-1  

NASA Astrophysics Data System (ADS)

This study documents the first example of in vitro solid-phase mineral oxide reduction by enzyme-containing membrane fractions. Previous in vitro studies have only reported the reduction of aqueous ions. Total membrane (TM) fractions from iron-grown cultures of Shewanella oneidensis MR-1 were isolated and shown to catalyze the reduction of goethite, hematite, birnessite, and ramsdellite/pyrolusite using formate. In contrast, nicotinamide adenine dinucleotide (NADH) and succinate cannot function as electron donors. The significant implications of observations related to this cell-free system are: (i) both iron and manganese mineral oxides are reduced by the TM fraction, but aqueous U(VI) is not; (ii) TM fractions from anaerobically grown, but not aerobically grown, cells can reduce the mineral oxides; (iii) electron shuttles and iron chelators are not needed for this in vitro reduction, documenting conclusively that reduction can occur by direct contact with the mineral oxide; (iv) electron shuttles and EDTA stimulate the in vitro Fe(III) reduction, documenting that exogenous molecules can enhance rates of enzymatic mineral reduction; and (v) multiple membrane components are involved in solid-phase oxide reduction. The membrane fractions, consisting of liposomes of cytoplasmic and outer membrane segments, contain at least 100 proteins including the enzyme that oxidizes formate, formate dehydrogenase. Mineral oxide reduction was inhibited by the addition of detergent Triton X-100, which solubilizes membranes and their associated proteins, consistent with the involvement of multiple electron carriers that are disrupted by detergent addition. In contrast, formate dehydrogenase activity was not inhibited by Triton X-100. The addition of anthraquinone-2,6-disulfonate (AQDS) and menaquinone-4 was unable to restore activity; however, menadione (MD) restored 33% of the activity. The addition of AQDS and MD to reactions without added detergent increased the rate of goethite reduction. The Michaelis-Menten Km values of 71 ± 22 m 2/L for hematite and 50 ± 16 m 2/L for goethite were calculated as a function of surface area of the two insoluble minerals. Vmax was determined to be 123 ± 14 and 156 ± 13 nmol Fe(II)/min/mg of TM protein for hematite and goethite, respectively. These values are consistent with in vivo rates of reduction reported in the literature. These observations are consistent with our conclusion that the enzymatic reduction of mineral oxides is an effective probe that will allow elucidation of molecular chemistry of the membrane-mineral interface where electron transfer occurs.

Ruebush, Shane S.; Icopini, Gary A.; Brantley, Susan L.; Tien, Ming

2006-01-01

40

ISOLATION OF PLASMA MEMBRANE FRAGMENTS FROM HELA CELLS  

PubMed Central

A method for isolating plasma membrane fragments from HeLa cells is described. The procedure starts with the preparation of cell membrane "ghosts," obtained by gentle rupture of hypotonically swollen cells, evacuation of most of the cell contents by repeated washing, and isolation of the ghosts on a discontinuous sucrose density gradient. The ghosts are then treated by minimal sonication (5 sec) at pH 8.6, which causes the ghost membranes to pinch off into small vesicles but leaves any remaining larger intracellular particulates intact and separable by differential centrifugation. The ghost membrane vesicles are then subjected to isopycnic centrifugation on a 20–50% w/w continuous sucrose gradient in tris-magnesium buffer, pH 8.6. A band of morphologically homogeneous smooth vesicles, derived principally from plasma membrane, is recovered at 30–33% (peak density = 1.137). The plasma membrane fraction contained a Na-K-activated ATPase activity of 1.5 µmole Pi/hr per mg, 3% RNA, and 13.8% of the NADH-cytochrome c reductase activity of a heavier fraction from the same gradient which contained mitochondria and rough endoplasmic vesicles. The plasma membranes of viable HeLa cells were marked with 125I-labeled horse antibody and followed through the isolation procedure. The specific antibody binding of the plasma membrane vesicle fraction was increased 49-fold over that of the original whole cells.

Boone, Charles W.; Ford, Lincoln E.; Bond, Howard E.; Stuart, Donald C.; Lorenz, Dianne

1969-01-01

41

Cryptococcus neoformans cryoultramicrotomy and vesicle fractionation reveals an intimate association between membrane lipids and glucuronxylomannan  

PubMed Central

Cryptococcus neoformans is an encapsulated pathogenic fungus. The cryptococcal capsule is composed of polysaccharides and is necessary for virulence. It has been previously reported that glucuronoxylomannan (GXM), the major capsular component, is synthesized in cytoplasmic compartments and transported to the extracellular space in vesicles, but knowledge on the organelles involved in polysaccharide synthesis and traffic is extremely limited. In this paper we report the GXM distribution in C. neoformans cells sectioned by cryoultramicrotomy and visualized by transmission electron microscopy (TEM) and polysaccharide immunogold staining. Cryosections of fungal cells showed high preservation of intracellular organelles and cell wall structure. Incubation of cryosections with an antibody to GXM revealed that cytoplasmic structures associated to vesicular compartments and reticular membranes are in close proximity to the polysaccharide. GXM was generally found in association with the membrane of intracellular compartments and within different layers of the cell wall. Analysis of extracellular fractions from cryptococcal supernatants by transmission electron microscopy in combination with serologic, chromatographic and spectroscopic methods revealed fractions containing GXM and lipids. These results indicate an intimate association of GXM and lipids in both intracellular and extracellular spaces consistent with polysaccharide synthesis and transport in membrane-associated structures.

Oliveira, Debora L.; Nimrichter, Leonardo; Miranda, Kildare; Frases, Susana; Faull, Kym F.; Casadevall, Arturo; Rodrigues, Marcio L.

2009-01-01

42

Stem Cells from Fetal Membranes  

Microsoft Academic Search

Stem cells that can be derived from fetal membranes represent an exciting field of research that bears tremendous potential for developmental biology and regenerative medicine. In this report we summarize contributions to a workshop in which newest insights into the characteristics, subtypes and molecular determinants of stem cells from trophoblast and endometrial tissues were presented.

M. Hemberger; W. Yang; D. Natale; Thomas L. Brown; C. Dunk; C. E. Gargett; S. Tanaka

2008-01-01

43

Pervaporation membranes in direct methanol fuel cells  

Microsoft Academic Search

The membranes in direct methanol fuel cells must both conduct protons and serve as a barrier for methanol. Nafion, the most common fuel cell membrane, is an excellent conductor but a poor barrier. Polyvinyl alcohol pervaporation membranes are good methanol barriers but poor conductors. These and most other pervaporation membranes offer no significant advantages over Nafion in methanol fuel cell

Bryan S. Pivovar; Yuxin Wang; E. L. Cussler

1999-01-01

44

A Major Fraction of Glycosphingolipids in Model and Cellular Cholesterol-containing Membranes Is Undetectable by Their Binding Proteins*  

PubMed Central

Glycosphingolipids (GSLs) accumulate in cholesterol-enriched cell membrane domains and provide receptors for protein ligands. Lipid-based “aglycone” interactions can influence GSL carbohydrate epitope presentation. To evaluate this relationship, Verotoxin binding its receptor GSL, globotriaosyl ceramide (Gb3), was analyzed in simple GSL/cholesterol, detergent-resistant membrane vesicles by equilibrium density gradient centrifugation. Vesicles separated into two Gb3/cholesterol-containing populations. The lighter, minor fraction (<5% total GSL), bound VT1, VT2, IgG/IgM mAb anti-Gb3, HIVgp120 or Bandeiraea simplicifolia lectin. Only IgM anti-Gb3, more tolerant of carbohydrate modification, bound both vesicle fractions. Post-embedding cryo-immuno-EM confirmed these results. This appears to be a general GSL-cholesterol property, because similar receptor-inactive vesicles were separated for other GSL-protein ligand systems; cholera toxin (CTx)-GM1, HIVgp120-galactosyl ceramide/sulfatide. Inclusion of galactosyl or glucosyl ceramide (GalCer and GlcCer) rendered VT1-unreactive Gb3/cholesterol vesicles, VT1-reactive. We found GalCer and GlcCer bind Gb3, suggesting GSL-GSL interaction can counter cholesterol masking of Gb3. The similar separation of Vero cell membrane-derived vesicles into minor “binding,” and major “non-binding” fractions when probed with VT1, CTx, or anti-SSEA4 (a human GSL stem cell marker), demonstrates potential physiological relevance. Cell membrane GSL masking was cholesterol- and actin-dependent. Cholesterol depletion of Vero and HeLa cells enabled differential VT1B subunit labeling of “available” and “cholesterol-masked” plasma membrane Gb3 pools by fluorescence microscopy. Thus, the model GSL/cholesterol vesicle studies predicted two distinct membrane GSL formats, which were demonstrated within the plasma membrane of cultured cells. Cholesterol masking of most cell membrane GSLs may impinge many GSL receptor functions.

Mahfoud, Radhia; Manis, Adam; Binnington, Beth; Ackerley, Cameron; Lingwood, Clifford A.

2010-01-01

45

The effect of passive mixing on pressure drop and oxygen mass fraction using opposing channel flow field design in a Proton Exchange Membrane Fuel Cell  

NASA Astrophysics Data System (ADS)

This study investigates a flow field with opposing channel design. Previous studies on flow field designs have been focused on improving fuel utilization which often leads to increased pressure drop. This increased pressure drop is typical because standard designs employ either a single flow channel to clear blockages or dead end condition to force the flow through the gas diffusion layer. The disadvantage with these designs is the increased resistance to the flow which requires higher pressure, which becomes a parasitic loss that lowers the system efficiency. For this study the focus was to reduce the pressure drop by providing a less resistive path to the flow. To achieve a less resistive path, the inlet channel was split into two opposing channels. These channels are then recombined only to be split again for the next leg. Therefore, the split channel design should reduce the pressure drop which reduces the parasitic load and ultimately contributes to higher system efficiency. In addition the recombining of the streams at each leg should induce mixing. Having opposing channels should also increase cross flow under the lands to reduce mass transfer loses. The cathode side of the fuel cell is especially sensitive to the mass transport losses since air (oxygen mixed with nitrogen) is used for supplying oxygen unlike the anode side which uses pure hydrogen. To test the hypothesis of having benefits from an opposing channel design, both an experimental and analytical approach was taken. For the experiment, a serpentine flow field and opposing channel flow field plates were compared over several flow rates with compressed air. To test the hypothesis of increased mass transfer, the two flow fields were modeled using a CFD software package, COMSOL. It was found that the opposing channel configuration for high flow rate with multiple entry and exit conditions exhibited significant improvement over the single serpentine channel. Pressure drop was ? less than the serpentine channel with similar conditions. Simulations for mass transfer show that recombining of the flow streams generate more uniform current density unlike the serpentine configuration where the current density was concentrated at the entrance of the flow stream. The background section provides a brief overview of the governing equations, the theory of flow field operation and previous bodies of work on flow field design. Recommendations are made for further verification of the design using a real working cell based on the results.

Singh, Anant Bir

46

Enzyme distributions in subcellular fractions of BHK cells infected with Semliki Forest virus: evidence for a major fraction of sphingomyelin synthase in the trans-Golgi network  

Microsoft Academic Search

BHK cells either untreated or infected with Semliki Forest virus have been fractionated on sucrose density gradients. Virus infection caused an increase in density of a membrane fraction enriched in sphingomyelin (SM), cholesterol, SM synthase and sialyltransferase activity. This increase in density was related to incorporation of viral proteins into this fraction, which is likely to contain trans-Golgi network (TGN)

David Allan; Maria Jesus Miro Obradors

1999-01-01

47

Cytosolic factors in bovine neutrophil oxidase activation. Partial purification and demonstration of translocation to a membrane fraction  

Microsoft Academic Search

The Oâ{sup {center dot}-}-generating oxidase of bovine neutrophils is activated in a cell-free system consisting of a particulate fraction enriched in plasma membrane and containing the dormant oxidase, a high-speed supernatant from neutrophil homogenate (cytosol), Mg ions, GTPγS, and arachidonic acid. The cytosolic components participating in the activation of the membrane-bound oxidase have been investigated. These components were resolved into

Jacques Doussiere; Marie Claire Pilloud; Pierre V. Vignais

1990-01-01

48

Electron transport systems of Nitrosomonas: isolation of a membrane-envelope fraction.  

PubMed

Freezing and thawing of Nitrosomonas, followed by centrifugation of the homogenate at 3,000 x g, resulted in a fraction which appeared to consist of an intact membrane-envelope complex and contained approximately 50% of the cell protein and more than 90% of the ubiquinone and cytochrome A-type mammalian cytochrome c oxidase activity. The supernatant fraction, resulting from subsequent centrifugation of the extract at 100,000 x g, contained approximately 50% of the cell protein and more than 80% of the B- and C-type cytochrome and P-463 and the enzymes glutamate dehydrogenase; hydroxylamine dehydrogenase; nitrite synthetase; nitrite reductase; and 2,6-dichlorophenolindophenol-, p-phenylenediamine-, pyrogallol-, and hydroquinone-oxidase. Data on the concentration of electron transport components in Nitrosomonas are presented. PMID:4336111

Hooper, A B; Erickson, R H; Terry, K R

1972-04-01

49

Wrinkled Membrane Morphology of Biological Cell  

NASA Astrophysics Data System (ADS)

Membranes of many biological cells possess a wrinkled surface topology that, in some instance, serves as a reservoir for providing large surface area and membrane expansion during osmotic swelling. We consider and model the development of the wrinkled morphology to result from buckling instabilities which occur during the membrane growth. In particular, we examine the wrinkled membrane morphology of white blood cell experimentally and numerically. Our results show that the deformation mismatch between the membrane and the cytoskeleton during membrane growth triggers buckling of the membrane. This behavior of the wrinkled topology enables expansion of the cell during swelling and reveals interesting details on the role of the membrane topology.

Wang, Lifeng; Castro, Carlos; Boyce, Mary

2010-03-01

50

Corrugated Membrane Fuel Cell Structures  

SciTech Connect

One of the most challenging aspects of traditional PEM fuel cell stacks is the difficulty achieving the platinum catalyst utilization target of 0.2 gPt/kWe set forth by the DOE. Good catalyst utilization can be achieved with state-of-the-art catalyst coated membranes (CCM) when low catalyst loadings (<0.3 mg/cm2) are used at a low current. However, when low platinum loadings are used, the peak power density is lower than conventional loadings, requiring a larger total active area and a larger bipolar plate. This results in a lower overall stack power density not meeting the DOE target. By corrugating the fuel cell membrane electrode structure, Ion Power?s goal is to realize both the Pt utilization targets as well as the power density targets of the DOE. This will be achieved by demonstrating a fuel cell single cell (50 cm2) with a twofold increase in the membrane active area over the geometric area of the cell by corrugating the MEA structure. The corrugating structure must be able to demonstrate the target properties of < 10 mOhm-cm2 electrical resistance at > 20 psi compressive strength over the active area, in combination with offering at least 80% of power density that can be achieved by using the same MEA in a flat plate structure. Corrugated membrane fuel cell structures also have the potential to meet DOE power density targets by essentially packaging more membrane area into the same fuel cell volume as compared to conventional stack constructions.

Grot, Stephen [President, Ion Power Inc.] President, Ion Power Inc.

2013-09-30

51

High-speed pectic enzyme fractionation by immobilised metal ion affinity membranes.  

PubMed

Immobilised metal ion affinity polysulfone hollow-fibre membranes, with a high capacity for protein adsorption, were prepared and their utilisation for commercial pectic enzyme fractionation was studied. The pass-through fraction containing pectinlyase is useful for fruit-juice clarification without methanol production on account of pectinesterase being retained by the IDA-Cu2+ membrane. PMID:11105247

Camperi, S A; Grasselli, M; Cascone, O

2000-01-01

52

Focus on membrane differentiation and membrane domains in the prokaryotic cell.  

PubMed

A summary is presented of membrane differentiation in the prokaryotic cell, with an emphasis on the organization of proteins in the plasma/cell membrane. Many species belonging to the Eubacteria and Archaea have special membrane domains and/or membrane proliferation, which are vital for different cellular processes. Typical membrane domains are found in bacteria where a specific membrane protein is abundantly expressed. Lipid rafts form another example. Despite the rareness of conventional organelles as found in eukaryotes, some bacteria are known to have an intricate internal cell membrane organization. Membrane proliferation can be divided into curvature and invaginations which can lead to internal compartmentalization. This study discusses some of the clearest examples of bacteria with such domains and internal membranes. The need for membrane specialization is highest among the heterogeneous group of bacteria which harvest light energy, such as photosynthetic bacteria and halophilic archaea. Most of the highly specialized membranes and domains, such as the purple membrane, chromatophore and chlorosome, are found in these autotrophic organisms. Otherwise the need for membrane differentiation is lower and variable, except for those structures involved in cell division. Microscopy techniques have given essential insight into bacterial membrane morphology. As microscopy will further contribute to the unraveling of membrane organization in the years to come, past and present technology in electron microscopy and light microscopy is discussed. Electron microscopy was the first to unravel bacterial morphology because it can directly visualize membranes with inserted proteins, which no other technique can do. Electron microscopy techniques developed in the 1950s and perfected in the following decades involve the thin sectioning and freeze fractioning of cells. Several studies from the golden age of these techniques show amazing examples of cell membrane morphology. More recently, light microscopy in combination with the use of fluorescent dyes has become an attractive technique for protein localization with the natural membrane. However, the resolution problem in light microscopy remains and overinterpretation of observed phenomena is a pitfall. Thus, light microscopy as a stand-alone technique is not sufficient to prove, for instance, the long-range helical distribution of proteins in membrane such as MinD spirals in Bacillus subtilis. Electron tomography is an emerging electron microscopy technique that can provide three-dimensional reconstructions of small, nonchemically fixed bacteria. It will become a useful tool for studying prokaryotic membranes in more detail and is expected to collect information complementary to those of advanced light microscopy. Together, microscopy techniques can meet the challenge of the coming years: to specify membrane structures in more detail and to bring them to the level of specific protein-protein interactions. PMID:23920497

Boekema, Egbert J; Scheffers, Dirk-Jan; van Bezouwen, Laura S; Bolhuis, Henk; Folea, I Mihaela

2013-01-01

53

Preparation and characterization of plasma membrane-enriched fractions from rat pancreatic islets  

PubMed Central

Methods have been developed for the isolation on a semi-micro scale of a plasma membrane-enriched fraction from rat islets of Langerhans. An important feature of these experiments is the use of 125I-labeled wheat germ agglutinin as a specific probe for plasma membrane-containing fractions. The partly purified plasma membrane fraction had a density in sucrose of about 1.10 and was enriched in the activities of 5'- nucleotidase, alkaline phosphatase, sodium-potassium, and magnesium- dependent ATPase and adenylate cyclase. It contained only very low levels of acid phosphatase, cytochrome c oxidase, insulin, and RNA. Further purification was hampered by the relatively small amounts of fresh plasma membrane material that could be obtained from 16-24 rats in each experiment. When islets were prelabeled with radioactive fucose, the plasma membrane-enriched fraction contained radioactivity at a four- to fivefold higher specific acivity than the whole islet homogenate. Sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis of plasma membrane-enriched fractions pooled from several experiments revealed a distinctive pattern of protein bands as compared with other less pure fractions. With respect to rapidity, apparent specificity, and easy reversibility of the labeling of the plasma membrane fraction, 125I-wheat germ agglutinin provides a highly useful tool for the detection of microgram quantities of plasma membrane components which should be applicable to many other systems as well.

1976-01-01

54

Artificial Red Cells with Polyhemoglobin Membranes.  

National Technical Information Service (NTIS)

Artificial red cells were prepared with polyhemoglobin membranes. Red-cell-size microdroplets containing 30% of hemoglobin were held in liquid membrane capsules and treated with glutaraldehyde that cross linked the hemoglobin at the surface of each microd...

T. A. Davis W. J. Asher G. T. Quinlan

1981-01-01

55

HPLC fractions of human uremic plasma inhibit the RBC membrane calcium pump.  

PubMed

We have reported that uremic plasma filtrates (UF) inhibit the red blood cell (RBC) membrane calcium pump. The inhibitor was dialyzable, smaller than 3,000 molecular weight, heat-stable, and protease-resistant. In the present study, we used reverse-phase preparative HPLC, analytical HPLC, and Sephadex G-25 elution to identify inhibitory fractions. Inhibition was confirmed in three different bioassays: (1) Sr2+ efflux in intact RBC, the primary bio-assay; (2) 45Ca efflux in intact RBC; and (3) calcium ATPase activity in isolated RBC membranes. Active fractions were analyzed by mass spectrometry, capillary electrophoresis, enzymatic analysis, gas chromatography-mass spectrometry, and nuclear magnetic resonance spectroscopy. These demonstrated a number of compounds, including: sugars, polyols, osmolytes like betaine and myoinositol, amino acids, and other metabolites, such as 3-D-hydroxybutyrate, dimethylglycine, trimethylamine-N-oxide, guanidinoacetic acid and glycine. Many individual compounds were then tested for an effect on the calcium pump. Thus, HPLC was able to separate a substantial number of compounds in inhibitory fractions. Efforts are under way for precise identification of the inhibitor, to advance our understanding of uremic toxicity and/or hypertension in CRF. PMID:9083269

Lindner, A; Vanholder, R; De Smet, R; Hinds, T R; Vogeleere, P; Sandra, P; Foxall, P; Ringoir, S

1997-04-01

56

Performances of ultrafiltration membranes for fractionating a fish protein hydrolysate: Application to the refining of bioactive peptidic fractions  

Microsoft Academic Search

Ultrafiltration membranes can be advantageously used to improve the bioactivity of a saithe protein hydrolysate containing peptides having a size lower than 7kDa, by fractionating or concentrating some specific molecular weight peptide classes. The aim of this study is to evaluate the behaviour of a 4kDa membrane in modified polyethersulfone in conditions close to industrial exploitation. This work consists in

A. Chabeaud; L. Vandanjon; P. Bourseau; P. Jaouen; M. Chaplain-Derouiniot; F. Guerard

2009-01-01

57

The distribution of ribosomal ribonucleic acids among subcellular fractions from bacteria and the adverse effect of the membrane fraction on the stability of ribosomes  

PubMed Central

1. The distributions of nucleic acids and protein among fractions obtained by differential centrifugation from species of Pseudomonas, Aerobacter, Escherichia, Proteus and Bacillus have been studied. 2. The DNA in a cell wall–membrane fraction obtained by low-speed centrifugation from the Gram-negative species could be removed by homogenizing and subsequent washing. About 7–14% of the total RNA remained firmly attached and resembled ribosomal RNA in base composition. A similar fraction from the Gram-positive B. subtilis contained about one-half of the total bacterial DNA and only 60% of this could be removed by homogenizing and subsequent washing. 3. A deposit obtained by high-speed centrifugation could be separated into a heavy ribosome layer and a light turbid layer. In E. coli B the latter contained about equal concentrations of RNA and DNA and accounted for about one-half of the total bacterial NADP-activated 6-phosphogluconate dehydrogenase. 4. The washed cell wall–membrane fraction from most species accelerated the degradation of ribosomes. In Pr. vulgaris the activity of this fraction was exceptionally high and resulted in the progressive degradation of ribosomes during their isolation from this species. 5. A possible connexion between ribosome degradation and the synthesis of flagella is discussed in the light of these results.

Wade, H. E.; Robinson, H. K.

1965-01-01

58

Effects of Normal and Activated Cell Fractions on the Growth of Tubercle Bacilli  

PubMed Central

Fractions prepared from normal and activated liver cells were tested for the antimycobacterial activity by the agar plate diffusion test. Results showed that lysosome extracts of normal and activated cells exerted no antibacterial activity, cell extracts and lysosome membranes exerted some activity, and cell membranes exerted the strongest activity. The active materials of activated cells exerted stronger antituberculous activity than the corresponding materials of normal cells. Degrees of the antimycobacterial activity of various cell fractions showed a close correlation with the amounts of nonesterified fatty acids. This correlation, as well as other data, suggested that the antimycobacterial activity of cell fractions was caused by toxic fatty acids which were produced during the hydrolysis of lipoporteins or phospholipids by the activity of tissue lipases. The relationship of these findings to the mechanism of cellular immunity is discussed. Images

Kochan, Ivan; Pellis, Neal R.; Pfohl, Dawn G.

1972-01-01

59

Isolation of a hemidesmosome-rich fraction from a human squamous cell carcinoma cell line.  

PubMed

Hemidesmosomes are cell-to-matrix adhesion complexes anchoring keratinocytes to basement membranes. For the first time, we present a method to prepare a fraction from human cultured cells that are highly enriched in hemidesmosomal proteins. Using DJM-1 cells derived from human squamous cell carcinoma, accumulation of hemidesmosomes was observed when these cells were cultured for more than 10 days in a commercial serum-free medium without supplemental calcium. Electron microscopy demonstrated that numerous electron-dense adhesion structures were present along the basal cell membranes of DJM-1 cells cultured under the aforementioned conditions. After removing cellular materials using an ammonia solution, hemidesmosomal proteins and deposited extracellular matrix were collected and separated by electrophoresis. There were eight major polypeptides, which were determined to be plectin, BP230, BP180, integrin ?6 and ?4 subunits, and laminin-332 by immunoblotting and mass spectrometry. Therefore, we designated this preparation as a hemidesmosome-rich fraction. This fraction contained laminin-332 exclusively in its unprocessed form, which may account for the promotion of laminin deposition, and minimal amounts of Lutheran blood group protein, a nonhemidesmosomal transmembrane protein. This hemidesmosome-rich fraction would be useful not only for biological research on hemidesmosomes but also for developing a serum test for patients with blistering skin diseases. PMID:24726610

Hirako, Yoshiaki; Yonemoto, Yuki; Yamauchi, Tomoe; Nishizawa, Yuji; Kawamoto, Yoshiyuki; Owaribe, Katsushi

2014-06-10

60

Fractional Cell Lethality Approach to Space Radiation Hazards.  

National Technical Information Service (NTIS)

A method of radiation hazard evaluation has been introduced in which the fractional number of inactivated cells of an organ is calculated. This fractional cell lethality (FCL) depends only on the particle energy spectrum and the probability of cell inacti...

S. B. Curtis D. L. Dye W. R. Sheldon

1964-01-01

61

Chemical composition of a purified membrane fraction from Sarcina aurantiaca in relation to growth phase  

Microsoft Academic Search

The effect of the age of a culture of Sarcina aurantiaca on the chemical composition of the total membrane fraction has been investigated. Whereas the protein content is constant and the carbohydrate increases with age, the lipid content decreases which may be a reflection of increased binding of the lipid to protein. The carbohydrates present in the membrane are mannose,

D. Thirkell; Elizabeth M. M. Gray

1974-01-01

62

Membrane heterogeneity in murine stem cells  

SciTech Connect

The cell membrane's role in hemopoietic differentiation and regulation is increasingly evident. Certain fluorescent molecules interact with membrane components at the molecular level. The organic dye merocyanine 540(MCN) is such a tool for studying hemopoietic stem cell membranes at the molecular level. This paper present work involving biologic interactions of MCN with CLL (chronic lymphocytic leukemia) and hemopoietic cell membranes. The authors work indicates that MNC staining reveals membrane differences that accompany cellular transformation and differentiation. Changes in MNC fluorescene indicate that hemopoietic cells are heterogeneous with respect to membrane properties, and sensitivity to MNC photoinactivation reveals that membrane properties may change as a function of differentiative state between pluripotent and committed granulocytic precursor cells.

Myers, C.P.; Kerk, D.K.; Norman, A.

1980-01-01

63

Co-localization of superoxide generation and NADP formation in plasma membrane fractions from human neutrophils  

Microsoft Academic Search

In order to resolve discrepancies in the literature concerning the subcellular localization of NADPH oxidase, we disrupted human neutrophils by nitrogen cavitation and fractionated the subcellular organelles on a discontinuous sucrose density gradient. The lightest fraction was 20- to 40-fold enriched for piasma membranes as determined by the marker enzymes alkaline phosphatase and phosphodiesterase I as well as by the

Pamela S. Shirley; David A. Bass; Cynthia J. Lees; J. Wallace Parce; B. Moseley Waite; Lawrence R. Dechatelet

1984-01-01

64

Membrane distribution of sodium-hydrogen and chloride-bicarbonate exchangers in crypt and villus cell membranes from rabbit ileum.  

PubMed Central

Present evidence suggests that in the small intestine, villus cells are primarily absorptive and crypt cells are primarily secretory. In order to further confirm that there are differences in transport properties between villus and crypt cells, we have separated villus from crypt cells, using calcium chelations techniques, and determined the distribution of Na:H and Cl:HCO3 exchange activity on brush border membrane and basolateral membrane preparations from these two cell populations. Separation of cells was determined utilizing alkaline phosphatase and maltase activity as a marker of villus cells and thymidine kinase activity as a marker of crypt cells. Utilizing these techniques, we were able to sequentially collect cells along the villus-crypt axis. Na-stimulated glucose and alanine uptake in brush border membrane vesicles diminished from the villus to the crypt region in the sequentially collected cells fractions, further suggesting separation of these cells. Brush border and basolateral membranes were then prepared from cells from the villus and crypt areas, utilizing a continuous sucrose gradient. In the villus cells, Na:H exchange activity was found associated with both the brush border and basolateral membrane, whereas, in crypt cells, Na:H exchange activity was only found on the basolateral membrane. Cl:HCO3 exchange activity was found only on the brush border membrane, in both villus and crypt cells. These studies suggest functional heterogeneity in ion transport between villus and crypt cells.

Knickelbein, R G; Aronson, P S; Dobbins, J W

1988-01-01

65

Membrane lipids and cell death: an overview  

Microsoft Academic Search

In this article we overview major aspects of membrane lipids in the complex area of cell death, comprising apoptosis and various forms of programmed cell death. We have focused here on glycerophospholipids, the major components of cellular membranes. In particular, we present a detailed appraisal of mitochondrial lipids that attract increasing interest in the field of cell death, while the

Ileana M. Cristea; Mauro Degli Esposti

2004-01-01

66

Cell membrane orientation visualized by polarized total internal reflection fluorescence.  

PubMed Central

In living cells, variations in membrane orientation occur both in easily imaged large-scale morphological features, and also in less visualizable submicroscopic regions of activity such as endocytosis, exocytosis, and cell surface ruffling. A fluorescence microscopic method is introduced here to visualize such regions. The method is based on fluorescence of an oriented membrane probe excited by a polarized evanescent field created by total internal reflection (TIR) illumination. The fluorescent carbocyanine dye diI-C(18)-(3) (diI) has previously been shown to embed in the lipid bilayer of cell membranes with its transition dipoles oriented nearly in the plane of the membrane. The membrane-embedded diI near the cell-substrate interface can be fluorescently excited by evanescent field light polarized either perpendicular or parallel to the plane of the substrate coverslip. The excitation efficiency from each polarization depends on the membrane orientation, and thus the ratio of the observed fluorescence excited by these two polarizations vividly shows regions of microscopic and submicroscopic curvature of the membrane, and also gives information regarding the fraction of unoriented diI in the membrane. Both a theoretical background and experimental verification of the technique is presented for samples of 1) oriented diI in model lipid bilayer membranes, erythrocytes, and macrophages; and 2) randomly oriented fluorophores in rhodamine-labeled serum albumin adsorbed to glass, in rhodamine dextran solution, and in rhodamine dextran-loaded macrophages. Sequential digital images of the polarized TIR fluorescence ratios show spatially-resolved time-course maps of membrane orientations on diI-labeled macrophages from which low visibility membrane structures can be identified and quantified. To sharpen and contrast-enhance the TIR images, we deconvoluted them with an experimentally measured point spread function. Image deconvolution is especially effective and fast in our application because fluorescence in TIR emanates from a single focal plane.

Sund, S E; Swanson, J A; Axelrod, D

1999-01-01

67

Polymer electrolyte membrane assembly for fuel cells  

NASA Technical Reports Server (NTRS)

An electrolyte membrane for use in a fuel cell can contain sulfonated polyphenylether sulfones. The membrane can contain a first sulfonated polyphenylether sulfone and a second sulfonated polyphenylether sulfone, wherein the first sulfonated polyphenylether and the second sulfonated polyphenylether sulfone have equivalent weights greater than about 560, and the first sulfonated polyphenylether and the second sulfonated polyphenylether sulfone also have different equivalent weights. Also, a membrane for use in a fuel cell can contain a sulfonated polyphenylether sulfone and an unsulfonated polyphenylether sulfone. Methods for manufacturing a membrane electrode assemblies for use in fuel cells can include roughening a membrane surface. Electrodes and methods for fabricating such electrodes for use in a chemical fuel cell can include sintering an electrode. Such membranes and electrodes can be assembled into chemical fuel cells.

Yen, Shiao-Ping S. (Inventor); Kindler, Andrew (Inventor); Yavrouian, Andre (Inventor); Halpert, Gerald (Inventor)

2000-01-01

68

Polymer electrolyte membrane assembly for fuel cells  

NASA Technical Reports Server (NTRS)

An electrolyte membrane for use in a fuel cell can contain sulfonated polyphenylether sulfones. The membrane can contain a first sulfonated polyphenylether sulfone and a second sulfonated polyphenylether sulfone, wherein the first sulfonated polyphenylether and the second sulfonated polyphenylether sulfone have equivalent weights greater than about 560, and the first sulfonated polyphenylether and the second sulfonated polyphenylether sulfone also have different equivalent weights. Also, a membrane for use in a fuel cell can contain a sulfonated polyphenylether sulfone and an unsulfonated polyphenylether sulfone. Methods for manufacturing a membrane electrode assemblies for use in fuel cells can include roughening a membrane surface. Electrodes and methods for fabricating such electrodes for use in a chemical fuel cell can include sintering an electrode. Such membranes and electrodes can be assembled into chemical fuel cells.

Yen, Shiao-Ping S. (Inventor); Kindler, Andrew (Inventor); Yavrouian, Andre (Inventor); Halpert, Gerald (Inventor)

2002-01-01

69

The isolation and fractionation of malaria-infected cells*  

PubMed Central

This paper is a critical review of procedures for the isolation of malarial parasites from host cells and their fractionation. The procedures are grouped according to the stage of parasite being isolated, and the procedures for isolation of the erythrocytic stages are further grouped by techniques used. Some types of procedure are described for isolation of all stages of the parasite, both those in the invertebrate and vertebrate hosts. The uses and limitations of the various procedures are described. It is concluded that all the procedures are useful for some purposes, but that from a morphological standpoint only natural release in culture and continuous flow oscillation provide large yields of intact erythrocytic parasites free of host cell membranes.

Kreier, Julius P.

1977-01-01

70

Understanding the ?-crystallin cell membrane conjunction  

PubMed Central

Purpose It is well established that levels of soluble ?-crystallin in the lens cytoplasm fall steadily with age, accompanied by a corresponding increase in the amount of membrane-bound ?-crystallin. Less well understood, is the mechanism driving this age-dependent membrane association. The aim of this study was to investigate the role of the membrane and its associated proteins and peptides in the binding of ?-crystallin. Methods Fiber cell membranes from human and bovine lenses were separated from soluble proteins by centrifugation. Membranes were stripped of associated proteins with successive aqueous, urea, and alkaline solutions. Protein constituents of the respective membrane isolates were examined by SDS–PAGE and western immunoblotting. Recombinant ?A- and ?B-crystallins were fluorescently-labeled with Alexa350® dye and incubated with the membrane isolates and the binding capacity of membrane for ?-crystallin was determined. Results The binding capacity of human membranes was consistently higher than that of bovine membranes. Urea- and alkali-treated membranes from the nucleus had similar binding capacities for ?A-crystallin, which were significantly higher than both cortical membrane extracts. ?B-Crystallin also had a higher affinity for nuclear membrane. However, urea-treated nuclear membrane had three times the binding capacity for ?B-crystallin as compared to the alkali-treated nuclear membrane. Modulation of the membrane-crystallin interaction was achieved by the inclusion of an NH2-terminal peptide of ?B-crystallin in the assays, which significantly increased the binding. Remarkably, following extraction with alkali, full length ?A- and ?B-crystallins were found to remain associated with both bovine and human lens membranes. Conclusions Fiber cell membrane isolated from the lens has an inherent capacity to bind ?-crystallin. For ?B-crystallin, this binding was found to be proportional to the level of extrinsic membrane proteins in cells isolated from the lens nucleus, indicating these proteins may play a role in the recruitment of ?B-crystallin. No such relationship was evident for ?A-crystallin in the nucleus, or for cortical membrane binding. Intrinsic lens peptides, which increase in abundance with age, may also function to modulate the interaction between soluble ?-crystallin and the membrane. In addition, the tight association between ?-crystallin and the lens membrane suggests that the protein may be an intrinsic component of the membrane structure.

Su, Shih-Ping; McArthur, Jason D.; Friedrich, Michael G.; Truscott, Roger J.W.

2011-01-01

71

Lipids as organizers of cell membranes  

PubMed Central

The 105th Boehringer Ingelheim Fonds International Titisee Conference ‘Lipids as Organizers of Cell Membranes' took place in March 2012, in Germany. Kai Simons and Gisou Van der Goot gathered cell biologists and biophysicists to discuss the interplay between lipids and proteins in biological membranes, with an emphasis on how technological advances could help fill the gap in our understanding of the lipid part of the membrane.

Kornmann, Benoit; Roux, Aurelien

2012-01-01

72

Effect of Phytoplankton Cell Geometry on Carbon Isotopic Fractionation  

NASA Astrophysics Data System (ADS)

The carbon isotopic compositions of the marine diatom Porosira glacialis and the marine cyanobacterium Synechococcus sp. were measured over a series of growth rates (?) in a continuous culture system in which the concentration and carbon isotopic composition of CO 2(aq) were determined. These data were compared with previously published isotopic results of growth rate experiments using the marine diatom Phaeodactylum tricornutum and the marine haptophyte Emiliania huxleyi. Systematic relationships were found to exist between ?/[CO 2(aq)] and carbon isotopic fractionation (? P) for each species. Maximum isotopic fractionation (? f) for P. glacialis, E. huxleyi, and P. tricornutum was ˜25‰, suggesting that this value may be typical for maximum fractionation associated with Rubisco and ?-carboxylases for marine eukaryotic algae. By contrast, ? f determined for Synechococcus clone CCMP838 was ˜7‰ lower. The slopes of the lines describing the relationship between ? P and ?/[CO 2(aq)] for eukaryotic algal species were different by a factor of more than 20. This result can be accounted for by differences in the surface area and cellular carbon content of the cells. Comparison of chemostat experimental results with calculated results using a diffusion based model imply that the algae in the experiments were actively transporting inorganic carbon across the cell membrane. Our results suggest that accurate estimates of paleo-[CO 2(aq)] from ? P measured in sediments will require knowledge of growth rate as well as cell surface area and either cell carbon quota or cell volume. Given growth rate estimates, our empirical relationship permits reliable calculations of paleo-[CO 2(aq)] using compound-specific isotopic analyses of C 37 alkadienones (select haptophytes) or fossilized frustules (diatoms).

Popp, Brian N.; Laws, Edward A.; Bidigare, Robert R.; Dore, John E.; Hanson, Kristi L.; Wakeham, Stuart G.

1998-01-01

73

Biofouling characteristics using flow field-flow fractionation: effect of bacteria and membrane properties.  

PubMed

In this study, membrane biofouling caused by bacteria that have different characteristics was evaluated using flow field-flow fractionation (FlFFF). Three different bacteria which differed from size and shape (Staphylococcus epidermidis, Escherichia coli, Flavobacterium lutescens) were investigated with GM ultrafiltration (UF, rough with a low negative surface charge and relatively high hydrophobicity) and NE70 nanofiltration (NF, smooth with a high negative surface charge and relatively low hydrophobicity) membranes. The FlFFF retention time of S. epidermidis, E. coli and F. lutescens was highly influenced by the ionic strength of the solution and the surface polarity of the membranes and bacteria. The NF membrane was found to have a higher potential of biofouling than the UF membrane with the bacteria tested in this study. E. coli was the most significant biofoulant among the bacteria tested on both membrane surfaces based on FlFFF retention times compared to other bacteria. PMID:19735999

Lee, Eunkyung; Shon, H K; Cho, Jaeweon

2010-03-01

74

Functional dynamics of cell surface membrane proteins  

NASA Astrophysics Data System (ADS)

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

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

2014-04-01

75

Differentiation of epithelial cells on microporous membranes  

Microsoft Academic Search

Summary Microporous membranes support the differentiation of a variety of cell types including the Madin-Darby canine kidney (MDCK) cell line and normal human epidermal keratinocytes (NHEK). Coincident with culturing these cells on microporous membranes is a change in the cytoskeletal architecture of the cells to a more in vivo-like morphology and the secretion of a basal lamina-like structure, the latter

Laura M. Patrone; Jeffrey R. Cook; Barbara E. Crute; Robert G. Van Buskirk

1992-01-01

76

Fractions!  

NSDL National Science Digital Library

Practice all of the activities to help you learn fractions! Go through all five levels of Fractions Review Activities Practice Naming Fractions Do you remember how to do Fraction Sets? Play these games when you have finished the top three activities: Cross the River Pizza Party Fractions Rescue Island Adding Subtracting Fractions SPLAT Mrs. Anderson's Fraction Games Action Fraction Soccer Shootout Fraction Multiplication Soccer Shootout Fraction Division Dirt Bike Fractions Comparisons ...

Lerdahl, Miss

2011-02-01

77

Imaging of membrane systems and membrane traffic in living cells.  

PubMed

Eukaryotic cells are composed of an intricate system of internal membranes that are organized into different compartments--including the endoplasmic reticulum (ER), the nuclear envelope, the Golgi complex (GC), lysosomes, endosomes, caveolae, mitochondria, and peroxisomes--that perform specialized tasks within the cell. The localization and dynamics of intracellular compartments are now being studied in living cells because of the availability of green fluorescent protein (GFP)-fusion proteins and recent advances in fluorescent microscope imaging systems. Results using these techniques are revealing how intracellular compartments maintain their steady-state organization and distributions, how they undergo growth and division, and how they transfer protein and lipid components between themselves through the formation and trafficking of membrane transport intermediates. This article describes methods using GFP-fusion proteins to visualize the behavior of organelles and to track membrane-bound transport intermediates moving between them. Practical issues related to the construction and expression of GFP-fusion proteins are discussed first. These are essential for optimizing the brightness and expression levels of GFP-fusion proteins so that intracellular membrane-bound structures containing these fusion proteins can be readily visualized. Next, techniques for performing time-lapse imaging using a confocal laser-scanning microscope (CLSM) are detailed, including the use of photobleaching to highlight organelles and transport intermediates. Methods for the acquisition and analysis of data are then discussed. Finally, commonly used and exciting new approaches for perturbing membrane traffic are outlined. PMID:22046036

Snapp, Erik Lee; Lajoie, Patrick

2011-11-01

78

Induction of Resistance by Listeria monocytogenes Cell Wall Fraction  

PubMed Central

A crude cell wall fraction of Listeria monocytogenes was prepared by sonic disruption and differential centrifugation of viable, washed cultures. When injected into mice, this sterile, crude cell wall fraction protected mice against an intraperitoneal challenge with 18 to 85 50% mean lethal dose of L. monocytogenes. Resistance was greatly enhanced when bacterial endotoxin (lipopolysaccharide) was injected along with the cell wall fraction. Resistance was measured both by enumerating the bacteria in the livers and spleens of vaccinated and control mice and by survival studies. Two major lines of evidence suggest that the resistance induced by cell wall fraction is at least in part specific. Unlike non-specific resistance, the cell wall fraction-induced resistance was relatively long-lived, (i.e., it was demonstrable 6 weeks after the last injection of cell wall fraction and lipopolysaccharide). In addition, cell wall fraction protected against challenge with L. monocytogenes, but not against challenge with S. typhimurium. Images

Rodriguez, Gilberto E.; McClatchy, J. Kenneth; Campbell, Priscilla A.

1974-01-01

79

Red cell membrane and cation deficiency in Rh null syndrome.  

PubMed

A 52-yr-old multiparous white female was found to have Rh null blood type. She had macrocytic anemia, with reticulocytosis (15%-20%), of long duration. Although stomatocytes in peripheral blood were numerous and osmotic fragility was increased, suggesting increased cell water, the RBC cation content, and thus cell water, was decreased. Cell dehydration was confirmed by an increased proportion of high density RBC on Stractan density gradients. The deformability of RBC from four gradient subpopulations was measured in the ektacytometer as a function of suspending medium osmolality. Analysis of these measurements showed an abnormal reduction in cell surface area with increasing cell density, thus explaining the increased osmotic fragility of whole blood. This was confirmed by a density-dependent reduction in cell cholesterol content, suggesting membrane instability in vivo. Rh null subpopulations showed a twofold increase in both ouabain-sensitive and -insensitive Na-K ATPase activity and 86Rb transport, even in the dense fraction with the fewest reticulocytes. No membrane protein or glycoprotein abnormality was detected by SDS-PAGE. The associated deficiencies of both membrane surface area and cation content in Rh null cells, as well as increased Na-K pump activity, suggest a pleiotropic functional interrelationship among Rh antigen, membrane stability, and cation regulation. PMID:6324926

Ballas, S K; Clark, M R; Mohandas, N; Colfer, H F; Caswell, M S; Bergren, M O; Perkins, H A; Shohet, S B

1984-05-01

80

Cowpea mosaic virus RNA replication in crude membrane fractions from infected cowpea and Chenopodium amaranticolor  

Microsoft Academic Search

SUMMARY The replication of cowpea mosaic virus (CPMV) RNA was studied in crude membrane fractions prepared from leaves of CPMV-infected cowpea and Chenopo- dium amaranticolor. In vitro replicase assays showed that in the cowpea extract only the replicative intermediate (RI) and replicative form (RF) were synthesized. In the C. amaranticolor extract however, single-stranded progeny RNA was produced in addition to

RIK EGGEN; ANITA KAAN; ROB GOLDBACH; AB VAN KAMMEN

1988-01-01

81

Fibronectin-degrading proteases from the membranes of transformed cells.  

PubMed

The local degradation of fibronectin substrata by Rous sarcoma virus-transformed chick embryonic fibroblasts requires cell-contact-related metalloendoprotease and serine-protease activities. Using fibronectin-containing SDS gels, two large proteases with apparent molecular weights of 120K and 150K were found only in the membrane fraction of transformed cells and were absent in normal cells. Both 120K and 150K proteases were active at neutral pH, but showed preferential inhibitor sensitivities of serine and metal proteases, respectively. The 150K protease appeared to account for most of the proteolytic activity since metalloendoprotease inhibitors completely blocked proteolytic activity of the 150K in fibronectin gels, more than 80% of the fibronectin-degrading activity of solubilized membranes, and largely suppressed the appearance of fibronectin degradation spots in cultures of transformed cells. PMID:3026634

Chen, J M; Chen, W T

1987-01-30

82

Membrane elastic properties and cell function.  

PubMed

Recent studies indicate that the cell membrane, interacting with its attached cytoskeleton, is an important regulator of cell function, exerting and responding to forces. We investigate this relationship by looking for connections between cell membrane elastic properties, especially surface tension and bending modulus, and cell function. Those properties are measured by pulling tethers from the cell membrane with optical tweezers. Their values are determined for all major cell types of the central nervous system, as well as for macrophage. Astrocytes and glioblastoma cells, which are considerably more dynamic than neurons, have substantially larger surface tensions. Resting microglia, which continually scan their environment through motility and protrusions, have the highest elastic constants, with values similar to those for resting macrophage. For both microglia and macrophage, we find a sharp softening of bending modulus between their resting and activated forms, which is very advantageous for their acquisition of phagocytic functions upon activation. We also determine the elastic constants of pure cell membrane, with no attached cytoskeleton. For all cell types, the presence of F-actin within tethers, contrary to conventional wisdom, is confirmed. Our findings suggest the existence of a close connection between membrane elastic constants and cell function. PMID:23844071

Pontes, Bruno; Ayala, Yareni; Fonseca, Anna Carolina C; Romão, Luciana F; Amaral, Rac?ele F; Salgado, Leonardo T; Lima, Flavia R; Farina, Marcos; Viana, Nathan B; Moura-Neto, Vivaldo; Nussenzveig, H Moysés

2013-01-01

83

Membrane Elastic Properties and Cell Function  

PubMed Central

Recent studies indicate that the cell membrane, interacting with its attached cytoskeleton, is an important regulator of cell function, exerting and responding to forces. We investigate this relationship by looking for connections between cell membrane elastic properties, especially surface tension and bending modulus, and cell function. Those properties are measured by pulling tethers from the cell membrane with optical tweezers. Their values are determined for all major cell types of the central nervous system, as well as for macrophage. Astrocytes and glioblastoma cells, which are considerably more dynamic than neurons, have substantially larger surface tensions. Resting microglia, which continually scan their environment through motility and protrusions, have the highest elastic constants, with values similar to those for resting macrophage. For both microglia and macrophage, we find a sharp softening of bending modulus between their resting and activated forms, which is very advantageous for their acquisition of phagocytic functions upon activation. We also determine the elastic constants of pure cell membrane, with no attached cytoskeleton. For all cell types, the presence of F-actin within tethers, contrary to conventional wisdom, is confirmed. Our findings suggest the existence of a close connection between membrane elastic constants and cell function.

Pontes, Bruno; Ayala, Yareni; Fonseca, Anna Carolina C.; Romao, Luciana F.; Amaral, Rac?ele F.; Salgado, Leonardo T.; Lima, Flavia R.; Farina, Marcos; Viana, Nathan B.; Moura-Neto, Vivaldo; Nussenzveig, H. Moyses

2013-01-01

84

A membrane bending model of outer hair cell electromotility.  

PubMed Central

We propose a new mechanism for outer hair cell electromotility based on electrically induced localized changes in the curvature of the plasma membrane (flexoelectricity). Electromechanical coupling in the cell's lateral wall is modeled in terms of linear constitutive equations for a flexoelectric membrane and then extended to nonlinear coupling based on the Langevin function. The Langevin function, which describes the fraction of dipoles aligned with an applied electric field, is shown to be capable of predicting the electromotility voltage displacement function. We calculate the electrical and mechanical contributions to the force balance and show that the model is consistent with experimentally measured values for electromechanical properties. The model rationalizes several experimental observations associated with outer hair cell electromotility and provides for constant surface area of the plasma membrane. The model accounts for the isometric force generated by the cell and explains the observation that the disruption of spectrin by diamide reduces force generation in the cell. We discuss the relation of this mechanism to other proposed models of outer hair cell electromotility. Our analysis suggests that rotation of membrane dipoles and the accompanying mechanical deformation may be the molecular mechanism of electromotility.

Raphael, R M; Popel, A S; Brownell, W E

2000-01-01

85

Alternate Fuel Cell Membranes for Energy Independence  

SciTech Connect

The overall objective of this project was the development and evaluation of novel hydrocarbon fuel cell (FC) membranes that possess high temperature performance and long term chemical/mechanical durability in proton exchange membrane (PEM) fuel cells (FC). The major research theme was synthesis of aromatic hydrocarbon polymers of the poly(arylene ether sulfone) (PAES) type containing sulfonic acid groups tethered to the backbone via perfluorinated alkylene linkages and in some cases also directly attached to the phenylene groups along the backbone. Other research themes were the use of nitrogen-based heterocyclics instead of acid groups for proton conduction, which provides high temperature, low relative humidity membranes with high mechanical/thermal/chemical stability and pendant moieties that exhibit high proton conductivities in the absence of water, and synthesis of block copolymers consisting of a proton conducting block coupled to poly(perfluorinated propylene oxide) (PFPO) blocks. Accomplishments of the project were as follows: 1) establishment of a vertically integrated program of synthesis, characterization, and evaluation of FC membranes, 2) establishment of benchmark membrane performance data based on Nafion for comparison to experimental membrane performance, 3) development of a new perfluoroalkyl sulfonate monomer, N,N-diisopropylethylammonium 2,2-bis(p-hydroxyphenyl) pentafluoropropanesulfonate (HPPS), 4) synthesis of random and block copolymer membranes from HPPS, 5) synthesis of block copolymer membranes containing high-acid-concentration hydrophilic blocks consisting of HPPS and 3,3'-disulfonate-4,4'-dichlorodiphenylsulfone (sDCDPS), 6) development of synthetic routes to aromatic polymer backbones containing pendent 1H-1,2,3-triazole moieties, 7) development of coupling strategies to create phase-separated block copolymers between hydrophilic sulfonated prepolymers and commodity polymers such as PFPO, 8) establishment of basic performance properties of experimental membranes, 9) fabrication and FC performance testing of membrane electrode assemblies (MEA) from experimental membranes, and 10) measurement of ex situ and in situ membrane durability of experimental membranes. Although none of the experimental hydrocarbon membranes that issued from the project displayed proton conductivities that met DOE requirements, the project contributed to our basic understanding of membrane structure-property relationships in a number of key respects. An important finding of the benchmark studies is that physical degradation associated with humidity and temperature variations in the FC tend to open new fuel crossover pathways and act synergistically with chemical degradation to accelerate overall membrane degradation. Thus, for long term membrane survival and efficient fuel utilization, membranes must withstand internal stresses due to humidity and temperature changes. In this respect, rigid aromatic hydrocarbon fuel cell membranes, e.g. PAES, offer an advantage over un-modified Nafion membranes. The benchmark studies also showed that broadband dielectric spectroscopy is a potentially powerful tool in assessing shifts in the fundamental macromolecular dynamics caused by Nafion chemical degradation, and thus, this technique is of relevance in interrogating proton exchange membrane durability in fuel cells and macromolecular dynamics as coupled to proton migration, which is of fundamental relevance in proton exchange membranes in fuel cells. A key finding from the hydrocarbon membrane synthesis effort was that rigid aromatic polymers containing isolated ion exchange groups tethered tightly to the backbone (short tether), such as HPPS, provide excellent mechanical and durability properties but do not provide sufficient conductivity, in either random or block configuration, when used as the sole ion exchange monomer. However, we continue to hypothesize that longer tethers, and tethered groups spaced more closely within the hydrophilic chain elements of the polymer, will yield highly conductive materials with excellent mech

Storey, Robson, F.; Mauritz, Kenneth, A.; Patton, Derek, L.; Savin, Daniel, A.

2012-12-18

86

Evaluating Mitochondrial Membrane Potential in Cells  

Microsoft Academic Search

Permeant cationic fluorescent probes are widely employed to monitor mitochondrial transmembrane potential and its changes.\\u000a The application of such potential-dependent probes in conjunction with both fluorescence microscopy and fluorescence spectroscopy\\u000a allows the monitoring of mitochondrial membrane potential in individual living cells as well as in large population of cells.\\u000a These approaches to the analysis of membrane potential is of extremely

Giancarlo Solaini; Gianluca Sgarbi; Giorgio Lenaz; Alessandra Baracca

2007-01-01

87

Identification of Glycan Structure Alterations on Cell Membrane Proteins in Desoxyepothilone B Resistant Leukemia Cells*  

PubMed Central

Resistance to tubulin-binding agents used in cancer is often multifactorial and can include changes in drug accumulation and modified expression of tubulin isotypes. Glycans on cell membrane proteins play important roles in many cellular processes such as recognition and apoptosis, and this study investigated whether changes to the glycan structures on cell membrane proteins occur when cells become resistant to drugs. Specifically, we investigated the alteration of glycan structures on the cell membrane proteins of human T-cell acute lymphoblastic leukemia (CEM) cells that were selected for resistance to desoxyepothilone B (CEM/dEpoB). The glycan profile of the cell membrane glycoproteins was obtained by sequential release of N- and O-glycans from cell membrane fraction dotted onto polyvinylidene difluoride membrane with PNGase F and ?-elimination respectively. The released glycan alditols were analyzed by liquid chromatography (graphitized carbon)-electrospray ionization tandem MS. The major N-glycan on CEM cell was the core fucosylated ?2–6 monosialo-biantennary structure. Resistant CEM/dEpoB cells had a significant decrease of ?2–6 linked sialic acid on N-glycans. The lower ?2–6 sialylation was caused by a decrease in activity of ?-galactoside ?2–6 sialyltransferase (ST6Gal), and decreased expression of the mRNA. It is clear that the membrane glycosylation of leukemia cells changes during acquired resistance to dEpoB drugs and that this change occurs globally on all cell membrane glycoproteins. This is the first identification of a specific glycan modification on the surface of drug resistant cells and the mechanism of this downstream effect on microtubule targeting drugs may offer a route to new interventions to overcome drug resistance.

Nakano, Miyako; Saldanha, Rohit; Gobel, Anja; Kavallaris, Maria; Packer, Nicolle H.

2011-01-01

88

Photothermal nanoblade for patterned cell membrane cutting  

PubMed Central

We report a photothermal nanoblade that utilizes a metallic nanostructure to harvest short laser pulse energy and convert it into a highly localized and specifically shaped explosive vapor bubble. Rapid bubble expansion and collapse punctures a lightly-contacting cell membrane via high-speed fluidic flows and induced transient shear stress. The membrane cutting pattern is controlled by the metallic nanostructure configuration, laser pulse polarization, and energy. Highly controllable, sub-micron sized circular hole pairs to half moon-like, or cat-door shaped, membrane cuts were realized in glutaraldehyde treated HeLa cells.

Wu, Ting-Hsiang; Teslaa, Tara; Teitell, Michael A.; Chiou, Pei-Yu

2010-01-01

89

Durability of PEM Fuel Cell Membranes  

NASA Astrophysics Data System (ADS)

Durability is still a critical limiting factor for the commercialization of polymer electrolyte membrane (PEM) fuel cells, a leading energy conversion technology for powering future hydrogen fueled automobiles, backup power systems (e.g., for base transceiver station of cellular networks), portable electronic devices, etc. Ionic conducting polymer (ionomer) electrolyte membranes are the critical enabling materials for the PEM fuel cells. They are also widely used as the central functional elements in hydrogen generation (e.g., electrolyzers), membrane cell for chlor-alkali production, etc. A perfluorosulfonic acid (PFSA) polymer with the trade name Nafion® developed by DuPont™ is the most widely used PEM in chlor-alkali cells and PEM fuel cells. Similar PFSA membranes have been developed by Dow Chemical, Asahi Glass, and lately Solvay Solexis. Frequently, such membranes serve the dual function of reactant separation and selective ionic conduction between two otherwise separate compartments. For some applications, the compromise of the "separation" function via the degradation and mechanical failure of the electrolyte membrane can be the life-limiting factor; this is particularly the case for PEM in hydrogen/oxygen fuel cells.

Huang, Xinyu; Reifsnider, Ken

90

Activated Membrane Patches Guide Chemotactic Cell Motility  

PubMed Central

Many eukaryotic cells are able to crawl on surfaces and guide their motility based on environmental cues. These cues are interpreted by signaling systems which couple to cell mechanics; indeed membrane protrusions in crawling cells are often accompanied by activated membrane patches, which are localized areas of increased concentration of one or more signaling components. To determine how these patches are related to cell motion, we examine the spatial localization of RasGTP in chemotaxing Dictyostelium discoideum cells under conditions where the vertical extent of the cell was restricted. Quantitative analyses of the data reveal a high degree of spatial correlation between patches of activated Ras and membrane protrusions. Based on these findings, we formulate a model for amoeboid cell motion that consists of two coupled modules. The first module utilizes a recently developed two-component reaction diffusion model that generates transient and localized areas of elevated concentration of one of the components along the membrane. The activated patches determine the location of membrane protrusions (and overall cell motion) that are computed in the second module, which also takes into account the cortical tension and the availability of protrusion resources. We show that our model is able to produce realistic amoeboid-like motion and that our numerical results are consistent with experimentally observed pseudopod dynamics. Specifically, we show that the commonly observed splitting of pseudopods can result directly from the dynamics of the signaling patches.

Hecht, Inbal; Skoge, Monica L.; Charest, Pascale G.; Ben-Jacob, Eshel; Firtel, Richard A.; Loomis, William F.; Levine, Herbert; Rappel, Wouter-Jan

2011-01-01

91

MYADM regulates Rac1 targeting to ordered membranes required for cell spreading and migration  

PubMed Central

Membrane organization into condensed domains or rafts provides molecular platforms for selective recruitment of proteins. Cell migration is a general process that requires spatiotemporal targeting of Rac1 to membrane rafts. The protein machinery responsible for making rafts competent to recruit Rac1 remains elusive. Some members of the MAL family of proteins are involved in specialized processes dependent on this type of membrane. Because condensed membrane domains are a general feature of the plasma membrane of all mammalian cells, we hypothesized that MAL family members with ubiquitous expression and plasma membrane distribution could be involved in the organization of membranes for cell migration. We show that myeloid-associated differentiation marker (MYADM), a protein with unique features within the MAL family, colocalizes with Rac1 in membrane protrusions at the cell surface and distributes in condensed membranes. MYADM knockdown (KD) cells had altered membrane condensation and showed deficient incorporation of Rac1 to membrane raft fractions and, similar to Rac1 KD cells, exhibited reduced cell spreading and migration. Results of rescue-of-function experiments by expression of MYADM or active Rac1L61 in cells knocked down for Rac1 or MYADM, respectively, are consistent with the idea that MYADM and Rac1 act on parallel pathways that lead to similar functional outcomes.

Aranda, Juan F.; Reglero-Real, Natalia; Kremer, Leonor; Marcos-Ramiro, Beatriz; Ruiz-Saenz, Ana; Calvo, Maria; Enrich, Carlos; Correas, Isabel; Millan, Jaime; Alonso, Miguel A.

2011-01-01

92

Fractional occurrence of defects in membranes and mechanically driven interleaflet phospholipid transport  

NASA Astrophysics Data System (ADS)

The picture of biological membranes as uniform, homogeneous bileaflet structures has been revised in recent times due to the growing recognition that these structures can undergo significant fluctuations both in local curvature and in thickness. In particular, evidence has been obtained that a temporary, localized disordering of the lipid bilayer structure (defects) may serve as a principal pathway for movement of lipid molecules from one leaflet of the membrane to the other. How frequently these defects occur and how long they remain open are important unresolved questions. In this report, we calculate the rate of molecular transport through a transient defect in the membrane and compare this result to measurements of the net transbilayer flux of lipid molecules measured in an experiment in which the lipid flux is driven by differences between the mechanical stress in the two leaflets of the membrane bilayer. Based on this comparison, we estimate the frequency of defect occurrence in the membrane. The occurrence of defects is rare: the probability of finding a defect in 1.0 ?m2 of a lecithin membrane is estimated to be ~6.0×10-6. Based on this fractional occurrence of defects, the free energy of defect formation is estimated to be ~1.0×10-19 J. The calculations provide support for a model in which interleaflet transport in membranes is accelerated by mechanically driven lipid flow.

Raphael, Robert M.; Waugh, Richard E.; Svetina, Saša.; Žekš, Boštjan

2001-11-01

93

Fractions!  

NSDL National Science Digital Library

On this page you will practice adding, subtracting, multiplying, and dividing fractions. 1. Practice identifying equivalent fractions. Finding equivalent fractions 2. Add and subtract fractions. The first levels have like denominators, but then the levels get harder with unlike denominators. Make sure you use some scratch paper. Get to the highest level you can! Fraction Race (adding and subtracting, like and unlike denominators) 3. Practice adding fractions with mixed numbers. Math splat -adding fractions using mixed numbers 4. Finally, get with a partner ...

Hbinggeli

2010-08-24

94

Interactions of VirB9, -10, and -11 with the membrane fraction of Agrobacterium tumefaciens: solubility studies provide evidence for tight associations.  

PubMed

The eleven predicted gene products of the Agrobacterium tumefaciens virB operon are believed to form a transmembrane pore complex through which T-DNA export occurs. The VirB10 protein is required for virulence and is a component of an aggregate associated with the membrane fraction of A. tumefaciens. Removal of the putative membrane-spanning domain (amino acids 22 through 55) disrupts the membrane topology of VirB10 (J. E. Ward, E. M. Dale, E. W. Nester, and A. N. Binns, J. Bacteriol. 172:5200-5210, 1990). Deletion of the sequences encoding amino acids 22 to 55 abolishes the ability of plasmid-borne virB10 to complement a null mutation in the virB10 gene, suggesting that the proper topology of VirB10 in the membrane may indeed play a crucial role in T-DNA transfer to the plant cell. Western blot (immunoblot) analysis indicated that the observed loss of virulence could not be attributed to a decrease in the steady-state levels of the mutant VirB10 protein. Although the deletion of the single transmembrane domain would be expected to perturb membrane association, VirB10 delta 22-55 was found exclusively in the membrane fraction. Urea extraction studies suggested that this membrane localization might be the result of a peripheral membrane association; however, the mutant protein was found in both inner and outer membrane fractions separated by sucrose density gradient centrifugation. Both wild-type VirB10 and wild-type VirB9 were only partially removed from the membranes by extraction with 1% Triton X-100, while VirB5 and VirB8 were Triton X-100 soluble. VirB11 was stripped from the membranes by 6 M urea but not by a more mild salt extraction. The fractionation patterns of VirB9, VirB10, and VirB11 were not dependent on each other or on VirB8 or VirD4. The observed tight association of VirB9, VirB10, and VirB11 with the membrane fraction support the notion that these proteins may exist as components of multiprotein pore complexes, perhaps spanning both the inner and outer membranes of Agrobacterium cells. PMID:7665464

Finberg, K E; Muth, T R; Young, S P; Maken, J B; Heitritter, S M; Binns, A N; Banta, L M

1995-09-01

95

Interactions of VirB9, -10, and -11 with the membrane fraction of Agrobacterium tumefaciens: solubility studies provide evidence for tight associations.  

PubMed Central

The eleven predicted gene products of the Agrobacterium tumefaciens virB operon are believed to form a transmembrane pore complex through which T-DNA export occurs. The VirB10 protein is required for virulence and is a component of an aggregate associated with the membrane fraction of A. tumefaciens. Removal of the putative membrane-spanning domain (amino acids 22 through 55) disrupts the membrane topology of VirB10 (J. E. Ward, E. M. Dale, E. W. Nester, and A. N. Binns, J. Bacteriol. 172:5200-5210, 1990). Deletion of the sequences encoding amino acids 22 to 55 abolishes the ability of plasmid-borne virB10 to complement a null mutation in the virB10 gene, suggesting that the proper topology of VirB10 in the membrane may indeed play a crucial role in T-DNA transfer to the plant cell. Western blot (immunoblot) analysis indicated that the observed loss of virulence could not be attributed to a decrease in the steady-state levels of the mutant VirB10 protein. Although the deletion of the single transmembrane domain would be expected to perturb membrane association, VirB10 delta 22-55 was found exclusively in the membrane fraction. Urea extraction studies suggested that this membrane localization might be the result of a peripheral membrane association; however, the mutant protein was found in both inner and outer membrane fractions separated by sucrose density gradient centrifugation. Both wild-type VirB10 and wild-type VirB9 were only partially removed from the membranes by extraction with 1% Triton X-100, while VirB5 and VirB8 were Triton X-100 soluble. VirB11 was stripped from the membranes by 6 M urea but not by a more mild salt extraction. The fractionation patterns of VirB9, VirB10, and VirB11 were not dependent on each other or on VirB8 or VirD4. The observed tight association of VirB9, VirB10, and VirB11 with the membrane fraction support the notion that these proteins may exist as components of multiprotein pore complexes, perhaps spanning both the inner and outer membranes of Agrobacterium cells.

Finberg, K E; Muth, T R; Young, S P; Maken, J B; Heitritter, S M; Binns, A N; Banta, L M

1995-01-01

96

The polysaccharide structure of potato cell walls: Chemical fractionation  

Microsoft Academic Search

Cell walls of potato tubers were fractionated by successive extraction with various reagents. A slightly degraded pectic fraction with 77% galacturonic acid was extracted in hot, oxalate-citrate buffer at pH 4. A further, major pectic fraction with 38% galacturonic acid was extracted in cold 0.1 M Na2CO3 with little apparent degradation. These two pectic fractions together made up 52% of

M. C. Jarvis; M. A. Hall; D. R. Threlfall; J. Friend

1981-01-01

97

Effect of Some Proteins on the Yeast Cell Membrane  

PubMed Central

Yeast cells, Candida utilis, in water suspension and in the absence of electrolytes were found to be very sensitive to several proteins of moderate size, including ribonuclease, protamine, lysozyme, bovine serum albumin, cytochrome c, and myoglobin. Viability ceases rapidly, and ultraviolet-absorbing compounds (260 m?) and the amino acid pool are released into the medium. The ultraviolet-absorbing material appears to be the nucleotide and coenzyme fraction usually extracted by 0.2 n perchloric acid at low temperature. The ribonucleic acid fraction remains in the cell ghosts and can be released by ribonuclease. The enzymatic properties of some of these proteins have no relation to their damaging effect on the cell membrane. Poly-l-lysine shows the same activity.

Yphantis, D. A.; Dainko, J. L.; Schlenk, F.

1967-01-01

98

Release of outer membrane fragments by exponentially growing Brucella melitensis cells.  

PubMed Central

Rough and smooth strains of Brucella melitensis released a membranous material that was devoid of detectable NADH oxidase and succinic dehydrogenase activity (cytoplasmic membrane markers) but that contained lipopolysaccharide, proteins, and phospholipids. This material was composed of two fractions that had similar chemical compositions but that were of different sizes which were separated by differential ultracentrifugation. Electron microscopy showed that both fractions are made of unit membrane structures. The membrane fragments were released during the exponential phase of growth, and no leakage of malic dehydrogenase activity (cytosol marker) was detected. Thus, the fragments were unlikely a result of cell lysis. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot analysis showed that, although group 2 Brucella outer membrane proteins and lipoprotein were not detected, the proteins in the membranous material were outer membrane proteins. Gas-liquid chromatography analysis showed a similar fatty acid profile for the cell envelope and the outer membrane fragments of the smooth strain B. melitensis 16M. In contrast, the outer membrane fragments from the rough 115 strain were enriched in palmitic and stearic acids. With respect to the unfractionated cell envelope, outer membrane fragments were enriched in phosphatidylcholine, a phospholipid that is unusual in bacterial membranes. Images

Gamazo, C; Moriyon, I

1987-01-01

99

Basement Membranes: Cell Scaffoldings and Signaling Platforms  

PubMed Central

Basement membranes are widely distributed extracellular matrices that coat the basal aspect of epithelial and endothelial cells and surround muscle, fat, and Schwann cells. These extracellular matrices, first expressed in early embryogenesis, are self-assembled on competent cell surfaces through binding interactions among laminins, type IV collagens, nidogens, and proteoglycans. They form stabilizing extensions of the plasma membrane that provide cell adhesion and that act as solid-phase agonists. Basement membranes play a role in tissue and organ morphogenesis and help maintain function in the adult. Mutations adversely affecting expression of the different structural components are associated with developmental arrest at different stages as well as postnatal diseases of muscle, nerve, brain, eye, skin, vasculature, and kidney.

Yurchenco, Peter D.

2011-01-01

100

Top down proteomics of human membrane proteins from enriched mitochondrial fractions.  

PubMed

The interrogation of intact integral membrane proteins has long been a challenge for biological mass spectrometry. Here, we demonstrate the application of top down mass spectrometry to whole membrane proteins below 60 kDa with up to 8 transmembrane helices. Analysis of enriched mitochondrial membrane preparations from human cells yielded identification of 83 integral membrane proteins, along with 163 membrane-associated or soluble proteins, with a median q value of 3 × 10(-10). An analysis of matching fragment ions demonstrated that significantly more fragment ions were found within transmembrane domains than would be expected based upon the observed protein sequence. In total, 46 proteins from the complexes of oxidative phosphorylation were identified which exemplifies the increasing ability of top down proteomics to provide extensive coverage in a biological network. PMID:23305238

Catherman, Adam D; Li, Mingxi; Tran, John C; Durbin, Kenneth R; Compton, Philip D; Early, Bryan P; Thomas, Paul M; Kelleher, Neil L

2013-02-01

101

Diffuse Charge Effects in Fuel Cell Membranes  

Microsoft Academic Search

It is commonly assumed that electrolyte membranes in fuel cells are electrically neutral, except in unsteady situations, when the double-layer capacitance is heuristically included in equivalent circuit calculations. Indeed, the standard model for electron transfer kinetics at the membrane\\/electrode interface is the Butler¿Volmer equation, where the interfacial overpotential is based on the total potential difference between the electrode and bulk

P. Maarten Biesheuvel; Alejandro A. Franco; Martin Z. Bazant

2009-01-01

102

Apoptosis-inducing activity of HPLC fraction from Voacanga globosa (Blanco) Merr. on the human colon carcinoma cell.  

PubMed

Voacanga globosa (Blanco), a plant endemic to the Philippines, is traditionally used especially by indigenous people of Bataan in the treatment of ulcers, wounds and tumorous growths. This study aimed to provide scientific evidence to therapeutic properties by determining cytotoxic and pro-apoptotic activity of HPLC fractions from leaves on HCT116 human colon carcinoma and A549 human lung carcinoma cell lines. Ethanolic extraction was performed on V globosa leaves followed by hexane and ethyl acetate partitioning. Silica gel column chromatography and high performance liquid chromatography (HPLC) produced MP1, MP2 and MP3 fractions. Cytotoxic activity of the fractions was determined through MTT assay against the cancer cell lines HCT116 and A549 and the non-cancer AA8 Chinese hamster ovarian cell line. Pro-apoptotic activities of the most active fractions were further assessed through DAPI staining, TUNEL assay and JC-1 mitochondrial membrane potential assay with HCT116 cells. While the MP1 fraction exerted no significant activity against all cell lines tested, MP2 and MP3 fractions demonstrated high toxicity against HCT116 and A549 cells. The MP3 fraction induced formation of apoptotic bodies, condensed DNA and other morphological changes consistent with apoptosis of HCT116 cells and TUNEL assay showed significant increase in DNA fragmentation over time. In these cells, the MP3 fraction also induced mitochondrial membrane destabilization, which is generally associated with the beginning of apoptosis. Phytochemical analysis demonstrated the presence only of saponins and terpenoids in the MP3 fraction. The results indicate that the MP3 fraction exerts cytotoxic activity on HCT116 cells via induction of apoptosis triggered by loss of mitochondrial membrane potential crucial for cell survival. PMID:24568467

Acebedo, Alvin Resultay; Amor, Evangeline Cancio; Jacinto, Sonia Donaldo

2014-01-01

103

Comparative Proteomics of Inner Membrane Fraction from Carbapenem-Resistant Acinetobacter baumannii with a Reference Strain  

PubMed Central

Acinetobacter baumannii has been identified by the Infectious Diseases Society of America as one of the six pathogens that cause majority of hospital infections. Increased resistance of A. baumannii even to the latest generation of ?-lactams like carbapenem is an immediate threat to mankind. As inner-membrane fraction plays a significant role in survival of A. baumannii, we investigated the inner-membrane fraction proteome of carbapenem-resistant strain of A. baumannii using Differential In-Gel Electrophoresis (DIGE) followed by DeCyder, Progenesis and LC-MS/MS analysis. We identified 19 over-expressed and 4 down-regulated proteins (fold change>2, p<0.05) in resistant strain as compared to reference strain. Some of the upregulated proteins in resistant strain and their association with carbapenem resistance in A. baumannii are: i) ?-lactamases, AmpC and OXA-51: cleave and inactivate carbapenem ii) metabolic enzymes, ATP synthase, malate dehydrogenase and 2-oxoglutarate dehydrogenase: help in increased energy production for the survival and iii) elongation factor Tu and ribosomal proteins: help in the overall protein production. Further, entry of carbapenem perhaps is limited by controlled production of OmpW and low levels of surface antigen help to evade host defence mechanism in developing resistance in A. baumannii. Present results support a model for the importance of proteins of inner-membrane fraction and their synergistic effect in the mediation of resistance of A. baumannii to carbapenem.

Tiwari, Vishvanath; Vashistt, Jitendraa; Kapil, Arti; Moganty, Rajeswari R.

2012-01-01

104

Specific binding of a fungal glucan phytoalexin elicitor to membrane fractions from soybean Glycine max  

PubMed Central

Treatment of soybean tissues with elicitors results in the production of phytoalexins, one of a number of inducible plant defense reactions against microbial infections. The present study uses a ?-1,3-[3H]glucan elicitor fraction from Phytophthora megasperma f. sp. glycinea, a fungal pathogen of soybean, to identify putative elicitor targets in soybean tissues. Use of the radiolabeled elicitor disclosed saturable high-affinity elicitor binding site(s) in membrane fractions of soybean roots. Highest binding activity is associated with a plasma membrane-enriched fraction. The apparent Kd value for ?-glucan elicitor binding is ?0.2 × 10-6 M and the maximum number of binding sites is 0.5 pmol per mg of protein. Competition studies with the [3H]glucan elicitor and a number of polysaccharides demonstrate that only polysaccharides of a branched ?-glucan type effectively displace the radiolabeled ligand from membrane binding. Differential displacing activity of the glucans on P. megasperma elicitor binding corresponds closely to their respective ability to elicit phytoalexin production in a cotyledon bioassay.

Schmidt, Walter E.; Ebel, Jurgen

1987-01-01

105

Specific binding of a fungal glucan phytoalexin elicitor to membrane fractions from soybean Glycine max  

SciTech Connect

Treatment of soybean tissues with elicitors results in the production of phytoalexins, one of a number of inducible plant defense reactions against microbial infections. The present study uses a ..beta..-1,3-(/sup 3/H) glucan elicitor fraction from Phytophthora megasperma f.sp. glycinea, a fungal pathogen of soybean, to identify putative elicitor targets in soybean tissues. Use of the radiolabeled elicitor disclosed saturable high-affinity elicitor binding site(s) in membrane fractions of soybean roots. Highest binding activity is associated with a plasma membrane-enriched fraction. The apparent K/sub d/ value for ..beta..-glucan elicitor binding is approx. = 0.2 x 10/sup -6/ M and the maximum number of binding sites is 0.5 pmol per mg of protein. Competition studies the (/sup 3/H)glucan elicitor and a number of polysaccharides demonstrate that only polysaccharides of a branched ..beta..-glucan type effectively displace the radiolabeled ligand from membrane binding. Differential displacing activity of the glucans on P. megasperma elicitor binding corresponds closely to their respective ability to elicit phytoalexin production in a cotyledon bioassay.

Schmidt, W.E.; Ebel, J.

1987-06-01

106

Tyrosine phosphorylation of membrane proteins mediates cellular invasion by transformed cells  

PubMed Central

Tyrosine phosphorylation of membrane-associated proteins is involved at two distinct sites of contact between cells and the extracellular matrix: adhesion plaques (cell adhesion and de-adhesion) and invadopodia (invasion into the extracellular matrix). Adhesion plaques from chicken embryonic fibroblasts or from cells transformed by Rous sarcoma virus contain low levels of tyrosine-phosphorylated proteins (YPPs) which were below the level of detection in 0.5-microns thin, frozen sections. In contrast, intense localization of YPPs was observed at invadopodia of transformed cells at sites of degradation and invasion into the fibronectin-coated gelatin substratum, but not in membrane extensions free of contact with the extracellular matrix. Local extracellular matrix degradation and formation of invadopodia were blocked by genistein, an inhibitor of tyrosine-specific kinases, but cells remained attached to the substratum and retained their free- membrane extensions. Invadopodia reduced or lost YPP labeling after treatment of the cells with genistein, but adhesion plaques retained YPP labeling. The plasma membrane contact fractions of normal and transformed cells have been isolated form cells grown on gelatin cross- linked substratum using a novel fractionation scheme, and analyzed by immunoblotting. Four major YPPs (150, 130, 81, and 77 kD) characterize invadopodial membranes in contact with the matrix, and are probably responsible for the intense YPP labeling associated with invadopodia extending into sites of matrix degradation. YPP150 may be an invadopodal-specific YPP since it is approximately 3.6-fold enriched in the invasive contact fraction relative to the cell body fraction and is not observed in normal contacts. YPP130 is enriched in transformed cell contacts but may also be present in normal contacts. The two major YPPs of normal contacts (130 and 71 kD) are much lower in abundance than the major tyrosine-phosphorylated bands associated with invadopodial membranes, and likely represent major adhesion plaque YPPs. YPP150, paxillin, and tensin appear to be enriched in the cell contact fractions containing adhesion plaques and invadopodia relative to the cell body fraction, but are also present in the soluble supernate fraction. However, vinculin, talin, and alpha-actinin that are localized at invadopodia, are equally concentrated in cell bodies and cell contacts as is the membrane-adhesion receptor beta 1 integrin. Thus, tyrosine phosphorylation of the membrane-bound proteins may contribute to the cytoskeletal and plasma membrane events leading to the formation and function of invadopodia that contact and proteolytically degrade the extracellular matrix; we have identified several candidate YPPs that may participate in the regulation of these processes.

1992-01-01

107

Membrane potential dynamics of grid cells  

PubMed Central

During navigation, grid cells increase their spike rates in firing fields arranged on a strikingly regular triangular lattice, while their spike timing is often modulated by theta oscillations. Oscillatory interference models of grid cells predict theta amplitude modulations of membrane potential during firing field traversals, while competing attractor network models predict slow depolarizing ramps. Here, using in-vivo whole-cell recordings, we tested these models by directly measuring grid cell intracellular potentials in mice running along linear tracks in virtual reality. Grid cells had large and reproducible ramps of membrane potential depolarization that were the characteristic signature tightly correlated with firing fields. Grid cells also exhibited intracellular theta oscillations that influenced their spike timing. However, the properties of theta amplitude modulations were not consistent with the view that they determine firing field locations. Our results support cellular and network mechanisms in which grid fields are produced by slow ramps, as in attractor models, while theta oscillations control spike timing.

Domnisoru, Cristina; Kinkhabwala, Amina A.; Tank, David W.

2014-01-01

108

Ehrlich cell plasma membrane redox system is modulated through signal transduction pathways involving c GMP and Ca 2+ as second messengers  

Microsoft Academic Search

Ehrlich cell plasma membrane ferricyanide reductase activity increased in the presence of mastoparan, a generic activator of G proteins, using either whole cells or isolated plasma membrane fractions. Agents that increase intracellularcAMP also increased the rate of ferricyanide reduction by Ehrlich cells. For the first time, evidence is shown on a modulation of plasma membrane redox system bycGMP. In fact,

Antonio del Castillo-Olivares; Alicia Esteban del Valle; Javier Márquez; Ignacio NÚñez de Castro; Miguel ángel Medina

1995-01-01

109

Modification and evaluation of fuel cell membranes  

NASA Astrophysics Data System (ADS)

The primary goals of this study were modification of existing NafionRTM membranes and characterization of newly developed hydrocarbon-based membranes for high temperature fuel cell applications. Various NafionRTM/silicate nanocomposites were formulated via in situ sol-gel reactions for tetraethylorthosilicate. Different silicate composition profiles generated across membrane cross-sections were investigated by EDAX/ESEM. Composite water uptake, proton conductivity and fuel cell performance were comparable to that of unmodified Nafion RTM. Tafel analysis showed better electrode kinetics for composites having more silicate in the middle and less or no silicate at electrolyte-electrode interfaces. All composites showed reduced fuel cross-over and superior mechanical as well as chemical durability than unmodified NafionRTM. Poly(cyclohexadiene) (PCHD) materials were characterized in the interest of developing alternative low-cost proton exchange membranes. All cross-linked sulfonated (xsPCHD) membranes showed significantly higher water uptake at 80 °C and higher proton conductivity at 120 °C at all relative humidities (RH), compared to the current benchmark membrane, NafionRTM. A xsPCHD-poly(ethylene glycol) (PEG) copolymer and a xsPCHD-PEG blend surpassed the DOE target by exhibiting proton conductivities of 141.44 and 322.40 mS/cm, respectively, at 50 % RH. Although the PCHD-based PEMs exhibited thermal stability up to 150 °C, they showed poor mechanical properties which would cause poor membrane durability during fuel cell operation. Atomic force microscopy studies demonstrated nanophase separated morphology of xsPCHD having a higher degree of connectedness of hydrophilic domains in the copolymer and blends relative to the xsPCHD homopolymer. Broadband dielectric spectroscopy (BDS) was used to study sub-Tg relaxations in annealed poly(2,5-benzimidazole) (ABPBI) fuel cell precursor materials. A trend in degree of connectivity of charge migration pathways and conductivity with annealing temperature and time was uncovered. Solid state 1H and 13C NMR studies showed hydrogen bonding group mobility while wide angle X-ray diffraction investigations indicated an increase in chain packing efficiency vs. temperature. BDS studies also investigated the effect of acid doping on poly(benzimidazole) (PBI) membrane macromolecular dynamics and sigmadc conductivity, sdc. High epsilon' values observed for acid doped samples in the low frequency regime could be due to membrane-electrode interfacial polarization. Distribution of relaxation time curves broadened while sigmadc increased with increase in acid doping level in the PBI membrane.

Nalawade, Amol Prataprao

110

Synthesis of cell wall xylans and glucans by golgi membranes  

SciTech Connect

We investigated the biosynthesis of mixed-linkage {beta}-D-glucan and glucuronoarabinoxylans which make up the hemicellulosic matrix of the primary cell walls of maize and other cereal grasses. The Golgi apparatus was enriched from plasma membrane and other organelles by flotation density gradient centrifugation. Glucan synthase I and II, which are established markers for Golgi and plasma membrane, respectively, displayed considerable overlap in conventional separations with sucrose density gradients. Flotation gradients improved separation of the membranes substantially, but the different synthases themselves also incorporated radioactivity from either 10 {mu}M or 1 mM UDP-({sup 14}C)-glucose into polymer. Relative incorporation of radioactivity into polymers from UDP-({sup 14}C)-xylose by the various membrane fractions was nearly identical to relative IDPase activities, indicating that combined xylosyl transferase-xylan synthase represents a new, unequivocal marker for the Golgi apparatus. We also have developed techniques of gas-liquid chromatography and radiogas proportional counting to achieve capillary quality separation of partially methylated alditol acetates with simultaneous determination of radioactivity in the derivatives. Digestion of polymeric products by specific endo-glycanohydrolases to diagnostic oligosaccharides also reveal specific kinds of polysaccharides synthesized by the Golgi membranes. A combination of these techniques provides unequivocal determination of the linkage structure of specific polymers synthesized by the purified Golgi apparatus.

Gibeaut, D.M.; Carpita, N.C. (Purdue Univ., West Lafayette, IN (USA))

1989-04-01

111

Effect of Diphenyl Ether Herbicides on Oxidation of Protoporphyrinogen to Protoporphyrin in Organellar and Plasma Membrane Enriched Fractions of Barley 1  

PubMed Central

In barley (Hordeum vulgare L.) root cells, activity for oxidizing protoporphyrinogen to protoporphyrin (protoporphyrinogen oxidase), a step in chlorophyll and heme synthesis, was found both in the crude mitochondrial fraction and in a plasma membrane enriched fraction separated by a sucrose gradient technique utilized for preparing plasma membranes. The specific activity (expressed as nanomoles of protoporphyrin formed per hour per milligram protein) in the mitochondrial fraction was 8 and in the plasma membrane enriched fraction was 4 to 6. The plasma membrane enriched fraction exhibited minimal cytochrome oxidase activity and no carotenoid content, indicating little contamination with mitochondrial or plastid membranes. Etioplasts from etiolated barley leaves exhibited a protoporphyrinogen oxidase specific activity of 7 to 12. Protoporphyrinogen oxidase activity in the barley root mitochondrial fraction and etioplast extracts was more than 90% inhibited by assay in the presence of the diphenyl ether herbicide acifluorfen methyl, but the activity in the plasma membrane enriched fraction exhibited much less inhibition by this herbicide (12 to 38% inhibition) under the same assay conditions. Acifluorfen-methyl inhibition of the organellar (mitochondrial or plastid) enzyme was maximal upon preincubation of the enzyme with 4 mm dithiothreitol, although a lesser degree of inhibition was noted if the organellar enzyme was preincubated in the presence of other reductants such as glutathione or ascorbate. Acifluorfen-methyl caused only 20% inhibition if the enzyme was preincubated in buffer without reductants. Incubation of barley etioplast extracts with the earlier tetrapyrrole precursor coproporphyrinogen and acifluorfen-methyl resulted in the accumulation of protoporphyrinogen, which could be converted to protoporphyrin even in the presence of the herbicide by the addition of the plasma membrane enriched fraction from barley roots. These findings have implications for the toxicity of diphenyl ether herbicides, whose light induced tissue damage is apparently caused by accumulation of the photoreactive porphyrin intermediate, protoporphyrin, when the organellar protoporphyrinogen oxidase enzyme is inhibited by herbicides. Our results suggest that the protoporphyrinogen that accumulates as a result of herbicide inhibition of the organellar enzyme can be oxidized to protoporphyrin by a protoporphyrinogen oxidizing activity that is located at sites such as the plasma membrane, which is much less sensitive to inhibition by diphenylether herbicides.

Jacobs, Judith M.; Jacobs, Nicholas J.; Sherman, Timothy D.; Duke, Stephen O.

1991-01-01

112

Electrophoretic characterization of a detergent-treated plasma membrane fraction from corn roots.  

PubMed

Experiments were conducted to determine conditions essential for electrophoretic characterization of a detergent-extracted plasma membrane fraction from corn (Zea mays L.) roots. Sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis (PAGE) initially gave poor resolution of polypeptides in the plasma membrane fraction and, upon detergent treatment for purification of the proton-pumping adenosine triphosphatase (ATPase), showed no enrichment for a 100 kilodalton catalytic subunit characteristic of the ATPase. In contrast to SDS-PAGE, phenol urea acetic acid (PAU)-PAGE clearly resolved two polypeptides in the 100 kilodalton region that were enriched during detergent treatment and indicated at least one polypeptide forms a phosphorylated intermediate characteristic of the ATPase. Problems with SDS-PAGE were found to be caused, in part, by a combination of endogenous proteases and heat-induced aggregation of high molecular weight proteins. The usually standard procedure of boiling the sample prior to SDS-PAGE caused the aggregation of the 100 kilodalton polypeptides. By controlling for proteases using chymostatin and/or phenylmethane sulfonyl floride, and not boiling the sample prior to electrophoresis, two polypeptides were clearly resolved by SDS-PAGE in the 100 kilodalton region of Triton X-114-extracted membranes from corn, oat, barley, and tomato. PMID:16665234

Gallagher, S R; Leonard, R T

1987-02-01

113

Membrane curvature and mechanisms of dynamic cell membrane remodelling  

Microsoft Academic Search

Membrane curvature is no longer seen as a passive consequence of cellular activity but an active means to create membrane domains and to organize centres for membrane trafficking. Curvature can be dynamically modulated by changes in lipid composition, the oligomerization of curvature scaffolding proteins and the reversible insertion of protein regions that act like wedges in membranes. There is an

Jennifer L. Gallop; Harvey T. McMahon

2005-01-01

114

Polyphosphoinositides Are Present in Plasma Membranes Isolated from Fusogenic Carrot Cells 1  

PubMed Central

Fusogenic carrot cells grown in suspension culture were labeled 12 hours with myo-[2-3H]inositol. Plasma membranes were isolated from the prelabeled fusogenic carrot cells by both aqueous polymer two-phase partitioning and Renografin density gradients. With both methods, the plasma membrane-enriched fractions, as identified by marker enzymes, were enriched in [3H]inositol-labeled phosphatidylinositol monophosphate (PIP) and phosphatidylinositol bisphosphate (PIP2). An additional [3H]inositol-labeled lipid, lysophosphatidylinositol monophosphate, which migrated between PIP and PIP2 on thin layer plates, was found primarily in the plasma membrane-rich fraction of the fusogenic cells. This was in contrast to lysophosphatidylinositol which is found primarily in the lower phase, microsomal/mitochrondrial-rich fraction.

Wheeler, Jeffery J.; Boss, Wendy F.

1987-01-01

115

Release of extracellular membrane vesicles from microvilli of epithelial cells is enhanced by depleting membrane cholesterol  

Microsoft Academic Search

We previously reported on the occurrence of prominin-1-carrying membrane vesicles that are released into body fluids from microvilli of epithelial cells. This release has been implicated in cell differentiation. Here we have characterized these vesicles released from the differentiated Caco-2 cells. We find that in these vesicles, prominin-1 directly interacts with membrane cholesterol and is associated with a membrane microdomain.

Anne-Marie Marzesco; Michaela Wilsch-Bräuninger; Véronique Dubreuil; Peggy Janich; Katja Langenfeld; Christoph Thiele; Wieland B. Huttner; Denis Corbeil

2009-01-01

116

Membrane electrode assembly for a fuel cell  

NASA Technical Reports Server (NTRS)

A catalyst ink for a fuel cell including a catalytic material and poly(vinylidene fluoride). The ink may be applied to a substrate to form an electrode, or bonded with other electrode layers to form a membrane electrode assembly (MEA).

Prakash, Surya (Inventor); Narayanan, Sekharipuram R. (Inventor); Atti, Anthony (Inventor); Olah, George (Inventor); Smart, Marshall C. (Inventor)

2006-01-01

117

Cell cycle regulation of Golgi membrane dynamics  

PubMed Central

The Golgi apparatus is a membranous organelle in the cell that plays essential roles in protein and lipid trafficking, sorting, processing and modification. Its basic structure is a stack of closely aligned flattened cisternae. In mammalian cells, dozens of Golgi stacks are often laterally linked into a ribbon-like structure. Biogenesis of the Golgi during cell division occurs through a sophisticated disassembly and reassembly process that can be divided into three distinct but cooperative steps, including the deformation and reformation of the Golgi cisternae, stacks and ribbon. Here, we review our current understanding of the protein machineries that control these three steps in the cycle of mammalian cell division: GRASP65 and GRASP55 in Golgi stack and ribbon formation; ubiquitin and AAA ATPases in post-mitotic Golgi membrane fusion; and golgins and cytoskeleton in Golgi ribbon formation.

Tang, Danming; Wang, Yanzhuang

2013-01-01

118

Model Assessment of Cell Membrane Breakdown in Clusters and Tissues Under High-Intensity Electric Pulsing  

Microsoft Academic Search

This paper presents a simulation study of cell membrane electroporation in clusters by high-intensity voltage pulses. The focus is on assessing effects associated with: 1) the variability in shape and randomness of the cells within clusters; 2) the density of clusters; 3) the effects in heterogeneous tissues; 4) the role of pulse width on fractional electroporation for given electrical characteristics;

Ravindra P. Joshi; Ashutosh Mishra; Karl H. Schoenbach

2008-01-01

119

Functional imaging of microdomains in cell membranes  

Microsoft Academic Search

The presence of microdomains or rafts within cell membranes is a topic of intense study and debate. The role of these structures\\u000a in cell physiology, however, is also not yet fully understood with many outstanding problems. This problem is partly based\\u000a on the small size of raft structures that presents significant problems to their in vivo study, i.e., within live

James Duggan; Ghadir Jamal; Mark Tilley; Ben Davis; Graeme McKenzie; Kelly Vere; Michael G. Somekh; Paul O’Shea; Helen Harris

2008-01-01

120

Control of cell membrane tension by myosin-I  

PubMed Central

All cell functions that involve membrane deformation or a change in cell shape (e.g., endocytosis, exocytosis, cell motility, and cytokinesis) are regulated by membrane tension. While molecular contacts between the plasma membrane and the underlying actin cytoskeleton are known to make significant contributions to membrane tension, little is known about the molecules that mediate these interactions. We used an optical trap to directly probe the molecular determinants of membrane tension in isolated organelles and in living cells. Here, we show that class I myosins, a family of membrane-binding, actin-based motor proteins, mediate membrane/cytoskeleton adhesion and thus, make major contributions to membrane tension. These studies show that class I myosins directly control the mechanical properties of the cell membrane; they also position these motor proteins as master regulators of cellular events involving membrane deformation.

Nambiar, Rajalakshmi; McConnell, Russell E.; Tyska, Matthew J.

2009-01-01

121

Influence of hydrophobic/hydrophilic fractions of extracellular organic matters of Microcystis aeruginosa on ultrafiltration membrane fouling.  

PubMed

Fouling is a major obstacle to maintain the efficiency of ultrafiltration-based drinking water treatment process. Algal extracellular organic matters (EOMs) are currently considered as one of the major sources of membrane fouling. The objective of this study was to investigate the influence of different hydrophobic/hydrophilic fractions of EOM extracted from Microcystis aeruginosa on ultrafiltration membrane fouling at lab scale. The experimental data indicated that EOM exhibited similar flux decline trends on polyethersulfone (PES) and regenerated cellulose (RC) membranes but caused greater irreversible fouling on PES membrane than RC membrane due to its hydrophobic property. It was also observed that charged hydrophilic (CHPI) and neutral hydrophilic (NHPI) fractions caused greater flux decline over hydrophobic (HPO) and transphilic (TPI) fractions. For PES membrane, the order of the irreversible fouling potentials for the four fractions was HPO>TPI>CHPI>NHPI, while the irreversible fouling potentials of RC membrane were tiny and could be ignored. Fluorescence excitation-emission matrix (EEM) spectra and Fourier transform infrared (FTIR) spectra suggested that protein-like, polysaccharide-like and humic-like substances were the major components responsible for membrane fouling. The results also indicated that the irreversible fouling increased as the pH decreased. The addition of calcium to feed solutions led to more severe flux decline and irreversible fouling. PMID:24140690

Zhou, Shiqing; Shao, Yisheng; Gao, Naiyun; Li, Lei; Deng, Jing; Tan, Chaoqun; Zhu, Mingqiu

2014-02-01

122

Subcellular localization of H(+)-ATPase from pumpkin hypocotyls (Cucurbita maxima L.) by membrane fractionation.  

PubMed

A new method of preparing sealed vesicles from membrane fractions of pumpkin hypocotyls in ethanolamine-containing buffers was used to investigate the subcellular localization of H(+)-ATPase measured as nigericin-stimulated ATPase. In a fluorescence-quench assay, the H(+) pump was directly demonstrated. The H(+) pump was substrate-specific for Mg·ATP and 0.1 mM diethylstilbestrol completely prevented the development of a ? pH. The presence of unsupecific phosphatase hampered the detection of nigericin-stimulated ATPase. Unspecific phosphatases could be demonstrated by comparing the broad substrate specificity of the hydrolytic activities of the fractions with the clear preference for Mg·ATP as the substrate for the proton pump. Inhibitor studies showed that neither orthovanadate nor molybdate are absolutely specific for ATPase or acid phosphatase, respectively. Diethylstilbestrol seemed to be a specific inhibitor of ATPase activity in fractions containing nigericin-stimulated ATPase, but it stimulated acid phosphatase which tended to obscure its effect on ATPase activity. Nigericin-stimulated ATPase had its optimum at pH 6.0 and the nigericin effect was K(+)-dependent. The combination of valinomycin and carbonylcyanide m-chlorophenylhydrazone had a similar effect to nigericin, but singly these ionophores were much less stimulatory. After prolonged centrifugation on linear sucrose gradients, nigericin-stimulated ATPase correlated in dense fractions with plasma membrane markers but a part of it remained at the interphase. This lessdense part of the nigericin-stimulated ATPase could be derived from tonoplast vesicles because ?-mannosidase, an enzyme of the vacuolar sap, remained in the upper part of the gradient. Nigericinstimulated ATPase did not correlate with the mitochondrial marker, cytochrome c oxidase, whereas azide inhibition of ATPase activity did. PMID:24258584

Scherer, G F

1984-03-01

123

Membrane isolation on polylysine-coated beads. Plasma membrane from HeLa cells  

PubMed Central

HeLa cell plasma membranes have been purified after binding cells to polylysine-coated polyacrylamide beads. Cell attachment to beads and membrane recovery were maximal in a sucrose-acetate buffer, pH 5.0, at 25 degrees C. Measurements of ouabain-sensitive NaK-adenosine triphosphatase, membrane-bound 125I-wheat germ agglutinin, and chemical analyses showed that membranes on beads were of comparable or greater purity than membranes isolated by conventional methods. Because the isolation procedure is rapid (approximately 2.5 h), and produces membranes whose protoplasmic surfaces are fully exposed, it should be a useful supplement to standard isolation techniques.

1977-01-01

124

Effects of chitosan on the protein profile of grape cell culture subcellular fractions.  

PubMed

Grapevine is a large source of healthy polyphenols for human diet, and red table-grapes and wines are the main source of stilbenes. These compounds are important both in the plant defence system and for human health. In the present study, Vitis vinifera cv. Barbera cell cultures were treated with 50 ?g/mL chitosan and proteomic analyses on soluble and membrane subcellular fractions were performed against suitable controls. Three soluble stilbene synthase protein spots, four stilbene synthase spots in the microsomal fraction and four spots of membrane ATPase subunits were identified, the accumulation of which was modulated in response to chitosan treatment. Present proteomic and immunolocalisation data seem to provide evidence supporting the hypothesis that a stilbene biosynthetic multi-enzyme complex is associated with the intracellular membrane. In addition, proteomic analyses showed a general decrease in the accumulation of proteins belonging to different primary metabolism pathways, both in the soluble and membrane fractions. In particular, energy, sugar and amino acid metabolisms were down-regulated as a consequence of chitosan and acetic acid treatments. These metabolic modifications could lead to a consistent change in the profile and amount of metabolites stored in grape berries, with consequent effects on taste, flavour, organoleptic and nutraceutical properties of derived food products. PMID:24590893

Ferri, Maura; Franceschetti, Marina; Naldrett, Michael J; Saalbach, Gerhard; Tassoni, Annalisa

2014-06-01

125

Catalytic membranes for fuel cells  

DOEpatents

A fuel cell of the present invention comprises a cathode and an anode, one or both of the anode and the cathode including a catalyst comprising a bundle of longitudinally aligned graphitic carbon nanotubes including a catalytically active transition metal incorporated longitudinally and atomically distributed throughout the graphitic carbon walls of said nanotubes. The nanotubes also include nitrogen atoms and/or ions chemically bonded to the graphitic carbon and to the transition metal. Preferably, the transition metal comprises at least one metal selected from the group consisting of Fe, Co, Ni, Mn, and Cr.

Liu, Di-Jia (Naperville, IL); Yang, Junbing (Bolingbrook, IL); Wang, Xiaoping (Naperville, IL)

2011-04-19

126

Molecular dynamics study of lipid bilayers modeling the plasma membranes of normal murine thymocytes and leukemic GRSL cells.  

PubMed

Molecular dynamics (MD) calculations for the plasma membranes of normal murine thymocytes and thymus-derived leukemic GRSL cells in water have been performed under physiological isothermal-isobaric conditions (310.15K and 1 atm) to investigate changes in membrane properties induced by canceration. The model membranes used in our calculations for normal and leukemic thymocytes comprised 23 and 25 kinds of lipids, respectively, including phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, sphingomyelin, lysophospholipids, and cholesterol. The mole fractions of the lipids adopted here were based on previously published experimental values. Our calculations clearly showed that the membrane area was increased in leukemic cells, and that the isothermal area compressibility of the leukemic plasma membranes was double that of normal cells. The calculated membranes of leukemic cells were thus considerably bulkier and softer in the lateral direction compared with those of normal cells. The tilt angle of the cholesterol and the conformation of the phospholipid fatty acid tails both showed a lower level of order in leukemic cell membranes compared with normal cell membranes. The lateral radial distribution function of the lipids also showed a more disordered structure in leukemic cell membranes than in normal cell membranes. These observations all show that, for the present thymocytes, the lateral structure of the membrane is considerably disordered by canceration. Furthermore, the calculated lateral self-diffusion coefficient of the lipid molecules in leukemic cell membranes was almost double that in normal cell membranes. The calculated rotational and wobbling autocorrelation functions also indicated that the molecular motion of the lipids was enhanced in leukemic cell membranes. Thus, here we have demonstrated that the membranes of thymocyte leukemic cells are more disordered and more fluid than normal cell membranes. PMID:23333323

Andoh, Yoshimichi; Okazaki, Susumu; Ueoka, Ryuichi

2013-04-01

127

Sputter-deposited fuel cell membranes and electrodes  

NASA Technical Reports Server (NTRS)

A method for preparing a membrane for use in a fuel cell membrane electrode assembly includes the steps of providing an electrolyte membrane, and sputter-depositing a catalyst onto the electrolyte membrane. The sputter-deposited catalyst may be applied to multiple sides of the electrolyte membrane. A method for forming an electrode for use in a fuel cell membrane electrode assembly includes the steps of obtaining a catalyst, obtaining a backing, and sputter-depositing the catalyst onto the backing. The membranes and electrodes are useful for assembling fuel cells that include an anode electrode, a cathode electrode, a fuel supply, and an electrolyte membrane, wherein the electrolyte membrane includes a sputter-deposited catalyst, and the sputter-deposited catalyst is effective for sustaining a voltage across a membrane electrode assembly in the fuel cell.

Narayanan, Sekharipuram R. (Inventor); Jeffries-Nakamura, Barbara (Inventor); Chun, William (Inventor); Ruiz, Ron P. (Inventor); Valdez, Thomas I. (Inventor)

2001-01-01

128

Interactions of Model Cell Membranes with Nanoparticles  

NASA Astrophysics Data System (ADS)

The same properties that give nanoparticles their enhanced function, such as high surface area, small size, and better conductivity, can also alter the cytotoxicity of nanomaterials. Ultimately, many of these nanomaterials will be released into the environment, and can cause cytotoxic effects to environmental bacteria, aquatic organisms, and humans. Previous results from our laboratory suggest that nanoparticles can have a detrimental effect on cells, depending on nanoparticle size. It is our goal to characterize the properties of nanomaterials that can result in membrane destabilization. We tested the effects of nanoparticle size and chemical functionalization on nanoparticle-membrane interactions. Gold nanoparticles at 2, 5,10, and 80 nm were investigated, with a concentration of 1.1x1010 particles/mL. Model cell membranes were constructed of of L-?-phosphatidylcholine (egg PC), which has negatively charged lipid headgroups. A quartz crystal microbalance with dissipation (QCM-D) was used to measure frequency changes at different overtones, which were related to mass changes corresponding to nanoparticle interaction with the model membrane. In QCM-D, a lipid bilayer is constructed on a silicon dioxide crystal. The crystals, oscillate at different harmonic frequencies depending upon changes in mass or energy dissipation. When mass is added to the crystal surface, such as through addition of a lipid vesicle solution, the frequency change decreases. By monitoring the frequency and dissipation, we could verify that a supported lipid bilayer (SLB) formed on the silica surface. After formation of the SLB, the nanoparticles can be added to the system, and the changes in frequency and dissipation are monitored in order to build a mechanistic understanding of nanoparticle-cell membrane interactions. For all of the smaller nanoparticles (2, 5, and 10 nm), nanoparticle addition caused a loss of mass from the lipid bilayer, which appears to be due to the formation of holes or pores in the cell membrane. The dissipation changes were small, which indicates that even with the membrane destabilization that occurs, the overall structure of the bilayer is not greatly perturbed. For the 80 nm nanoparticles, we initially saw the same pattern as the smaller nanoparticles with a mass loss from the membrane, but eventually we saw a large decrease in frequency, representing an increase in mass. This addition of mass may be attributed to adsorption of the gold nanoparticles onto the bilayer. The 80 nm particles also created a change in the energy dissipation, which suggests that the formation of the bilayer was altered with the adsorbed particles. This study suggests that nanoparticle size controls the mechanism by which nanoparticles interact with model cell membranes. We are extending this work to other types of gold nanoparticles. We are interested in examining the role of nanoparticle hydrophobicity and type of chemical functionalization on the interactions of the nanoparticle with a model membrane. We are also conducting studies on environmental bacteria, to correlate the mechanisms of nanoparticle cytoxicity with killing data on bacterial cells.

D'Angelo, S. M.; Camesano, T. A.; Nagarajan, R.

2011-12-01

129

Improved Recovery and Identification of Membrane Proteins from Rat Hepatic Cells using a Centrifugal Proteomic Reactor*  

PubMed Central

Despite their importance in many biological processes, membrane proteins are underrepresented in proteomic analysis because of their poor solubility (hydrophobicity) and often low abundance. We describe a novel approach for the identification of plasma membrane proteins and intracellular microsomal proteins that combines membrane fractionation, a centrifugal proteomic reactor for streamlined protein extraction, protein digestion and fractionation by centrifugation, and high performance liquid chromatography-electrospray ionization-tandem MS. The performance of this approach was illustrated for the study of the proteome of ER and Golgi microsomal membranes in rat hepatic cells. The centrifugal proteomic reactor identified 945 plasma membrane proteins and 955 microsomal membrane proteins, of which 63 and 47% were predicted as bona fide membrane proteins, respectively. Among these proteins, >800 proteins were undetectable by the conventional in-gel digestion approach. The majority of the membrane proteins only identified by the centrifugal proteomic reactor were proteins with ?2 transmembrane segments or proteins with high molecular mass (e.g. >150 kDa) and hydrophobicity. The improved proteomic reactor allowed the detection of a group of endocytic and/or signaling receptor proteins on the plasma membrane, as well as apolipoproteins and glycerolipid synthesis enzymes that play a role in the assembly and secretion of apolipoprotein B100-containing very low density lipoproteins. Thus, the centrifugal proteomic reactor offers a new analytical tool for structure and function studies of membrane proteins involved in lipid and lipoprotein metabolism.

Zhou, Hu; Wang, Fangjun; Wang, Yuwei; Ning, Zhibin; Hou, Weimin; Wright, Theodore G.; Sundaram, Meenakshi; Zhong, Shumei; Yao, Zemin; Figeys, Daniel

2011-01-01

130

Proliferation of Schwann cells induced by axolemmal and myelin membranes  

SciTech Connect

Purified Schwann Cells were cultured from neonatal rat sciatic nerve using a modification of the method of Brockes. Schwann cells and contaminating fibroblasts were unambiguously identified using fluorescent antibodies of 2'3' cyclic nucleotide 3'-phosphodiesterase and the thy 1.1 antigen respectively. The Schwann cells were quiescent unless challenged with mitogens. They proliferated rapidly in response to the soluble mitogen, cholera toxin, or to membrane fractions from rat CNS or PNS, prepared by the method of DeVries. Mitogenic activity was present in both axolemmal and myelin enriched fractions and promoted a 10-15 fold increase in the rate of /sup 3/H-thymidine uptake. The axolemmal mitogen was sensitive to heat (80/sup 0/C for 10 minutes), trypsin digestion (0.05% x 30 mins) or to treatment with endoglycosidase D, suggesting that it could be a glycoprotein. Fifty percent of the axolemmal mitogenic activity was solubilized in 1% octyl-glucoside. The solubilized material, however, was very unstable and further purification was not possible. The myelin associated mitogenic activity was markedly different. It was resistant to freeze thaw cycles, trypsin digestion of endoglycosidase treatment and the activity was actually enhanced by heating at 100/sup 0/C for two hours. It is proposed that the axolemmal activity is responsible for Schwann cell proliferation during development and that the myelin associated activity promotes Schwann cell proliferation during Wallerian degeneration.

Dinneen, M..

1985-01-01

131

Fuel cell membranes and crossover prevention  

DOEpatents

A membrane electrode assembly for use with a direct organic fuel cell containing a formic acid fuel includes a solid polymer electrolyte having first and second surfaces, an anode on the first surface and a cathode on the second surface and electrically linked to the anode. The solid polymer electrolyte has a thickness t:.gtoreq..times..times..times..times. ##EQU00001## where C.sub.f is the formic acid fuel concentration over the anode, D.sub.f is the effective diffusivity of the fuel in the solid polymer electrolyte, K.sub.f is the equilibrium constant for partition coefficient for the fuel into the solid polymer electrolyte membrane, I is Faraday's constant n.sub.f is the number of electrons released when 1 molecule of the fuel is oxidized, and j.sub.f.sup.c is an empirically determined crossover rate of fuel above which the fuel cell does not operate.

Masel, Richard I. (Champaign, IL); York, Cynthia A. (Newington, CT); Waszczuk, Piotr (White Bear Lake, MN); Wieckowski, Andrzej (Champaign, IL)

2009-08-04

132

Molecular Basis of Red Cell Membrane Disorders  

Microsoft Academic Search

We will consider an array of genetic disorders of the red cell membrane. Some affect well-known genes. The mutations of most cases of hereditary spherocytosis (HS) are located in the following genes: ANK1, SPTB, SLC4A1, EPB42 and SPTA1, which encode ankyrin, spectrin ?-chain, the anion exchanger 1 (band 3), protein 4.2 and spectrin ?-chain, respectively. A dominant form of distal

Jean Delaunay

2002-01-01

133

Cell Membrane Diversity in Noncovalent Protein Transduction  

Microsoft Academic Search

Crossing of the plasma membrane for all macromolecules without energy, receptors or any artificial methods was thought to\\u000a be difficult. Our previous studies demonstrated that arginine-rich intracellular delivery (AID) peptides are able to deliver\\u000a macromolecules, such as proteins, RNAs and DNAs, into either animal or plant cells. Cellular internalization could be mediated\\u000a by effective and nontoxic AID peptides in either

Betty Revon Liu; Jyh-Ching Chou; Han-Jung Lee

2008-01-01

134

New membranes for direct methanol fuel cells  

Microsoft Academic Search

The performance of direct methanol fuel cells (DMFC) is limited by the cross-over of methanol through the electrolyte. Electrolyte membranes prepared by blending of sulfonated arylene main-chain polymers like sulfonated PEEK Victrex (sPEEK) or sulfonated PSU Udel (sPSU) with basic polymers like poly(4-vinylpyridine) (P4VP) or polybenzimidazole (PBI) show excellent chemical and thermal stability, good proton-conductivity, and good performance in H2

L. Jörissen; V. Gogel; J. Kerres; J. Garche

2002-01-01

135

Bioluminescence Assay for Detecting Cell Surface Membrane Protein Expression  

PubMed Central

Abstract We have developed a method to measure the amounts of cell surface-expressed membrane proteins with bioluminescence. Dinoflagellate luciferase was expressed on the surface of a mammalian cell as a chimeric fusion protein with a membrane protein of interest. Using a membrane-impermeable substrate to quantify the membrane-displayed luciferase, the expression of the membrane protein on the cell surface was determined. By inclusion of a quenching step for the luminescent activity of luciferase on the cell surface, we were able to monitor the membrane protein expression kinetics by measuring the luminescence recovery from the cell surface after quenching. The reported methods provide a convenient way to monitor the kinetics of expression and transport of membrane proteins to the cell surface. It is applicable to the high-throughput analysis of drugs or drug candidates concerning their effects on membrane protein expression.

Kato, Mieko; Chiba, Tomoki; Li, Min

2011-01-01

136

Outer membrane proteins of Bartonella henselae and their interaction with human endothelial cells  

Microsoft Academic Search

Members of the genusBartonellaare unique in that they are bacteria which cause proliferation of microvascular endothelial cells and neovascularization (angiogenesis). The mechanisms by whichBartonella henselaecauses these processes are unknown. Given the importance of surface-exposed determinants in the pathogenesis of many organisms, outer membrane proteins (OMPs) ofB. henselaewere identified. Enrichment of the outer membrane fraction ofB. henselaeby sarkosyl treatment of total

Andrew W. O Burgess; Burt E Anderson

1998-01-01

137

Detection of urease in the cell wall and membranes from leaf tissues of bromeliad species.  

PubMed

Urea is an important nitrogen source for some bromeliad species, and in nature it is derived from the excretion of amphibians, which visit or live inside the tank water. Its assimilation is dependent on the hydrolysis by urease (EC: 3.5.1.5), and although this enzyme has been extensively studied to date, little information is available about its cellular location. In higher plants, this enzyme is considered to be present in the cytoplasm. However, there is evidence that urease is secreted by the bromeliad Vriesea gigantea, implying that this enzyme is at least temporarily located in the plasmatic membrane and cell wall. In this article, urease activity was measured in different cell fractions using leaf tissues of two bromeliad species: the tank bromeliad V. gigantea and the terrestrial bromeliad Ananas comosus (L.) Merr. In both species, urease was present in the cell wall and membrane fractions, besides the cytoplasm. Moreover, a considerable difference was observed between the species: while V. gigantea had 40% of the urease activity detected in the membranes and cell wall fractions, less than 20% were found in the same fractions in A. comosus. The high proportion of urease found in cell wall and membranes in V. gigantea was also investigated by cytochemical detection and immunoreaction assay. Both approaches confirmed the enzymatic assay. We suggest this physiological characteristic allows tank bromeliads to survive in a nitrogen-limited environment, utilizing urea rapidly and efficiently and competing successfully for this nitrogen source against microorganisms that live in the tank water. PMID:19508368

Aguetoni Cambuí, Camila; Gaspar, Marília; Mercier, Helenice

2009-05-01

138

AC field induced cell membrane temperature gradients  

NASA Astrophysics Data System (ADS)

While generally inducing minimal heating in many biomedical applications, electric fields may still induce significant temperature gradients, particularly for pulses of short duration and AC (sinusoidal) fields of high frequency, such as microwaves. This paper extends a recent analysis of temperature gradients across a biological cell and membrane for single pulses [(A. L. Garner, et al., J. Appl. Phys. 113, 214701 (2013).] to multiple pulses or AC fields where the time between the two pulses, or the period for AC signals, is shorter than the thermal diffusion time. We calculate profiles of the induced temperature changes and gradients across a biological cell for AC wave of different frequencies and show that the location of the peak temperature and gradient shifts toward the center of the cell during subsequent half-waves. Higher frequency fields induce higher temperature gradients with the temperature gradient shifts toward the center of the cell for subsequent cycles.

Garner, Allen L.; Deminsky, Maxim; Neculaes, V. Bogdan; Potapkin, Boris

2014-03-01

139

Fuel cell membrane hydration and fluid metering  

DOEpatents

A hydration system includes fuel cell fluid flow plate(s) and injection port(s). Each plate has flow channel(s) with respective inlet(s) for receiving respective portion(s) of a given stream of reactant fluid for a fuel cell. Each injection port injects a portion of liquid water directly into its respective flow channel. This serves to hydrate at least corresponding part(s) of a given membrane of the corresponding fuel cell(s). The hydration system may be augmented by a metering system including flow regulator(s). Each flow regulator meters an injecting at inlet(s) of each plate of respective portions of liquid into respective portion(s) of a given stream of fluid by corresponding injection port(s).

Jones, Daniel O. (Glenville, NY); Walsh, Michael M. (Fairfield, CT)

2003-01-01

140

Fuel cell membrane hydration and fluid metering  

DOEpatents

A hydration system includes fuel cell fluid flow plate(s) and injection port(s). Each plate has flow channel(s) with respective inlet(s) for receiving respective portion(s) of a given stream of reactant fluid for a fuel cell. Each injection port injects a portion of liquid water directly into its respective flow channel in order to mix its respective portion of liquid water with the corresponding portion of the stream. This serves to hydrate at least corresponding part(s) of a given membrane of the corresponding fuel cell(s). The hydration system may be augmented by a metering system including flow regulator(s). Each flow regulator meters an injecting at inlet(s) of each plate of respective portions of liquid into respective portion(s) of a given stream of fluid by corresponding injection port(s).

Jones, Daniel O. (Glenville, NY); Walsh, Michael M. (Fairfield, CT)

1999-01-01

141

Fluorescence imaging of membrane dynamics in living cells  

NASA Astrophysics Data System (ADS)

Methods of wide-field fluorescence microscopy for measuring membrane dynamics of living cells are described, including spectral imaging as well as anisotropy imaging of the membrane marker 6-dodecanoyl-2-dimethylamino naphthalene (laurdan). Plasma membranes are selected by illumination with an evanescent electromagnetic field and distinguished from intracellular membranes assessed by whole-cell illumination. While fluorescence spectra of laurdan appeared red-shifted with decreasing membrane stiffness, fluorescence anisotropy and rotational correlation times were reduced with increasing membrane fluidity. Membrane stiffness was found to increase with decreasing temperature and increasing amounts of cholesterol and was always higher for the plasma membrane than for intracellular membranes. These effects may have some clinical relevance in the research of drug resistance or cell aging.

Weber, Petra; Wagner, Michael; Schneckenburger, Herbert

2010-07-01

142

Plasma membrane micro domains regulate TACE-dependent TNFR1 shedding in human endothelial cells  

PubMed Central

Upon stimulation by histamine, human vascular endothelial cells (EC) shed a soluble form of TNFR1 (sTNFR1) that binds up free TNF, dampening the inflammatory response. Shedding occurs through proteolytic cleavage of plasma membrane-expressed TNFR1 catalyzed by TNF-? converting enzyme (TACE). Surface expressed TNFR1 on EC is largely sequestered into specific plasma membrane micro domains, the lipid rafts/caveolae. The purpose of this study was to determine the role of these domains in TACE-mediated TNFR1 shedding in response to histamine. Human Umbilical Vein Endothelial Cells (HUVEC)-derived EA.hy926 cells respond to histamine via H1 receptors to shed TNFR1. Both depletion of cholesterol by methyl-?-cyclodextrin (MBCD) and siRNA knockdown of the scaffolding protein caveolin-1 (cav-1), treatments that disrupt caveolae, reduce histamine-induced shedding of membrane-bound TNFR1. Moreover, immunoblotting of discontinuous sucrose gradient fractions show that TACE, like TNFR1, is present within low density membrane fractions, concentrated within caveolae, in unstimulated EA.hy926 endothelial cells and co-immunoprecipitates with cav-1. Silencing of cav-1 reduces the levels of both TACE and TNFR1 protein and displaces TACE, from low density membrane fractions where TNFR1 remains. In summary, we show that endothelial lipid rafts/caveolae co-localize TACE to surface expressed TNFR1, promoting efficient shedding of sTNFR1 in response to histamine.

D'Alessio, Alessio; Esposito, Bianca; Giampietri, Claudia; Ziparo, Elio; Pober, Jordan S.; Filippini, Antonio

2011-01-01

143

Polymer synthesis toward fuel cell membrane materials  

NASA Astrophysics Data System (ADS)

Fuel cells are a promising technology that will be part of the future energy landscape. New membranes for alkaline and proton exchange membrane fuel cells are needed to improve the performance, simplify the system, and reduce cost. Polymer chemistry can be applied to develop new polymers and to assemble polymers into improved membranes that need less water, have increased performance and are less expensive, thereby removing the deficiencies of current membranes. Nucleophilic aromatic substitution polymerization typically produces thermally stable engineering polymers that can be easily functionalized. New functional monomers were developed to explore new routes to novel functional polymers. Sulfonamides were discovered as new activating groups for polymerization of high molecular weight thermooxidatively stable materials with sulfonic acid latent functionality. While the sulfonamide functional polymers could be produced, the sulfonamide group proved to be too stable to convert into a sulfonic acid after reaction. The reactivity of 2-aminophenol was investigated to search for a new class of ion conducting polymer materials. Both the amine and the phenol groups are found to be reactive in a nucleophilic aromatic substitution, however not to the extent to allow the formation of high molecular weight polymer materials. Layer-by-layer films were assembled from aqueous solutions of poly(styrene sulfonate) and trimethylammonium functionalized poly(phenylene oxide). The deposition conditions were adjusted to increase the free charge carrier content, and chloride conductivites reached almost 30 mS/cm for the best films. Block and random poly(phenylene oxide) copolymers were produced from 2,6-dimethylphenol and 2,6-diphenylphenol and the methyl substituted repeat units were functionalized with trimethylammonium bromide. The block copolymers displayed bromide conductivities up to 26 mS/cm and outperformed the random copolymers, indicating that morphology has an effect on ion transport.

Rebeck, Nathaniel T.

144

The chemical composition of wool. XV. The cell membrane couplex.  

PubMed

The cell membrane complex of wool has been examined by electron microscopy of stained cross sections after immersion of the wool in formic acid. The cell membrane complex of the cortex is considerably modified by the treatment, but that of the cuticle appears unchanged. Resistant membranes from cuticle cells, cortical cells and wool have been prepared by treatment with performic acid-ammonia. Amino acid analyses show that the resistant membranes from the cuticle contain citrulline but those from cortical cells do not. It is concluded that the cell membrane complex of the cuticle differs from that of the cortex. Because of the high lysine content of the resistant membranes, their resistance to chemical attack, the hydrophobicity of epicuticle and the observation of a small amount of epsilon-(gamma-glutamyl)lysine, it is postulated that the resistant membranes may contain an appreciable amount of epsilon-(gamma-glutamyl)lysine cross links. PMID:60989

Peters, D E; Bradbury, J H

1976-03-01

145

Biodegradation characteristics and size fractionation of landfill leachate for integrated membrane treatment.  

PubMed

The fate of organics and nitrogen during the biological treatment with MBR and subsequent membrane filtration processes (nano filtration, NF; reverse osmosis, RO) were investigated for a landfill leachate. The chemical oxygen demand (COD) and total Kjeldahl nitrogen (TKN) removal performances of membrane bioreactor (MBR) were obtained to be around 89% and 85%, respectively. The effluent COD of MBR was measured to be 1935 mg/L (30 kDa) which is much lower than experimentally determined soluble inert COD of 3200 mg/L using 0.45 ?m filter. The readily and slowly biodegradable COD fractions were estimated to be 17% and 52% of raw influent COD, respectively. The respirometry based modeling test performed on raw leachate exhibited much slower degradation kinetics compared to municipal wastewater. A unique subset of model parameters was extracted from batch respirometry by using acclimated MBR sludge. The sequential ultrafiltration (UF) experiments (particle size distribution, PSD) revealed that most of the organics was below 2 nm filter mesh size. In addition, NF/RO post treatment after MBR system was required to increase COD and total nitrogen (TN) removal performances up to 99%. Relatively lower salt rejection rates around 94% was obtained for RO system as a post treatment of MBR system. PMID:23856313

Insel, Güçlü; Dagdar, Mina; Dogruel, Serdar; Dizge, Nadir; Ubay Cokgor, Emine; Keskinler, Bülent

2013-09-15

146

Magnetically responsive microflaps reveal cell membrane boundaries from multiple angles.  

PubMed

A microflap system to incline adherent cells in the desired orientation is described. Inclination angles of cell-laden microflaps are precisely controlled by the applied magnetic field, enabling us to observe cell-membrane boundaries from multiple angles. This system is equipped with conventional microscopes, allowing clear focused images of cell-membrane boundaries to be obtained with high magnification. PMID:24677083

Teshima, Tetsuhiko; Onoe, Hiroaki; Aonuma, Hiroka; Kuribayashi-Shigetomi, Kaori; Kamiya, Koki; Tonooka, Taishi; Kanuka, Hirotaka; Takeuchi, Shoji

2014-05-01

147

Fractionation of protein hydrolysates of fish and chicken using membrane ultrafiltration: investigation of antioxidant activity.  

PubMed

In this work, chicken and fish peptides were obtained using the proteolytic enzymes ?-Chymotrypsin and Flavourzyme. The muscle was hydrolyzed for 4 h, and the resulting peptides were evaluated. Hydrolysates were produced from Argentine croaker (Umbrina canosai) with a degree of hydrolysis (DH) of 25.9 and 27.6% and from chicken (Gallus domesticus) with DH of 17.8 and 20.6% for Flavourzyme and ?-Chymotrypsin, respectively. Membrane ultrafiltration was used to separate fish and chicken hydrolysates from Flavourzyme and ?-Chymotrypsin based on molecular weight cutoff of >1,000, <1,000 and >500, and <500 Da, to produce fractions (F1,000, F1,000-500, and F500) with antioxidant activity. Fish hydrolysates produced with Flavourzyme (FHF) and ?-Chymotrypsin showed 60.8 and 50.9% of peptides with a molecular weight of <3 kDa in its composition, respectively. To chicken hydrolysates produced with Flavourzyme and ?-Chymotrypsin (CHC) was observed 83 and 92.4% of peptides with a molecular weight of <3 kDa. The fraction that showed, in general, higher antioxidant potential was F1,000 from FHF. When added 40 mg/mL of FHF and CHC, 93 and 80% of lipid oxidation in ground beef homogenates was inhibited, respectively. The composition of amino acids indicated higher amino acids hydrophobic content and amino acids containing sulfuric residues for FHF, which showed antioxidant potential. PMID:24449375

Centenaro, Graciela Salete; Salas-Mellado, Myriam; Pires, Carla; Batista, Irineu; Nunes, Maria L; Prentice, Carlos

2014-03-01

148

Triton X-114 phase fractionation of an integral membrane surface protein mediating monoclonal antibody killing of Mycoplasma hyorhinis.  

PubMed Central

A previously defined immunoglobulin M(kappa) monoclonal antibody reacting with a surface epitope of Mycoplasma hyorhinis is shown in this report to mediate specific, complement-dependent mycoplasmacidal activity. Immunoblot analysis of mycoplasma components and their tryptic cleavage products showed that the epitope recognized was present on a protein with an apparent molecular weight of 23,000 (p23) and on a limit tryptic fragment of this protein with an apparent molecular weight of 18,000 (p18). Both p23 and p18 are shown by Triton X-114 phase fractionation to partition efficiently into the hydrophobic detergent phase. Other antigens bearing epitopes not expressed at the cell surface were present among the numerous hydrophilic proteins found in the aqueous phase. The external orientation and membrane association of the p23 antigen were further established by demonstrating that trypsin treatment of intact mycoplasmas generated the antigenic p18 fragment, which remained tightly associated with the organism. These results localize an epitope responsible for antibody-mediated mycoplasma killing onto a specific, surface-exposed region of an integral membrane protein of this organism. Since the monoclonal antibody used in this study does not bind to the surface of all strains of M. hyorhinis, the epitope identified also defines a structural marker of antigenic surface variation within this species, a feature previously observed during serological classification of the organism. Analysis of the antigenic and structural features of the p23 surface antigen may therefore be useful in establishing mechanisms of surface antigen variation among integral membrane proteins of mycoplasmas that could dictate important antigenic characteristics recognized during chronic disease caused by these agents. Images

Riethman, H C; Boyer, M J; Wise, K S

1987-01-01

149

Immunization Against Coccidioidomycosis by Killed Cell and Cell Fraction Vaccines.  

National Technical Information Service (NTIS)

Aerosol and subcutaneous vaccination with killed arthrosporess and subcutaneous vaccination using a boivin-type fraction were compared for their efficacy in protecting rhesus monkeys against lethal aerosol challenge with C. immitis. Complete protection ag...

J. T. Sinski E. P. Lowe N. F. Conant H. F. Hardin M. W. Castleberry

1964-01-01

150

Membrane disorder and phospholipid scrambling in electropermeabilized and viable cells.  

PubMed

Membrane electropermeabilization relies on the transient permeabilization of the plasma membrane of cells submitted to electric pulses. This method is widely used in cell biology and medicine due to its efficiency to transfer molecules while limiting loss of cell viability. However, very little is known about the consequences of membrane electropermeabilization at the molecular and cellular levels. Progress in the knowledge of the involved mechanisms is a biophysical challenge. As a transient loss of membrane cohesion is associated with membrane permeabilization, our main objective was to detect and visualize at the single-cell level the incidence of phospholipid scrambling and changes in membrane order. We performed studies using fluorescence microscopy with C6-NBD-PC and FM1-43 to monitor phospholipid scrambling and membrane order of mammalian cells. Millisecond permeabilizing pulses induced membrane disorganization by increasing the translocation of phosphatidylcholines according to an ATP-independent process. The pulses induced the formation of long-lived permeant structures that were present during membrane resealing, but were not associated with phosphatidylcholine internalization. These pulses resulted in a rapid phospholipid flip/flop within less than 1s and were exclusively restricted to the regions of the permeabilized membrane. Under such electrical conditions, phosphatidylserine externalization was not detected. Moreover, this electrically-mediated membrane disorganization was not correlated with loss of cell viability. Our results could support the existence of direct interactions between the movement of membrane zwitterionic phospholipids and the electric field. PMID:24583083

Escoffre, Jean-Michel; Bellard, Elisabeth; Faurie, Cécile; Sébaï, Sarra C; Golzio, Muriel; Teissié, Justin; Rols, Marie-Pierre

2014-07-01

151

Water and methanol uptakes in Nafion membranes and membrane effects on direct methanol cell performance  

Microsoft Academic Search

This paper compares direct methanol fuel cells (DMFCs) employing two types of Nafion{reg{underscore}sign} (E.I.DuPont de Nemours and Company) membranes of different equivalent weight (EW). Methanol and water uptakes in 1,100 and 1,200 EW Nafion membranes were determined by weighing PâOâ-dried and methanol solution-equilibrated membranes. Both methanol and water uptakes in the 1,200 EW membrane were about 70--74% of those in

X. Ren; T. E. Springer; S. Gottesfeld

2000-01-01

152

Variation in the lipid and fatty acid composition in purified membrane fractions from Sarcina aurantiaca in relation to growth phase  

Microsoft Academic Search

S. aurantiaca membrane lipid contains both branched and straight-chain fatty acids from C9 to C22 with the saturated branched C15 predominating in almost all of the lipid fractions studied. Unsaturated fatty acids are only present in low concentrations. Significant amounts of straight-chain, even-numbered acids, more common in gram-negative and gram-variable bacteria, are also present. All lipid fractions show a marked

D. Thirkell; Elizabeth M. M. Gray

1974-01-01

153

Membrane Purification Cell for Aluminum Recycling  

SciTech Connect

Recycling mixed aluminum scrap usually requires adding primary aluminum to the scrap stream as a diluent to reduce the concentration of non-aluminum constituents used in aluminum alloys. Since primary aluminum production requires approximately 10 times more energy than melting scrap, the bulk of the energy and carbon dioxide emissions for recycling are associated with using primary aluminum as a diluent. Eliminating the need for using primary aluminum as a diluent would dramatically reduce energy requirements, decrease carbon dioxide emissions, and increase scrap utilization in recycling. Electrorefining can be used to extract pure aluminum from mixed scrap. Some example applications include producing primary grade aluminum from specific scrap streams such as consumer packaging and mixed alloy saw chips, and recycling multi-alloy products such as brazing sheet. Electrorefining can also be used to extract valuable alloying elements such as Li from Al-Li mixed scrap. This project was aimed at developing an electrorefining process for purifying aluminum to reduce energy consumption and emissions by 75% compared to conventional technology. An electrolytic molten aluminum purification process, utilizing a horizontal membrane cell anode, was designed, constructed, operated and validated. The electrorefining technology could also be used to produce ultra-high purity aluminum for advanced materials applications. The technical objectives for this project were to: - Validate the membrane cell concept with a lab-scale electrorefining cell; - Determine if previously identified voltage increase issue for chloride electrolytes holds for a fluoride-based electrolyte system; - Assess the probability that voltage change issues can be solved; and - Conduct a market and economic analysis to assess commercial feasibility. The process was tested using three different binary alloy compositions (Al-2.0 wt.% Cu, Al-4.7 wt.% Si, Al-0.6 wt.% Fe) and a brazing sheet scrap composition (Al-2.8 wt.% Si-0.7 wt.% Fe-0.8 wt.% Mn),. Purification factors (defined as the initial impurity concentration divided by the final impurity concentration) of greater than 20 were achieved for silicon, iron, copper, and manganese. Cell performance was measured using its current and voltage characteristics and composition analysis of the anode, cathode, and electrolytes. The various cells were autopsied as part of the study. Three electrolyte systems tested were: LiCl-10 wt. % AlCl3, LiCl-10 wt. % AlCl3-5 wt.% AlF3 and LiF-10 wt.% AlF3. An extended four-day run with the LiCl-10 wt.% AlCl3-5 wt.% AlF3 electrolyte system was stable for the entire duration of the experiment, running at energy requirements about one third of the Hoopes and the conventional Hall-Heroult process. Three different anode membranes were investigated with respect to their purification performance and survivability: a woven graphite cloth with 0.05 cm nominal thickness & > 90 % porosity, a drilled rigid membrane with nominal porosity of 33%, and another drilled rigid graphite membrane with increased thickness. The latter rigid drilled graphite was selected as the most promising membrane design. The economic viability of the membrane cell to purify scrap is sensitive to primary & scrap aluminum prices, and the cost of electricity. In particular, it is sensitive to the differential between scrap and primary aluminum price which is highly variable and dependent on the scrap source. In order to be economically viable, any scrap post-processing technology in the U.S. market must have a total operating cost well below the scrap price differential of $0.20-$0.40 per lb to the London Metal Exchange (LME), a margin of 65%-85% of the LME price. The cost to operate the membrane cell is estimated to be < $0.24/lb of purified aluminum. The energy cost is estimated to be $0.05/lb of purified aluminum with the remaining costs being repair and maintenance, electrolyte, labor, taxes and depreciation. The bench-scale work on membrane purification cell process has demonstrated technological advantages and subs

David DeYoung; James Wiswall; Cong Wang

2011-11-29

154

Fractions  

NSDL National Science Digital Library

This is an accessible, easy-to-read book introducing fractions. It can be downloaded in PowerPoint, Impress, and Flash formats. For struggling or non-readers the book can be read aloud in a variety of voices. All of the books on the Tar Heel Reader site can be used with the Intellikeys keyboard with a custom overlay, a touch screen, and/or 1-3 switches. The text and background colors can be modified for students with visual impairments.

Cowley, K.

2011-05-09

155

Estimation of fractional changes in peak g\\/sub Na\\/, g\\/sub Na\\/, E\\/sub Na\\/, and h\\/sub infinity \\/ (V) of cardiac cells from V\\/sub max\\/ of the propagating action potential  

Microsoft Academic Search

Fractional changes in the peak sodium conductances of the cardiac cell membrane during the action potential are often estimated from fractional changes in the minimum time derivative of the action potential upstroke (V max). The present model study shows that this approach is valid for propagating action potentials provided that the membrane capacitance does not change and that the nonsodium

FERNAND A. ROBERGE; LUC BOUCHER

1990-01-01

156

Binding and labeling of omega-conotoxin GVIA in crude membranes from subfractionated fractions and various areas of chick brain  

Microsoft Academic Search

Specific binding and specific labeling of125I-?-CgTX were investigated in crude membranes from both subfractionated fractions and various brain areas in chick whole brain.\\u000a The specific activities of the marker enzymes 2?,3?-cyclic nucleotide 3?-phosphorylase, Na\\/K ATPase and succinic dehydrogenase\\u000a in the subfractionated fractions were three- to five-fold higher than those in the P2 fraction. However, the amount of specific [125I]?-CgTX binding

Seiji Ichida; Tetsuyuki Wada; Kiyo Hashimoto; Yasunari Kasamatsu; Takafumi Akimoto; Miki Tahara

1996-01-01

157

Polarizability of red blood cells with an anisotropic membrane  

Microsoft Academic Search

We predict the complex polarizability of a realistic model of a red blood cell (RBC), with an inhomogeneous dispersive and anisotropic membrane. In this model, the frequency-dependent complex electrical parameters of the individual cell layers are described by the Debye equation while the dielectric anisotropy of the cell membrane is taken into account by the different permittivities along directions normal

José Luis Sebastián; Sagrario Muñoz; Miguel Sancho; Genoveva Martínez; Karan V. I. S. Kaler

2010-01-01

158

Anticancer effects of human amniotic membrane and its epithelial cells.  

PubMed

Anticancer property of the amniotic membrane, the innermost layer of fetal membrane, was previously hypothesized. The recent reports confirmed the published hypotheses and developed new hypotheses in this context. Based on some evidences, it is hypothesized that inducing of apoptosis in cancer cells is originated from amniotic epithelial cells and cancer cell cycle arrest arises from amniotic mesenchymal cells. We also hypothesized here that apoptosis and cell cycle arrest in cancer cells induced by amniotic membrane arise from release of soluble factors from amniotic cells. PMID:24556192

Niknejad, Hassan; Yazdanpanah, Ghasem

2014-04-01

159

Humidification studies on polymer electrolyte membrane fuel cell  

Microsoft Academic Search

Two methods of humidifying the anode gas, namely, external and membrane humidification, for a polymer electrolyte membrane fuel (PEMFC) cell are explained. It is found that the water of solvation of protons decreases with increase in the current density and the electrode area. This is due to insufficient external humidification. In a membrane-based humidification, an optimum set of parameters, such

P Sridhar; Ramkumar Perumal; N Rajalakshmi; M Raja; K. S Dhathathreyan

2001-01-01

160

HTO electrolysis method by using proton exchange membrane fuel cell  

Microsoft Academic Search

The application of a proton exchange membrane fuel cell (PEMFC) system to electrolysis for HTO recovered from a solid breeder blanket is discussed in this report. The amount of gas permeation through the membrane in this system and the tritium inventory in the membrane were calculated, and it was found that the effect of these phenomena on the performance of

Hiroki Takata; Masabumi Nishikawa; Takayuki Egawa; Nobukazu Mizuno

2007-01-01

161

Isolation of plasma membranes from purified mouse spermatogenic cells.  

PubMed

Plasma membranes have been prepared from purified pachytene spermatocytes, round spermatids and residual bodies of the adult mouse testis using procedures modified from other authors'. Isolated membranes have been examined using electron microscopy, lectin binding and enzymic assays. Ultrastructural observation reveals smooth unit-membrane vesicles from 0.4-1.7 micrometer diameter. No contamination by nuclei, mitochondria or lysosomes is detected microscopically. Radiolabelled lectin-binding experiments [125I-RCAI, 125I-green pea lectin] indicate that cell surface label cofractionates with material identified morphologically as plasma membrane. Estimates of total recovery of membrane, based upn the lectin data, average 33%. Biochemical analysis of subcellular markers reveal that no detectable DNA and only 1.2% of the total cellular RNA cofractionate with membranes. A variety of enzyme assays suggests little contamination by cytosol enzymes, Golgi material or mitochondria. Assays of 5'-nucleotidase (E.C. 3.1.3.5) indicate that this enzyme is not a major component of developing mouse spermatogenic cell membranes. Instead, Sertoli cells represent the most important source of this enzyme in the adult seminiferous tubule. Polyacrylamide gel analysis of membranes isolated from purified germ cells reveals significant differences in the protein compositions of pachytene spermatocyte and round spermatid membranes. The preparation of highly purified plasma membranes from homogeneous populations of spermatogenic cells should facilitate the biochemical characterization of cell surface antigens specific to developing male germ cells. PMID:7419622

Millette, C F; O'Brien, D A; Moulding, C T

1980-06-01

162

Xyloglucan biosynthesis by Golgi membranes from suspension-cultured sycamore (Acer pseudoplatanus) cells  

SciTech Connect

Xyloglucan is a major hemicellulose polysaccharide in plant cell walls. Biosynthesis of such cell wall polysaccharides is closely linked to the process of plant cell growth and development. Xyloglucan polysaccharides consist of a {beta}-1,4 glucan backbone synthesized by xyloglucan synthase and sidechains of xylose, galactose, and fucose added by other transferase enzymes. Most plant Golgi and plasma membranes also contain glucan synthases I II, which make {beta}-1,4 and {beta}-1,3 glucans, respectively. All of these enzymes have very similar activities. Cell walls on suspension-cultured cells from Acer pseudoplatanus (sycamore maple) were enzymatically softened prior to cell disruption by passing through a 30 {mu}m nylon screen. Cell membranes from homogenates were separated by ultracentrifugation on top-loaded or flotation sucrose density gradients. Samples were collected by gradient fractionation and assayed for membrane markers and xyloglucan and glucan synthase activities. Standard marker assays (cyt. c reductase for eR, IDPase UDPase for Golgi, and eosin 5{prime}-malelmide binding for plasma membrane) showed partial separation of these three membrane types. Golgi and plasma membrane markers overlapped in most gradients. Incorporation of {sup 14}C-labeled sugars from UDP-glucose and UDP-xylose was used to detect xyloglucan synthase, glucan synthases I II, and xylosyl transferase in Golgi membrane fractions. These activities overlapped, although distinct peaks of xyloglucan synthase and xylosyl transferase were found. Ca{sup ++} had a stimulatory effect on glucan synthases I II, while Mn{sup ++} had an inhibitory effect on glucan synthase I in the presence of Ca{sup ++}. The similarity of these various synthase activities demonstrates the need for careful structural characterization of newly synthesized polysaccharides.

White, A.R.; Xin, Yi (North Dakota State Univ., Fargo (USA))

1990-05-01

163

How helminths use excretory secretory fractions to modulate dendritic cells  

PubMed Central

It is well known that helminth parasites have immunomodulatory effects on their hosts. They characteristically cause a skew toward TH2 immunity, stimulate Treg cells while simultaneously inhibiting TH1 and TH17 responses. Additionally, they induce eosinophilia and extensive IgE release. The exact mechanism of how the worms achieve this effect have yet to be fully elucidated; however, parasite-derived secretions and their interaction with antigen presenting cells have been centrally implicated. Herein, we will review the effects of helminth excretory-secretory fractions on dendritic cells and discuss how this interaction is crucial in shaping the host response.

White, Rhiannon R.; Artavanis-Tsakonas, Katerina

2012-01-01

164

Nonhumidified High-Temperature Membranes Developed for Proton Exchange Membrane Fuel Cells  

NASA Technical Reports Server (NTRS)

Fuel cells are being considered for a wide variety of aerospace applications. One of the most versatile types of fuel cells is the proton-exchange-membrane (PEM) fuel cell. PEM fuel cells can be easily scaled to meet the power and space requirements of a specific application. For example, small 100-W PEM fuel cells are being considered for personal power for extravehicular activity suit applications, whereas larger PEM fuel cells are being designed for primary power in airplanes and in uninhabited air vehicles. Typically, PEM fuel cells operate at temperatures up to 80 C. To increase the efficiency and power density of the fuel cell system, researchers are pursuing methods to extend the operating temperature of the PEM fuel cell to 180 C. The most widely used membranes in PEM fuel cells are Nafion 112 and Nafion 117--sulfonated perfluorinated polyethers that were developed by DuPont. In addition to their relatively high cost, the properties of these membranes limit their use in a PEM fuel cell to around 80 C. The proton conductivity of Nafion membranes significantly decreases above 80 C because the membrane dehydrates. The useful operating range of Nafion-based PEM fuel cells can be extended to over 100 C if ancillary equipment, such as compressors and humidifiers, is added to maintain moisture levels within the membrane. However, the addition of these components reduces the power density and increases the complexity of the fuel cell system.

Kinder, James D.

2005-01-01

165

Structural Analysis of Sterol Distributions in the Plasma Membrane of Living Cells  

Microsoft Academic Search

Although plasma membrane (PM) cholesterol-rich and -poor domains have been isolated by subcellular fractionation, the real-time arrangement of cholesterol in such domains in living cells is still unclear. Therefore, dehydroergosterol (DHE), a naturally occurring fluorescent sterol, was incorporated into cultured L-cell fibroblasts. Two PM markers, the enhanced cyan fluorescent protein (ECFP-Mem) and 3'-dioctadecyloxacarbocyanine perchlorate (DiOC18(3)), were used to distinguish DHE

Weimin Zhang; Avery L. McIntosh; Hai Xu; Di Wu; Todd Gruninger; Barbara Atshaves; J. C. Steve Liu; Friedhelm Schroeder

2005-01-01

166

The application of Dow Chemical's perfluorinated membranes in proton-exchange membrane fuel cells  

NASA Technical Reports Server (NTRS)

Dow Chemical's research activities in fuel cells revolve around the development of perfluorosulfonic acid membranes useful as the proton transport medium and separator. Some of the performance characteristics which are typical for such membranes are outlined. The results of tests utilizing a new experimental membrane useful in proton-exchange membrane fuel cells are presented. The high voltage at low current densities can lead to higher system efficiencies while, at the same time, not sacrificing other critical properties pertinent to membrane fuel cell operation. A series of tests to determine response times indicated that on-off cycles are on the order of 80 milliseconds to reach 90 percent of full power. The IR free voltage at 100 amps/sq ft was determined and the results indicating a membrane/electrode package resistance to be .15 ohm-sq cm at 100 amps/sq ft.

Eisman, G. A.

1989-01-01

167

Organic\\/inorganic hybrid membranes for direct methanol fuel cells  

Microsoft Academic Search

Organic\\/inorganic hybrid membrane with silica supported heteropoly acid was applied to reduce methanol cross-over with keeping ionic conductivity high for direct methanol fuel cells (DMFCs). In situ micro-emulsion impregnation of silica supported heteropoly acid into a proton exchange membrane (PEM), Nafion®, resulted in a homogeneous distribution of proton conducting inorganic networks in the hybrid membrane. The hybrid membranes showed 50–80%

HaeKyoung Kim; Hyuk Chang

2007-01-01

168

Distribution of Intact and Core Membrane Lipids of Archaeal Glycerol Dialkyl Glycerol Tetraethers among Size-Fractionated Particulate Organic Matter in Hood Canal, Puget Sound  

PubMed Central

There is great interest in the membrane lipids of archaea (glycerol dialkyl glycerol tetraethers [GDGTs]) as tracers of archaeal biomass because of their utility as paleoproxies and because of the biogeochemical importance of archaea. While core GDGTs (formed by hydrolysis of polar head groups of intact GDGTs after cell death) are appropriate for paleostudies, they have also been used to trace archaeal populations. Also, despite the small size (0.2 by 0.7 ?m) of cultivated marine archaea, 0.7-?m glass-fiber filters (GFFs) are typically used to collect GDGTs from natural waters. We quantified both core and intact GDGTs in free-living (0.2- to 0.7-?m), suspended (0.7- to 60-?m), and aggregate (>60-?m) particle size fractions in Puget Sound (Washington State). On average, the free-living fraction contained 36% of total GDGTs, 90% of which were intact. The intermediate-size fraction contained 62% of GDGTs, and 29% of these were intact. The aggregate fraction contained 2% of the total GDGT pool, and 29% of these were intact. Our results demonstrate that intact GDGTs are largely in the free-living fraction. Because only intact GDGTs are present in living cells, protocols that target this size fraction and analyze the intact GDGT pool are necessary to track living populations in marine waters. Core GDGT enrichment in larger-size fractions indicates that archaeal biomass may quickly become attached or entrained in particles once the archaea are dead or dying. While the concentrations of the two pools were generally not correlated, the similar sizes of the core and intact GDGT pools suggest that core GDGTs are removed from the water column on timescales similar to those of cell replication, on timescales of days to weeks.

Huguet, Carme; Truxal, Laura T.

2012-01-01

169

Chemicals and energy co-generation from direct hydrocarbons/oxygen proton exchange membrane fuel cell  

NASA Astrophysics Data System (ADS)

A proton exchange membrane fuel cell for chemicals and energy co-generation was set up with hydrocarbons ethane, propane and butane as fuels, and the electrochemical performance of the cell was studied by using linear potential sweep, alternating current impedance and gas chromatography. The cell performance can be improved to a great extent by increasing the platinum load in the catalyst, by treating the membrane with phosphoric acid and by elevating temperature. The improvement of cell performance by the increase of platinum load is ascribed to the increase of reaction sites for hydrocarbon oxidation, that by phosphoric acid treatment to the increase of proton conductivity in Nafion membrane, and that by elevating temperature to the improvement in thermodynamic as well as kinetic aspects. Only a small fraction of the hydrocarbon is converted to carbon dioxide in this cell during its power generation. The current efficiency is 5% for the conversion of ethane to carbon dioxide in the ethane/oxygen fuel cell with 20% carbon-supported platinum as catalyst and phosphoric acid-treated membrane as proton exchange membrane at 0.2 V, 80 °C and ambient pressure. The reaction activity of hydrocarbons at the anode is in the order of propane, butane and ethane. The possible chemicals produced from the cell were hydrocarbons with more than six carbons, which are inactive at the anode under cell conditions.

Li, W. S.; Lu, D. S.; Luo, J. L.; Chuang, K. T.

170

Separation of Human Skin Cells by Velocity Sedimentation into Functionally Distinct Fractions  

Microsoft Academic Search

The epidermis consists of a heterogeneous population of cells including Langerhans cells, Merkel cells, melanocytes, and keratinocytes in various stages of differentiation. The current study was undertaken to determine if skin cell suspensions can be separated into morphologically and\\/or functionally distinct fractions. Skin cells were suspended by trypsinization and separated into multiple fractions by velocity sedimentation. Certain fractions reproducibly stimulated

Vera B. Morhenn; Eric D. Starr; Candace Terrell; Alvin J. Cox; Edgar G. Engleman

1982-01-01

171

Conductivity Measurements of Synthesized Heteropoly Acid Membranes for Proton Exchange Membrane Fuel Cells  

SciTech Connect

Fuel cell technology is receiving attention due to its potential to be a pollution free method of electricity production when using renewably produced hydrogen as fuel. In a Proton Exchange Membrane (PEM) fuel cell H2 and O2 react at separate electrodes, producing electricity, thermal energy, and water. A key component of the PEM fuel cell is the membrane that separates the electrodes. DuPont’s Nafion® is the most commonly used membrane in PEM fuel cells; however, fuel cell dehydration at temperatures near 100°C, resulting in poor conductivity, is a major hindrance to fuel cell performance. Recent studies incorporating heteropoly acids (HPAs) into membranes have shown an increase in conductivity and thus improvement in performance. HPAs are inorganic materials with known high proton conductivities. The primary objective of this work is to measure the conductivity of Nafion, X-Ionomer membranes, and National Renewable Energy Laboratory (NREL) Developed Membranes that are doped with different HPAs at different concentrations. Four-point conductivity measurements using a third generation BekkTech? conductivity test cell are used to determine membrane conductivity. The effect of multiple temperature and humidification levels is also examined. While the classic commercial membrane, Nafion, has a conductivity of approximately 0.10 S/cm, measurements for membranes in this study range from 0.0030 – 0.58 S/cm, depending on membrane type, structure of the HPA, and the relative humidity. In general, the X-ionomer with H6P2W21O71 HPA gave the highest conductivity and the Nafion with the 12-phosphotungstic (PW12) HPA gave the lowest. The NREL composite membranes had conductivities on the order of 0.0013 – 0.025 S/cm.

Record, K.A.; Haley, B.T.; Turner, J.

2006-01-01

172

Chromatin fractionation analysis of licensing factors in Mammalian cells.  

PubMed

ORC, Cdc6, Cdt1, and MCM2-7 are replication-licensing factors, which play a central role in the once-per-cell cycle control of DNA replication. ORC, Cdc6, and Cdt1 collaborate to load MCM2-7 onto replication origins in order to license them for replication. MCM2-7 is a DNA helicase directly involved in DNA replication and dissociates from DNA as S phase progresses and each replicon is replicated. In the cell cycle, the loading of MCM2-7 is restricted during the end of mitosis and the G1 phase. Thus, the levels of chromatin-bound MCM2-7 and its loaders oscillate during the cell cycle. Chromatin association of these factors can be analyzed by separating a cell lysate into soluble and chromatin-enriched insoluble fractions in mammalian cells. PMID:24906333

Nishitani, Hideo; Morino, Masayuki; Murakami, Yusuke; Maeda, Takeshi; Shiomi, Yasushi

2014-01-01

173

Water and methanol uptakes in Nafion membranes and membrane effects on direct methanol cell performance  

SciTech Connect

This paper compares direct methanol fuel cells (DMFCs) employing two types of Nafion{reg{underscore}sign} (E.I.DuPont de Nemours and Company) membranes of different equivalent weight (EW). Methanol and water uptakes in 1,100 and 1,200 EW Nafion membranes were determined by weighing P{sub 2}O{sub 5}-dried and methanol solution-equilibrated membranes. Both methanol and water uptakes in the 1,200 EW membrane were about 70--74% of those in the 1,100 EW membrane. The methanol crossover rate corresponding to that in a DMFC at open circuit was measured using a voltammetric method in the DMFC configuration and under the same cell operating conditions. After accounting for the thickness difference between the membrane samples, the methanol crossover rate through a 1,200 EW membrane was 52% of that through an 1,100 EW membrane. To resolve the cathode and anode performances in an operating DMFC, a dynamic hydrogen electrode was used as a reference electrode. Results show that in an operating DMFC the cathode can be easily flooded, as shown in a DMFC using 1,100 EW membrane. An increase in methanol crossover rate decreases the DMFC cathode potential at open circuit. At a high cell current density, the DMFC cathode potential can approach that of a H{sub 2}/air cell.

Ren, X.; Springer, T.E.; Gottesfeld, S.

2000-01-01

174

New ETFE-based membrane for direct methanol fuel cell  

Microsoft Academic Search

The investigated membranes are based on 35?? m thick commercial poly(ethylene-alt-tetrafluoroethylene) (ETFE) films. The films were made proton conductive by means of irradiation treatment followed by sulfonation. These membranes have exceptionally low water uptake and excellent dimensional stability. The new membranes are investigated widely in a laboratory-scale direct methanol fuel cell (DMFC). The temperature range used in the fuel cell

V. Saarinen; T. Kallio; M. Paronen; P. Tikkanen; E. Rauhala; K. Kontturi

2005-01-01

175

BIOCHEMICAL AND MORPHOLOGICAL COMPARISON OF PLASMA MEMBRANE AND MILK FAT GLOBULE MEMBRANE FROM BOVINE MAMMARY GLAND  

PubMed Central

Purified plasma membrane fractions from lactating bovine mammary glands and membranes of milk fat globules from the same source were similar in distribution and fatty acid composition of phospholipids. The sphingomyelin content of the phospholipid fraction of both membranes was higher than in these fractions from other cell components, ?-carotene, a constituent characteristic of milk fat, was present in the lipid fraction of the plasma membrane. Cholesterol esters of plasma membrane were similar in fatty acid composition to those of milk fat globule membranes. Disc electrophoresis of either membrane preparation on polyacrylamide gels revealed a single major protein component characteristic of plasma membrane from other sources. Distinct morphological differences between plasma membrane and milk fat globule membranes were observed in both thin sections and in negatively stained material. Plasma membrane was vesicular in appearance while milk fat globule membranes had a platelike aspect. These observations are consistent with derivation of fat globule membrane from plasma membrane accompanied by structural rearrangement of membrane constituents.

Keenan, T. W.; Morre, D. James; Olson, Diane E.; Yunghans, W. N.; Patton, Stuart

1970-01-01

176

In situ proton exchange membrane fuel cell durability of poly(vinylidene fluoride)\\/polyelectrolyte blend Arkema M43 membrane  

Microsoft Academic Search

A typical perfluorosulfonic acid (PFSA) polymer electrolyte membrane is composed of a single type of polymer in order to meet the strict requirements for a fuel cell membrane. The Arkema Inc. membrane technology provides a simple and lower cost route to the design of durable membrane materials. The membrane employs two intimately mixed polymers: Kynar® PVDF, which provides excellent mechanical

Tao Zhang; Wensheng He; James Goldbach; David Mountz; Jung Yi

177

Electron-beam direct processing on living cell membrane  

SciTech Connect

We demonstrated a direct processing on a living Hep G2 cell membrane in conventional cultivation conditions using an electron beam. Electron beam-induced deposition from liquid precursor 3,4-ethylenedioxythiophene and ablation was performed on the living cells. The 2.5-10 keV electron beam which was irradiated through a 100-nm-thick SiN nanomembrane could induce a deposition pattern and a ablation on a living cell membrane. This electron beam direct processing can provide simple in-situ cell surface modification for an analytical method of living cell membrane dynamic.

Hoshino, Takayuki [Department of Bio-Applications and Systems Engineering, Tokyo University of Agriculture and Technology, 2-24-16 Naka-cho, Koganei, Tokyo 184-8588 (Japan); Morishima, Keisuke [Department of Bio-Applications and Systems Engineering, Tokyo University of Agriculture and Technology, 2-24-16 Naka-cho, Koganei, Tokyo 184-8588 (Japan); Department of Mechanical Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871 Japan (Japan)

2011-10-24

178

Hemoglobin s polymerization and red cell membrane changes.  

PubMed

Different pathways lead from the simple point mutation in hemoglobin to the membrane changes that characterize the altered interaction of the sickle red blood cell with its environment, including endothelial cells, white blood cells, and platelets. Polymerization and oxidation-induced damage to both lipid and protein components of the red cell membrane, as well as the generation of bioreactive membrane material (microparticles), has a profound effect on all tissues and organs, and defines the vasculopathy of the patient with sickle cell disease. PMID:24589260

Kuypers, Frans A

2014-04-01

179

Cytolytic peptides induce biphasic permeability changes in mammalian cell membranes.  

PubMed

The cytolytic peptides melittin and gramicidin S are naturally occurring agents that provide a comparative model for studies of complement, immunotoxin and cell-mediated membrane permeability. Most attempts to characterize cytolytic peptides have used model membrane systems including phospholipid vesicles or erythrocytes. Membrane vesicles permit the use of self-quenching concentrations of fluorescent permeability markers, while erythrocytes release measurable hemoglobin. Attempts at measuring early membrane permeability changes in nucleated mammalian cells have been limited. To measure the kinetics of mammalian cell membrane permeability changes induced by cytolytic peptides, we developed a 96-well fluorescence cytolysis assay using the cytoplasmic fluorescent dye calcein as the membrane permeability marker. To facilitate rapid assessment of membrane permeability, trypan blue was added to the assay solution to quench (a) released fluorescence and (b) retained intracellular fluorescence. Trypan blue also provided a complementary visual assessment of cell viability. Using this assay, a detailed kinetic analysis demonstrated permeability of the cell membranes within seconds of exposure to the cytolytic peptides. The rapid permeabilization of the cell membranes was confirmed by flow cytometry using the calcium indicator dye fluo-3. The assay also demonstrated a second slower phase of marker release over the next several hours. The fluorescence cytolysis assay was able to reliably detect the biphasic permeability changes associated with the melittin and gramicidin S peptides suggesting the potential utility of this assay in the assessment of other cytolytic agents. PMID:11334966

Su, M; He, C; West, C A; Mentzer, S J

2001-06-01

180

Self-humidifying polymer electrolyte membranes for fuel cells  

Microsoft Academic Search

Polymer electrolyte fuel cells have attracted enormous interest as a primary power source for electric vehicles. Water management in the electrolyte is one of the complicated problems to be overcome. A new self-humidifying electrolyte membrane is proposed to solve this problem. Self-humidification allows the use of very thin membranes, simultaneously allowing high performance of the cell. Use of the new,

Masahiro Watanabe; Hiroyuki Uchida; Yasuhiro Seki; Masaomi Emori; P. Stonehart

1996-01-01

181

Mechanisms of cell membrane permeabilization with ultrasound and contrast microbubbles  

Microsoft Academic Search

New clinical applications of ultrasound contrast agents extend beyond imaging and diagnostic towards therapeutic applications. A number of experimental findings have now demonstrated evidence of increased cell membrane permeability through sonoporation process. To explore the mechanisms by which the activation of microbubbles with ultrasound waves breach cell membranes, an electrophysiological experimental method is set up. The method consists of measuring

T. A. Tran; S. Roger; J. Y. Le Guennec; F. Tranquart; A. Bouakaz

2005-01-01

182

Effect of EMP fields on cell membrane potentials  

SciTech Connect

A simple model is presented for cell membrane potentials induced during exposure to electromagnetic pulse (EMP). Using calculated values of internal electric field strength induced during EMP exposure, the model predicts that cell membrane potentials of about 100 mV may be induced for time frames on the order of 10 ns. Possible biological effects of these potentials including electroporation area discussed.

Gailey, P.C.; Easterly, C.E.

1993-06-01

183

The functional activity of cell membranes in multiple sclerosis  

Microsoft Academic Search

The approach of the present study was determined by the marked interest in recent years in the problems of the pathogenesis of a number of diseases from the perspectives of the pathological chemistry of cell membranes. Studies devoted to the investigation of the structure and function of cell membranes in progressive muscular dystrophies [2, 15], disturbances in cerebral circulation [8,

P. V. Predtechenskaya; A. P. Ierusalimskii

1993-01-01

184

THE ENZYMATIC IODINATION OF THE RED CELL MEMBRANE  

Microsoft Academic Search

An enzymatic iodination procedure utilizing lactoperoxidase (LPO), radioactive iodide, and hydrogen peroxide generated by a glucose oxidase-glucose system has been described and utilized for a study of the red cell membrane . 97 % of the incorporated isotope is in the erythrocyte ghost and 3 % is associated with hemoglobin . No significant labeling of the red cell membrane occurs

ANN L. HUBBARD; ZANVIL A. COHN

1972-01-01

185

Finite element analysis of microelectrotension of cell membranes  

PubMed Central

Electric fields can be focused by micropipette-based electrodes to induce stresses on cell membranes leading to tension and poration. To date, however, these membrane stress distributions have not been quantified. In this study, we determine membrane tension, stress, and strain distributions in the vicinity of a microelectrode using finite element analysis of a multiscale electro-mechanical model of pipette, media, membrane, actin cortex, and cytoplasm. Electric field forces are coupled to membranes using the Maxwell stress tensor and membrane electrocompression theory. Results suggest that micropipette electrodes provide a new non-contact method to deliver physiological stresses directly to membranes in a focused and controlled manner, thus providing the quantitative foundation for micreoelectrotension, a new technique for membrane mechanobiology.

Bae, Chilman

2011-01-01

186

Preparation and performance of nano silica\\/Nafion composite membrane for proton exchange membrane fuel cells  

Microsoft Academic Search

Composite membranes made from Nafion ionomer with nano phosphonic acid-functionalised silica and colloidal silica were prepared and evaluated for proton exchange membrane fuel cells (PEMFCs) operating at elevated temperature and low relative humidity (RH). The phosphonic acid-functionalised silica additive obtained from a sol–gel process was well incorporated into Nafion membrane. The particle size determined using transmission electron microscope (TEM) had

Keping Wang; Scott McDermid; Jing Li; Natalia Kremliakova; Paul Kozak; Chaojie Song; Yanghua Tang; Jianlu Zhang; Jiujun Zhang

2008-01-01

187

Membrane Nanowaves in Single and Collective Cell Migration  

PubMed Central

We report the characterization of three-dimensional membrane waves for migrating single and collective cells and describe their propagation using wide-field optical profiling technique with nanometer resolution. We reveal the existence of small and large membrane waves the amplitudes of which are in the range of ?3–7 nm to ?16–25 nm respectively, through the cell. For migrating single-cells, the amplitude of these waves is about 30 nm near the cell edge. Two or more different directions of propagation of the membrane nanowaves inside the same cell can be observed. After increasing the migration velocity by BMP-2 treatment, only one wave direction of propagation exists with an increase in the average amplitude (more than 80 nm near the cell edge). Furthermore for collective-cell migration, these membrane nanowaves are attenuated on the leader cells and poor transmission of these nanowaves to follower cells was observed. After BMP-2 treatment, the membrane nanowaves are transmitted from the leader cell to several rows of follower cells. Surprisingly, the vast majority of the observed membrane nanowaves is shared between the adjacent cells. These results give a new view on how single and collective-cells modulate their motility. This work has significant implications for the therapeutic use of BMPs for the regeneration of skin tissue.

Zouani, Omar F.; Gocheva, Veronika; Durrieu, Marie-Christine

2014-01-01

188

Systematic Design of Polymer Electrolyte Membranes for Fuel Cells Using a Pore-Filling Membrane Concept  

NASA Astrophysics Data System (ADS)

In this chapter, systematic membrane design and development using our pore-filling membrane concept is described. Pore-filling electrolyte membranes for use as electrolyte membranes for polymer electrolyte membrane fuel cells (PEMFCs) or direct methanol fuel cells (DMFCs) are described. The pores of a porous substrate are filled with a polymer electrolyte and the membrane swelling is suppressed by the substrate matrix. Proton conductivity is achieved through the impregnated electrolyte polymer. Fuel crossover is reduced by suppression of the electrolyte polymer swelling and mechanical strength is maintained by the substrate. Using this concept, high proton conductivity has been shown to exist with reduced membrane fuel crossover and good dimensional stability. In this chapter, high performance pore-filling electrolyte membranes and their DMFC or PEMFC performances are shown. To achieve a high energy density DMFC device, we must use a high concentration of methanol aqueous solution as the fuel and crossover must be reduced. The membrane also showed almost no dimensional change with variation in humidity. The DMFC and PEMFC performances are also described with several varieties of pore-filling membranes for each application.

Yamaguchi, Takeo

189

HTO electrolysis method by using proton exchange membrane fuel cell  

NASA Astrophysics Data System (ADS)

The application of a proton exchange membrane fuel cell (PEMFC) system to electrolysis for HTO recovered from a solid breeder blanket is discussed in this report. The amount of gas permeation through the membrane in this system and the tritium inventory in the membrane were calculated, and it was found that the effect of these phenomena on the performance of electrolysis system can be disregarded. However, the trapped tritium in the membrane will damage the structure of the membrane, and we need to replace it about once every 10 days. This duration is so short that we should prepare other backup plans to convert HTO to HT in the blanket purge gas.

Takata, Hiroki; Nishikawa, Masabumi; Egawa, Takayuki; Mizuno, Nobukazu

2007-08-01

190

Electrolyte Composition of Mink (Mustela vison) Erythrocytes and Active Cation Transporters of the Cell Membrane  

PubMed Central

Red blood cells from mink (Mustela vison) were characterized with respect to their electrolyte content and their cell membranes with respect to enzymatic activity for cation transport. The intra- and extracellular concentrations of Na+, K+, Cl-, Ca2+ and Mg2+ were determined in erythrocytes and plasma, respectively. Plasma and red cell water content was determined, and molal electrolyte concentrations were calculated. Red cells from male adult mink appeared to be of the low-K+, high-Na+ type as seen in other carnivorous species. The intracellular K+ concentration is slightly higher than the extracellular one and the plasma-to-cell chemical gradient for Na+ is weak, though even the molal concentrations may differ significantly. Consistent with the high intracellular Na+ and low K+ concentrations, a very low or no ouabain-sensitive Na+,K+-ATPase activity and no K+-activated pNPPase activity were found in the plasma membrane fraction from red cells. The Cl- and Mg2+ concentrations expressed per liter cell water were significantly higher in red cells than in plasma whereas the opposite was the case with Ca2+. The distribution of Cl- thus does not seem compatible with an inside-negative membrane potential in mink erythrocytes. In spite of a steep calcium gradient across the red cell membrane, neither a calmodulin-activated Ca2+-ATPase activity nor an ATP-activated Ca2+-pNPPase activity were detectable in the plasma membrane fraction. The origin of a supposed primary Ca2+ gradient for sustaining of osmotic balance thus seems uncertain.

Hansen, O; Clausen, TN

2001-01-01

191

Electrolyte composition of mink (Mustela vison) erythrocytes and active cation transporters of the cell membrane.  

PubMed

Red blood cells from mink (Mustela vison) were characterized with respect to their electrolyte content and their cell membranes with respect to enzymatic activity for cation transport. The intra- and extracellular concentrations of Na+, K+, Cl-, Ca2+ and Mg2+ were determined in erythrocytes and plasma, respectively. Plasma and red cell water content was determined, and molal electrolyte concentrations were calculated. Red cells from male adult mink appeared to be of the low-K+, high-Na+ type as seen in other carnivorous species. The intracellular K+ concentration is slightly higher than the extracellular one and the plasma-to-cell chemical gradient for Na+ is weak, though even the molal concentrations may differ significantly. Consistent with the high intracellular Na+ and low K+ concentrations, a very low or no ouabain-sensitive Na+, K(+)-ATPase activity and no K(+)-activated pNPPase activity were found in the plasma membrane fraction from red cells. The Cl- and Mg2+ concentrations expressed per liter cell water were significantly higher in red cells than in plasma whereas the opposite was the case with Ca2+. The distribution of Cl- thus does not seem compatible with an inside-negative membrane potential in mink erythrocytes. In spite of a steep calcium gradient across the red cell membrane, neither a calmodulin-activated Ca(2+)-ATPase activity nor an ATP-activated Ca(2+)-pNPPase activity were detectable in the plasma membrane fraction. The origin of a supposed primary Ca2+ gradient for sustaining of osmotic balance thus seems uncertain. PMID:11503371

Hansen, O; Clausen, T N

2001-01-01

192

Studying the Nucleated Mammalian Cell Membrane by Single Molecule Approaches  

PubMed Central

The cell membrane plays a key role in compartmentalization, nutrient transportation and signal transduction, while the pattern of protein distribution at both cytoplasmic and ectoplasmic sides of the cell membrane remains elusive. Using a combination of single-molecule techniques, including atomic force microscopy (AFM), single molecule force spectroscopy (SMFS) and stochastic optical reconstruction microscopy (STORM), to study the structure of nucleated cell membranes, we found that (1) proteins at the ectoplasmic side of the cell membrane form a dense protein layer (4 nm) on top of a lipid bilayer; (2) proteins aggregate to form islands evenly dispersed at the cytoplasmic side of the cell membrane with a height of about 10–12 nm; (3) cholesterol-enriched domains exist within the cell membrane; (4) carbohydrates stay in microdomains at the ectoplasmic side; and (5) exposed amino groups are asymmetrically distributed on both sides. Based on these observations, we proposed a Protein Layer-Lipid-Protein Island (PLLPI) model, to provide a better understanding of cell membrane structure, membrane trafficking and viral fusion mechanisms.

Wang, Feng; Wu, Jiazhen; Gao, Jing; Liu, Shuheng; Jiang, Junguang; Jiang, Shibo; Wang, Hongda

2014-01-01

193

Axially resolved polarisation microscopy of membrane dynamics in living cells  

NASA Astrophysics Data System (ADS)

Membrane dynamics has a large impact on cellular uptake and release of various metabolites or pharmaceutical agents. For a deeper understanding of the cellular processes involved, we used U373-MG human glioblastoma cells as a model system. As conventional microscopy does not permit to investigate individual layers in living cells, we used structured illumination techniques and total internal reflection fluorescence microscopy (TIRFM) to analyse the plasma membrane and intracellular membranes of living cells selectively. Optical image sections provide a high resolution and the possibility of 3D reconstruction. Membranes of living cells were characterized by the membrane marker 6-dodecanoyl-2-dimethylamino naphthalene (laurdan). Due to its spectral and kinetic properties this fluorescence marker appears appropriate for measuring membrane stiffness and fluidity. After excitation with linearly polarized laser pulses, membrane fluidity of human glioblastoma cells was determined by measurements of steady-state and time-resolved fluorescence anisotropy r(t), since with increasing viscosity of the environment, the rotation of an excited molecule is impeded. The corresponding time constant ?r of molecular relaxation decreased with temperature and increased with the amount of cholesterol. In addition, fluorescence anisotropy r(t) values of the plasma membrane were larger than the values of intracellular membranes for all temperatures in the range of 16°C<=T<=41°C.

Wagner, Michael; Weber, Petra; Schneckenburger, Herbert

2007-06-01

194

Analysis of proton exchange membrane fuel cell performance with alternate membranes  

Microsoft Academic Search

Renewed interest in proton exchange membrane fuel cell technology for space and terrestrial (particularly electric vehicles) was stimulated by the demonstration, in the mid 1980s, of high energy efficiencies and high power densities. One of the most vital components of the PEMFC is the proton conducting membrane. In this paper, an analysis is made of the performances of PEMFCs with

Masanobu Wakizoe; Omourtag A. Velev; Supramaniam Srinivasan

1995-01-01

195

Enantiomeric fraction determination of 2-arylpropionic acids in a package plant membrane bioreactor.  

PubMed

Enantiomeric compositions of three 2-arylpropionic acid (2-APA) drugs, ibuprofen, naproxen, and ketoprofen, were monitored in a membrane bioreactor (MBR) treating municipal effluent in a small rural town in Australia. Specific enantiomers were determined as amide diastereomers using the chiral derivatizing reagent, (R)-1-phenylethylamine (PEA), followed by gas chromatography-tandem mass spectrometry (GC-MS/MS). The six individual enantiomers were quantified by isotope dilution and the enantiomeric fractions (EFs) were determined. Over four separate sampling events, ibuprofen EF ranged from 0.88 to 0.94 (median 0.93) in the influent and 0.38 to 0.40 (median 0.39) in the effluent. However, no significant change in ketoprofen EF was observed, with influent EFs of 0.56-0.60 (median 0.58) and effluent EFs 0.54-0.68 (median 0.56). This is the first report of enantiospecific analysis of ketoprofen in municipal wastewater and it is not yet clear why such different behavior was observed compared to ibuprofen. Naproxen EF was consistently measured at 0.99 in the influent and ranged from 0.86 to 0.94 (median 0.91) in the effluent. This study demonstrates that EF is a relatively stable parameter and does not fluctuate according to concentration or other short-term variables introduced by sampling limitations. The enantiospecific analysis of chiral chemicals presents a promising approach to elucidate a more thorough understanding of biological treatment processes and a potential tool for monitoring the performance of key biological pathways. PMID:23620266

Hashim, Nor H; Stuetz, Richard M; Khan, Stuart J

2013-05-01

196

Development and characterization of proton conductive membranes and membrane electrode assemblies for fuel cells  

NASA Astrophysics Data System (ADS)

Polymer electrolyte membrane fuel cells (PEMFCs), including hydrogen fuel cells (PEMFCs) and direct methanol fuel cells (DMFCs), are considered as attractive electrical power sources. However, there are some technical obstacles that impede the commercialization of PEMFCs. For instance, in H 2-PEMFCs, carbon monoxide (CO) poisoning of the anode catalyst causes serious performance loss; in DMFCs, methanol crossover through the membrane reduces the overall fuel cell efficiency. This work focused on: (1) developing high performance membrane electrode assemblies (MEAs) and investigating their behavior at higher temperature H2-PEMFC with H2+CO as the fuel; (2) improving DMFCs efficiency by preparing low methanol crossover/good proton conductivity membranes based on NafionRTM matrix; (3) synthesizing and modifying low cost sulfonated hydrocarbon (SPEEK) membranes for both H2-PEMFCs and DMFCs applications. High performance membrane electrode assemblies (MEAs) with composite NafionRTM-TeflonRTM-Zr(HPO 4)2 membranes were prepared, optimized and characterized at higher temperature (> 100°C)/lower relative humidity (< 100% RH) condition, using H2 or H2+CO as the fuel. Effects of CO concentration, temperature, relative humidity to the CO poisoning on H 2-PEMFC were studied by applying various electrochemical techniques. The electrochemical oxidation mechanism of H2/CO in higher temperature PEMFC was investigated and simulated. Two type of membranes based on NafionRTM matrix were prepared: silica/NafionRTM membrane and palladium impregnated NafionRTM (Pd-NafionRTM) membrane. The composite silica/NafionRTM membrane was developed by in-situ sol-gel reaction followed by solution casting, while the Pd-NafionRTM was fabricated via a supercritical fluid CO2 (scCO 2) route. Reduced methanol crossover and enhanced efficiency was observed by applying each of the two membranes to DMFCs. In addition, the research demonstrated that scCO2 is a promising technique for modifying membranes or depositing nano-particle electrocatalysts onto electrolyte. Sulfonated poly(ether ether ketone) (SPEEK) was synthesized by a sulfonation reaction using poly(ether ether ketone) (PEEK). Multilayer structure SPEEK membranes with methanol barriers were fabricated and showed enhanced membrane stability in DMFCs. Improved MEA performance was obtained due to lower methanol crossover and the presence of a good membrane/electrode interface for facilitating proton transfer.

Jiang, Ruichun

197

Cell Membranes Under Hydrostatic Pressure Subjected to Micro-Injection  

NASA Astrophysics Data System (ADS)

The work is concerned with the determination of the mechanical behaviour of cell membranes under uniform hydrostatic pressure subject to micro-injections. For that purpose, assuming that the shape of the deformed cell membrane is axisymmetric a variational statement of the problem is developed on the ground of the so-called spontaneous curvature model. In this setting, the cell membrane is regarded as an axisymmetric surface in the three-dimensional Euclidean space providing a stationary value of the shape energy functional under the constraint of fixed total area and fixed enclosed volume. The corresponding Euler-Lagrange equations and natural boundary conditions are derived, analyzed and used to express the forces and moments in the membrane. Several examples of such surfaces representing possible shapes of cell membranes under pressure subjected to micro injection are determined numerically.

Vassilev, Vassil M.; Kostadinov, Kostadin G.; Mladenov, Ivaïlo M.; Shulev, Assen A.; Stoilov, Georgi I.; Djondjorov, Peter A.

2011-04-01

198

Detecting Nanodomains in Living Cell Membrane by Fluorescence Correlation Spectroscopy  

NASA Astrophysics Data System (ADS)

Cell membranes actively participate in numerous cellular functions. Inasmuch as bioactivities of cell membranes are known to depend crucially on their lateral organization, much effort has been focused on deciphering this organization on different length scales. Within this context, the concept of lipid rafts has been intensively discussed over recent years. In line with its ability to measure diffusion parameters with great precision, fluorescence correlation spectroscopy (FCS) measurements have been made in association with innovative experimental strategies to monitor modes of molecular lateral diffusion within the plasma membrane of living cells. These investigations have allowed significant progress in the characterization of the cell membrane lateral organization at the suboptical level and have provided compelling evidence for the in vivo existence of raft nanodomains. We review these FCS-based studies and the characteristic structural features of raft nanodomains. We also discuss the findings in regards to the current view of lipid rafts as a general membrane-organizing principle.

He, Hai-Tao; Marguet, Didier

2011-05-01

199

Composite Membranes for Medium-Temperature Pem Fuel Cells  

NASA Astrophysics Data System (ADS)

The main obstacles to greater commercialization of polymer electrolyte fuel cells are mostly related to the low-proton conductivity at low-relative humidity of the known ionomeric membranes, to their high methanol permeability and poor mechanical properties above ~130oC. A possible solution for these problems has been found in the development of composite membranes, where particles of suitable fillers are dispersed in the ionomer matrix. The preparation methods for obtaining composite membranes are described, and recent work dealing with composite ionomeric membranes containing silica, heteropolyacids, layered metal phosphates, and phosphonates is reviewed. Finally, new strategies for the preparation of nano-composite membranes and for the filling of porous polymeric membranes with highly conductive zirconium phosphonates are described. The expected influence of size and orientation of these particles on membrane properties, such as conductivity and permeability to methanol, is also discussed.

Alberti, G.; Casciola, M.

2003-08-01

200

Measurement of the nonlinear elasticity of red blood cell membranes  

NASA Astrophysics Data System (ADS)

The membranes of human red blood cells (RBCs) are a composite of a fluid lipid bilayer and a triangular network of semiflexible filaments (spectrin). We perform cellular microrheology using the dynamic membrane fluctuations of the RBCs to extract the elastic moduli of this composite membrane. By applying known osmotic stresses, we measure the changes in the elastic constants under imposed strain and thereby determine the nonlinear elastic properties of the membrane. We find that the elastic nonlinearities of the shear modulus in tensed RBC membranes can be well understood in terms of a simple wormlike chain model. Our results show that the elasticity of the spectrin network can mostly account for the area compression modulus at physiological osmolality, suggesting that the lipid bilayer has significant excess area. As the cell swells, the elastic contribution from the now tensed lipid membrane becomes dominant.

Park, Yongkeun; Best, Catherine A.; Kuriabova, Tatiana; Henle, Mark L.; Feld, Michael S.; Levine, Alex J.; Popescu, Gabriel

2011-05-01

201

Time-Resolved Molecular Transport across Living Cell Membranes  

PubMed Central

It is shown that the nonlinear optical phenomenon known as second-harmonic generation can be used for label-free, time-resolved study of the transport of molecules through living cell membranes. The adsorption and transport of a 300-Da molecular-mass hydrophobic ion at the Escherichia coli membrane is observed. Remarkably, at low ion concentrations, the second-harmonic generation technique clearly exposes a multistep molecular transport process: Transport of the molecular ion across the outer and cytoplasmic membranes of the Gram-negative bacteria is recorded, in sequence, in time. Fitting of the data to a multiprocess kinematic model reveals that the transport of this hydrophobic ion through the outer membrane is much faster than through the cytoplasmic membrane, likely reflecting the effectiveness of ion transport porins. The observations illustrate an experimental means for studying the interactions of small molecules with cell membranes.

Zeng, Jia; Eckenrode, Heather M.; Dounce, Susan M.; Dai, Hai-Lung

2013-01-01

202

Polymer membranes for high temperature proton exchange membrane fuel cell: Recent advances and challenges  

Microsoft Academic Search

Proton-exchange membrane fuel cells (PEMFCs) are considered to be a promising technology for efficient power generation in the 21st century. Currently, high temperature proton exchange membrane fuel cells (HT-PEMFC) offer several advantages, such as high proton conductivity, low permeability to fuel, low electro-osmotic drag coefficient, good chemical\\/thermal stability, good mechanical properties and low cost. Owing to the aforementioned features, high

Saswata Bose; Tapas Kuila; Thi Xuan Hien Nguyen; Nam Hoon Kim; Kin-tak Lau; Joong Hee Lee

2011-01-01

203

Alternative Sources of Adult Stem Cells: Human Amniotic Membrane  

Microsoft Academic Search

\\u000a Human amniotic membrane is a highly promising cell source for tissue engineering. The cells thereof, human amniotic epithelial\\u000a cells (hAEC) and human amniotic mesenchymal stromal cells (hAMSC), may be immunoprivileged, they represent an early developmental\\u000a status, and their application is ethically uncontroversial. Cell banking strategies may use freshly isolated cells or involve\\u000a in vitro expansion to increase cell numbers. Therefore,

Susanne Wolbank; Martijn van Griensven; Regina Grillari-Voglauer; Anja Peterbauer-Scherb

2010-01-01

204

Human hepatocytes and endothelial cells in organotypic membrane systems.  

PubMed

The realization of organotypic liver model that exhibits stable phenotype is a major challenge in the field of liver tissue engineering. In this study we developed liver organotypic co-culture systems by using synthetic and biodegradable membranes with primary human hepatocytes and human umbilical vein endothelial cells (HUVEC). Synthetic membranes prepared by a polymeric blend constituted of modified polyetheretherketone (PEEK-WC) and polyurethane (PU) and biodegradable chitosan membranes were developed by phase inversion technique and used in homotypic and organotypic culture systems. The morphological and functional characteristics of cells in the organotypic co-culture membrane systems were evaluated in comparison with homotypic cultures and traditional systems. Hepatocytes in the organotypic co-culture systems exhibit compact polyhedral cells with round nuclei and well demarcated cell-cell borders like in vivo, as a result of heterotypic interaction with HUVECs. In addition HUVECs formed tube-like structures directly through the interactions with the membranes and hepatocytes and indirectly through the secretion of ECM proteins which secretion improved in the organotypic co-culture membrane systems. The heterotypic cell-cell contacts have beneficial effect on the hepatocyte albumin production, urea synthesis and drug biotransformation. The developed organotypic co-culture membrane systems elicit liver specific functions in vitro and could be applied for the realization of engineered liver tissues to be used in tissue engineering, drug metabolism studies and bioartificial liver devices. PMID:21871658

Salerno, Simona; Campana, Carla; Morelli, Sabrina; Drioli, Enrico; De Bartolo, Loredana

2011-12-01

205

Membrane fusion in cells: molecular machinery and mechanisms.  

PubMed

Membrane fusion is a sine qua non process for cell physiology. It is critical for membrane biogenesis, intracellular traffic, and cell secretion. Although investigated for over a century, only in the last 15 years, the molecular machinery and mechanism of membrane fusion has been deciphered. The membrane fusion event elicits essentially three actors on stage: anionic phospholipids - phosphatidylinositols, phosphatidyl serines, specific membrane proteins, and the calcium ions, all participating in a well orchestrated symphony. Three soluble N-ethylmaleimide-sensitive factor (NSF)-attachment protein receptors (SNAREs) have been implicated in membrane fusion. Target membrane proteins, SNAP-25 and syntaxin (t- SNARE) and secretory vesicle-associated membrane protein (v-SNARE) or VAMPwere discovered in the 1990's and suggested to be the minimal fusion machinery. Subsequently, the molecular mechanism of SNARE-induced membrane fusion was discovered. It was demonstrated that when t-SNARE-associated lipid membrane is exposed to v-SNARE-associated vesicles in the presence of Ca(2+), the SNARE proteins interact in a circular array to form conducting channels, thus establishing continuity between the opposing bilayers. Further it was proved that SNAREs bring opposing bilayers close to within a distance of 2-3 Angstroms, allowing Ca(2+) to bridge them. The bridging of bilayers by Ca(2+) then leads to the expulsion of water between the bilayers at the contact site, allowing lipid mixing and membrane fusion. Calcium bridging of opposing bilayers leads to the release of water, both from the water shell of hydrated Ca(2+) ions, as well as the displacement of loosely coordinated water at the phosphate head groups in the lipid membrane. These discoveries provided for the first time, the molecular mechanism of SNARE-induced membrane fusion in cells. Some of the seminal discoveries are briefly discussed in this minireview. PMID:16796809

Leabu, M

2006-01-01

206

Immunological studies of an organic anion-binding protein isolated from rat liver cell plasma membrane.  

PubMed Central

The mechanism of organic anion uptake by hepatocytes has kinetics that suggest facilitated diffusion, and carrier-mediated membrane transport has been postulated. In previous studies, we purified a 55,000-mol wt organic anion-binding protein (OABP) by affinity chromatography on sulfobromophthalein (BSP)-Sepharose of deoxycholate solubilized liver cell plasma membrane preparations. Using specific goat and rabbit antibodies to OABP, we have now investigated the distribution of this protein in liver fractions and other tissues by an enzyme-linked immunosorbent assay and by the immunoblot (Western blot) procedure. These studies indicated that OABP is present in significant amounts in all tissues examined except for blood. Although OABP has not as yet been isolated from each of these tissues and characterized, OABP in heart retained the ability to bind organic anions, and was purified by affinity chromatography on BSP-sepharose. In liver, OABP was membrane bound and remained so after extraction with 0.9 M NaCl, which suggests that it is an intrinsic membrane protein. OABP did not have a ubiquitous subcellular distribution within the hepatocyte. Preparation of subfractions of liver cell plasma membrane revealed that OABP is present in the sinusoidal and absent from the canalicular membrane. Immunofluorescence studies performed in short-term cultured hepatocytes suggest that OABP is associated with the surface of these cells and does not have a significant intracellular distribution. Images

Wolkoff, A W; Sosiak, A; Greenblatt, H C; Van Renswoude, J; Stockert, R J

1985-01-01

207

Peptides in common bean fractions inhibit human colorectal cancer cells.  

PubMed

The aim of this study was to characterize peptides present in common bean non-digestible fractions (NDF) produced after enzymatic digestion and determine their antiproliferative action on human colorectal cancer cells. Five NDF peptides represented 70% of total protein (GLTSK, LSGNK, GEGSGA, MPACGSS and MTEEY) with antiproliferative activity on human colon cancer cells. Based on the antiproliferative effect, HCT116 cell line was most sensitive to bean Azufrado Higuera (IC50=0.53mg/ml) and RKO to Bayo Madero (IC50=0.51mg/ml) peptide extracts. Both cultivars increased significantly (p<0.05) the expression of p53 in HCT116 by 76% and 68%, respectively. Azufrado Higuera modified the expression of cell cycle regulation proteins p21 and cyclin B1. Bayo Madero modified the expression of mitochondrial activated apoptotic proteins BAD, cytC, c-casp3, Survivin, BIRC7. Results suggest that peptides present in common bean NDF contributed to the antiproliferative effect on human colorectal cancer cells by modifying molecules involved in either cell cycle arrest or apoptosis. PMID:24679790

Luna Vital, Diego A; González de Mejía, Elvira; Dia, Vermont P; Loarca-Piña, Guadalupe

2014-08-15

208

Improved Membrane Materials for PEM Fuel Cell Application  

SciTech Connect

The overall goal of this project is to collect and integrate critical structure/property information in order to develop methods that lead to significant improvements in the durability and performance of polymer electrolyte membrane fuel cell (PEMFC) materials. This project is focused on the fundamental improvement of PEMFC membrane materials with respect to chemical, mechanical and morphological durability as well as the development of new inorganically-modified membranes.

Kenneth A. Mauritz; Robert B. Moore

2008-06-30

209

Regulatory Aspects of Membrane Microdomain (Raft) Dynamics in Live Cells  

Microsoft Academic Search

Most vertebrate cells display a considerable microheterogeneity in their plasma membranes, often termed microdomain structure.\\u000a Some of these microdomains are enriched in glycosphingolipids and cholesterol and are resistant to solubilization with nonionic\\u000a detergents; they are therefore called detergent-insoluble-glycolipid enriched membrane (DIG) or glycosphingolipid enriched\\u000a membrane (GEM). These domains, also called “lipid rafts” (Simons and Ikonen, 1997), may form at the

János Matkó; János Szöll?si

210

Adaptive changes in pancreatic beta cell fractional area and beta cell turnover in human pregnancy  

PubMed Central

Aims/hypothesis We sought to establish the extent and basis for adaptive changes in beta cell numbers in human pregnancy. Methods Pancreas was obtained at autopsy from women who had died while pregnant (n?=?18), post-partum (n?=?6) or were not pregnant at or shortly before death (controls; n?=?20). Pancreases were evaluated for fractional pancreatic beta cell area, islet size and islet fraction of beta cells, beta cell replication (Ki67) and apoptosis (TUNEL), and indirect markers of beta cell neogenesis (insulin-positive cells in ducts and scattered beta cells in pancreas). Results The pancreatic fractional beta cell area was increased by ?1.4-fold in human pregnancy, with no change in mean beta cell size. In pregnancy there were more small islets rather than an increase in islet size or beta cells per islet. No increase in beta cell replication or change in beta cell apoptosis was detected, but duct cells positive for insulin and scattered beta cells were increased with pregnancy. Conclusions/interpretation The adaptive increase in beta cell numbers in human pregnancy is not as great as in most reports in rodents. This increase in humans is achieved by increased numbers of beta cells in apparently new small islets, rather than duplication of beta cells in existing islets, which is characteristic of pregnancy in rodents. Electronic supplementary material The online version of this article (doi:10.1007/s00125-010-1809-6) contains supplementary material, which is available to authorised users.

Butler, A. E.; Cao-Minh, L.; Galasso, R.; Rizza, R. A.; Corradin, A.; Cobelli, C.

2010-01-01

211

Membrane-particle interactions in an asymmetric flow field flow fractionation channel studied with titanium dioxide nanoparticles.  

PubMed

Asymmetric flow field flow fractionation operated in a multidetector approach (A4F-MDA) is a powerful tool to perform size-classified nanoparticle analysis. Recently several publications mentioned insufficient recovery rates and even retention time shifts attributed to unspecific membrane-particle interactions. One hypothesis to explain this phenomenon is based on the surface charge (zeta-potential) of the membrane material and the particle. In this study, we investigated in how far the ?-potential of A4F membrane and particles would determine the outcome of A4F in terms of feasibility, separation efficiency, retention time, and recovery rate, or whether other factors such as membrane morphology and particle size were equally important. We systematically studied the influence of the ?-potential on the interactions between the most commonly used A4F membrane materials and two representative types of titanium dioxide nanoparticles (TiO2 NP). Furthermore the effect of different carrier media and additional surfactants on the surface charge of membranes and particles was investigated and the influence of the particle size and the particle concentration on the recovery rate was evaluated. We found that the eligibility of an A4F method can be predicted based on the ?-potential of the NPs and the A4F membrane. Furthermore knowing the ?-potential allows to tuning the separation efficiency of an A4F method. On the other hand we observed significant shifts in retention time for different membrane materials that impede the determination of particle size based on the classical A4F theory. These shifts cannot be attributed to the ?-potential. Also the ?-potential does not account for varying recovery rates of different particle types, instead the particle size seems to be the limiting factor. Therefore, the proper characterization of a polydisperse sample remains a challenge. PMID:24556173

Bendixen, Nina; Losert, Sabrina; Adlhart, Christian; Lattuada, Marco; Ulrich, Andrea

2014-03-21

212

Effect of hydroperoxides on red blood cell membrane mechanical properties.  

PubMed

We investigate the effect of oxidative stress on red blood cell membrane mechanical properties in vitro using detailed analysis of the membrane thermal fluctuation spectrum. Two different oxidants, the cytosol-soluble hydrogen peroxide and the membrane-soluble cumene hydroperoxide, are used, and their effects on the membrane bending elastic modulus, surface tension, strength of confinement due to the membrane skeleton, and 2D shear elastic modulus are measured. We find that both oxidants alter significantly the membrane elastic properties, but their effects differ qualitatively and quantitatively. While hydrogen peroxide mainly affects the elasticity of the membrane protein skeleton (increasing the membrane shear modulus), cumene hydroperoxide has an impact on both membrane skeleton and lipid bilayer mechanical properties, as can be seen from the increased values of the shear and bending elastic moduli. The biologically important implication of these results is that the effects of oxidative stress on the biophysical properties, and hence the physiological functions, of the cell membrane depend on the nature of the oxidative agent. Thermal fluctuation spectroscopy provides a means of characterizing these different effects, potentially in a clinical milieu. PMID:22004746

Hale, John P; Winlove, C Peter; Petrov, Peter G

2011-10-19

213

Effect of Hydroperoxides on Red Blood Cell Membrane Mechanical Properties  

PubMed Central

We investigate the effect of oxidative stress on red blood cell membrane mechanical properties in vitro using detailed analysis of the membrane thermal fluctuation spectrum. Two different oxidants, the cytosol-soluble hydrogen peroxide and the membrane-soluble cumene hydroperoxide, are used, and their effects on the membrane bending elastic modulus, surface tension, strength of confinement due to the membrane skeleton, and 2D shear elastic modulus are measured. We find that both oxidants alter significantly the membrane elastic properties, but their effects differ qualitatively and quantitatively. While hydrogen peroxide mainly affects the elasticity of the membrane protein skeleton (increasing the membrane shear modulus), cumene hydroperoxide has an impact on both membrane skeleton and lipid bilayer mechanical properties, as can be seen from the increased values of the shear and bending elastic moduli. The biologically important implication of these results is that the effects of oxidative stress on the biophysical properties, and hence the physiological functions, of the cell membrane depend on the nature of the oxidative agent. Thermal fluctuation spectroscopy provides a means of characterizing these different effects, potentially in a clinical milieu.

Hale, John P.; Winlove, C. Peter; Petrov, Peter G.

2011-01-01

214

Existence of a Flat Phase in Red Cell Membrane Skeletons  

NASA Astrophysics Data System (ADS)

Biomolecular membranes display rich statistical mechanical behavior. They are classified as liquid in the absence of shear elasticity in the plane of the membrane and tethered (solid) when the neighboring molecules or subunits are connected and the membranes exhibit solid-like elastic behavior in the plane of the membrane. The spectrin skeleton of red blood cells was studied as a model tethered membrane. The static structure factor of the skeletons, measured by small-angle x-ray and light scattering, was fitted with a structure factor predicted with a model calculation. The model describes tethered membrane sheets with free edges in a flat phase, which is a locally rough but globally flat membrane configuration. The fit was good for large scattering vectors. The membrane roughness exponent, zeta, defined through h propto L^zeta, where h is the average amplitude of out-of-plane fluctuations and L is the linear membrane dimension, was determined to be 0.65 ± 0.10. Computer simulations of model red blood cell skeletons also showed this flat phase. The value for the roughness exponent, which was determined from the scaling properties of membranes of different sizes, was consistent with that from the experiments.

Schmidt, Christoph F.; Svoboda, Karel; Lei, Ning; Petsche, Irena B.; Berman, Lonny E.; Safinya, Cyrus R.; Grest, Gary S.

1993-02-01

215

Cytoskeletal control of the red-blood cell membrane  

NASA Astrophysics Data System (ADS)

We have shown (Physical Review Letters, 90, 228101 (2003)) that the thermal fluctuations of red blood cells can be accounted for by a model of a nearly-free, but confined bilayer membrane with a finite tension; both the confinement and tension arise from the coupling of the membrane with the cytoskeleton. Recently, we have shown that these relatively gentle effects of the cytoskeleton-membrane couplings on the membrane fluctuations are due to the dilute nature of the coupling molecules. To quantify this, we predict the fluctuation amplitude for a microscopic model of the inhomogeneous coupling of a fluid membrane and a fixed cytoskeleton. The coupling is modeled as periodic and harmonic, and we consider the linear response of the membrane. We find that there is indeed, an effective surface tension and confinement of such a membrane, in accord with our phenomenological model, and relate these quantities to the strength and periodicity of the microscopic coupling. We also find, surprisingly, that the membrane can develop a spontaneous breaking of the cytoskeleton symmetry, at low confinements. Finally we address the role of ATP activity on the cytoskeleton-driven fluctuations and the equilibrium shape of the cell. We examine in detail the role of spectrin disconnections as the main ATP-activated network defects on the global cell shape and membrane fluctuations.

Gov, Nir; Safran, Sam

2004-03-01

216

Layer-by-layer Cell Membrane Assembly  

PubMed Central

Eukaryotic subcellular membrane systems, such as the nuclear envelope or endoplasmic reticulum, present a rich array of architecturally and compositionally complex supramolecular targets that are yet inaccessible. Here we describe layer-by-layer phospholipid membrane assembly on microfluidic droplets, a route to structures with defined compositional asymmetry and lamellarity. Starting with phospholipid-stabilized water-in-oil droplets trapped in a static droplet array, lipid monolayer deposition proceeds as oil/water phase boundaries pass over the droplets. Unilamellar vesicles assembled layer-by-layer support functional insertion of both purified and in situ expressed membrane proteins. Synthesis and chemical probing of asymmetric unilamellar and double bilayer vesicles demonstrate the programmability of both membrane lamellarity and lipid leaflet composition during assembly. The immobilized vesicle arrays are a pragmatic experimental platform for biophysical studies of membranes and their associated proteins, particularly complexes that assemble and function in multilamellar contexts in vivo.

Matosevic, Sandro; Paegel, Brian M.

2014-01-01

217

Radiation Interaction with Therapeutic Drugs and Cell Membranes  

SciTech Connect

This transient permeabilized state of the cell membrane, named the 'cell electroporation' (CE) can be used to increase cells uptake of drugs that do not readily pass cell membrane, thus enabling their cytotoxicity. The anticancer drugs, such as bleomycin (BL) and cisplatin, are the most candidates for the combined use with ionizing and non-ionizing radiation fields. The methods and installations for the cell electroporation by electron beam (EB) and microwave (MW) irradiation are presented. The viability tests of the human leukocytes under EB and MW exposure with/without the BL in the cell cultures are discussed.

Martin, Diana I.; Manaila, Elena N.; Matei, Constantin I.; Iacob, Nicusor I.; Ighigeanu, Daniel I.; Craciun, Gabriela D. [Accelerators Laboratory, National Institute for Lasers, Plasma and Radiation Physics, 409, Atomistilor St., 077125 Magurele (Romania); Moisescu, Mihaela I.; Savopol, Tudor D.; Kovacs, Eugenia A. [Carol Davila University of Medicine and Pharmacy, 8, Eroii Sanitari St., 050474 Bucharest (Romania); Cinca, Sabin A. [Oncology Institute Prof. A. Trestioreanu, 252, Fundeni St., 022328 Bucharest (Romania); Margaritescu, Irina D. [Military Clinical Hospital, 88, Mircea Vulcanescu St., 010825 Bucharest (Romania)

2007-04-23

218

Single cell wound generates electric current circuit and cell membrane potential variations that requires calcium influx.  

PubMed

Breaching of the cell membrane is one of the earliest and most common causes of cell injury, tissue damage, and disease. If the compromise in cell membrane is not repaired quickly, irreversible cell damage, cell death and defective organ functions will result. It is therefore fundamentally important to efficiently repair damage to the cell membrane. While the molecular aspects of single cell wound healing are starting to be deciphered, its bio-physical counterpart has been poorly investigated. Using Xenopus laevis oocytes as a model for single cell wound healing, we describe the temporal and spatial dynamics of the wound electric current circuitry and the temporal dynamics of cell membrane potential variation. In addition, we show the role of calcium influx in controlling electric current circuitry and cell membrane potential variations. (i) Upon wounding a single cell: an inward electric current appears at the wound center while an outward electric current is observed at its sides, illustrating the wound electric current circuitry; the cell membrane is depolarized; calcium flows into the cell. (ii) During cell membrane re-sealing: the wound center current density is maintained for a few minutes before decreasing; the cell membrane gradually re-polarizes; calcium flow into the cell drops. (iii) In conclusion, calcium influx is required for the formation and maintenance of the wound electric current circuitry, for cell membrane re-polarization and for wound healing. PMID:24801267

Luxardi, Guillaume; Reid, Brian; Maillard, Pauline; Zhao, Min

2014-07-24

219

An experimental and theoretical analysis of ultrasound-induced permeabilization of cell membranes.  

PubMed

Application of ultrasound transiently permeabilizes cell membranes and offers a nonchemical, nonviral, and noninvasive method for cellular drug delivery. Although the ability of ultrasound to increase transmembrane transport has been well demonstrated, a systematic dependence of transport on ultrasound parameters is not known. This study examined cell viability and cellular uptake of calcein using 3T3 mouse cell suspension as a model system. Cells were exposed to varying acoustic energy doses at four different frequencies in the low frequency regime (20-100 kHz). At all frequencies, cell viability decreased with increasing acoustic energy dose, while the fraction of cells exhibiting uptake of calcein showed a maximum at an intermediate energy dose. Acoustic spectra under various ultrasound conditions were also collected and assessed for the magnitude of broadband noise and subharmonic peaks. While the cell viability and transport data did not show any correlation with subharmonic (f/2) emission, they correlated with the broadband noise, suggesting a dominant contribution of transient cavitation. A theoretical model was developed to relate reversible and irreversible membrane permeabilization to the number of transient cavitation events. The model showed that nearly every stage of transient cavitation, including bubble expansion, collapse, and subsequent shock waves may contribute to membrane permeabilization. For each mechanism, the volume around the bubble within which bubbles induce reversible and irreversible membrane permeabilization was determined. Predictions of the model are consistent with experimental data. PMID:12719239

Sundaram, Jagannathan; Mellein, Berlyn R; Mitragotri, Samir

2003-05-01

220

Ultrafiltration by a compacted clay membrane. I - Oxygen and hydrogen isotopic fractionation. II - Sodium ion exclusion at various ionic strengths.  

NASA Technical Reports Server (NTRS)

Laboratory experiments were carried out to determine the magnitude of the isotopic fractionation of distilled water and of 0.01N NaCl forced to flow at ambient temperature under a hydraulic pressure drop of 100 bars across a montmorillonite disk compacted to a porosity of 35% by a pressure of 330 bars. The ultrafiltrates in both experiments were depleted in D by 2.5% and in O-18 by 0.8% relative to the residual solution. No additional isotopic fractionation due to a salt-filtering mechanism was observed at NaCl concentrations up to 0.01N. Adsorption is most likely the principal mechanism which produces isotopic fractionation, but molecular diffusion may play a minor role. The results suggest that oxygen and hydrogen isotopic fractionation of ground water during passage through compacted clayey sediments should be a common occurrence, in accord with published interpretations of isotopic data from the Illinois and Alberta basins. It is shown how it is possible to proceed from the ion exchange capacity of clay minerals and, by means of the Donnan membrane equilibrium concept and the Teorell-Meyer-Siever theory, develop a theory to explain why and to what extent ultrafiltration occurs when solutions of known concentration are forced to flow through a clay membrane.

Coplen, T. B.; Hanshaw, B. B.

1973-01-01

221

Nafion-layered sulfonated polysulfone fuel cell membranes  

NASA Astrophysics Data System (ADS)

Sulfonated polysulfone (SPSU) with high ion exchange capacity (IEC) and ion conductivity was synthesized through deep sulfonation of polysulfone using the trimethysilyl chlorosulfonate as sulfonation agent. The silicon-containing compounds formed during the synthesis of SPSU were completely removed from the SPSU by simple evaporation. Water swelling, ion exchange capacity, conductivity and fuel cell performance were measured for the SPSU membranes. SPSU membranes with IEC twice that of Nafion-115 were prepared. The conductivity of the SPSU increased exponentially with the relative humidity (RH), achieving conductivities of 0.1 S cm -1 for RH > 70%. A Nafion-layered SPSU PEM fuel cell membrane was synthesized through pressing a thin Nafion-115 layer on to both sides of SPSU membrane. The Nafion layers on the SPSU prevented the water-soluble SPSU from being washed out of MEA, and the membrane was stable during the fuel cell operation up to 120 °C.

Chen, Sheng-Li; Bocarsly, A. B.; Benziger, J.

222

The Mode of Segregation of the Bacterial Cell Membrane  

PubMed Central

The distribution of phospholipids from labeled parental membranes to progeny cells was studied by autoradiography and a minicell system. The minicell experiments showed that, during growth, the parental membrane is diluted at the same rate in cells and in minicells, which indicates that ends of cells are not different from the cylindrical portions with regard to the distribution of parental molecules. The same result was obtained after labeling heme-containing proteins with ?-aminolevulinic acid. The autoradiographic experiments indicate that the membrane segregates in about 250 subunits 4 × 104 nm2 in size. These subunits appear to be conserved during growth. Images

Green, Elizabeth Weaver; Schaechter, Moselio

1972-01-01

223

Use of semipermeable membrane devices (SPMDs) to characterize dissolved hydrocarbon fractions of both dispersed and undispersed oil.  

PubMed

Crude oil contamination remains a problem along coastal California and its impacts on pelagic organisms are of concern. Previous crude and dispersed oil studies showed a decrease in fish toxicity when Corexit 9500 dispersant was applied. However, observed sublethal metabolic effects were similar for both oil conditions, suggesting fish were accumulating similar dissolved hydrocarbons. This study aimed to characterize the bioavailable fraction of the water-accommodated fraction (WAF) and the chemically-enhanced WAF (CEWAF) of Prudhoe Bay Crude Oil (PBCO), using semi-permeable membrane devices (SPMDs) as fish models. Seven accumulated PAHs were identified (naphthalene, 2-methylnaphthalene, 1-methylnapthalene, biphenyl, fluorene, dibenzothiophene and phenanthrene) from 24 h static exposures. Although WAF and CEWAF oil loadings differed by eight-fold, accumulated dissolved concentrations among the seven PAHs differed by some three-fold. Overall, the use of SPMDs in characterizing the dissolved fraction of PBCO, has provided a better understanding of the bioavailability of crude and dispersed oil. PMID:24056734

Van Scoy, April R; Voorhees, Jennifer; Anderson, Brian S; Philips, Bryn M; Tjeerdema, Ronald S

2013-10-23

224

Heterogeneity of Arabinogalactan-Proteins on the Plasma Membrane of Rose Cells.  

PubMed Central

Arabinogalactan-proteins (AGPs) have been purified from the plasma membrane of suspension-cultured Paul's Scarlet rose (Rosa sp.) cells. The two most abundant and homogeneous plasma membrane AGP fractions were named plasma membrane AGP1 (PM-AGP1) and plasma membrane AGP2 (PM-AGP2) and had apparent molecular masses of 140 and 217 kD, respectively. Both PM-AGP1 and PM-AGP2 had [beta]-(1-3)-, [beta]-(1,6)-, and [beta]-(1,3,6)-galactopyranosyl residues, predominantly terminal [alpha]-arabinofuranosyl residues, and (1,4)- and terminal glucuronopyranosyl residues. The protein moieties of PM-AGP1 and PM-AGP2 were both rich in hydroxyproline, alanine, and serine, but differed in the abundance of hydroxyproline, which was 1.6 times higher in PM-AGP2 than in PM-AGP1. Another difference was the overall protein content, which was 3.7% (w/w) in PM-AGP1 and 15% in PM-AGP2. As judged by their behavior on reverse-phase chromatography, PM-AGP1 and PM-AGP2 were not more hydrophobic than AGPs from the cell wall or culture medium. In contrast, a minor plasma membrane AGP fraction eluted later on reverse-phase chromatography and was more negatively charged at pH 5 than either PM-AGP1 or PM-AGP2. The more negatively charged fraction contained molecules with a glycosyl composition characteristic of AGPs and included at least two different macromolecules. The results of this investigation indicate that Rosa plasma membrane contains at least four distinct AGPs or AGP-like molecules. These molecules differed from each other in size, charge, hydrophobicity, amino-acyl composition, and/or protein content.

Serpe, M. D.; Nothnagel, E. A.

1996-01-01

225

Reactivation of Escherichia coli cells, inactivated by ultraviolet rays, with cell extracts of propionic acid bacteria: Fractionation of the extract  

SciTech Connect

Separation of Propionibacterium shermanii extract into fractions and testing them for their reactivating effect on UV-inactivated Escherichia coli AB-1157 cells showed that the activity was associated with the fraction of soluble proteins. The activity was not demonstrated in the fractions of RNA, DNA, ribosomes, or cell walls. Fractional salting out and subsequent testing of the fractions showed two active protein fractions: fraction I (20-40% of ammonium sulfate saturating concentration) and fraction II (60-80%). These fractions were separated by HPLC into seven and eight subfractions, respectively. Reactivating activity was showed in subfraction 4 (fraction I) and subfractions 5 and 6 (fraction II). Electrophoresis showed five and four polypeptides in subfractions 4 and 5, respectively. Subfraction 6 (fraction II) contained one protein with a molecular mass of about 30 kDa. This protein, apparently, was responsible for the protective properties of fraction II. 9 refs., 2 figs., 4 tabs.

Vorob`eva, L.I.; Khodzhaev, E.Yu.; Ponomareva, G.M.; Ambrosov, I.V. [M.V. Lomonosov Moscow State Univ. (Russian Federation)

1995-01-01

226

Adaptive changes in pancreatic beta cell fractional area and beta cell turnover in human pregnancy  

Microsoft Academic Search

Aims\\/hypothesis  We sought to establish the extent and basis for adaptive changes in beta cell numbers in human pregnancy.\\u000a \\u000a \\u000a \\u000a Methods  Pancreas was obtained at autopsy from women who had died while pregnant (n?=?18), post-partum (n?=?6) or were not pregnant at or shortly before death (controls; n?=?20). Pancreases were evaluated for fractional pancreatic beta cell area, islet size and islet fraction of beta

A. E. Butler; L. Cao-Minh; R. Galasso; R. A. Rizza; A. Corradin; C. Cobelli; P. C. Butler

2010-01-01

227

Band 3 and glycophorin are progressively aggregated in density-fractionated sickle and normal red blood cells. Evidence from rotational and lateral mobility studies.  

PubMed Central

Band 3 aggregation in the plane of the red blood cell (RBC) membrane is postulated to be important in the pathophysiology of hemolysis of dense sickle and normal RBCs. We used the fluorescence photobleaching recovery and polarized fluorescence depletion techniques to measure the lateral and rotational mobility of band 3, glycophorins, and phospholipid analogues in membranes of density-separated intact RBCs from seven patients with sickle cell disease and eight normal controls. The fractions of laterally mobile band 3 and glycophorin decreased progressively as sickle RBC density increased. Normal RBCs also showed a progressive decrease in band 3 fractional mobility with increasing buoyant density. Rapidly rotating, slowly rotating, and rotationally immobile forms of band 3 were observed in both sickle and normal RBC membranes. The fraction of rapidly rotating band 3 progressively decreased and the fraction of rotationally immobile band 3 progressively increased with increasing sickle RBC density. Changes in the fraction of rotationally immobile band 3 were not reversible upon hypotonic swelling of dense sickle RBCs, and normal RBCs osmotically shrunken in sucrose buffers failed to manifest band 3 immobilization at median cell hemoglobin concentration values characteristic of dense sickle RBCs. We conclude that dense sickle and normal RBCs acquire irreversible membrane abnormalities that cause transmembrane protein immobilization and band 3 aggregation. Band 3 aggregates could serve as cell surface sites of autologous antibody binding and thereby lead to removal of dense sickle and normal (senescent) RBCs from the circulation.

Corbett, J D; Golan, D E

1993-01-01

228

Water transport in polymer electrolyte membrane fuel cells  

Microsoft Academic Search

Polymer electrolyte membrane fuel cell (PEMFC) has been recognized as a promising zero-emission power source for portable, mobile and stationary applications. To simultaneously ensure high membrane proton conductivity and sufficient reactant delivery to reaction sites, water management has become one of the most important issues for PEMFC commercialization, and proper water management requires good understanding of water transport in different

Kui Jiao; Xianguo Li

2011-01-01

229

Shape Dynamics of Nearly Spherical Membrane Bounded Fluid Cells  

Microsoft Academic Search

This paper considers the hydrodynamics of low Reynolds number shape fluctuations of incompressible fluid cells with incompressible membrane boundaries. It is assumed that the membrane can be characterized by a shear modulus ?, a bending modulus kc, an intrinsic mean curvature Co, and a surface viscosity ?M. The problem is formulated for the general case, and is solved analytically in

Mark A. Peterson

1985-01-01

230

Membrane fluidity measurements in peripheral cells from Huntington's disease patients  

PubMed Central

The primary gene defect of Huntington's disease is believed to involve the membrane of some peripheral cells. The membrane fluidity of skin fibroblasts, erythrocytes and leucocytes was measured by fluorescence polarisation of a lipid specific probe, 1,6 diphenyl, 1,3,5 hexatriene. No significant difference between controls and Huntington's disease patients could be demonstrated.

Beverstock, G C; Pearson, P L

1981-01-01

231

Membrane organization and cell fusion during mating in fission yeast requires multipass membrane protein Prm1.  

PubMed

The involvement of Schizosaccharomyces pombe prm1(+) in cell fusion during mating and its relationship with other genes required for this process have been addressed. S. pombe prm1? mutant exhibits an almost complete blockade in cell fusion and an abnormal distribution of the plasma membrane and cell wall in the area of cell-cell interaction. The distribution of cellular envelopes is similar to that described for mutants devoid of the Fig1-related claudin-like Dni proteins; however, prm1(+) and the dni(+) genes act in different subpathways. Time-lapse analyses show that in the wild-type S. pombe strain, the distribution of phosphatidylserine in the cytoplasmic leaflet of the plasma membrane undergoes some modification before an opening is observed in the cross wall at the cell-cell contact region. In the prm1? mutant, this membrane modification does not take place, and the cross wall between the mating partners is not extensively degraded; plasma membrane forms invaginations and fingers that sometimes collapse/retract and that are sometimes strengthened by the synthesis of cell-wall material. Neither prm1? nor prm1? dni? zygotes lyse after cell-cell contact in medium containing and lacking calcium. Response to drugs that inhibit lipid synthesis or interfere with lipids is different in wild-type, prm1?, and dni1? strains, suggesting that membrane structure/organization/dynamics is different in all these strains and that Prm1p and the Dni proteins exert some functions required to guarantee correct membrane organization that are critical for cell fusion. PMID:24514900

Curto, M-Ángeles; Sharifmoghadam, Mohammad Reza; Calpena, Eduardo; De León, Nagore; Hoya, Marta; Doncel, Cristina; Leatherwood, Janet; Valdivieso, M-Henar

2014-04-01

232

Membrane fouling of submerged membrane bioreactors: impact of mean cell residence time and the contributing factors.  

PubMed

In this study, four bench-scale pre-anoxic submerged membrane bioreactors (MBR) were operated simultaneously at different mean cell residence times (MCRTs) (3, 5, 10, and 20 days) to systematically elucidate the contributing factors of membrane fouling. Severe membrane fouling was first observed in the 3-day followed by the 5-day MCRT MBRs. Minimal membrane fouling was detected in the 10 and 20-day MCRT MBRs. The fouling of microfiltration membrane was not controlled by mixed liquor suspended solids concentration or zeta potential of biomass. Instead, membrane fouling rate increased with increasing soluble microbial products and extracellular polymeric substances concentrations, which both increased with decreasing MCRT. Total organic carbon, protein, carbohydrate, and UV254 absorbance in the mixed liquor supernatant increased with decreasing MCRT and were consistently higher than those of the effluent. Accumulation of carbohydrates rather than proteins in the mixed liquor supernatant was found to decrease with increasing MCRT. Normalized capilliary suction time value rather than the capilliary suction time value would indicate membrane fouling potential of a mixed liquor. Image analysis of the fouled membrane using scanning electron microscope and confocal laser scanning microscope showed that biofilm formation was the cause of membrane fouling. PMID:16683612

Ng, How Y; Tan, Teck Wee; Ong, Say Leong

2006-04-15

233

Design of efficient methanol impermeable membranes for fuel cell applications.  

PubMed

In this paper, the design of efficient composite membranes based on sulfonated polysulfone and acidic silica material with characteristics and properties such as methanol barrier, high proton conductivity and suitable fuel cells performance is presented. A positive influence of nanosized acidic silica powders, used as an additive filler in the preparation of composite membranes, due to an efficient hydrophilic inter-distribution inside the membrane when compared to pure silica, is found. A series of different techniques such as XRF, FT-IR, TGA, DSC, IEC and conductivity measurements are used to highlight the properties of acidic silica material and composite membranes. The composite membrane based on acidic silica (SPSf-SiO(2)-S) shows the lowest crossover current (only 8 mA cm(-2)), which is 43% lower than that of a pure SPSf membrane and 33% lower compared to a composite membrane based on bare silica (SPSf-SiO(2)). These significant differences are attributed to the increasing diffusion path length of MeOH/H(2)O clusters in the composite membranes. The maximum DMFC performance at 30 °C is achieved with the SPSf-SiO(2)-S membrane (23 mW cm(-2)), whereas the MEAs based on SPSf-SiO(2) and pure SPSf membranes reached 21 and 16 mW cm(-2), respectively. These significant results of the composite SPSf-SiO(2)-S membrane are ascribed at a good compromise among high proton conductivity, low swelling and low methanol crossover compared to pure SPSf and (unmodified silica)-SPSf membranes. A preliminary short durability test of 100 h performed in a cell with the composite SPSf-SiO(2)-S membrane shows remarkable performance stability during chrono-voltammetric measurements (60 mA cm(-2)) at 30 °C. PMID:22274611

Lufrano, F; Baglio, V; Di Blasi, O; Staiti, P; Antonucci, V; Aricò, A S

2012-02-28

234

Correlations between the Dielectric Properties and Exterior Morphology of Cells Revealed by Dielectrophoretic Field-Flow Fractionation  

PubMed Central

Although dielectrophoresis (DEP) has great potential for addressing clinical cell isolation problems based on cell dielectric differences, a biological basis for predicting the DEP behavior of cells has been lacking. Here, the dielectric properties of the NCI-60 panel of tumor cell types have been measured by dielectrophoretic (DEP) field-flow fractionation, correlated with the exterior morphologies of the cells during growth, and compared with the dielectric and morphological characteristics of the subpopulations of peripheral blood. In agreement with earlier findings, cell total capacitance varied with both cell size and plasma membrane folding and the dielectric properties of the NCI-60 cell types in suspension reflected the plasma membrane area and volume of the cells at their growth sites. Therefore, the behavior of cells in DEP-based manipulations is largely determined by their exterior morphological characteristics prior to release into suspension. As a consequence, DEP is able to discriminate between cells of similar size having different morphological origins, offering a significant advantage over size-based filtering for isolating circulating tumor cells, for example. The findings provide a framework for anticipating cell dielectric behavior on the basis of structure-function relationships and suggest that DEP should be widely applicable as a surface marker-independent method for sorting cells.

Gascoyne, Peter R. C.; Shim, Sangjo; Noshari, Jamileh; Becker, Frederick F.; Stemke-Hale, Katherine

2013-01-01

235

Subcellular fractionation and morphology of calf aortic smooth muscle cells: studies on whole aorta, aortic explants, and subcultures grown under  

PubMed Central

A comparative biochemical and morphological study was made of calf aortic smooth muscle cells found in situ and grown in vitro under various conditions. Striking alterations in enzyme contents, physical properties, and morphological appearances of lysosomes, endoplasmic reticulum, plasma membranes and, to a lesser extent, mitochondria were observed upon culturing of calf aortic smooth muscle cells. These changes first appeared in cells growing out of tissue explants. They developed further upon subculturing of the cells and depended greatly on the culture conditions used. The alterations included increases in specific activities of some 5- to 25-fold of four acid hydrolases, an average ninefold increase in 5' -nucleotidase, sevenfold increase in cytochrome oxidase, and fourfold increase in neutral ?-glucosidase in subcultured smooth muscle cells compared to aortic cells in situ. Cell fractionation studies showed significant shifts in the equilibrium densities of plasma membranes, microsomes, and lysosomes, but not of mitochondria, in smooth muscle cells growing out from explants and in subcultured cells, compared to cells isolated from intact aortas. Although the cells grown in vitro exhibited typical phenotypic features of smooth muscle cells such as abundant myofilaments and surface vesicles, alterations in the morphological appearance of the endoplasmic reticulum, Golgi apparatus, and, especially, lysosomes were observed. These results demonstrate significant differences in specific cellular characteristics and functions of aortic smooth muscle cells grown in vitro compared to aortic cells in situ.

Fowler, S; Shio, H; Wolinsky, H

1977-01-01

236

Adaptation of yeast cell membranes to ethanol  

SciTech Connect

A highly ethanol-tolerant Saccharomyces wine strain is able, after growth in the presence of ethanol, to efficiently improve the ethanol tolerance of its membrane. A less-tolerant Saccharomyces laboratory strain, however, is unable to adapt its membrane to ethanol. Furthermore, after growth in the presence of ethanol, the membrane of the latter strain becomes increasingly sensitive, although this is a reversible process. Reversion to a higher tolerance occurs only after the addition of an energy source and does not take place in the presence of cycloheximide.

Jimenez, J.; Benitez, T.

1987-05-01

237

Membrane glycoprotein changes associated with anthracycline resistance in HL60 cells  

Microsoft Academic Search

The glycoproteins on the surface of HL-60\\/S wild-type, drug-sensitive human leukemia cells and HL-60\\/AR anthracycline-resistant cells which do not overexpress the P-glycoprotein, were characterized by labeling with [35S]-methionine, NaB[3H4], phosphorus 32, or sodium iodide I 125. HL-60\\/S and HL-60\\/AR cell lysates and membrane fractions tagged with [35S]-methionine or phosphorus 32 showed no significant differences in their protein patterns as analyzed

James E. Gervasoni Jr; Robert N. Taub; Michelle Rosado; Sindu Krishna; Valerie J. Stewart; Daniel M. Knowles; Kapil Bhalla; Douglas D. Ross; Michael A. Baker; Jose Lutzky; Alexander A. Hindenburg

1991-01-01

238

The Anti-angiogenic Peptide Anginex Disrupts the Cell Membrane  

PubMed Central

Anginex is a synthetic beta-sheet peptide with anti-angiogenic and anti-tumor activity. When added to cultured endothelial cells at concentrations ranging from 2.5 ?M to 25 ?M, anginex induced cell death, which was reflected by a strong increase of subdiploid cells and fragments, loss of cellular ATP, and LDH release. Cytotoxicity remained the same whether cells were treated with anginex at 4 °C or at 37 °C. At low temperatures, fluorescein-conjugated anginex accumulated on the endothelial surface, but did not reach into the cytoplasm, indicating that the cell membrane is the primary target for the peptide. Within minutes of treatment, anginex caused endothelial cells to take up propidium iodide and undergo depolarization, both parameters characteristic for permeabilization of the cell membrane. This process was amplified when cells were activated with hydrogen peroxide. Red blood cell membranes were essentially unaffected by anginex. Anginex bound lipid bilayers with high affinity and with a clear preference for anionic over zwitterionic phospholipids. Structural studies by circular dichroism and solid-state nuclear magnetic resonance showed that anginex forms a beta-sheet and adopts a unique and highly ordered conformation upon binding to lipid membranes. This is consistent with lipid micellization or the formation of pore-forming beta-barrels. The data suggest that the cytotoxicity of anginex stems from its ability to target and disrupt the endothelial cell membrane, providing a possible explanation for the angiostatic activity of the peptide.

Pilch, Jan; Franzin, Carla M.; Knowles, Lynn M.; Ferrer, Fernando J.; Marassi, Francesca M.; Ruoslahti, Erkki

2010-01-01

239

Low Crossover Polymer Electrolyte Membranes for Direct Methanol Fuel Cells  

NASA Technical Reports Server (NTRS)

Direct Methanol Fuel Cells (DMFC's) using polymer electrolyte membranes are promising power sources for portable and vehicular applications. State of the art technology using Nafion(R) 117 membranes (Dupont) are limited by high methanol permeability and cost, resulting in reduced fuel cell efficiencies and impractical commercialization. Therefore, much research in the fuel cell field is focused on the preparation and testing of low crossover and cost efficient polymer electrolyte membranes. The University of Southern California in cooperation with the Jet Propulsion Laboratory is focused on development of such materials. Interpenetrating polymer networks are an effective method used to blend polymer systems without forming chemical links. They provide the ability to modify physical and chemical properties of polymers by optimizing blend compositions. We have developed a novel interpenetrating polymer network based on poly (vinyl - difluoride)/cross-linked polystyrenesulfonic acid polymer composites (PVDF PSSA). Sulfonation of polystyrene accounts for protonic conductivity while the non-polar, PVDF backbone provides structural integrity in addition to methanol rejection. Precursor materials were prepared and analyzed to characterize membrane crystallinity, stability and degree of interpenetration. USC JPL PVDF-PSSA membranes were also characterized to determine methanol permeability, protonic conductivity and sulfur distribution. Membranes were fabricated into membrane electrode assemblies (MEA) and tested for single cell performance. Tests include cell performance over a wide range of temperatures (20 C - 90 C) and cathode conditions (ambient Air/O2). Methanol crossover values are measured in situ using an in-line CO2 analyzer.

Prakash, G. K. Surya; Smart, Marshall; Atti, Anthony R.; Olah, George A.; Narayanan, S. R.; Valdez, T.; Surampudi, S.

1996-01-01

240

Membrane processes and biophysical characterization of living cells decorated with chromatic polydiacetylene vesicles  

Microsoft Academic Search

The structural complexity of the cell membrane makes analysis of membrane processes in living cells, as compared to model membrane systems, highly challenging. Living cells decorated with surface-attached colorimetric\\/fluorescent polydiacetylene patches might constitute an effective platform for analysis and visualization of membrane processes in situ. This work examines the biological and chemical consequences of plasma membrane labeling of promyelocytic leukemia

Natalie Groysman; Zulfiya Orynbayeva; Marina Katz; Sofiya Kolusheva; Marina Khanin; Michael Danilenko; Raz Jelinek

2008-01-01

241

Affinity driven molecular transfer from erythrocyte membrane to target cells.  

PubMed

A wide variety of antimicrobial peptides are known to bind to - and disrupt microbial plasma membranes. Recently, derivatives of the antimicrobial peptide dermaseptin S4 were shown to selectively disrupt the plasma membrane of the intracellular parasite Plasmodium falciparum without harming that of the mammalian host cell. The resulting antimalarial activity is allegedly exerted after the harmless peptide binding to the membrane of the host cell, followed by peptide translocation across a number of intracellular membrane systems and interaction with that of the intraerythrocyte parasite. In this study, we present evidence in support of the ability of a membrane-bound peptide, the dermaseptin S4 derivative K(4)-S4(1-13)a, to transfer from red blood cells (RBCs) to another distant membrane. Binding of K(4)-S4(1-13)a to the plasma membrane of RBCs was assessed in vitro and in vivo, and found to be rapid, spontaneous and receptor independent, as was the transfer of the RBC-bound peptide to the plasma membrane of microorganisms. The present study further provides a basis for the possible use of RBCs as a transport vehicle to deliver drugs to distant targets. This drug delivery system involves the transient "loading" of RBCs with a lipophilic "hook" peptide. Such a peptide has enough affinity for the RBC's plasma membrane to bind to the membrane, but given the opportunity, the peptide will exit its position and transfer to another (target) cell for which it has a greater affinity. The efficacy of such an affinity driven transfer system was demonstrated experimentally by the transfer of K(4)-S4(1-13)a from pre-loaded RBCs to bacteria, yeast and protozoan target cells. PMID:11587797

Feder, R; Nehushtai, R; Mor, A

2001-10-01

242

Diffusion Coefficient of Fluorescent Phosphatidylinositol 4,5-bisphosphate in the Plasma Membrane of Cells  

PubMed Central

Phosphatidylinositol 4,5-bisphosphate (PIP2) controls a surprisingly large number of processes in cells. Thus, many investigators have suggested that there might be different pools of PIP2 on the inner leaflet of the plasma membrane. If a significant fraction of PIP2 is bound electrostatically to unstructured clusters of basic residues on membrane proteins, the PIP2 diffusion constant, D, should be reduced. We microinjected micelles of Bodipy TMR-PIP2 into cells, and we measured D on the inner leaflet of fibroblasts and epithelial cells by using fluorescence correlation spectroscopy. The average ± SD value from all cell types was D = 0.8 ± 0.2 ?m2/s (n = 218; 25°C). This is threefold lower than the D in blebs formed on Rat1 cells, D = 2.5 ± 0.8 ?m2/s (n = 26). It is also significantly lower than the D in the outer leaflet or in giant unilamellar vesicles and the diffusion coefficient for other lipids on the inner leaflet of these cell membranes. The simplest interpretation is that approximately two thirds of the PIP2 on inner leaflet of these plasma membranes is bound reversibly.

Golebiewska, Urszula; Nyako, Marian; Woturski, William; Zaitseva, Irina

2008-01-01

243

Membrane and actin reorganization in electropulse-induced cell fusion.  

PubMed

When cells of Dictyostelium discoideum are exposed to electric pulses they are induced to fuse, yielding motile polykaryotic cells. By combining electron microscopy and direct recording of fluorescent cells, we have studied the emergence of fusion pores in the membranes and the localization of actin to the cell cortex. In response to electric pulsing, the plasma membranes of two contiguous cells are turned into tangles of highly bent and interdigitated membranes. Live-imaging of cells double-labeled for membranes and filamentous actin revealed that actin is induced to polymerize in the fusion zone to temporarily bridge the gaps in the vesiculating membrane. The diffusion of green fluorescent protein (GFP) from one fusion partner to the other was scored using spinning disc confocal microscopy. Fusion pores that allowed intercellular exchange of GFP were formed after a delay, which lasted up to 24 seconds after exposure of the cells to the electric field. These data indicate that the membranes persist in a fusogenic state before pores of about 3 nm diameter are formed. PMID:23447671

Gerisch, Günther; Ecke, Mary; Neujahr, Ralph; Prassler, Jana; Stengl, Andreas; Hoffmann, Max; Schwarz, Ulrich S; Neumann, Eberhard

2013-05-01

244

Cell-free system for synthesizing membrane proteins cell free method for synthesizing membrane proteins  

DOEpatents

The invention provides an in vitro method for producing proteins, membrane proteins, membrane-associated proteins, and soluble proteins that interact with membrane-associated proteins for assembly into an oligomeric complex or that require association with a membrane for proper folding. The method comprises, supplying intracytoplasmic membranes from organisms; modifying protein composition of intracytoplasmic membranes from organism by modifying DNA to delete genes encoding functions of the organism not associated with the formation of the intracytoplasmic membranes; generating appropriate DNA or RNA templates that encode the target protein; and mixing the intracytoplasmic membranes with the template and a transcription/translation-competent cellular extract to cause simultaneous production of the membrane proteins and encapsulation of the membrane proteins within the intracytoplasmic membranes.

Laible, Philip D; Hanson, Deborah K

2013-06-04

245

Decreasing Outer Hair Cell Membrane Cholesterol Increases Cochlear Electromechanics  

NASA Astrophysics Data System (ADS)

The effect of decreasing membrane cholesterol on the mechanical response of the cochlea to acoustic and/or electrical stimulation was monitored using laser interferometry. In contrast to pharmacological interventions that typically decrease cochlear electromechanics, reducing membrane cholesterol increased the response. The electromechanical response in untreated preparations was asymmetric with greater displacements in response to positive currents and cholesterol depletion increased the asymmetry. The results confirm that outer hair cell electromotility is enhanced by low membrane cholesterol. The asymmetry of the response indicates the outer hair cell resting membrane potential is hyperpolarized relative to the voltage of maximum gain for the outer hair cell voltage-displacement function. The magnitude of the response increase suggests a non-uniform distribution of cholesterol along the lateral wall of normal adult outer hair cells.

Brownell, William E.; Jacob, Stefan; Hakizimana, Pierre; Ulfendahl, Mats; Fridberger, Anders

2011-11-01

246

Strategies for the cell-free expression of membrane proteins.  

PubMed

Cell-free expression offers an interesting alternative method to produce membrane proteins in high amounts. Elimination of toxicity problems, reduced proteolytic degradation and a nearly unrestricted option to supply potentially beneficial compounds like cofactors, ligands or chaperones into the reaction are general advantages of cell-free expression systems. Furthermore, the membrane proteins may be translated directly into appropriate hydrophobic and membrane-mimetic surrogates, which might offer significant benefits for the functional folding of the synthesized proteins. Cell-free expression is a rapidly developing and highly versatile technique and several systems of both, prokaryotic and eukaryotic origins, have been established. We provide protocols for the cell-free expression of membrane proteins in different modes including their expression as precipitate as well as their direct synthesis into detergent micelles or lipid bilayers. PMID:20204858

Reckel, Sina; Sobhanifar, Solmaz; Durst, Florian; Löhr, Frank; Shirokov, Vladimir A; Dötsch, Volker; Bernhard, Frank

2010-01-01

247

Evaluation of composite membranes for direct methanol fuel cells  

NASA Astrophysics Data System (ADS)

The performance of direct methanol fuel cells (DMFCs) can be significantly affected by the transport of methanol through the membrane, depolarising the cathode. In this paper, the literature on composite membranes that have been developed for reduction of methanol crossover in DMFCs is reviewed. While such membranes can be effective in reducing methanol permeability, this is usually combined with a reduction in proton conductivity. Measurements of methanol permeability and proton conductivity are relatively straightforward, and these parameters (or a membrane 'selectivity' based on the ratio between them) are often used to characterize DMFC membranes. However, we have carried out one-dimensional simulations of DMFC performance for a wide range of membrane properties, and the results indicate that DMFC performance is normally either limited by methanol permeability or proton conductivity. Thus use of a 'selectivity' is not appropriate for comparison of membrane materials, and results from the model can be used to compare different membranes. The results also show that Nafion ® 117 has an optimum thickness, where DMFC performance is equally limited by both methanol permeability and proton conductivity. The model also indicates that new composite membranes based on Nafion ® can only offer significant improvement in DMFC performance by enabling operation with increased methanol concentration in the fuel. A number of composite membrane materials that have been reported in the literature are shown to deliver significant reduction in DMFC performance due to reduced proton conductivity, although improved performance at high methanol concentration may be possible.

Li, X.; Roberts, E. P. L.; Holmes, S. M.

248

Modeling of interactions between nanoparticles and cell membranes  

NASA Astrophysics Data System (ADS)

Rapid development of nanotechnology and ability to manufacture materials and devices with nanometer feature size leads to exciting innovations in many areas including the medical and electronic fields. However, the possible health and environmental impacts of manufactured nanomaterials are not fully known. Recent experimental reports suggest that some of the manufactured nanomaterials, such as fullerenes and carbon nanotubes, are highly toxic even in small concentrations. The goal of the current work is to understand the mechanisms responsible for the toxicity of nanomaterials. In the current study coarse-grained molecular dynamics simulations are employed to investigate the interactions between NPs and cellular membranes at a molecular level. One of the possible toxicity mechanisms of the nanomaterials is membrane disruption. Possibility of membrane disruption exposed to the manufactured nanomaterials are examined by considering chemical reactions and non-reactive physical interactions as chemical as well as physical mechanisms. Mechanisms of transport of carbon-based nanoparticles (fullerene and its derivative) across a phospholipid bilayer are investigated. The free energy profile is obtained using constrained simulations. It is shown that the considered nanoparticles are hydrophobic and therefore they tend to reside in the interior of the lipid bilayer. In addition, the dynamics of the membrane fluctuations is significantly affected by the nanoparticles at the bilayer-water interface. The hydrophobic interaction between the particles and membrane core induces the strong coupling between the nanoparticle motion and membrane deformation. It is observed that the considered nanoparticles affect several physical properties of the membrane. The nanoparticles embedded into the membrane interior lead to the membrane softening, which becomes more significant with increase in CNT length and concentration. The lateral pressure profile and membrane energy in the membrane containing the nanoparticles exhibit localized perturbation around the nanoparticle. The nanoparticles are not likely to affect membrane protein function by the weak perturbation of the internal stress in the membrane. Due to the short-ranged interactions between the nanoparticles, the nanoparticles would not form aggregates inside membranes. The effect of lipid peroxidation on cell membrane deformation is assessed. The peroxidized lipids introduce a perturbation to the internal structure of the membrane leading to higher amplitude of the membrane fluctuations. Higher concentration of the peroxidized lipids induces more significant perturbation. Cumulative effects of lipid peroxidation caused by nanoparticles are examined for the first time. The considered amphiphilic particle appears to reduce the perturbation of the membrane structure at its equilibrium position inside the peroxidized membrane. This suggests a possibility of antioxidant effect of the nanoparticle.

Ban, Young-Min

249

A Flow Cell for Use with an Oxygen Membrane Electrode  

Microsoft Academic Search

A flow cell was constructed from plexiglass. The cell was designed to allow insertion and removal of a Clark-type oxygen membrane electrode. It was used in a flow system to amperometrically determine glucose and glucose oxidase via oxygen depletion. Hydrogen peroxide was determined by oxidation at +0.9 V vs. the silver\\/silver chloride electrode by removing the electrode membrane. Alternatively, a

E. Lawrence Gulberg; Gary D. Christian

1979-01-01

250

Basement Membrane Matrix (BME) has Multiple Uses with Stem Cells  

Microsoft Academic Search

The utilization of basement membrane matrix has helped to overcome many of the obstacles associated with stem cell research.\\u000a Initially, there were several problems with investigating stem cells, including difficult extraction from tissues, the need\\u000a for feeder layers, poor survival, minimal proliferation, limited differentiation in vitro, and inadequate survival when injected\\u000a or transplanted in vivo. Given that the basement membrane

Irina Arnaoutova; Jay George; Hynda K. Kleinman; Gabriel Benton

251

Fibronectin coating of oxygenator membranes enhances endothelial cell attachment  

PubMed Central

Background Extracorporeal membrane oxygenation (ECMO) can replace the lungs’ gas exchange capacity in refractory lung failure. However, its limited hemocompatibility, the activation of the coagulation and complement system as well as plasma leakage and protein deposition hamper mid- to long-term use and have constrained the development of an implantable lung assist device. In a tissue engineering approach, lining the blood contact surfaces of the ECMO device with endothelial cells might overcome these limitations. As a first step towards this aim, we hypothesized that coating the oxygenator’s gas exchange membrane with proteins might positively influence the attachment and proliferation of arterial endothelial cells. Methods Sheets of polypropylene (PP), polyoxymethylpentene (TPX) and polydimethylsiloxane (PDMS), typical material used for oxygenator gas exchange membranes, were coated with collagen, fibrinogen, gelatin or fibronectin. Tissue culture treated well plates served as controls. Endothelial cell attachment and proliferation were analyzed for a period of 4 days by microscopic examination and computer assisted cell counting. Results Endothelial cell seeding efficiency is within range of tissue culture treated controls for fibronectin treated surfaces only. Uncoated membranes as well as all other coatings lead to lower cell attachment. A confluent endothelial cell layer develops on fibronectin coated PDMS and the control surface only. Conclusions Fibronectin increases endothelial cells’ seeding efficiency on different oxygenator membrane material. PDMS coated with fibronectin shows sustained cell attachment for a period of four days in static culture conditions.

2013-01-01

252

Association of pp60src and src protein kinase activity with the plasma membrane of nonpermissive and permissive avian sarcoma virus-infected cells.  

PubMed Central

The intracellular localization of pp60src and src protein kinase activity in avian sarcoma virus (ASV)-infected chicken embryo fibroblasts and transformed and morphologically reverted field vole cells was examined by subcellular fractionation procedures. Fractionation by differential centrifugation of Dounce-homogenized cellular extracts prepared from vole cells showed that 83 to 91% of pp60src sedimented with particulate subcellular components from both transformed and revertant vole cells. A slightly lesser amount (60 to 70%) of pp60src was found associated with the particulate fraction from ASV-infected chicken embryo fibroblasts. The distribution of src protein kinase activity in the cytosol and particulate cell fractions was identical to that of pp60src, indicating no detectable differences in the activity of cytosol- and particulate-associated pp60src. When subcellular components of the cell were fractionated by discontinuous sucrose gradient centrifugation, similar amounts of both pp60src and src protein kinase activity cosedimented with the plasma membrane fractions from both transformed and revertant vole cells, as well as from ASV-infected chicken embryo fibroblasts. src protein kinase activity associated with plasma membrane fractions prepared from vole cells and ASV-infected chicken embryo fibroblasts was resistant to extraction with high salt concentrations, but partial elution was achieved with nonionic detergent. Thus, in both transformed and morphologically reverted vole cells, pp60src is intimately associated with the plasma membrane. Since transforming virus can be rescued from revertant vole cells by fusion to chicken embryo fibroblasts, revertant vole cell pp60src is capable of inducing morphological transformation. Thus, although the data presented herein suggest that transformation requires the association of pp60src with the plasma membrane, the binding of pp60src to the plasma membrane per se is insufficient to induce morphological transformation and requires the additional interaction with a specific target membrane protein which appears to be defective in revertant vole cells. Images

Krzyzek, R A; Mitchell, R L; Lau, A F; Faras, A J

1980-01-01

253

Comparative lipidomics analysis of HIV-1 particles and their producer cell membrane in different cell lines.  

PubMed

Human immunodeficiency virus type 1 (HIV-1) is a retrovirus that obtains its lipid envelope by budding through the plasma membrane of infected host cells. Various studies indicated that the HIV-1 membrane differs from the producer cell plasma membrane suggesting virus budding from pre-existing subdomains or virus-mediated induction of a specialized budding membrane. To perform a comparative lipidomics analysis by quantitative mass spectrometry, we first evaluated two independent methods to isolate the cellular plasma membrane. Subsequent lipid analysis of plasma membranes and HIV-1 purified from two different cell lines revealed a significantly different lipid composition of the viral membrane compared with the host cell plasma membrane, independent of the cell type investigated. Virus particles were significantly enriched in phosphatidylserine, sphingomyelin, hexosylceramide and saturated phosphatidylcholine species when compared with the host cell plasma membrane of the producer cells; they showed reduced levels of unsaturated phosphatidylcholine species, phosphatidylethanolamine and phosphatidylinositol. Cell type-specific differences in the lipid composition of HIV-1 and donor plasmamembranes were observed for plasmalogen-phosphatidylethanolamine and phosphatidylglycerol, which were strongly enriched only in HIV-1 derived from MT-4 cells. MT-4 cell-derived HIV-1 also contained dihydrosphingomyelin as reported previously, but this lipid class was also enriched in the host cell membrane. Taken together, these data strongly support the hypothesis that HIV-1 selects a specific lipid environment for its morphogenesis. PMID:23279151

Lorizate, Maier; Sachsenheimer, Timo; Glass, Bärbel; Habermann, Anja; Gerl, Mathias J; Kräusslich, Hans-Georg; Brügger, Britta

2013-02-01

254

Computational modeling and optimization of proton exchange membrane fuel cells  

Microsoft Academic Search

Improvements in performance, reliability and durability as well as reductions in production costs, remain critical prerequisites for the commercialization of proton exchange membrane fuel cells. In this thesis, a computational framework for fuel cell analysis and optimization is presented as an innovative alternative to the time consuming trial-and-error process currently used for fuel cell design. The framework is based on

Marc Secanell Gallart

2007-01-01

255

Development of a proton exchange membrane fuel cell cogeneration system  

Microsoft Academic Search

A proton exchange membrane fuel cell (PEMFC) cogeneration system that provides high-quality electricity and hot water has been developed. A specially designed thermal management system together with a microcontroller embedded with appropriate control algorithm is integrated into a PEM fuel cell system. The thermal management system does not only control the fuel cell operation temperature but also recover the heat

Jenn Jiang Hwang; Meng Lin Zou

2010-01-01

256

Concomitant Alterations of Sodium Flux and Membrane Phospholipid Metabolism in Red Blood Cells: Studies in Hereditary Spherocytosis*  

PubMed Central

The role of membrane phosphatides in transport processes has been investigated in red cells from splenectomized patients with hereditary spherocytosis (HS). Incorporation of inorganic 32phosphate into the membrane phosphatides of HS red cells was approximately twice normal, coinciding with the nearly twofold increment in flux of sodium ions in the cells. A consistent, inordinate increase in specific activity of a chromatographic fraction containing phosphatidylserine provided the bulk of the over-all increase in labeling of HS red cell phosphatides. The specific activity of phosphatidic acid was increased but not consistently. Radioactivity of the “acidic phosphatides” (phosphatidylserine and phosphatidic acid fractions) decreased, in general, when the sodium flux was low, i.e., when the cells were suspended in media of low sodium content. When the cation flux was elevated (hypotonic media), there was a marked (ca. 35%) increase in the labeling of phosphatidylserine fractions. Normal red cells whose permeability to cations was increased by exposure to 0.5 N butanol also exhibited increased labeling of acidic phosphatides. Considerations of the stoichiometry of cation transport and phosphatide labeling make it unlikely that phospholipids act directly as carrier molecules for cations in red cell membranes. On the other hand, the involvement of these lipid substances in cation movements is substantiated by correlating several different states of sodium flux with the labeling of the phosphatidic acid and phosphatidylserine fractions. Images

Jacob, Harry S.; Karnovsky, Manfred L.

1967-01-01

257

Controlled permeation of cell membrane by single bubble acoustic cavitation.  

PubMed

Sonoporation is the membrane disruption generated by ultrasound and has been exploited as a non-viral strategy for drug and gene delivery. Acoustic cavitation of microbubbles has been recognized to play an important role in sonoporation. However, due to the lack of adequate techniques for precise control of cavitation activities and real-time assessment of the resulting sub-micron process of sonoporation, limited knowledge has been available regarding the detail processes and correlation of cavitation with membrane disruption at the single cell level. In the current study, we developed a combined approach including optical, acoustical, and electrophysiological techniques to enable synchronized manipulation, imaging, and measurement of cavitation of single bubbles and the resulting cell membrane disruption in real-time. Using a self-focused femtosecond laser and high frequency ultrasound (7.44MHz) pulses, a single microbubble was generated and positioned at a desired distance from the membrane of a Xenopus oocyte. Cavitation of the bubble was achieved by applying a low frequency (1.5MHz) ultrasound pulse (duration 13.3 or 40?s) to induce bubble collapse. Disruption of the cell membrane was assessed by the increase in the transmembrane current (TMC) of the cell under voltage clamp. Simultaneous high-speed bright field imaging of cavitation and measurements of the TMC were obtained to correlate the ultrasound-generated bubble activities with the cell membrane poration. The change in membrane permeability was directly associated with the formation of a sub-micrometer pore from a local membrane rupture generated by bubble collapse or bubble compression depending on ultrasound amplitude and duration. The impact of the bubble collapse on membrane permeation decreased rapidly with increasing distance (D) between the bubble (diameter d) and the cell membrane. The effective range of cavitation impact on membrane poration was determined to be D/d=0.75. The maximum mean radius of the pores was estimated from the measured TMC to be 0.106±0.032?m (n=70) for acoustic pressure of 1.5MPa (duration 13.3?s), and increased to 0.171±0.030?m (n=125) for acoustic pressure of 1.7MPa and to 0.182±0.052?m (n=112) for a pulse duration of 40?s (1.5MPa). These results from controlled cell membrane permeation by cavitation of single bubbles revealed insights and key factors affecting sonoporation at the single cell level. PMID:21945682

Zhou, Y; Yang, K; Cui, J; Ye, J Y; Deng, C X

2012-01-10

258

[Characteristics of red cell membrane disorders in the Japanese population].  

PubMed

The characteristic features of the incidence of hereditary red cell membrane disorders in the Japanese population are described, based on our studies on 610 patients from 353 kindreds during 20 years since 1975. These patients were screened by a protocol on red cell morphology (scanning and transmission electron microscopy), red cell membrane proteins (sodium dodecylsulfate polyacrylamide gel electrophoresis, and kinetics of membrane proteins), membrane lipids, biophysical studies (ektacytometry, mechanical stability, and fluorescence recovery after photobleaching method), and membrane transport (sodium influx and efflux, and anion transport). Hereditary spherocytosis (HS) is most frequent (308 patients from 156 kindreds), hereditary elliptocytosis (HE) is the second (98 patients from 47 kindreds) followed by hereditary stomatocytosis (57 patients from 40 kindreds). Among the molecular abnormalities detected, alpha-spectrin mutation in the Japanese HE patients appeared extremely rare (only one family with spectrin alpha 1/74), despite three novel beta-spectrin mutations were found out of nine world-wide cases. Most of the Japanese HE patients were associated with partial protein 4.1 deficiencies. Ankyrin abnormalities in the Japanese HS patients appeared less common than those in the Western countries. Complete protein 4.2 deficiencies (34 patients from 20 kindreds) were unique in the Japanese population. Membrane lipid abnormalities included hereditary high red cell membrane phosphatidylcholine hemolytic anemia (30 patients from 18 kindreds), congenital beta-lipoprotein deficiency (acanthocytosis: seven patients from five kindreds), and each one patient of congenital lecithin: cholesterol acyltransferase deficiency and of congenital alpha-lipoprotein deficiency (Tangier disease). PMID:9136602

Yawata, Y

1997-04-01

259

Infrared nonlinear optical measurements of membrane potential in photoreceptor cells.  

PubMed Central

In the past it has not been possible to measure optically the membrane potential of cells and collections of cells that are either naturally photosensitive or that can be activated by photolyzable caged transmitter molecules. This paper reports on a unique application of nonlinear optics that can monitor the potential of cellular membranes with a near-infrared source. Among many other singular advantages, this nonlinear optical approach to measuring membrane potential does not activate light sensitive cells or cell suspensions and cellular networks surrounded with photolyzable molecules. To demonstrate this capability we show that the technique can be applied to living photoreceptor cells that are very sensitive to visible light. These cells are ideal for characterizing such a new technique, not only because of their unmatched sensitivity to light, but also because their electrical responses have been extensively characterized (Minks and Selinger, 1992). Images FIGURE 2

Ben-Oren, I; Peleg, G; Lewis, A; Minke, B; Loew, L

1996-01-01

260

Proton conducting membranes for high temperature fuel cells with solid state water free membranes  

NASA Technical Reports Server (NTRS)

A water free, proton conducting membrane for use in a fuel cell is fabricated as a highly conducting sheet of converted solid state organic amine salt, such as converted acid salt of triethylenediamine with two quaternized tertiary nitrogen atoms, combined with a nanoparticulate oxide and a stable binder combined with the converted solid state organic amine salt to form a polymeric electrolyte membrane. In one embodiment the membrane is derived from triethylenediamine sulfate, hydrogen phosphate or trifiate, an oxoanion with at least one ionizable hydrogen, organic tertiary amine bisulfate, polymeric quaternized amine bisulfate or phosphate, or polymeric organic compounds with quaternizable nitrogen combined with Nafion to form an intimate network with ionic interactions.

Narayanan, Sekharipuram R. (Inventor); Yen, Shiao-Pin S. (Inventor)

2006-01-01

261

Actin-propelled Invasive Membrane Protrusions Promote Fusogenic Protein Engagement During Cell-Cell Fusion  

PubMed Central

Cell-cell fusion is critical for the conception, development and physiology of multicellular organisms. Although cellular fusogenic proteins and the actin cytoskeleton are implicated in cell-cell fusion, whether and how they coordinate to promote plasma membrane fusion remain unclear. Here, we reconstituted a high-efficiency, inducible cell-fusion culture system in the normally non-fusing Drosophila S2R+ cells. Both fusogenic proteins and actin cytoskeletal rearrangements were necessary for cell fusion, and, in combination, were sufficient to impart fusion competence. Localized actin polymerization triggered by specific cell-cell or cell-matrix adhesion molecules propelled invasive cell membrane protrusions, which, in turn, promoted fusogenic protein engagement and plasma membrane fusion. This de novo cell-fusion culture system reveals a general role for actin-propelled invasive membrane protrusions in driving fusogenic protein engagement during cell-cell fusion.

Shilagardi, Khurts; Li, Shuo; Luo, Fengbao; Marikar, Faiz; Duan, Rui; Jin, Peng; Kim, Ji Hoon; Murnen, Katherine; Chen, Elizabeth H.

2013-01-01

262

Toward mechanical manipulations of cell membranes and membrane proteins using an atomic force microscope  

Microsoft Academic Search

Recent advances in the use of the atomic force microscope (AFM) for manipulating cell membranes and membrane proteins are\\u000a reviewed. Early pioneering work on measurements of the magnitude of the force required to create indentations with defined\\u000a depth on their surfaces and to separate interacting pairs of avidin-biotin, antigen-antibody, and complementary DNA pairs\\u000a formed the basis of this field. The

Atsushi Ikai; Rehana Afrin

2003-01-01

263

Characterization of Lens Fiber Cell Triton Insoluble Fraction Reveals ERM (Ezrin, Radixin, Moesin) proteins as Major Cytoskeletal-associated Proteins  

PubMed Central

To understand lens fiber cell elongation- and differentiation-associated cytoskekeletal remodeling, here we identified and characterized the major protein components of lens fiber cell triton X-100 insoluble fraction by mass spectrometry and immunoblot analysis. This analysis identified spectrin, filensin, vimentin, tubulin, phakinin and ?-actin as major cytoskeletal proteins in the lens fibers. Importantly, ezrin, radixin and moesin (ERM), heat-shock cognate protein 70, and ?/?-crystallins were identified as major cytoskeletal-associated proteins. ERM proteins were confirmed to exist as active phosphorylated forms that exhibited intense distribution in the organelle free-zone fibers. Furthermore, ERM protein phosphorylation was found to be dramatically reduced in Rho GTPase-targeted transgenic mouse lenses. These data identify the ERM proteins, which crosslink the plasma membrane and actin, as major and stable cytoskeletal-associated proteins in lens fibers, and indicate a potential role(s) for the ERMs in fiber cell actin cytoskeletal and membrane organization.

Vasantha Rao, P.; Ho, Tammy; Skiba, Nikolai P.; Maddala, Rupalatha

2008-01-01

264

Functional ryanodine receptors in the plasma membrane of RINm5F pancreatic beta-cells.  

PubMed

Ryanodine receptors (RyR) are Ca(2+) channels that mediate Ca(2+) release from intracellular stores in response to diverse intracellular signals. In RINm5F insulinoma cells, caffeine, and 4-chloro-m-cresol (4CmC), agonists of RyR, stimulated Ca(2+) entry that was independent of store-operated Ca(2+) entry, and blocked by prior incubation with a concentration of ryanodine that inactivates RyR. Patch-clamp recording identified small numbers of large-conductance (gamma(K) = 169 pS) cation channels that were activated by caffeine, 4CmC or low concentrations of ryanodine. Similar channels were detected in rat pancreatic beta-cells. In RINm5F cells, the channels were blocked by cytosolic, but not extracellular, ruthenium red. Subcellular fractionation showed that type 3 IP(3) receptors (IP(3)R3) were expressed predominantly in endoplasmic reticulum, whereas RyR2 were present also in plasma membrane fractions. Using RNAi selectively to reduce expression of RyR1, RyR2, or IP(3)R3, we showed that RyR2 mediates both the Ca(2+) entry and the plasma membrane currents evoked by agonists of RyR. We conclude that small numbers of RyR2 are selectively expressed in the plasma membrane of RINm5F pancreatic beta-cells, where they mediate Ca(2+) entry. PMID:19116207

Rosker, Christian; Meur, Gargi; Taylor, Emily J A; Taylor, Colin W

2009-02-20

265

Oxidative events in neuronal and glial cell-enriched fractions of rat cerebral cortex  

Microsoft Academic Search

The aim of this work was to investigate how neurons and glial cells separated from rat brain cortex respond to “in vitro” oxidative stress induced by incubation of the cellular fractions in the presence of prooxidant mixtures; in addition, the endogenous enzymatic antioxidant capacity of the purified fractions was investigated. Neuronal and glial cell-enriched fractions were obtained from rat cerebral

Carla Café; Carla Torri; Laura Bertorelli; Fulvio Tartara; Flavio Tancioni; Paolo Gaetani; Riccardo Rodriguez Y Baena; Fulvio Marzatico

1995-01-01

266

Heterotypic binding between neuronal membrane vesicles and glial cells is mediated by a specific cell adhesion molecule  

PubMed Central

By means of a multistage quantitative assay, we have identified a new kind of cell adhesion molecule (CAM) on neuronal cells of the chick embryo that is involved in their adhesion to glial cells. The assay used to identify the binding component (which we name neuron-glia CAM or Ng-CAM) was designed to distinguish between homotypic binding (e.g., neuron to neuron) and heterotypic binding (e.g., neuron to glia). This distinction was essential because a single neuron might simultaneously carry different CAMs separately mediating each of these interactions. The adhesion of neuronal cells to glial cells in vitro was previously found to be inhibited by Fab' fragments prepared from antisera against neuronal membranes but not by Fab' fragments against N-CAM, the neural cell adhesion molecule. This suggested that neuron-glia adhesion is mediated by specific cell surface molecules different from previously isolated CAMs . To verify that this was the case, neuronal membrane vesicles were labeled internally with 6-carboxyfluorescein and externally with 125I-labeled antibodies to N-CAM to block their homotypic binding. Labeled vesicles bound to glial cells but not to fibroblasts during a 30-min incubation period. The specific binding of the neuronal vesicles to glial cells was measured by fluorescence microscopy and gamma spectroscopy of the 125I label. Binding increased with increasing concentrations of both glial cells and neuronal vesicles. Fab' fragments prepared from anti-neuronal membrane sera that inhibited binding between neurons and glial cells were also found to inhibit neuronal vesicle binding to glial cells. The inhibitory activity of the Fab' fragments was depleted by preincubation with neuronal cells but not with glial cells. Trypsin treatment of neuronal membrane vesicles released material that neutralized Fab' fragment inhibition; after chromatography, neutralizing activity was enriched 50- fold. This fraction was injected into mice to produce monoclonal antibodies; an antibody was obtained that interacted with neurons, inhibited binding of neuronal membrane vesicles to glial cells, and recognized an Mr = 135,000 band in immunoblots of embryonic chick brain membranes. These results suggest that this molecule is present on the surfaces of neurons and that it directly or indirectly mediates adhesion between neurons and glial cells. Because the monoclonal antibody as well as the original polyspecific antibodies that were active in the assay did not bind to glial cells, we infer that neuron- glial interaction is heterophilic, i.e., it occurs between Ng-CAM on neurons and an as yet unidentified CAM present on glial cells.

1984-01-01

267

Protein diffusion in plant cell plasma membranes: the cell-wall corral  

PubMed Central

Studying protein diffusion informs us about how proteins interact with their environment. Work on protein diffusion over the last several decades has illustrated the complex nature of biological lipid bilayers. The plasma membrane contains an array of membrane-spanning proteins or proteins with peripheral membrane associations. Maintenance of plasma membrane microstructure can be via physical features that provide intrinsic ordering such as lipid microdomains, or from membrane-associated structures such as the cytoskeleton. Recent evidence indicates, that in the case of plant cells, the cell wall seems to be a major player in maintaining plasma membrane microstructure. This interconnection / interaction between cell-wall and plasma membrane proteins most likely plays an important role in signal transduction, cell growth, and cell physiological responses to the environment.

Martiniere, Alexandre; Runions, John

2013-01-01

268

New insights in the visualization of membrane permeabilization and DNA\\/membrane interaction of cells submitted to electric pulses  

Microsoft Academic Search

Electropermeabilization designates the use of electric pulses to overcome the barrier of the cell membrane. This physical method is used to transfer anticancer drugs or genes into living cells. Its mechanism remains to be elucidated. A position-dependent modulation of the membrane potential difference is induced, leading to a transient and reversible local membrane alteration. Electropermeabilization allows a fast exchange of

Emilie Phez; Cécile Faurie; Muriel Golzio; Justin Teissié; Marie-Pierre Rols

2005-01-01

269

Leea indica Ethyl Acetate Fraction Induces Growth-Inhibitory Effect in Various Cancer Cell Lines and Apoptosis in Ca Ski Human Cervical Epidermoid Carcinoma Cells  

PubMed Central

The anticancer potential of Leea indica, a Chinese medicinal plant was investigated for the first time. The crude ethanol extract and fractions (ethyl acetate, hexane, and water) of Leea indica were evaluated their cytotoxicity on various cell lines (Ca Ski, MCF 7, MDA-MB-435, KB, HEP G2, WRL 68, and Vero) by MTT assay. Leea indica ethyl acetate fraction (LIEAF) was found showing the greatest cytotoxic effect against Ca Ski cervical cancer cells. Typical apoptotic morphological changes such as DNA fragmentation and chromatin condensation were observed in LIEAF-treated cells. Early signs of apoptosis such as externalization of phosphatidylserine and disruption of mitochondrial membrane potential indicated apoptosis induction. This was further substantiated by dose- and time-dependent accumulation of sub-G1 cells, depletion of intracellular glutathione, and activation of caspase-3. In conclusion, these results suggested that LIEAF inhibited cervical cancer cells growth by inducing apoptosis and could be developed as potential anticancer drugs.

Yau Hsiung, Wong; Abdul Kadir, Habsah

2011-01-01

270

Leea indica Ethyl Acetate Fraction Induces Growth-Inhibitory Effect in Various Cancer Cell Lines and Apoptosis in Ca Ski Human Cervical Epidermoid Carcinoma Cells.  

PubMed

The anticancer potential of Leea indica, a Chinese medicinal plant was investigated for the first time. The crude ethanol extract and fractions (ethyl acetate, hexane, and water) of Leea indica were evaluated their cytotoxicity on various cell lines (Ca Ski, MCF 7, MDA-MB-435, KB, HEP G2, WRL 68, and Vero) by MTT assay. Leea indica ethyl acetate fraction (LIEAF) was found showing the greatest cytotoxic effect against Ca Ski cervical cancer cells. Typical apoptotic morphological changes such as DNA fragmentation and chromatin condensation were observed in LIEAF-treated cells. Early signs of apoptosis such as externalization of phosphatidylserine and disruption of mitochondrial membrane potential indicated apoptosis induction. This was further substantiated by dose- and time-dependent accumulation of sub-G(1) cells, depletion of intracellular glutathione, and activation of caspase-3. In conclusion, these results suggested that LIEAF inhibited cervical cancer cells growth by inducing apoptosis and could be developed as potential anticancer drugs. PMID:21423690

Yau Hsiung, Wong; Abdul Kadir, Habsah

2011-01-01

271

Anhydrous Proton-Conducting Membranes for Fuel Cells  

NASA Technical Reports Server (NTRS)

Polymeric electrolyte membranes that do not depend on water for conduction of protons are undergoing development for use in fuel cells. Prior polymeric electrolyte fuel-cell membranes (e.g., those that contain perfluorosulfonic acid) depend on water and must be limited to operation below a temperature of 125 C because they retain water poorly at higher temperatures. In contrast, the present developmental anhydrous membranes are expected to function well at temperatures up to 200 C. The developmental membranes exploit a hopping-and-reorganization proton- conduction process that can occur in the solid state in organic amine salts and is similar to a proton-conduction process in a liquid. This process was studied during the 1970s, but until now, there has been no report of exploiting organic amine salts for proton conduction in fuel cells.

Narayanan, Sekharipuram; Yen, Shiao-Pin S.

2005-01-01

272

Turgor Pressure Sensing in Plant Cell Membranes 1  

PubMed Central

Experimental evidence is reviewed which shows that the cell membrane is compressible by both mechanical and electrical forces. Calculations are given which show that significant changes in the thickness of cell membranes can occur as a result of (a) direct compression due to the turgor pressure; (b) indirect effects due to the stretching of the cell wall; and (c) the stresses induced by the electric field in the membrane. Such changes in the membrane thickness may provide the pressure-transducing mechanism required for osmoregulation and growth. An important feature of the model is that this pressure transduction can occur not only in the plasmalemma (where there is a pressure gradient), but also in the tonoplast.

Coster, Hans G. L.; Steudle, Ernst; Zimmermann, Ulrich

1976-01-01

273

Simultaneous manipulation and detection of living cell membrane dynamics  

NASA Astrophysics Data System (ADS)

We report a novel optical-tweezers-based method to study the membrane motion at the leading edge of biological cells with nanometer spatial and microsecond temporal resolution. A diffraction-limited laser spot was positioned at the leading edge of a cell, and the forward scattered light was imaged on a quadrant photodiode that served as a position sensitive device. The universality of this technique is demonstrated with different cell types. We investigated the membrane motion at the leading edge of red blood cells in detail and showed that this technique can achieve simultaneous manipulation and detection of cellular edge dynamics with unprecedented precision.

Gögler, Michael; Betz, Timo; Alfons Käs, Josef

2007-07-01

274

Membrane Targeting of P-type ATPases in Plant Cells  

SciTech Connect

How membrane proteins are targeted to specific subcellular locations is a very complex and poorly understood area of research. Our long-term goal is to use P-type ATPases (ion pumps), in a model plant system Arabidopsis, as a paradigm to understand how members of a family of closely related membrane proteins can be targeted to different subcellular locations. The research is divided into two specific aims. The first aim is focused on determining the targeting destination of all 10 ACA-type calcium pumps (Arabidopsis Calcium ATPase) in Arabidopsis. ACAs represent a plant specific-subfamily of plasma membrane-type calcium pumps. In contrast to animals, the plant homologs have been found in multiple membrane systems, including the ER (ACA2), tonoplast (ACA4) and plasma membrane (ACA8). Their high degree of similarity provides a unique opportunity to use a comparative approach to delineate the membrane specific targeting information for each pump. One hypothesis to be tested is that an endomembrane located ACA can be re-directed to the plasma membrane by including targeting information from a plasma membrane isoform, ACA8. Our approach is to engineer domain swaps between pumps and monitor the targeting of chimeric proteins in plant cells using a Green Fluorescence Protein (GFP) as a tag. The second aim is to test the hypothesis that heterologous transporters can be engineered into plants and targeted to the plasma membrane by fusing them to a plasma membrane proton pump. As a test case we are evaluating the targeting properties of fusions made between a yeast sodium/proton exchanger (Sod2) and a proton pump (AHA2). This fusion may potentially lead to a new strategy for engineering salt resistant plants. Together these aims are designed to provide fundamental insights into the biogenesis and function of plant cell membrane systems.

Jeffrey F. Harper, Ph.D.

2004-06-30

275

Ultrafiltration by a compacted clay membrane-I. Oxygen and hydrogen isotopic fractionation  

USGS Publications Warehouse

Laboratory experiments were carried out to determine the magnitude of the isotopic fractionation of distilled water and of 0.01 N NaCl forced to flow at ambient temperature under a hydraulic pressure drop of 100 bars across a montmorillonite disc compacted to a porosity of 35 per cent by a pressure of 330 bars. The ultrafiltrates in both experiments were depleted in D by 2.5%. and in O18 by 0.8%. relative to the residual solution. No additional isotopic fractionation due to a salt filtering mechanism was observed at NaCl concentrations up to 0.01 N. Adsorption is most likely the principal mechanism which produces isotopic fractionation, but molecular diffusion may play a minor role. The results suggest that oxygen and hydrogen isotopic fractionation of ground water during passage through compacted clayey sediments should be a common occurrence, in accord with published interpretations of isotopic data from the Illinois and Alberta basins. ?? 1973.

Coplen, T. B.; Hanshaw, B. B.

1973-01-01

276

Evidence for copurification of micronuclei in sucrose density gradient-enriched plasma membranes from cell lines  

Microsoft Academic Search

Sucrose density gradient-enriched membrane preparations and membrane fraction enrichment through affinity purification techniques are commonly used in proteomic analysis. However, published proteomic profiles characterized by the above methods show the presence of nuclear proteins in addition to membrane proteins. While shuttling of nuclear proteins across cellular compartments and their transient residency at membrane interfaces could explain some of these observations,

Sambasivarao Damaraju; Nancy Zhang; Nan Li; Lidan Tao; Vijaya L. Damaraju; Jennifer Dufour; Cheryl Santos; Xue-Jun Sun; John Mackey; David S. Wishart; Carol E. Cass; Liang Li

2010-01-01

277

Utrophin, the autosomal homologue of dystrophin, is widely-expressed and membrane-associated in cultured cell lines.  

PubMed

Utrophin, the autosomal dystrophin-related protein (DRP), is expressed in HeLa cells, smooth muscle-like BC3H1 cells from mouse brain, COS monkey kidney cells, the P388D1 monocyte-macrophage cell line and untransformed human skin fibroblasts, as well as in rat C6 glioma and Schwannoma cells. It was undetectable, however, in the Sp2/O mouse myeloma cell line and in hybridoma lines derived from it. Dystrophin was not detected in any of these cell lines. Although all utrophin-containing cells were capable of forming monolayers in culture, no major effects of either attachment to substratum or length of time in culture (2-17 days) on utrophin levels were observed. After subcellular fractionation of BC3H1 or glioma cells, nearly all of the utrophin was found in the Triton-soluble fraction, suggesting an association with cell membranes. PMID:1426262

Nguyen, T M; Le, T T; Blake, D J; Davies, K E; Morris, G E

1992-11-16

278

Fuel cell electrolyte membrane with basic polymer  

DOEpatents

The present invention is an electrolyte membrane comprising an acid and a basic polymer, where the acid is a low-volatile acid that is fluorinated and is either oligomeric or non-polymeric, and where the basic polymer is protonated by the acid and is stable to hydrolysis.

Larson, James M. (Saint Paul, MN); Pham, Phat T. (Little Canada, MN); Frey, Matthew H. (Cottage Grove, MN); Hamrock, Steven J. (Stillwater, MN); Haugen, Gregory M. (Edina, MN); Lamanna, William M. (Stillwater, MN)

2010-11-23

279

Fuel cell electrolyte membrane with basic polymer  

DOEpatents

The present invention is an electrolyte membrane comprising an acid and a basic polymer, where the acid is a low-volatile acid that is fluorinated and is either oligomeric or non-polymeric, and where the basic polymer is protonated by the acid and is stable to hydrolysis.

Larson, James M.; Pham, Phat T.; Frey, Matthew H.; Hamrock, Steven J.; Haugen, Gregory M.; Lamanna, William M.

2012-12-04

280

A Comparative Spin-Label Study of Isolated Plasma Membranes and Plasma Membranes of Whole Cells and Protoplasts from Cold-Hardened and Nonhardened Winter Rye  

PubMed Central

Lipid-lipid and lipid-protein interactions in the plasma membranes of whole cells and protoplasts and an isolated plasma membrane fraction from winter rye (Secale cereale L. cv Puma) have been studied by spin labeling. Spectra were recorded between ?40°C and 40°C using the freely diffusing spin-label, 16-doxyl stearic acid, as a midbilayer membrane probe. The probe was reduced by the whole cells and protoplasts and reoxidized by external potassium ferricyanide. The reoxidized probe was assumed to be localized in the plasma membrane. The spectra consisted of the superposition of a narrow and a broad component indicating that both fluid and immobilized lipids were present in the plasma membrane. The two components were separated by digital subtraction of the immobilized component. Temperature profiles of the membranes were developed using the percentage of immobilized lipid present at each temperature and the separation between the outermost hyperfine lines for the fluid lipid component. Lipid immobilization was attributed to lipid-protein interactions, lipid-cell wall interactions, and temperature-induced lipid phase transitions to the gel-state. Temperature profiles were compared for both cold-hardened and nonhardened protoplasts, plasma membranes, and plasma membrane lipids, respectively. Although cold-hardening extended the range of lipid fluidity by 5°C, it had no effect on lipid-protein interactions or activation energies of lipid mobility. Differences were found, however, between the temperature profiles for the different samples, suggesting that alterations in the plasma membrane occurred as a consequence of the isolation methods used.

Windle, John J.

1988-01-01

281

A review of polymer electrolyte membranes for direct methanol fuel cells  

Microsoft Academic Search

This review describes the polymer electrolyte membranes (PEM) that are both under development and commercialized for direct methanol fuel cells (DMFC). Unlike the membranes for hydrogen fuelled PEM fuel cells, among which perfluorosulfonic acid based membranes show complete domination, the membranes for DMFC have numerous variations, each has its advantages and disadvantages. No single membrane is emerging as absolutely superior

Vladimir Neburchilov; Jonathan Martin; Haijiang Wang; Jiujun Zhang

2007-01-01

282

Role of Rab GTPases in Membrane Traffic and Cell Physiology  

PubMed Central

Intracellular membrane traffic defines a complex network of pathways that connects many of the membrane-bound organelles of eukaryotic cells. Although each pathway is governed by its own set of factors, they all contain Rab GTPases that serve as master regulators. In this review, we discuss how Rabs can regulate virtually all steps of membrane traffic from the formation of the transport vesicle at the donor membrane to its fusion at the target membrane. Some of the many regulatory functions performed by Rabs include interacting with diverse effector proteins that select cargo, promoting vesicle movement, and verifying the correct site of fusion. We describe cascade mechanisms that may define directionality in traffic and ensure that different Rabs do not overlap in the pathways that they regulate. Throughout this review we highlight how Rab dysfunction leads to a variety of disease states ranging from infectious diseases to cancer.

HUTAGALUNG, ALEX H.; NOVICK, PETER J.

2013-01-01

283

Incorporation of Photosynthetic Reaction Centers in the Membrane of Human Cells: Toward a New Tool for Optical Control of Cell Activity  

SciTech Connect

The Photosystem I (PSI) reaction center is a photosynthetic membrane complex in which light-induced charge separation is accompanied by the generation of an electric potential. It has been recently proposed as a means to confer light sensitivity to cells possessing voltage-activated ion channels, but the feasibility of heterologous incorporation has not been demonstrated. In this work, methods of delivery and detection of PSI in the membrane of human cells are presented. Purified fractions of PSI were reconstituted in proteoliposomes that were used as vehicles for the membrane incorporation. A fluorescent impermeable dye was entrapped in the vesicles to qualitatively analyze the nature of the vesicle cell interaction. After incorporation, the localization and orientation of the complexes in the membrane was studied using immuno-fluorescence microscopy. The results showed complexes oriented as in native membranes, which were randomly distributed in clusters over the entire surface of the cell. Additionally, analysis of cell viability showed that the incorporation process does not damage the cell membrane. Taken together, the results of this work suggest that the mammalian cellular membrane is a reasonable environment for the incorporation of PSI complexes, which opens the possibility of using these molecular photovoltaic structures for optical control of cell activity.

Pennisi, Cristian P. [Aalborg University, Aalborg, Denmark; Jensen, Poul Erik [VKR Research Center, University of Copenhagen, Denmark; Zachar, Vladimir [Laboratory for Stem Cell Research, Aalborg University, Aalborg, Denmark; Greenbaum, Elias [ORNL; Yoshida, Ken [Aalborg University, Aalborg, Denmark

2009-01-01

284

Modulated red blood cell survival by membrane protein clustering  

Microsoft Academic Search

Human and murine blood cells treated with ZnCl2 and bis(sulfosuccinimidyl)suberate (BS3) (a cross linking agent) undergo band 3 clustering and binding of hemoglobin to red blood cell membrane proteins. These clusters induce autologous IgG binding and complement fixation, thus favouring the phagocytosis of ZnCl2\\/BS3 treated cells by macrophages. The extension of red blood cell opsonization can be easily modulated by

Laura Chiarantini; Luigia Rossi; Alessandra Fraternale; Mauro Magnani

1995-01-01

285

Microencapsulation of yeast cells in the calcium alginate membrane  

Microsoft Academic Search

Cells of Saccharomyces cerevisiae (ATCC 24858) were encapsulated in the calcium alginate membrane and cultured. Swelling of the capsule was prevented by adding 0.2 g CaCl2 to 1 L growth medium. The dry cell concentration based on the inner volume of the capsule reached 309 g\\/L, which was much higher than could be obtained by cell entrapment. All the cells

Soo Hwan Cheong; Joong Kon Park; Beom Soo Kim; Ho Nam Chang

1993-01-01

286

Anion selective membrane. [ion exchange resins and ion exchange membrane electrolytes for electrolytic cells  

NASA Technical Reports Server (NTRS)

Experimental anion permselective membranes were prepared and tested for their suitability as cell separators in a chemical redox power storage system being developed at NASA-Lewis Research Center. The goals of long-term (1000 hr) oxidative and thermal stability at 80 C in FeCl3 and CrCl3 electrolytes were met by most of the weak base and strong base amino exchange groups considered in the program. Good stability is exhibited by several of the membrane substrate resins. These are 'styrene' divinylbenzene copolymer and PVC film. At least four membrane systems produce strong flexible films with electrochemical properties (resistivity, cation transfer) superior to those of the 103QZL, the most promising commercial membrane. The physical and chemical properties of the resins are listed.

Alexander, S. S.; Geoffroy, R. R.; Hodgdon, R. B.

1975-01-01

287

Cytotoxic effects of Kingella kingae outer membrane vesicles on human cells  

PubMed Central

Kingella kingae is an emerging pathogen causing osteoarticular infections in pediatric patients. Electron microscopy of K. kingae clinical isolates revealed the heterogeneously-sized membranous structures blebbing from the outer membrane that were classified as outer membrane vesicles (OMVs). OMVs purified from the secreted fraction of a septic arthritis K. kingae isolate were characterized. Among several major proteins, K. kingae OMVs contained virulence factors RtxA toxin and PilC2 pilus adhesin. RtxA was also found secreted as a soluble protein in the extracellular environment indicating that the bacterium may utilize different mechanisms for the toxin delivery. OMVs were shown to be hemolytic and possess some leukotoxic activity while high leukotoxicity was detected in the non-hemolytic OMV-free component of the secreted fraction. OMVs were internalized by human osteoblasts and synovial cells. Upon interaction with OMVs, the cells produced increased levels of human granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleuskin 6 (IL-6) suggesting that these cytokines might be involved in the signaling response of infected joint and bone tissues during natural K. kingae infection. This study is the first report of OMV production by K. kingae and demonstrates that OMVs are a complex virulence factor of the organism causing cytolytic and inflammatory effects on host cells.

Maldonado, R; Wei, R; Kachlany, SC; Kazi, M; Balashova, NV

2011-01-01

288

Nonlinear electro-mechanobiological behavior of cell membrane during electroporation  

NASA Astrophysics Data System (ADS)

A nonlinear electroporation (EP) model is proposed to study the electro-mechanobiological behavior of cell membrane during EP, by taking the nonlinear large deformation of the membrane into account. The proposed model predicts the critical transmembrane potential and the activation energy for EP, the equilibrium pore size, and the resealing process of the pore. Single-cell EP experiments using a micro EP chip were conducted on chicken red blood cells at different temperatures to determine the activation energy and the critical transmembrane potential for EP. The experimental results are in good agreement with the theoretical predictions.

Deng, Peigang; Lee, Yi-Kuen; Lin, Ran; Zhang, Tong-Yi

2012-07-01

289

Selective binding of Trypanosoma cruzi to host cell membrane polypeptides.  

PubMed Central

An adaptation of the immunoblotting technique was used to investigate binding interactions between Trypanosoma cruzi and mammalian host cells at the molecular level. A specific binding interaction was observed between T. cruzi and two host cell membrane polypeptides with molecular masses of approximately 32 and 34 kilodaltons. This molecular interaction was observed with antigen extracts of T. cruzi and with live, infective trypomastigote stages of the parasite, suggesting that the observed phenomenon may have relevance to the initial attachment of the parasite to the host cell membrane before invasion. Images

Davis, C D; Kuhn, R E

1990-01-01

290

Reconstitution in vitro of neurotoxin-responsive ion efflux by using membrane glycoproteins of neuroblastoma cells.  

PubMed Central

Glycoproteins were purified from a clonal cell line of mouse neuroblastoma, N-18, labeled metabolically with L-[3H]fucose. The purified radioactive glycoproteins were reconstituted into artificial phosphatidylcholine vesicles. When the vesicles were preloaded with cesium acetate and treated with neurotoxins to activate the Na+ channel, a shift in intravesicular density was observed to a less dense position after centrifugation on sucrose gradients. This shift was partially inhibited by tetrodotoxin, which prevents the activation of the Na+ channel. A similarly derived fraction of [14C]fucose-containing glycoproteins from a neuroblastoma cell line that does not possess excitable membranes, N1A-103, was reconstituted into phospholipid vesicles, and, after preloading with cesium ions, the fraction was combined with those of the 3H-labeled glycoproteins of the differentiated cells, N-18, which have excitable membranes. Only the 3H-labeled glycoprotein-containing vesicles were responsive to the neurotoxins, as shown by a shift in intravesicular density on sucrose gradients. These results are interpreted as a demonstration of the reconstitution of glycoproteins to form the activated Na+ channel. Comparison of the radioactive glycoprotein profiles after polyacrylamide gel electrophoresis showed that glycoproteins of Mr 200,000, Mr 165,000, and Mr 65,000 were common to the reconstituted fractions that were biologically active.

Giovanni, M Y; Glick, M C

1983-01-01

291

A Novel Virus-Host Cell Membrane Interaction  

PubMed Central

Studies on the virus–cell interactions have proven valuable in elucidating vital cellular processes. Interestingly, certain virus–host membrane interactions found in eukaryotic systems seem also to operate in prokaryotes (Bamford, D.H., M. Romantschuk, and P.J. Somerharju, 1987. EMBO (Eur. Mol. Biol. Organ.) J. 6:1467–1473; Romantschuk, M., V.M. Olkkonen, and D.H. Bamford. 1988. EMBO (Eur. Mol. Biol. Organ.) J. 7:1821–1829). ?6 is an enveloped double-stranded RNA virus infecting a gram-negative bacterium. The viral entry is initiated by fusion between the virus membrane and host outer membrane, followed by delivery of the viral nucleocapsid (RNA polymerase complex covered with a protein shell) into the host cytosol via an endocytic-like route. In this study, we analyze the interaction of the nucleocapsid with the host plasma membrane and demonstrate a novel approach for dissecting the early events of the nucleocapsid entry process. The initial binding of the nucleocapsid to the plasma membrane is independent of membrane voltage (??) and the K+ and H+ gradients. However, the following internalization is dependent on plasma membrane voltage (??), but does not require a high ATP level or K+ and H+ gradients. Moreover, the nucleocapsid shell protein, P8, is the viral component mediating the membrane–nucleocapsid interaction.

Poranen, Minna M.; Daugelavicius, Rimantas; Ojala, Paivi M.; Hess, Michael W.; Bamford, Dennis H.

1999-01-01

292

Membrane Nanotubes in Urothelial Cell Line T24  

Microsoft Academic Search

Membrane nanotubes (also referred as tunnelling nanotubes—TNTs, nanotu- bules, cytonemes), that directly connect separated neighboring cells, may offer a very specific and effective way of intercellular transport and communication. Our experiments on T24 cell line show that TNTs can be divided into two types with respect to their biochemical and biophysical characteristics and the nature of their formation. As type

CHAPTER T HREE; Marusa Lokar; Sarka Perutkova; Veronika Kralj-Iglic; Peter Veranic

293

Chapter 3 Membrane Nanotubes in Urothelial Cell Line T24  

Microsoft Academic Search

Membrane nanotubes (also referred as tunnelling nanotubes—TNTs, nanotubules, cytonemes), that directly connect separated neighboring cells, may offer a very specific and effective way of intercellular transport and communication. Our experiments on T24 cell line show that TNTs can be divided into two types with respect to their biochemical and biophysical characteristics and the nature of their formation. As type I

Maruša Lokar; Šárka Perutková; Veronika Kralj-Igli?; Aleš Igli?; Peter Verani?

2009-01-01

294

Regulation of membrane trafficking in polarized epithelial cells  

PubMed Central

Polarized epithelial cells continuously sort transmembrane proteins to either apical or basolateral plasma membrane domains. Research in recent years has made tremendous progress in understanding the molecular mechanisms of the major pathways to either basolateral or apical domain. This understanding will help us elucidating how these pathways are interconnected in ensuring maintenance of cell polarity and integrity of epithelial monolayers.

Folsch, Heike

2008-01-01

295

Light-sensitive membrane potentials in onion guard cells  

Microsoft Academic Search

INTRACELLULAR electrical recordings in onion guard cells show that they maintain a membrane potential difference (MPD), inside negative. The MPD is light-sensitive; cells subjected to short light and dark cycles depolarise in the dark and hyperpolarise in the light. The swiftness of the electrical changes makes them among the fastest known stomatal responses, suggesting a causal relationship between the reception

E. Zeiger; W. Moody; P. Hepler; F. VARELA

1977-01-01

296

Stem Cells from Fetal Membranes – A Workshop Report  

Microsoft Academic Search

Stem cells that can be derived from fetal membranes represent an exciting field of research that bears tremendous potential for developmental biology and regenerative medicine. In this report we summarize contributions to a workshop in which newest insights into the characteristics, subtypes and molecular determinants of stem cells from trophoblast and endometrial tissues were presented.

M. Hemberger; W. Yang; D. Natale; T. L. Brown; C. Dunk; C. E. Gargett; S. Tanaka

2008-01-01

297

Electrical coupling in proton exchange membrane fuel cell stacks  

Microsoft Academic Search

A mathematical model describing the effects of electrical coupling of proton exchange membrane unit fuel cells through shared bipolar plates is developed. Here, the unit cells are described by simple, steady-state, 1D models appropriate for straight reactant gas channel designs. A linear asymptotic version of the model is used to give analytic insight into the effect of the coupling, including

Peter Berg; Atife Caglar; Keith Promislow; Jean St-Pierre; Brian Wetton

2006-01-01

298

Numerical modeling transport phenomena in proton exchange membrane fuel cells  

NASA Astrophysics Data System (ADS)

To study the coupled phenomena occurring in proton exchange membrane fuel cells, a two-phase, one-dimensional, non-isothermal model is developed in the chapter 1. The model includes water phase change, proton transport in the membrane and electro-osmotic effect. The thinnest, but most complex layer in the membrane electrode assembly, catalyst layer, is considered an interfacial boundary between the gas diffusion layer and the membrane. Mass and heat transfer and electro-chemical reaction through the catalyst layer are formulated into equations, which are applied to boundary conditions for the gas diffusion layer and the membrane. Detail accounts of the boundary equations and the numerical solving procedure used in this work are given. The polarization curve is calculated at different oxygen pressures and compared with the experimental results. When the operating condition is changed along the polarization curve, the change of physicochemical variables in the membrane electrode assembly is studied. In particular, the over-potential diagram presents the usage of the electrochemical energy at each layer of the membrane electrode assembly. Humidity in supplying gases is one of the most important factors to consider for improving the performance of PEMFE. Both high and low humidity conditions can result in a deteriorating cell performance. The effect of humidity on the cell performance is studied in the chapter 2. First, a numerical model based on computational fluid dynamics is developed. Second, the cell performances are simulated, when the relative humidity is changed from 0% to 100% in the anode and the cathode channel. The simulation results show how humidity in the reactant gases affects the water content distribution in the membrane, the over-potential at the catalyst layers and eventually the cell performance. In particular, the rapid enhancement in the cell performance caused by self-hydrating membrane is captured by the simulation. Fully humidifying either H2 in the anode or air in the cathode makes the membrane conductive enough for proton transfer. Water supplied more than that can hinder mass transfer of O2 and degrade the cell performance. At low output voltage, the limiting current density is increased by reducing the humidity at the cathode.

Suh, DongMyung

299

Degradation process of fuel cell membrane observed by positron  

NASA Astrophysics Data System (ADS)

To investigate degradation process due to radicals in fuel cell membrane by means of positron annihilation spectroscopy, three kinds of radicals, HO•, H• and O2•- are produced through water radiolysis. The results show that the cluster structure and proton conductivity was greatly affected by reductive radicals. This is because the oxidative radical is responsible for the dissociation of sulfonic group, whereas the reductive radical breaks down the cluster in the membrane and disrupts proton conduction, which is consistent with solution analysis.

Honda, Y.; Aoyagi, Y.; Tojo, S.; Watanabe, G.; Akiyama, Y.; Nishijima, S.

2013-06-01

300

Microbial fuel cells (MFCs) with interpolymer cation exchange membranes  

Microsoft Academic Search

Interpolymer cation exchange membranes have been prepared from the system polyethylene\\/poly(styrene-co-divinylbenzene) [PE\\/poly(St-co-DVB)] by their sulfonation with a solution of chlorosulfonic acid in 1,2-dichloroethane. The membranes have been characterized electrochemically in mediator-less microbial fuel cell (MFC). MFC has worked on Escherichia coli bacteria with ferrocyanide as an oxidizer. Real-time voltage and current measurement gave us reliable results. Consequently, it was found

Micha? Grzebyk; Gryzelda Po?niak

2005-01-01

301

Nanodomain stabilization dynamics in plasma membranes of biological cells  

NASA Astrophysics Data System (ADS)

We discover that a synergistically amplifying role of stabilizing membrane proteins and continuous lipid recycling can explain the physics governing the stability, polydispersity, and dynamics of lipid raft domains in plasma membranes of biological cells. We establish the conjecture using a generalized order parameter based on theoretical formalism, endorsed by detailed scaling arguments and domain mapping. Quantitative agreements with morphological distributions of raft complexes, as obtained from Förster resonance energy transfer based visualization, support the present theoretical conjecture.

Das, Tamal; Maiti, Tapas K.; Chakraborty, Suman

2011-02-01

302

Overexpression of membrane proteins in mammalian cells for structural studies  

PubMed Central

The number of structures of integral membrane proteins from higher eukaryotes is steadily increasing due to a number of innovative protein engineering and crystallization strategies devised over the last few years. However, it is sobering to reflect that these structures represent only a tiny proportion of the total number of membrane proteins encoded by a mammalian genome. In addition, the structures determined to date are of the most tractable membrane proteins, i.e., those that are expressed functionally and to high levels in yeast or in insect cells using the baculovirus expression system. However, some membrane proteins that are expressed inefficiently in these systems can be produced at sufficiently high levels in mammalian cells to allow structure determination. Mammalian expression systems are an under-used resource in structural biology and represent an effective way to produce fully functional membrane proteins for structural studies. This review will discuss examples of vertebrate membrane protein overexpression in mammalian cells using a variety of viral, constitutive or inducible expression systems.

Andrell, Juni

2013-01-01

303

Influence of the phosphoric acid-doping level in a polybenzimidazole membrane on the cell performance of high-temperature proton exchange membrane fuel cells  

Microsoft Academic Search

The acid migration in phosphoric acid-doped polybenzimidazole (PBI) membrane high-temperature proton exchange membrane fuel cells (HT-PEMFC) during operation is experimentally evaluated to clarify the influence of the acid balance between the membrane and electrodes on cell performance. A method for controlling the amount of phosphoric acid doped in PBI membranes is investigated, and PBI membranes with various amounts of phosphoric

Yuka Oono; Atsuo Sounai; Michio Hori

2009-01-01

304

Understanding the transport processes in polymer electrolyte membrane fuel cells  

NASA Astrophysics Data System (ADS)

Polymer electrolyte membrane (PEM) fuel cells are energy conversion devices suitable for automotive, stationary and portable applications. An engineering challenge that is hindering the widespread use of PEM fuel cells is the water management issue, where either a lack of water (resulting in membrane dehydration) or an excess accumulation of liquid water (resulting in fuel cell flooding) critically reduces the PEM fuel cell performance. The water management issue is addressed by this dissertation through the study of three transport processes occurring in PEM fuel cells. Water transport within the membrane is a combination of water diffusion down the water activity gradient and the dragging of water molecules by protons when there is a proton current, in a phenomenon termed electro-osmotic drag, EOD. The impact of water diffusion and EOD on the water flux across the membrane is reduced due to water transport resistance at the vapor/membrane interface. The redistribution of water inside the membrane by EOD causes an overall increase in the membrane resistance that regulates the current and thus EOD, thereby preventing membrane dehydration. Liquid water transport in the PEM fuel cell flow channel was examined at different gas flow regimes. At low gas Reynolds numbers, drops transitioned into slugs that are subsequently pushed out of the flow channel by the gas flow. The slug volume is dependent on the geometric shape, the surface wettability and the orientation (with respect to gravity) of the flow channel. The differential pressure required for slug motion primarily depends on the interfacial forces acting along the contact lines at the front and the back of the slug. At high gas Reynolds number, water is removed as a film or as drops depending on the flow channel surface wettability. The shape of growing drops at low and high Reynolds number can be described by a simple interfacial energy minimization model. Under flooding conditions, the fuel cell local current can be significantly reduced due to diffusional limitation of the transport of gaseous reactants through inerts such as water vapor and nitrogen gas. A non-uniform current distribution across the membrane electrode assembly can cause pinhole formation and ultimately, fuel cell failure.

Cheah, May Jean

305

Effect of Micropolyspora faeni cells and cell wall fractions on rabbit alveolar macrophages.  

PubMed Central

The reactivity of alveolar macrophages (AM) to cells and cell wall fractions (CWF) of Micropolyspora faeni was investigated. Exposure of cultured AM to M. faeni and its CWF caused the AM to form clumps or aggregates which remained attached to the culture dish surface. Other gram-positive and gram-negative bacteria as well as yeast, zymosan, latex microspheres, and isolated peptidoglycan from Listeria monocytogenes did not cause this response. The response was independent of species source and antibody content of the serum used in culture. The use of heat-inactivated sera negated the role of complement activation in the aggregation of AM. AM cultures required a period of culture before exposure to cells or CWF for this response to occur. This response was both time and dose dependent. Rabbit peritoneal macrophages also exhibited the clumping response. Degradation of a purified CWF, fraction 3, with lysozyme greatly diminished the clumping response. Chemical purification of fraction 3 with periodate, formamide, or trichloracetic acid also decreased this activity. These data suggest that the major active component causing this response is peptidoglycan but that other materials associated with the cell wall may also be important. A soluble-factor chemotactic for normal rabbit AM was found in the culture fluid of AM exposed to fraction 3. M. faeni cells and CWF also caused normal rabbit AM to chemiluminesce. Images

Learn, D B; Snyder, I S

1983-01-01

306

Polymer-zeolite nanocomposite membranes for proton exchange membrane fuel cells  

NASA Astrophysics Data System (ADS)

Proton exchange membrane fuel cells (PEMFCs) have recently received a great deal of attention for their potential as compact, high efficiency power sources for portable, distributed generation, and transportation applications. Unfortunately, current proton exchange membrane (PEM) technology hinders fuel cell performance by limiting fuel cell operation temperature and methanol feed concentration in direct methanol fuel cells (DMFCs). Nafion-zeolite nanocomposite membranes that take advantage of the hydrophilicity, selectivity, and proton conductivity of zeolite nanocrystals have been developed to address these problems. All known zeolite topologies were evaluated as potential additives to Nafion proton exchange membranes. Zeolites Y and beta were determined to have great potential as additives due to their low framework density, three dimensional pore structure, and high hydrophilicity. Zeolite Y nanocrystal syntheses were optimized to enhance yield and produce smaller crystal size. Significant improvement of the acid stability of the zeolite Y nanocrystals was not achieved with both ammonium hexafluorosilicate treatments and direct high silica nanocrystal synthesis. However, control of zeolite Y nanocrystal framework Si/Al ratio was demonstrated in the range of SiO2/Al2O3 = 4.38 to 5.84 by manipulating the tetramethylammonium structure directing agent hydroxide content. Zeolite beta nanocrystals were investigated due to their inherent high silica content and high acid stability. Zeolite beta nanocrystals were hydrothermally synthesized with and without phenethyl (called PE-BEA and BEA respectively) organic functional groups. Sulfonic acid functionalized zeolite beta (SAPE-BEA) was generated by treating the PE-BEA nanocrystals with a concentrated sulfuric acid post synthesis treatment. SAPE-BEA samples demonstrated proton conductivities up to 0.01 S/cm at room temperature under water-saturated conditions using a newly developed characterization technique. With optimization, acid functionalized zeolite materials could possibly perform as competent stand-alone proton conducting materials with the proper engineering. BEA and SAPE-BEA zeolite nanocrystals mixed with suspensions of Nafion were cast into nanocomposite membranes. DMFC membrane electrode assemblies (MEAs) prepared with a 2.5wt% SAPE-BEA nanocomposite membrane delivered twice the peak power of a MEA with a commercial Nafion 117 membrane. Membrane performance improvements of this magnitude could ultimately lead to DMFC cost and size reductions that make the technology commercially viable for a variety of applications.

Holmberg, Brett Anderson

307

Plasma membrane reorganization induced by tumor promoters in an epithelial cell line  

SciTech Connect

The effects of phorbol ester tumor promoters on the lateral diffusion in plasma membrane lipid environments were examined by the technique of fluorescence recovery after photobleaching. To this end, the probe collarein, a fluorescent lipid analog that has the property of exclusive localization in the plasma membrane, was synthesized. Measured decreases in three parameters [percentage of fluorescence bleached (30%), percentage of recovery (52%), and half-time for recovery (52%)] connoted the appearance of an immobile fraction upon exposure to tumor promoters. These data are consistent with lipid reorganization in response to a reorganization of the intra- and perimembranous macromolecular scaffolding upon the interaction of cells with tumor promoters. The idea of induced reorganization is supported by experiments in which cell shape change, brought about by either exposure to cytochalasin B or growth on matrices of collagen, fibronectin, or laminin, resulted in values in the fluorescence recovery after photobleaching technique similar to those with active phorbol esters.

PACKARD, BEVERLY S.; SAXTON, MICHAEL J.; BISSELL, MINA J.; KLEIN, MELVIN P.

1984-01-01

308

Computational analysis of polarizations in membrane-electrode-assembly for proton exchange membrane fuel cells  

Microsoft Academic Search

The effects of platinum loadings and thickness of catalyst layer on the overall polarization of proton exchange membrane fuel cell (PEMFC) were investigated based on flooded agglomerate model. To obtain accurately simulated data on anode hydrogen oxidation reaction (HOR), the well-known dual-pathway kinetic model was taken into account. The presented model was validated with experimental data taken from the literatures.

Chi-Young Jung; Wha-Jung Kim; Sung-Chul Yi

2009-01-01

309

Corona discharge in electroporation of cell membranes  

NASA Astrophysics Data System (ADS)

The objective of the present work is to demonstrate that electrical corona discharge is very efficient in cellular membrane electroporation due to current pulses with sharp front (2-5 ns) and to the fact that corona discharge is associated with UV radiation and micro particles emission. A comparison between DC and AC at 800 Hz and a special waveform to corona application is presented. The comparison is analyzed by means of applying all these in the maceration process (electroplasmolysis) of red wine production and in the processes of different types of the microbes.

Cramariuc, R.; Tudorache, A.; Popa, M. E.; Branduse, E.; Nisiparu, L.; Mitelut, A.; Turtoi, M. O.; Fotescu, L.

2008-12-01

310

Optimization of a proton exchange membrane fuel cell membrane electrode assembly  

Microsoft Academic Search

A computational framework for fuel cell analysis and optimization is presented as an innovative alternative to the time consuming\\u000a trial-and-error process currently used for fuel cell design. The framework is based on a two-dimensional through-the-channel\\u000a isothermal, isobaric and single phase membrane electrode assembly (MEA) model. The model input parameters are the manufacturing\\u000a parameters used to build the MEA: platinum loading,

Marc Secanell; Ron Songprakorp; Ned Djilali; Afzal Suleman

2010-01-01

311

Basement membranes and artificial substrates in cell transplantation  

Microsoft Academic Search

This article will concentrate largely on the current developments in the area of cell transplantations presented at the 1st Workshop for Cell Transplantation in Age-related Macular Degeneration. In particular, this brief review will address our current understanding of the role of cell–matrix interactions by covering the pathobiology of normal ageing Bruch’s membrane; some of the problems faced at the time

Carl Sheridan; Rachel Williams; Ian Grierson

2004-01-01

312

Electrophoresis of concanavalin A receptors along embryonic muscle cell membrane  

Microsoft Academic Search

Fluorescent concanavalin A (con A)-labelling showed that an electric field of 4 V cm-1 grossly redistributed con A receptors along the plasma membranes of living muscle cells within 4 h. This field produced a voltage drop of 12 mV across these 30 µm-wide cells. The movement of receptors was independent of cell metabolism and seemed to be electrophoretic in nature.

Mu-Ming Poo; Kenneth R. Robinson

1977-01-01

313

Calix[4]arene C-90 Selectively Inhibits Ca2+,Mg2+-ATPase of Myometrium Cell Plasma Membrane.  

PubMed

The supramolecular compound calix[4]arene C-90 (5,11,17,23-tetra(trifluoro)methyl(phenylsulfonylimino)-methylamino-25,26,27,28-tetrapropoxycalix[4]arene) is shown to efficiently inhibit the ATP hydrolase activity of Ca2+,Mg2+-ATPase in the myometrium cell plasma membrane fraction and also in a preparation of the purified enzyme solubilized from this subcellular fraction. The inhibition coefficient I0.5 values were 20.2 ± 0.5 and 58.5 ± 6.4 µM for the membrane fraction and the solubilized enzyme, respectively. The inhibitory effect of calix[4]arene C-90 was selective comparatively to other ATPases localized in the plasma membrane: calix[4]arene C-90 did not influence the activities of Na+,K+-ATPase and "basal" Mg2+-ATPase. The inhibitory effect of calix[4]arene C-90 on the Ca2+,Mg2+-ATPase activity was associated with the cooperative action of four trifluoromethylphenylsulfonylimine (sulfonylamidine) groups oriented similarly on the upper rim of the calix[4]arene macrocycle (the calix[4]arene "bowl"). The experimental findings seem to be of importance for studies, using calix[4]arene C-90, of membrane mechanisms of regulation of calcium homeostasis in smooth muscle cells and also for investigation of the participation of the plasma membrane Ca2+-pump in control of electro- and pharmacomechanical coupling in myocytes. PMID:24954592

Veklich, T A; Shkrabak, A A; Slinchenko, N N; Mazur, I I; Rodik, R V; Boyko, V I; Kalchenko, V I; Kosterin, S A

2014-05-01

314

Specific binding of a fungal glucan phytoalexin elicitor to membrane fractions from soybean Glycine max  

Microsoft Academic Search

Treatment of soybean tissues with elicitors results in the production of phytoalexins, one of a number of inducible plant defense reactions against microbial infections. The present study uses a ..beta..-1,3-(³H) glucan elicitor fraction from Phytophthora megasperma f.sp. glycinea, a fungal pathogen of soybean, to identify putative elicitor targets in soybean tissues. Use of the radiolabeled elicitor disclosed saturable high-affinity elicitor

W. E. Schmidt; J. Ebel

1987-01-01

315

A novel self-humidifying membrane electrode assembly with water transfer region for proton exchange membrane fuel cells  

Microsoft Academic Search

A novel self-humidifying membrane electrode assembly (MEA) with the active electrode region surrounded by a unactive “water transfer region (WTR)” was proposed to achieve effective water management and high performance for proton exchange membrane fuel cells (PEMFCs). By this configuration, excess water in the cathode was transferred to anode through Nafion membrane to humidify hydrogen. Polarization curves and power curves

Er-Dong Wang; Peng-Fei Shi; Chun-Yu Du

2008-01-01

316

Potential growth inhibitory effect of maitake D-fraction on canine cancer cells.  

PubMed

The postulated anticancer effect of D-fraction, the bioactive extract of maitake mushroom, on three types (CF33, CF21, and CL-1) of canine cancer cells was evaluated. The effect of D-fraction on several human cancer cells was also investigated. The effect of other beta-glucan products was likewise examined. D-fraction was highly effective on the canine cancer cells, either potently inhibiting cell growth or directly killing cells. Similar effects were also demonstrated in certain human cancer cells. However, other beta-glucan products relevant to D-fraction had no such effects on canine cancer cells. Therefore, D-fraction is a potent natural agent that could be useful in treating canine cancers as well as other veterinary cancers. PMID:15719326

Konno, Sensuke

2004-01-01

317

Enhancement of cytotoxicity of NK cells by D-Fraction, a polysaccharide from Grifola frondosa.  

PubMed

In innate immunity, activated natural killer (NK) cells attack and damage pathogens such as bacteria and virus without restriction by the MHC antigen. NK cells activated by IL-12 have been reported to recognize and kill tumor cells in perforin-mediated apoptosis. We have reported that D-Fraction, a polysaccharide extracted from the maitake mushroom (Grifola frondosa), activates macrophages, dendritic cells, and T cells and inhibits the growth of tumor cells. However, the effects of D-Fraction on NK cell function in the innate immune response are not well known. In the present study, we administered D-Fraction to MM-46 mammary tumor-bearing C3H/HeJ mice intraperitoneally for 3 consecutive days and investigated its effects on the activation and cytotoxicity of NK cells. D-Fraction significantly enhanced the cytotoxicity against NK-sensitive YAC-1 cells and the expression of CD223 on NK cells. D-Fraction also increased the expression of CD86 on macrophages. In addition, the levels of IL-12 in the culture supernatant of whole spleen cells and in serum increased, compared with the control corresponding to an increase in expression of IL-12 receptor betaI on NK cells. These results suggest that D-Fraction enhances the cytotoxicity of NK cells through the production of IL-12 by macrophages activated by D-Fraction. PMID:15706424

Kodama, Noriko; Asakawa, Akihiro; Inui, Akio; Masuda, Yuki; Nanba, Hiroaki

2005-03-01

318

Membrane tension of red blood cells pairwisely interacting in simple shear flow.  

PubMed

Flow-induced membrane tension contributes to the release of molecules by red blood cells (RBCs), and extremely high tension may cause haemolysis. Here, we investigated the membrane tension of RBCs during pairwise interactions in simple shear flow, given that pairwise interactions form the basis of many-body interactions. RBCs were modelled as capsules with a two-dimensional hyperelastic membrane, and large deformations were solved by the finite element method. Due to the small size of the RBCs, surrounding fluid motion was estimated as a Stokes flow and solved by the boundary element method. The results showed that the maximum isotropic tension appeared around the dimple of the biconcave surface and not around the rim. A comparison of the results with solitary cases indicated that the maximum principal tension and isotropic tension were significantly increased by cell-cell interaction effects. As the volume fraction of RBCs is large under physiological conditions, as well as in blood flow in vitro, cell-cell interactions must be analysed carefully when considering mechanotransduction and haemolysis in blood flow. PMID:23102822

Omori, Toshihiro; Ishikawa, Takuji; Imai, Yohsuke; Yamaguchi, Takami

2013-02-01

319

Proton-conductive nanochannel membrane for fuel-cell applications.  

PubMed

Novel design of proton conductive membrane for direct methanol fuel cells is based on proton conductivity of nanochannels, which is acquired due to the electric double layer overlap. Proton conductivity and methanol permeability of an array of nanochannels were studied. Anodic aluminum oxide with pore diameter of 20 nm was used as nanochannel matrix. Channel surfaces of an AAO template were functionalized with sulfonic groups to increase proton conductivity of nanochannels. This was done in two steps; at first -SH groups were attached to walls of nanochannels using (3-Mercaptopropyl)-trimethyloxysilane and then they were converted to -SO3H groups using hydrogen peroxide. Treatment steps were analyzed by Fourier Transform Infrared spectroscopy and X-ray Photoelectron Spectroscopy. Proton conductivity and methanol permeability were measured. The data show methanol permeability of membrane to be an order of magnitude lower, than that measured of Nafion. Ion conductivity of functionalized AAO membrane was measured by an impedance analyzer at frequencies ranging from 1 Hz to 100 kHz and voltage 50 mV to be 0.15 Scm(-1). Measured ion conductivity of Nafion membrane was 0.05 Scm(-1). Obtained data show better results in comparison with commonly used commercial available proton conductive membrane Nafion, thus making nanochannel membrane very promising for use in fuel cell applications. PMID:19441568

Oleksandrov, Sergiy; Lee, Jeong-Woo; Jang, Joo-Hee; Haam, Seungjoo; Chung, Chan-Hwa

2009-02-01

320

Microstructured Electrolyte Membranes to Improve Fuel Cell Performance  

NASA Astrophysics Data System (ADS)

Fuel cells, with the advantages of high efficiency, low greenhouse gas emission, and long lifetime are a promising technology for both portable power and stationary power sources. The development of efficient electrolyte membranes with high ionic conductivity, good mechanical durability and dense structure at low cost remains a challenge to the commercialization of fuel cells. This thesis focuses on exploring novel composite polymer membranes and ceramic electrolytes with the microstructure engineered to improve performance in direct methanol fuel cells (DMFCs) and solid oxide fuel cells (SOFCs), respectively. Polymer/particle composite membranes hold promise to meet the demands of DMFCs at lower cost. The structure of composite membranes was controlled by aligning proton conducting particles across the membrane thickness under an applied electric field. The field-induced structural changes caused the membranes to display an enhanced water uptake, proton conductivity, and methanol permeability in comparison to membranes prepared without an applied field. Although both methanol permeability and proton conductivity are enhanced by the applied field, the permeability increase is relatively lower than the proton conductivity improvement, which results in enhanced proton/methanol selectivity and improved DMFC performance. Apatite ceramics are a new class of fast ion conductors being studied as alternative SOFC electrolytes in the intermediate temperature range. An electrochemical/hydrothermal deposition method was developed to grow fully dense apatite membranes containing well-developed crystals with c-axis alignment to promote ion conductivity. Hydroxyapatite seed crystals were first deposited onto a metal substrate electrochemically. Subsequent ion substitution during the hydrothermal growth process promoted the formation of dense, fully crystalline films with microstructure optimal for ion transport. The deposition parameters were systematically investigated, such as reactant type, reagent concentration, solution pH, and reaction time. Dense apatite films were formed on palladium substrates that can serve as intermediate temperature fuel cell anodes. The novel apatite membrane structure is promising for fuel cell applications, as well as in improving the biocompatibility of orthopedic implants when coated on stainless steel or titanium substrates.

Wei, Xue

321

Dual network model for red blood cell membranes  

NASA Astrophysics Data System (ADS)

A two-component network is studied by Monte Carlo simulation to model the lipid/spectrin membrane of red blood cells. The model predicts that the shear modulus decreases rapidly with the maximum length of the model spectrin and should be in the 10-7 J/m2 range for human red blood cells. A simplified model for the isolated spectrin network shows a negative Lamé coefficient ?. Transverse fluctuations of the dual membrane are found to be fluidlike over the range of wavelengths investigated.

Boal, David H.; Seifert, Udo; Zilker, Andreas

1992-12-01

322

Immunologically Induced Alterations of Airway Smooth Muscle Cell Membrane  

NASA Astrophysics Data System (ADS)

Active and passive sensitization, both in vivo and in vitro, caused significant hyperpolarization of airway smooth muscle cell preparations isolated from guinea pigs. An increase in the contribution of the electrogenic Na+ pump to the resting membrane potential was responsible for this change. Hyperpolarization, as induced by passive sensitization, was not prevented by agents that inhibit specific mediators of anaphylaxis but was abolished when serum from sensitized animals was heated. The heat-sensitive serum factor, presumably reaginic antibodies, appears to be responsible for the membrane hyperpolarization of airway smooth muscle cells after sensitization.

Souhrada, M.; Souhrada, J. F.

1984-08-01

323

Contrasting molecular dynamics in red and purple membrane fractions of the Halobacterium halobium  

SciTech Connect

/sup 2/H-nuclear magnetic resonance (NMR) has been used to study the dynamics of amino acid residues in bacteriorhodopsin with results that depend on the method of sample preparation. We show here that in (/sup 2/H)-leucine-labeled samples the intensity of the isotropic signal varies according to the degree of residual contamination of the sample with red membrane. We conclude that few of the surface leucine residues of bacteriorhodopsin are moving isotropically on the /sup 2/H-NMR time scale.

Herzfeld, J.; Mulliken, C.M.; Siminovitch, D.J.; Griffin, R.G.

1987-11-01

324

New High-Temperature Membranes Developed for Proton Exchange Membrane Fuel Cells  

NASA Technical Reports Server (NTRS)

Fuel cells are receiving a considerable amount of attention for potential use in a variety of areas, including the automotive industry, commercial power generation, and personal electronics. Research at the NASA Glenn Research Center has focused on the development of fuel cells for use in aerospace power systems for aircraft, unmanned air vehicles, and space transportation systems. These applications require fuel cells with higher power densities and better durability than what is required for nonaerospace uses. In addition, membrane cost is a concern for any fuel cell application. The most widely used membrane materials for proton exchange membrane (PEM) fuel cells are based on sulfonated perfluorinated polyethers, typically Nafion 117, Flemion, or Aciplex. However, these polymers are costly and do not function well at temperatures above 80 C. At higher temperatures, conventional membrane materials dry out and lose their ability to conduct protons, essential for the operation of the fuel cell. Increasing the operating temperature of PEM fuel cells from 80 to 120 C would significantly increase their power densities and enhance their durability by reducing the susceptibility of the electrode catalysts to carbon monoxide poisoning. Glenn's Polymers Branch has focused on developing new, low-cost membranes that can operate at these higher temperatures. A new series of organically modified siloxane (ORMOSIL) polymers were synthesized for use as membrane materials in a high-temperature PEM fuel cell. These polymers have an organic portion that can allow protons to transport through the polymer film and a cross-linked silica network that gives the polymers dimensional stability. These flexible xerogel polymer films are thermally stable, with decomposition onset as high as 380 C. Two types of proton-conducting ORMOSIL films have been produced: (1) NASA-A, which can coordinate many highly acid inorganic salts that facilitate proton conduction and (2) NASA-B, which has been produced and which incorporates strongly acidic (proton donating) functional groups into the polymer backbone. Both of these polymer films have demonstrated significantly higher proton conductivity than Nafion at elevated temperatures and low relative humidities. An added advantage is that these polymers are very inexpensive to produce because their starting materials are commodity chemicals that are commercially available in large volumes.

Kinder, James D.

2004-01-01

325

Predicting membrane flux decline from complex mixtures using flow-field flow fractionation measurements and semi-empirical theory.  

PubMed

Flow-Field Flow Fractionation (FI-FFF) is an idealization of the cross flow membrane filtration process in that, (1) the filtration flux and crossflow velocity are constant from beginning to end of the device, (2) the process is a relatively well-defined laminar-flow hydrodynamic condition, and (3) the solutes are introduced as a pulse-input that spreads due to interactions with each other and the membrane in the dilute-solution limit. We have investigated the potential for relating FI-FFF measurements to membrane fouling. An advection-dispersion transport model was used to provide 'ideal' (defined as spherical, non-interacting solutes) solute residence time distributions (RTDs) for comparison with 'real' RTDs obtained experimentally at different cross-field velocities and solution ionic strength. An RTD moment analysis based on a particle diameter probability density function was used to extract "effective" characteristic properties, rather than uniquely defined characteristics, of the standard solute mixture. A semi-empirical unsteady-state, flux decline model was developed that uses solute property parameters. Three modes of flux decline are included: (1) concentration polarization, (2) cake buildup, and (3) adsorption on/in pores, We have used this model to test the hypothesis-that an analysis of a residence time distribution using FI-FFF can describe 'effective' solute properties or indices that can be related to membrane flux decline in crossflow membrane filtration. Constant flux filtration studies included the changes of transport hydrodynamics (solvent flux to solute back diffusion (J/k) ratios), solution ionic strength, and feed water composition for filtration using a regenerated cellulose ultrafiltration membrane. Tests of the modeling hypothesis were compared with experimental results from the filtration measurements using several correction parameters based on the mean and variance of the solute RTDs. The corrections used to modify the boundary layer mass transfer coefficient and the specific resistance of cake or adsorption layers demonstrated that RTD analysis is potentially useful technique to describe colloid properties but requires improvements. PMID:16003965

Pellegrino, J; Wright, S; Ranvill, J; Amy, G

2005-01-01

326

Membrane with internal passages to permit fluid flow and an electrochemical cell containing the same  

NASA Technical Reports Server (NTRS)

The invention provides an improved proton exchange membrane for use in electrochemical cells having internal passages parallel to the membrane surface, an apparatus and process for making the membrane, membrane and electrode assemblies fabricated using the membrane, and the application of the membrane and electrode assemblies to a variety of devices, both electrochemical and otherwise. The passages in the membrane extend from one edge of the membrane to another and allow fluid flow through the membrane and give access directly to the membrane for purposes of hydration.

Cisar, Alan J. (Inventor); Gonzalez-Martin, Anuncia (Inventor); Hitchens, G. Duncan (Inventor); Murphy, Oliver J. (Inventor)

1997-01-01

327

Lactic acid fermentation in cell-recycle membrane bioreactor.  

PubMed

Traditional lactic acid fermentation suffers from low productivity and low product purity. Cell-recycle fermentation has become one of the methods to obtain high cell density, which results in higher productivity. Lactic acid fermentation was investigated in a cell-recycle membrane bioreactor at higher substrate concentrations of 100 and 120 g/dm3. A maximum cell density of 145 g/dm3 and a maximum productivity of 34 g/(dm3.h) were achieved in cell-recycle fermentation. In spite of complete consumption of substrate, there was a continuous increase in cell density in cell-recycle fermentation. Control of cell density in cell-recycle fermentation was attempted by cell bleeding and reduction in yeast extract concentration. PMID:16484726

Choudhury, B; Swaminathan, T

2006-02-01

328

The development of PTFE\\/Nafion\\/TEOS membranes for application in moderate and high temperature proton exchange membrane fuel cells  

Microsoft Academic Search

PTFE\\/Nafion (PN) and PTFE\\/Nafion\\/TEOS (PNS) membranes were fabricated for the application of moderate and high temperature proton exchange membrane fuel cells (PEMFCs), respectively. Membrane electrode assemblies (MEAs) were fabricated by PTFE\\/Nafion (and PTFE\\/Nafion\\/TEOS) membranes with commercially available low and high temperature gas diffusion electrodes (GDEs). The effects of relative humidity, operation temperature, and back pressure on the performance and durability

Guo-Bin Jung; Feng-Bor Weng; Chao-Chun Peng; Ting-Chu Jao

2011-01-01

329

Radiation effects on membranes - 1. Cellular permeability and cell survival  

SciTech Connect

The effect of various doses of ..gamma.. radiation (5-60 krad) on the membrane permeability and cell survival of Candida albicans, a pathogenic yeast, was investigated. A reduction in the cell survival and in the accumulation of amino acids (proline, glycine, lysine, and glutamic acid) was observed following irradiation. The rate of oxygen uptake, which is often associated with transport, was also reduced. There was no damage to available sulfhydryl groups following the exposure of cells to various doses of ..gamma.. radiation. The membrane lipid composition of C. albicans cells can be altered by growing them in alkanes of varying chain lengths. The effects of such altered lipid composition on radiosensitivity was examined. It was observed that C. albicans cells with altered lipid content acquire resistance to ..gamma.. radiation.

Khare, S.; Jayakumar, A.; Trivedi, A.; Kesavan, P.C.; Prasad, R.

1982-05-01

330

Photobleaching regions of living cells to monitor membrane traffic.  

PubMed

Eukaryotic cells are composed of an intricate system of internal membranes that are organized into different compartments--including the endoplasmic reticulum (ER), the nuclear envelope, the Golgi complex (GC), lysosomes, endosomes, caveolae, mitochondria, and peroxisomes--that perform specialized tasks within the cell. The localization and dynamics of intracellular compartments are now being studied in living cells because of the availability of green fluorescent protein (GFP)-fusion proteins and recent advances in fluorescent microscope imaging systems. This protocol outlines two methods for photobleaching living cells to monitor membrane traffic. The first method involves selective photobleaching using a confocal laser-scanning microscope (CLSM) that can bleach discrete selected regions of interest. As outlined in the second method, photobleaching can also be performed with older CLSMs that lack the capacity for selective photobleaching. In this case, photobleaching is accomplished by zooming into a small region of the cell and scanning with full laser power. PMID:22046038

Snapp, Erik Lee; Lajoie, Patrick

2011-11-01

331

Evidence for Bidirectional Endocannabinoid Transport across Cell Membranes*  

PubMed Central

Despite extensive research on the trafficking of anandamide (AEA) across cell membranes, little is known about the membrane transport of other endocannabinoids, such as 2-arachidonoylglycerol (2-AG). Previous studies have provided data both in favor and against a cell membrane carrier-mediated transport of endocannabinoids, using different methodological approaches. Because AEA and 2-AG undergo rapid and almost complete intracellular hydrolysis, we employed a combination of radioligand assays and absolute quantification of cellular and extracellular endocannabinoid levels. In human U937 leukemia cells, 100 nm AEA and 1 ?m 2-AG were taken up through a fast and saturable process, reaching a plateau after 5 min. Employing differential pharmacological blockage of endocannabinoid uptake, breakdown, and interaction with intracellular binding proteins, we show that eicosanoid endocannabinoids harboring an arachidonoyl chain compete for a common membrane target that regulates their transport, whereas other N-acylethanolamines did not interfere with AEA and 2-AG uptake. By combining fatty acid amide hydrolase or monoacyl glycerol lipase inhibitors with hydrolase-inactive concentrations of the AEA transport inhibitors UCM707 (1 ?m) and OMDM-2 (5 ?m), a functional synergism on cellular AEA and 2-AG uptake was observed. Intriguingly, structurally unrelated AEA uptake inhibitors also blocked the cellular release of AEA and 2-AG. We show, for the first time, that UCM707 and OMDM-2 inhibit the bidirectional movement of AEA and 2-AG across cell membranes. Our findings suggest that a putative endocannabinoid cell membrane transporter controls the cellular AEA and 2-AG trafficking and metabolism.

Chicca, Andrea; Marazzi, Janine; Nicolussi, Simon; Gertsch, Jurg

2012-01-01

332

The role of cell membranes in the regulation of lignification in pine cells  

NASA Technical Reports Server (NTRS)

The identity of pine cell membranes bearing PAL enzyme activity, the isolation of a plasma membrane preparation from pine cells for testing as a regulatory barrier in lignification, and the measurement of the geopotential effect in pine stems are presented. A model to describe and predict the interaction of gravity and lignification of higher plants was developed.

Hendrix, D. L.

1978-01-01

333

The Lipid Raft Microdomain-Associated Protein Reggie-1\\/ Flotillin-2 is Expressed in Human B Cells and Localized at the Plasma Membrane and Centrosome in PBMCs  

Microsoft Academic Search

Reggie-1\\/flotillin-2 is a plasma membrane-associated cytoplasmic protein, which defines non-caveolar raft microdomains. Reggie-1\\/flotillin-2 is enriched in detergent insoluble (TX100) membrane fractions (DIG), co-localizes with activated GPI-linked proteins and the fyn-kinase in neurons and T cells, and thus apparently participates in the assembly of protein complexes essential for signal transduction. In T cells activated by crosslinking the GPI-linked protein Thy-1 or

Samuel Solomon; Madhan Masilamani; Lawrence Rajendran; Martin Bastmeyer; Claudia A. O. Stuermer; Harald Illges

2002-01-01

334

Inorganic Nanoporous Membranes for Immunoisolated Cell-Based Drug Delivery  

PubMed Central

Materials advances enabled by nanotechnology have brought about promising approaches to improve the encapsulation mechanism for immunoisolated cell-based drug delivery. Cell-based drug delivery is a promising treatment for many diseases but has thus far achieved only limited clinical success. Treatment of insulin dependent diabetes mellitus (IDDM) by transplantation of pancreatic ?-cells represents the most anticipated application of cell-based drug delivery technology. This review outlines the challenges involved with maintaining transplanted cell viability and discusses how inorganic nanoporous membranes may be useful in achieving clinical success.

Mendelsohn, Adam; Desai, Tejal

2014-01-01

335

Ethyl acetate fraction of Garcina epunctata induces apoptosis in human promyelocytic cells (HL-60) through the ROS generation and G0/G1 cell cycle arrest: a bioassay-guided approach.  

PubMed

Number of deaths due to cancer diseases is increasing in the world. There is an urgent need to develop alternative therapeutic measures against the disease. Our study reports the cytotoxicity activity of Garcina epunctata (gutifferae) in human promyelocytic leukemia cells (HL-60) and prostate cancer cells (PC-3) was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). Changes in mitochondrial membrane potential (MMP), reactive oxygen species (ROS) and morphological changes associated with apoptosis were examined by flow cytometry and Hoescht staining respectively. The results of in vitro antiproliferative screening of fractions and extract from G. epunctata indicated that three fractions inhibited the viability of PC-3 cells with IC?? varied from 50 to 88 ?/ml while two fractions inhibited the proliferation of HL-60 cells with IC?? range between 47.5 and 12 ?g/ml. Among the entire fraction tested, Hex-EtOAc (75:25) showed cytotoxic effects on the two cell lines and EtOAc fraction was most active only HL-60 cells (12 ?g/ml). Treatment of HL-60 cells with G. epunctata (20, 50, 100 ?g/ml) for 24 h led to a significant dose-dependent increase in the percentage of cells in sub-G1 phase by analysis of the content of DNA in cells, and a number of apoptotic bodies containing nuclear fragments were observed in cells treated with 100 ?g/ml. The EtOAc fraction of G. epunctata treatment significantly arrested HL-60 cells at the G0/G1 phase (p<0.05) and ROS was significantly elevated as well as the loss of membrane mitochondrial potential in a concentration dependant manner. The results demonstrated that the EtOAc fraction of G. epunctata inhibited the proliferation of HL-60 cells, leading to cell cycle arrest and programmed cell death, which was confirmed to occur through the mitochondrial pathway. PMID:23981377

Constant Anatole, Pieme; Guru, Santoh Kumar; Bathelemy, Ngamegni; Jeanne, Ngogang; Bhushan, Shashi; Murayama, Tetsuya; Saxena, Ajit Kumar

2013-11-01

336

Sequential CD34 cell fractionation by magnetophoresis in a magnetic dipole flow sorter.  

PubMed

Cell separation and fractionation based on fluorescent and magnetic labeling procedures are common tools in contemporary research. These techniques rely on binding of fluorophores or magnetic particles conjugated to antibodies to target cells. Cell surface marker expression levels within cell populations vary with progression through the cell cycle. In an earlier work we showed the reproducible magnetic fractionation (single pass) of the Jurkat cell line based on the population distribution of CD45 surface marker expression. Here we present a study on magnetic fractionation of a stem and progenitor cell (SPC) population using the established acute myelogenous leukemia cell line KG-1a as a cell model. The cells express a CD34 cell surface marker associated with the hematopoietic progenitor cell activity and the progenitor cell lineage commitment. The CD34 expression level is approximately an order of magnitude lower than that of the CD45 marker, which required further improvements of the magnetic fractionation apparatus. The cells were immunomagnetically labeled using a sandwich of anti-CD34 antibody-phycoerythrin (PE) conjugate and anti-PE magnetic nanobead and fractionated into eight components using a continuous flow dipole magnetophoresis apparatus. The CD34 marker expression distribution between sorted fractions was measured by quantitative PE flow cytometry (using QuantiBRITE PE calibration beads), and it was shown to be correlated with the cell magnetophoretic mobility distribution. A flow outlet addressing scheme based on the concept of the transport lamina thickness was used to control cell distribution between the eight outlet ports. The fractional cell distributions showed good agreement with numerical simulations of the fractionation based on the cell magnetophoretic mobility distribution in the unsorted sample. PMID:20024182

Schneider, Thomas; Karl, Stephan; Moore, Lee R; Chalmers, Jeffrey J; Williams, P Stephen; Zborowski, Maciej

2010-01-01

337

Fractional Proliferation: A method to deconvolve cell population dynamics from single-cell data  

PubMed Central

We present an integrated method that exploits extended time-lapse automated imaging to quantify dynamics of cell proliferation. Cell counts are fit with a Quiescence-Growth model that estimates rates of cell division, entry into quiescence and death. The model is constrained with rates extracted experimentally from the behavior of tracked single cells over time. We visualize the output of the analysis in Fractional Proliferation graphs, which deconvolve dynamic proliferative responses to perturbations into the relative contributions of dividing, quiescent (non-dividing) and dead cells. The method reveals that the response of “oncogene-addicted” human cancer cells to tyrosine kinase inhibitors is a composite of altered rates of division, death and entry into quiescence, challenging the notion that such cells simply ‘die’ in response to oncogene-targeted therapy.

Tyson, Darren R.; Garbett, Shawn P.; Frick, Peter L.; Quaranta, Vito

2012-01-01

338

Brefeldin A-dependent Membrane Tubule Formation Reconstituted In Vitro Is Driven by a Cell Cycle-regulated Microtubule Motor  

PubMed Central

Treatment of cultured cells with brefeldin A (BFA) induces the formation of extensive membrane tubules from the Golgi apparatus, trans-Golgi network, and early endosomes in a microtubule-dependent manner. We have reconstituted this transport process in vitro using Xenopus egg cytosol and a rat liver Golgi-enriched membrane fraction. The presence of BFA results in the formation of an intricate, interconnected tubular membrane network, a process that, as in vivo, is inhibited by nocodazole, the H1 anti-kinesin monoclonal antibody, and by membrane pretreatment with guanosine 5?-O-(3-thiotriphosphate). Surprisingly, membrane tubule formation is not due to the action of conventional kinesin or any of the other motors implicated in Golgi membrane dynamics. Two candidate motors of ?100 and ?130 kDa have been identified using the H1 antibody, both of which exhibit motor properties in a biochemical assay. Finally, BFA-induced membrane tubule formation does not occur in metaphase cytosol, and because membrane binding of both candidate motors is not altered after incubation in metaphase compared with interphase cytosol, these results suggest that either the ATPase or microtubule-binding activity of the relevant motor is cell cycle regulated.

Robertson, Alasdair M.; Allan, Victoria J.

2000-01-01

339

Nanoceramic oxide hybrid electrolyte membranes for proton exchange membrane fuel cells.  

PubMed

This review reports on the functions and applications of nanoceramic oxides in proton exchange membrane fuel cells (PEMFCs). Such materials are mainly used as fillers to enhance the water uptake and proton conductivity of polymeric matrices at high temperatures under low relative humidity. To further enhance the mechanical property of proton exchange membranes (PEMs), the functionalized ceramic oxides with organic groups are introduced. Furthermore, the inorganic PEMs are developed to improve their proton conductivities at elevated temperatures. Due to the inherent disadvantages of polymeric PEMs, it is believed that the inorganic PEMs based on porous ceramic oxides are a promising new candidate as solid electrolyte membranes in PEMFCs at high temperatures and with low relative humidity. PMID:24749420

Xu, Feng; Mu, Shichun

2014-02-01

340

Nitrogen Isotope Fractionation Increases with the Cell-Specific Dissimilatory Nitrate Reduction Rate  

NASA Astrophysics Data System (ADS)

The use of the nitrogen (N) isotopes to estimate the impacts and rates of different N transformations depends on knowledge of their extent of isotope fractionation under environmentally relevant physico-chemical conditions. Though the extent of N isotope fractionation during denitrification by pure cultures of bacteria has been determined in the past, relatively large variation in the isotope effect during apparently replicate experiments has been perplexing and the values that should be most relevant for environmental applications have not been clear. We measured the extent of N and O isotope fractionation during nitrate reduction by two bacterial denitrifiers, Pseudomonas chlororaphis ATCC 43928 and Paracoccus denitrificans ATCC 19367 that were grown in 1L batch reactors in the presence of differing carbon sources that included complex organic (e.g, bactopeptone and casein) or defined (e.g., glucose and acetate) carbon compounds and varying concentrations of dissolved oxygen (0 - 4 mM) and nitrate (25 - 800 mM) in the assay medium. For P. denitrificans and P. Chlororaphis , the total range of the N isotope effect (15?) varied from 22.3 to 9.3 ‰ and 34.3 to 15.6 ‰, respectively. Despite this large variation, the O-to-N isotope effect ratio centered around 1, consistent with our previous work. A systematic pattern that has emerged from these studies is that the N and O isotope effect during denitrification increases with increasing cell specific nitrate reduction (CSNR) rate. This sense of variation runs counter to expectations from studies of carbon and sulfur isotope effects during methanogenesis and sulfate reduction, respectively, in which higher substrate consumption rates are associated with lower isotope effects. As with many multi-step microbial processes, variability in the dissimilatory nitrate reduction isotope effect may arise from variation in the “relative” rate and reversibility of (1) nitrate uptake into the denitrifying cell, and/or (2) nitrate binding by dissimilatory nitrate reductase. As an example of a plausible explanation for the isotope effect/CSNR rate relationship that involves cellular uptake, an increase in the CSNR rate may be matched by an even greater increase in nitrate uptake across the cell membrane, leading to more nitrate efflux from the cell and thus more complete expression of the nitrate reductase isotope effect in the media. In addition to discussing such mechanistic possibilities and their physiological implications, we will consider the significance of our findings for the interpretation of oceanic nitrate isotope data.

Kritee, K.; Sigman, D. M.; Granger, J.

2009-12-01

341

CD2 Promotes Human Natural Killer Cell Membrane Nanotube Formation  

PubMed Central

Membrane nanotubes are thin membranous projections that physically connect two cells. While nanotubes have been studied in human natural killer (NK) cells and are implicated in aiding NK cell cytotoxic function, requirements for their formation to susceptible target cells remain incompletely understood. Here we demonstrate that the CD2-CD58/48 receptor-ligand interaction promotes and is required for nanotube formation in human NK cells. In the CD2? NK cell line YTS, a stable CD2 expression variant enabled effective nanotube formation, and was associated with better cytotoxic function. Importantly, only interactions between an NK cell and a susceptible target cell were associated with multiple nanotubes and the number of nanotubes was inversely correlated with their length. Quantitative live cell fluorescence microscopy of CD2 nanotubes revealed time-dependent enrichment and localization of CD2 to the nanotube tip, and blocking CD2 receptor-ligand interactions prevented nanotube formation. Increased nanotube formation was not simply a feature of receptor-ligand pairing, as a KIR-MHC interaction in the same cell line system failed to promote nanotube formation. Additionally, blocking LFA-1-ICAM and 2B4-CD48 receptor-ligand interactions failed to inhibit nanotube formation. Thus only specific receptor-ligand pairs promote nanotubes. CD2 also promoted nanotube formation in ex vivo NK cells suggesting that CD2 plays a crucial role in the generation of nanotubes between an NK cell and its target.

Comerci, Colin J.; Mace, Emily M.; Banerjee, Pinaki P.; Orange, Jordan S.

2012-01-01

342

Properties of electrophoretic fractions of human embryonic kidney cells separated on space shuttle flight STS-8  

NASA Technical Reports Server (NTRS)

Suspensions of cultured primary human embryonic kidney cells were subjected to continuous flow electrophoresis on Space Shuttle flight STS-8. The objectives of the experiments were to obtain electrophoretically separated fractions of the original cell populations and to test these fractions for the amount and kind of urokinase (a kidney plasminogen activator that is used medically for digesting blood clots), the morphologies of cells in the individual fractions, and their cellular electrophoretic mobilities after separation and subsequent proliferation. Individual fractions were successfully cultured after return from orbit, and they were found to differ substantially from one another and from the starting sample with respect to all of these properties.

Morrison, D. R.; Lewis, M. L.; Barlow, G. H.; Todd, P. W.; Kunze, M. E.; Sarnoff, B. E.; Li, Z. K.

1985-01-01

343

Deoxygenation affects fluorescence photobleaching recovery measurements of red cell membrane protein lateral mobility.  

PubMed Central

We have used the fluorescence photobleaching recovery technique to study the dependence on oxygen tension of the lateral mobility of fluorescently labeled band 3, the phospholipid analogue fluorescein phosphatidylethanolamine, and glycophorins in normal red blood cell membranes. Band 3 protein and sialic acid moieties on glycophorins were labeled specifically with eosin maleimide and fluorescein thiosemicarbazide, respectively. The band 3 diffusion rate increased from 1.7 x 10(-11) cm2 s-1 to 6.0 x 10(-11) cm2 s-1 as oxygen tension was decreased from 156 to 2 torr, and a further increase to 17 x 10(-11) cm2 s-1 occurred as oxygen tension was decreased from 2 to 0 torr. The fractional mobility of band 3 decreased from 58 to 32% as oxygen tension was decreased from 156 to 0 torr. The phospholipid diffusion coefficient remained constant as oxygen tension was decreased from 156 to 20 torr, but increased from 2.3 x 10(-9) cm2 s-1 to 7.1 x 10(-9) cm2 s-1 as oxygen tension was decreased from 20 to 0 torr. Neither the diffusion coefficient nor the fractional mobility of glycophorins changed significantly at low oxygen tension. Under non-bleaching excitation conditions, intensities of fluorescence emission were identical for oxygenated and deoxygenated eosin-labeled RBCs. Deoxygenated eosin-labeled RBCs required 160-fold greater laser intensities than did oxygenated RBCs to achieve comparable extents of photobleaching, however. Oxygen seems to act as a facilitator of fluorophore photobleaching and may thereby protect the fluorescently labeled red cell membrane from photodamage. Removal of oxygen may allow excited state fluorophores in close proximity to the plasma membrane to react with neighboring proteins or lipids during photobleaching. This effect has important implications for the ability of the fluorescence photobleaching recovery technique to report accurate lateral mobilities of cell membrane molecules under hypoxic conditions.

Corbett, J. D.; Cho, M. R.; Golan, D. E.

1994-01-01

344

Extracellular Heme Uptake and the Challenges of Bacterial Cell Membranes  

PubMed Central

In bacteria, the fine balance of maintaining adequate iron levels while preventing the deleterious effects of excess iron has led to the evolution of sophisticated cellular mechanisms to obtain, store, and regulate iron. Iron uptake provides a significant challenge given its limited bioavailability and need to be transported across the bacterial cell wall and membranes. Pathogenic bacteria have circumvented the iron-availability issue by utilizing the hosts' heme-containing proteins as a source of iron. Once internalized, iron is liberated from the porphyrin enzymatically for cellular processes within the bacterial cell. Heme, a lipophilic and toxic molecule, poses a significant challenge in terms of transport given its chemical reactivity. As such, pathogenic bacteria have evolved sophisticated membrane transporters to coordinate, sequester, and transport heme. Recent advances in the biochemical and structural characterization of the membrane-bound heme transport proteins are discussed in the context of ligand coordination, protein–protein interaction, and heme transfer.

Smith, Aaron D.; Wilks, Angela

2013-01-01

345

Durable, Low-cost, Improved Fuel Cell Membranes  

SciTech Connect

The development of low cost, durable membranes and membranes electrode assemblies (MEAs) that operate under reduced relative humidity (RH) conditions remain a critical challenge for the successful introduction of fuel cells into mass markets. It was the goal of the team lead by Arkema, Inc. to address these shortages. Thus, this project addresses the following technical barriers from the fuel cells section of the Hydrogen Fuel Cells and Infrastructure Technologies Program Multi-Year Research, Development and Demonstration Plan: (A) Durability (B) Cost Arkema’s approach consisted of using blends of polyvinylidenefluoride (PVDF) and proprietary sulfonated polyelectrolytes. In the traditional approach to polyelectrolytes for proton exchange membranes (PEM), all the required properties are “packaged” in one macromolecule. The properties of interest include proton conductivity, mechanical properties, durability, and water/gas transport. This is the case, for example, for perfluorosulfonic acid-containing (PFSA) membranes. However, the cost of these materials is high, largely due to the complexity and the number of steps involved in their synthesis. In addition, they suffer other shortcomings such as mediocre mechanical properties and insufficient durability for some applications. The strength and originality of Arkema’s approach lies in the decoupling of ion conductivity from the other requirements. Kynar® PVDF provides an exceptional combination of properties that make it ideally suited for a membrane matrix (Kynar® is a registered trademark of Arkema Inc.). It exhibits outstanding chemical resistance in highly oxidative and acidic environments. In work with a prior grant, a membrane known as M41 was developed by Arkema. M41 had many of the properties needed for a high performance PEM, but had a significant deficiency in conductivity at low RH. In the first phase of this work, the processing parameters of M41 were explored as a means to increase its proton conductivity. Optimizing the processing of M41 was found to increase its proton conductivity by almost an order of magnitude at 50% RH. Characterization of the membrane morphology with Karren More at Oak Ridge National Laboratory showed that the membrane morphology was complex. This technology platform was dubbed M43 and was used as a baseline in the majority of the work on the project. Although its performance was superior to M41, M43 still showed proton conductivity an order of magnitude lower than that of a PFSA membrane at 50% RH. The MEA performance of M43 could be increased by reducing the thickness from 1 to 0.6 mils. However, the performance of the thinner M43 still did not match that of a PFSA membrane.

Chris Roger; David Mountz; Wensheng He; Tao Zhang

2011-03-17

346

Function recovery after chemobleaching (FRAC): evidence for activity silent membrane receptors on cell surface.  

PubMed

Membrane proteins represent approximately 30% of the proteome of both prokaryotes and eukaryotes. Unique to cell surface receptors is their biogenesis pathway, which involves vesicular trafficking from the endoplasmic reticulum through the Golgi apparatus and to the cell surface. Increasing evidence suggests specific regulation of biogenesis for different membrane receptors, hence affecting their surface expression. We report the development of a pulse-chase assay to monitor function recovery after chemobleaching (FRAC) to probe the transit time of the Kir2.1 K+ channel to reach the cell surface. Our results reveal that the channel activity is contributed by a small fraction of channel protein, providing evidence of activity-silent "sleeping" molecules on the cell surface. This method distinguishes molecular density from functional density, and the assay strategy is generally applicable to other membrane receptors. The ability of the reported method to access the biogenesis pathways in a high-throughput manner facilitates the identification and evaluation of molecules affecting receptor trafficking. PMID:15548608

Sun, Haiyan; Shikano, Sojin; Xiong, Qiaojie; Li, Min

2004-11-30

347

Mass spectrometric characterization of proteins extracted from Jurkat T cell detergent-resistant membrane domains.  

PubMed

Plasma membranes of most cell types are thought to contain microdomains commonly referred to as lipid rafts, biochemically distinct from bulk plasma membrane, apparently enriched for proteins involved in signal transduction. In T cells, it is believed that lipid rafts aggregate at the site of T cell receptor engagement and act as foci for initiation of the signaling process. In order to gain insight into the possible functioning of lipid rafts, we applied microcapillary liquid chromatography electrospray ionization tandem mass spectrometry (microLC-ESI-MS/MS) methodologies to the identification of proteins which copurified with lipid rafts. Following isolation of lipid rafts as Triton-insoluble, low-density membrane fractions from Jurkat T cells, tryptic digests were generated of individual protein bands resolved electrophoretically. Alternatively, cysteine-containing peptides were isolated from total tryptic digests of unseparated lipid raft proteins following labeling with a cysteine-specific biotinylation reagent and avidin affinity purification. In both cases, protein identifications were made by comparison of tandem MS spectra generated by microLC-ESI-MS/MS to both protein and DNA sequence databases using Sequest software. Proteins identified essentially fell into two groups: cytoskeletal proteins, and proteins involved in signal transduction. These findings are discussed in the light of the current understanding of both lipid raft biology and signal transduction. PMID:11683502

von Haller, P D; Donohoe, S; Goodlett, D R; Aebersold, R; Watts, J D

2001-08-01

348

Activating photoactivatable proteins with laser light to visualize membrane systems and membrane traffic in living cells.  

PubMed

Eukaryotic cells are composed of an intricate system of internal membranes that are organized into different compartments--including the endoplasmic reticulum (ER), the nuclear envelope, the Golgi complex (GC), lysosomes, endosomes, caveolae, mitochondria, and peroxisomes--that perform specialized tasks within the cell. The localization and dynamics of intracellular compartments are now being studied in living cells because of the availability of green fluorescent protein (GFP)-fusion proteins and recent advances in fluorescent microscope imaging systems, such as the confocal laser-scanning microscope (CLSM). This protocol describes the steps for activating one of the first photoactivatable proteins, PA-GFP. PMID:22046039

Snapp, Erik Lee; Lajoie, Patrick

2011-11-01

349

Remodeling the cell surface distribution of membrane proteins during the development of epithelial cell polarity  

PubMed Central

The development of polarized epithelial cells from unpolarized precursor cells follows induction of cell-cell contacts and requires resorting of proteins into different membrane domains. We show that in MDCK cells the distributions of two membrane proteins, Dg-1 and E- cadherin, become restricted to the basal-lateral membrane domain within 8 h of cell-cell contact. During this time, however, 60-80% of newly synthesized Dg-1 and E-cadherin is delivered directly to the forming apical membrane and then rapidly removed, while the remainder is delivered to the basal-lateral membrane and has a longer residence time. Direct delivery of greater than 95% of these proteins from the Golgi complex to the basal-lateral membrane occurs greater than 48 h later. In contrast, we show that two apical proteins are efficiently delivered and restricted to the apical cell surface within 2 h after cell-cell contact. These results provide insight into mechanisms involved in the development of epithelial cell surface polarity, and the establishment of protein sorting pathways in polarized cells.

1992-01-01

350

Single cell electric impedance topography: Mapping membrane capacitance  

PubMed Central

Single-cell electric impedance topography (sceTopo), a technique introduced here, maps the spatial distribution of capacitance (i.e. displacement current) associated with the membranes of isolated, living cells. Cells were positioned in the center of a circular recording chamber surrounded by eight electrodes. Electrodes were evenly distributed on the periphery of the recording chamber. Electric impedance measured between adjacent electrode pairs (10 kHz–5 MHz) was used to construct topographical maps of the spatial distribution of membrane capacitance. Xenopus Oocytes were used as a model cell to develop sceTopo because these cells consist of two visually distinguishable hemispheres, each with distinct membrane composition and structure. Results showed significant differences in the imaginary component of the impedance between the two oocyte hemispheres. In addition, the same circumferential array was used to map the size of the extracellular electrical shunt path around the cell, providing a means to estimate the location and shape of the cell in the recording chamber.

Dharia, Sameera; Ayliffe, Harold E.

2010-01-01

351

Electrospun fiber membranes enable proliferation of genetically modified cells  

PubMed Central

Polycaprolactone (PCL) and its blended composites (chitosan, gelatin, and lecithin) are well-established biomaterials that can enrich cell growth and enable tissue engineering. However, their application in the recovery and proliferation of genetically modified cells has not been studied. In the study reported here, we fabricated PCL-biomaterial blended fiber membranes, characterized them using physicochemical techniques, and used them as templates for the growth of genetically modified HCT116-19 colon cancer cells. Our data show that the blended polymers are highly miscible and form homogenous electrospun fiber membranes of uniform texture. The aligned PCL nanofibers support robust cell growth, yielding a 2.5-fold higher proliferation rate than cells plated on standard plastic plate surfaces. PCL-lecithin fiber membranes yielded a 2.7-fold higher rate of proliferation, while PCL-chitosan supported a more modest growth rate (1.5-fold higher). Surprisingly, PCL-gelatin did not enhance cell proliferation when compared to the rate of cell growth on plastic surfaces.

Borjigin, Mandula; Eskridge, Chris; Niamat, Rohina; Strouse, Bryan; Bialk, Pawel; Kmiec, Eric B

2013-01-01

352

Electrospun fiber membranes enable proliferation of genetically modified cells.  

PubMed

Polycaprolactone (PCL) and its blended composites (chitosan, gelatin, and lecithin) are well-established biomaterials that can enrich cell growth and enable tissue engineering. However, their application in the recovery and proliferation of genetically modified cells has not been studied. In the study reported here, we fabricated PCL-biomaterial blended fiber membranes, characterized them using physicochemical techniques, and used them as templates for the growth of genetically modified HCT116-19 colon cancer cells. Our data show that the blended polymers are highly miscible and form homogenous electrospun fiber membranes of uniform texture. The aligned PCL nanofibers support robust cell growth, yielding a 2.5-fold higher proliferation rate than cells plated on standard plastic plate surfaces. PCL-lecithin fiber membranes yielded a 2.7-fold higher rate of proliferation, while PCL-chitosan supported a more modest growth rate (1.5-fold higher). Surprisingly, PCL-gelatin did not enhance cell proliferation when compared to the rate of cell growth on plastic surfaces. PMID:23467983

Borjigin, Mandula; Eskridge, Chris; Niamat, Rohina; Strouse, Bryan; Bialk, Pawel; Kmiec, Eric B

2013-01-01

353

Impact of Prolonged Fraction Delivery Times Simulating IMRT on Cultured Nasopharyngeal Carcinoma Cell Killing  

Microsoft Academic Search

Purpose: To determine the impact of prolonged fraction delivery times (FDTs) simulating intensity-modulated radiotherapy (IMRT) on cultured nasopharyngeal carcinoma (NPC) cell killing. Methods and Material: Cultured NPC cell lines CNE1 and CNE2 were used in this study. The biological effectiveness of fractionated irradiation protocols simulating conventional external beam radiotherapy and IMRT (FDT of 15, 36, and 50 minutes) was estimated

Xiao-Kang Zheng; Long-Hua Chen; Wen-Jun Wang; Feng Ye; Jia-Bing Liu; Qi-Sheng Li; Hen-Wen Sun

2010-01-01

354

An adhesion-based method for plasma membrane isolation: evaluating cholesterol extraction from cells and their membranes.  

PubMed

A method to isolate large quantities of directly accessible plasma membrane from attached cells is presented. The method is based on the adhesion of cells to an adsorbed layer of polylysine on glass plates, followed by hypotonic lysis with ice-cold distilled water and subsequent washing steps. Optimal conditions for coating glass plates and time for cell attachment were established. No additional chemical or mechanical treatments were used. Contamination of the isolated plasma membrane by cell organelles was less than 5%. The method uses inexpensive, commercially available polylysine and reusable glass plates. Plasma membrane preparations can be made in 15 min. Using this method, we determined that methyl-beta-cyclodextrin differentially extracts cholesterol from fibroblast cells and their plasma membranes and that these differences are temperature dependent. Determination of the cholesterol/phospholipid ratio from intact cells does not reflect methyl-beta-cyclodextrin plasma membrane extraction properties. PMID:19631189

Bezrukov, Ludmila; Blank, Paul S; Polozov, Ivan V; Zimmerberg, Joshua

2009-11-15

355

Neuroprotective properties of Loranthus parasiticus aqueous fraction against oxidative stress-induced damage in NG108-15 cells.  

PubMed

Loranthus parasiticus, a Chinese folk medicine, has been widely used for the treatment of brain diseases, particularly in southwest China. Hence, the present neuroprotection model was designed to investigate its neuroprotective properties against H(2)O(2)-induced oxidative stress in NG108-15 cells. L. parasiticus aqueous fraction (LPAF), which was selected in the present study, had proved to be the most active fraction among the other tested extracts and fractions in our previous screening. The restoration of depleted intracellular glutathione (GSH), a major endogenous antioxidant, by LPAF was observed after H(2)O(2) insult. Pretreatment with LPAF substantially reduced the production of intracellular reactive oxygen species generated from H(2)O(2). Apoptotic features such as externalization of phosphatidylserine and disruption of mitochondrial membrane potential were significantly attenuated by LPAF. In addition, cell cycle analysis revealed a prominent decrease in the H(2)O(2)-induced sub-G(1) population by LPAF. Moreover, apoptotic morphological analysis by DAPI nuclear staining demonstrated that NG108-15 cells treated with H(2)O(2) exhibited apoptotic features, while such changes were greatly reduced in cells pretreated with LPAF. Taken together, these findings confirmed that LPAF exerts marked neuroprotective activity, which raises the possibility of potential therapeutic application of LPAF for managing oxidative stress-related neurological disorders and supports the traditional use of L. parasiticus in treating brain-related diseases. PMID:22318341

Wong, Daniel Zin Hua; Kadir, Habsah Abdul; Lee, Choy Long; Goh, Bey Hing

2012-07-01

356

Effect of Chemicals on the Cell Membrane Transport of Nucleosides.  

National Technical Information Service (NTIS)

An apparatus and methodology for a high speed kinetic assay of purine efflux has been developed. The procedure is based on a flow system with a membrane filter to remove preloaded L5178Y cells and a sensitive rapid detector of the fluorescence emission of...

P. W. Wigler

1983-01-01

357

Basolateral membrane K+ channels in renal epithelial cells  

PubMed Central

The major function of epithelial tissues is to maintain proper ion, solute, and water homeostasis. The tubule of the renal nephron has an amazingly simple structure, lined by epithelial cells, yet the segments (i.e., proximal tubule vs. collecting duct) of the nephron have unique transport functions. The functional differences are because epithelial cells are polarized and thus possess different patterns (distributions) of membrane transport proteins in the apical and basolateral membranes of the cell. K+ channels play critical roles in normal physiology. Over 90 different genes for K+ channels have been identified in the human genome. Epithelial K+ channels can be located within either or both the apical and basolateral membranes of the cell. One of the primary functions of basolateral K+ channels is to recycle K+ across the basolateral membrane for proper function of the Na+-K+-ATPase, among other functions. Mutations of these channels can cause significant disease. The focus of this review is to provide an overview of the basolateral K+ channels of the nephron, providing potential physiological functions and pathophysiology of these channels, where appropriate. We have taken a “K+ channel gene family” approach in presenting the representative basolateral K+ channels of the nephron. The basolateral K+ channels of the renal epithelia are represented by members of the KCNK, KCNJ, KCNQ, KCNE, and SLO gene families.

Devor, Daniel C.

2012-01-01

358

Membrane electrolytic cell for minimizing hypochlorite and chlorate formation  

Microsoft Academic Search

An electrolytic cell for the electrolysis of an alkali metal chloride brine is comprised of an anode compartment and a cathode compartment separated by a cation exchange membrane. The anode is comprised of an unflattened expanded structure of a valve metal selected from the group consisting of titanium, tantalum, niobium, and alloys thereof. At least one side of the anode

D. L. Fair; D. D. Justice; K. E. Woodard Jr

1985-01-01

359

Apoptosis method for biomimetic artificial cell membranes employing nanophotonic theranostics  

NASA Astrophysics Data System (ADS)

Colloidal biomimetic disc shaped metallic gold shells with a uniform size distribution were synthesized using red blood cells as sacrificial templates. Red blood cells do not reproduce by dividing; hence they are truly colloidal particles. They are almost completely filled with hemoglobin allowing for an extremely dynamic work cycle with long intercellular vacations separated by self-destructive workloads on the cell surface. This method of exchange is emulated in the presented research. The colloidal disc shaped gold shells were coated with multiple layers of 50nm fluorescent polystyrene spheres followed by chemical removal of the gold core. This process yielded hollow synthetic biomimetic membranes with a strong optical signature that are diffusely permeable to water and impervious to particles larger than a few nanometers. Currently, the most successful synthetic intravascular oxygen carrying materials are perfluorocarbons; however, they break down quickly in roughly 50 hours from overexposure to their in vivo workload. The meso-porous membrane cages will be filled with hundreds of fibrous spheroid conglomerates composed of perfluorocarbon chains that can protrude through the meso-porous membrane as they thermally jostle about the cage. This is to statistically limit the exposure time of individual polymer strands to the self-destructive work at the surface and hopefully will greatly increase the effective functioning lifetime of the perfluorocarbon-based synthetic red blood cell. The artificial membranes are intentionally designed to be weak allowing them to flex under normal pressures and to hopefully burst under more extreme conditions such as blockage.

Gilleland, Cody L.; Waters, Brian D.; Jarvis, Brandon; Schaefers, Justin K.; Renfro, Tim; Gutierrez, Jose; Ussery, Geoffrey; Cavanah, Taylor; Glosser, R.; Landon, Preston B.

2005-08-01

360

Sulfonated Nanoplates in Proton Conducting Membranes for Fuel Cells  

SciTech Connect

Surface-functionalized nanoplates are synthesized by anchoring sulfonic acid containing siloxanes on zirconium phosphate, and in turn blended with Nafion to fabricate proton conducting membranes. The effects of these sulfonated nanoplates on proton conduction, hydro-characteristics and fuel cell performance are reported.

Chen, W.F.; Ni’mah, H.; Yu-Cheng Shen, Y.-C.; Kuo, P.-L.

2011-09-29

361

Carbon monoxide poisoning of proton exchange membrane fuel cells  

Microsoft Academic Search

SUMMARY Proton exchange membrane fuel cell (PEMFC) performance degrades when carbon monoxide (CO) is present in the fuel gas; this is referred to as CO poisoning. This paper investigates CO poisoning of PEMFCs by reviewing work on the electrochemistry of CO and hydrogen, the experimental performance of PEMFCs exhibiting CO poisoning, methods to mitigate CO poisoning and theoretical models of

J. J. Baschuk; Xianguo Li

2001-01-01

362

Electroporating Fields Target Oxidatively Damaged Areas in the Cell Membrane  

Microsoft Academic Search

Reversible electropermeabilization (electroporation) is widely used to facilitate the introduction of genetic material and pharmaceutical agents into living cells. Although considerable knowledge has been gained from the study of real and simulated model membranes in electric fields, efforts to optimize electroporation protocols are limited by a lack of detailed understanding of the molecular basis for the electropermeabilization of the complex

P. Thomas Vernier; Zachary A. Levine; Yu-Hsuan Wu; Vanessa Joubert; Matthew J. Ziegler; Lluis M. Mir; D. Peter Tieleman; Boris Rubinsky

2009-01-01

363

The Appropriateness of Unbiased Optical Fractionators to Assess Cell Proliferation in the Adult Hippocampus  

PubMed Central

Optical fractionators have dominated the field of neural cell counting for two decades. These unbiased stereological techniques are often used for the quantification of hippocampal cell proliferation in neurogenesis experiments. However, the heterogeneous distribution of labeled cells, especially in the form of clusters, confounds the application of these techniques. A critical evaluation of the applicability of the optical fractionator suggests that absolute counting achieves higher efficiency in the quantification of cell proliferation than unbiased estimations.

Noori, Hamid R.; Fornal, Casimir A.

2011-01-01

364

Exploring red blood cell membrane dynamics with digital holographic microscopy  

NASA Astrophysics Data System (ADS)

Digital Holographic Microscopy (DHM) has been used to investigate the spontaneous cell membrane fluctuations (CMF) of the Red Blood Cell. DHM as an interferometric technique is able to accurately provide the wavefront deformation induced by a transparent specimen, including living cells in a transmission configuration. From a numerical reconstruction of a single hologram, quantitative phase contrast images are obtained. The local phase shift is proportional to the specimen thickness with accuracy of 5-10 nm. As a non invasive full field technique DHM is particularly well suited to assess and study membrane fluctuations of a large number of cells simultaneously. In our analysis we show that CMF amplitudes are unhomogenously distributed on the cellular surface and seem to correlate with the biconcave equilibrium shape of erythrocytes. A mean fluctuation amplitude of 47 nm is measured in a group of 198 erythrocytes.

Boss, Daniel; Kuehn, Jonas; Depeursinge, Christian; Magistretti, Pierre J.; Marquet, Pierre

2010-04-01

365

Osmotically induced membrane tension facilitates the triggering of living cell electropermeabilization  

Microsoft Academic Search

Very little is known about the molecular mechanisms supporting living cell membrane electropermeabilization. This concept is based on the local membrane permeability induced by cell exposure to brief and intense external electric field pulses. During the electric field application, an electro-induced membrane electric potential difference is created that is locally associated with the dielectric properties of the plasma membrane. When

C. Barrau; J. Teissié; B. Gabriel

2004-01-01

366

Effect of acetaminophen on the membrane anchoring of Na+, K+ATPase of rat renal cortical cells.  

PubMed

In previous works we reported that the administration of a toxic dose of acetaminophen (APAP) induces acute renal failure (ARF) and promotes changes on Na(+), K(+)ATPase distribution in renal proximal plasma membranes. In the present work, we analyzed if APAP could promote the dissociation of Na(+), K(+)ATPase from its membrane anchorage. The participation of calpain activation was also evaluated. We analyzed the Triton X-100 extractability of Na(+), K(+)ATPase in freshly isolated cortical cell suspensions incubated with different APAP concentrations (0.1, 1, 10 and 100 mM). Both alpha(1) and beta(1) subunits were studied by Western blot. APAP promoted the increment of both subunits abundance in the Triton-soluble fraction. Calpain activation was detected in the membrane fractions of cells incubated with APAP. Incubation with APAP 0.1, 1 and 10 mM did not promote an increment in LDH release compared with controls, while APAP 100 mM promoted an increased LDH release. Our results show that incubation of proximal cells with sublethal and lethal APAP concentrations promotes the detachment of Na(+), K(+)ATPase from its membrane anchoring. Inhibition of calpain activation by SJA 7029 protected against APAP-induced membrane damage but not against APAP-induced increase of the Triton X-100 extractability of Na(+), K(+)ATPase. PMID:15949700

Trumper, Laura; Coux, Gabriela; Monasterolo, Liliana A; Molinas, Sara; García, Verónica M C; Elías, M Mónica

2005-06-10

367

The nature of the membrane sites controlling anion permeability of human red blood cells as determined by studies with disulfonic stilbene derivatives  

Microsoft Academic Search

Summary The disulfonic acid stilbene derivative SITS reported to be covalently bonded to the membrane of the red blood cell, was found to be largely reversibly bound. Reversal of its specific inhibitory effect on anion permeability was attained by washing the cells with buffer containing albumin. The small fraction of covalently bonded SITS could be increased by prolonging the time

Z. I. Cabantchik; A. Rothstein

1972-01-01

368

Adhesively-Tensed Cell Membranes: Lysis Kinetics and Atomic Force Microscopy Probing  

Microsoft Academic Search

Membrane tension underlies a range of cell physiological processes. Strong adhesion of the simple red cell is used as a simple model of a spread cell with a finite membrane tension—a state which proves useful for studies of both membrane rupture kinetics and atomic force microscopy (AFM) probing of native structure. In agreement with theories of strong adhesion, the cell

Alina Hategan; Richard Law; Samuel Kahn; Dennis E. Discher

2003-01-01

369

Distinct Role of Rab27a in Granule Movement at the Plasma Membrane and in the Cytosol of NK Cells  

PubMed Central

Protocols were developed to automate image analysis and to track the movement of thousands of vesicular compartments in live cells. Algorithms were used to discriminate among different types of movement (e.g. random, caged, and directed). We applied these tools to investigate the steady-state distribution and movement of lytic granules (LG) in live natural killer (NK) cells by high-speed 3-dimensional (3D) spinning disc confocal and 2-dimensional total internal reflection fluorescence microscopy. Both mouse NK cells and a human NK cell line deficient in the small GTPase Rab27a were examined. The unbiased analysis of large datasets led to the following observations and conclusions. The majority of LG in the cytosol and at the plasma membrane of unstimulated NK cells are mobile. The use of inhibitors indicated that movement in the cytosol required microtubules but not actin, whereas movement at the plasma membrane required both. Rab27a deficiency resulted in fewer LG, and in a reduced fraction of mobile LG, at the plasma membrane. In contrast, loss of Rab27a increased the fraction of mobile LG and the extent of their movement in the cytosol. Therefore, in addition to its documented role in LG delivery to the plasma membrane, Rab27a may restrict LG movement in the cytosol.

Long, Eric O.

2010-01-01

370

MG53 regulates membrane budding and exocytosis in muscle cells.  

PubMed

Membrane recycling and remodeling contribute to multiple cellular functions, including cell fusion events during myogenesis. We have identified a tripartite motif (TRIM72) family member protein named MG53 and defined its role in mediating the dynamic process of membrane fusion and exocytosis in striated muscle. MG53 is a muscle-specific protein that contains a TRIM motif at the amino terminus and a SPRY motif at the carboxyl terminus. Live cell imaging of green fluorescent protein-MG53 fusion construct in cultured myoblasts showed that although MG53 contains no transmembrane segment it is tightly associated with intracellular vesicles and sarcolemmal membrane. RNA interference-mediated knockdown of MG53 expression impeded myoblast differentiation, whereas overexpression of MG53 enhanced vesicle trafficking to and budding from sarcolemmal membrane. Co-expression studies indicated that MG53 activity is regulated by a functional interaction with caveolin-3. Our data reveal a new function for TRIM family proteins in regulating membrane trafficking and fusion in striated muscles. PMID:19029292

Cai, Chuanxi; Masumiya, Haruko; Weisleder, Noah; Pan, Zui; Nishi, Miyuki; Komazaki, Shinji; Takeshima, Hiroshi; Ma, Jianjie

2009-01-30

371

Cholesterol-mediated membrane surface area dynamics in neuroendocrine cells.  

PubMed

How cholesterol, a key membrane constituent, affects membrane surface area dynamics in secretory cells is unclear. Using methyl-beta-cyclodextrin (MbetaCD) to deplete cholesterol, we imaged melanotrophs from male Wistar rats in real-time and monitored membrane capacitance (C(m)), fluctuations of which reflect exocytosis and endocytosis. Treatment with MbetaCD reduced cellular cholesterol and caused a dose-dependent attenuation of the Ca(2+)-evoked increase in C(m) (IC50 = 5.3 mM) vs. untreated cells. Cytosol dialysis of MbetaCD enhanced the attenuation of C(m) increase (IC50 = 3.3 mM), suggesting cholesterol depletion at intracellular membrane sites was involved in attenuating exocytosis. Acute extracellular application of MbetaCD resulted in an immediate C(m) decline, which correlated well with the cellular surface area decrease, indicating the involvement of cholesterol in the regulation of membrane surface area dynamics. This decline in C(m) was three-fold slower than MbetaCD-mediated fluorescent cholesterol decay, implying that exocytosis is the likely physiological means for plasma membrane cholesterol replenishment. MbetaCD had no effect on the specific C(m) and the blockade of endocytosis by Dyngo 4a, confirmed by inhibition of dextran uptake, also had no effect on the time-course of MbetaCD-induced C(m) decline. Thus acute exposure to MbetaCD evokes a C(m) decline linked to the removal of membrane cholesterol, which cannot be compensated for by exocytosis. We propose that the primary contribution of cholesterol to surface area dynamics is via its role in regulated exocytosis. PMID:24046863

Rituper, Bostjan; Chowdhury, Helena Haque; Jorgacevski, Jernej; Coorssen, Jens R; Kreft, Marko; Zorec, Robert

2013-07-01

372

Externally cooled high temperature polymer electrolyte membrane fuel cell stack  

Microsoft Academic Search

One key issue in high temperature polymer electrolyte membrane fuel cell (HT-PEMFC) stack development is heat removal at the operating temperature of 140–180°C. Conventionally, this process is done using coolants such as thermooil, steam or pressurized water. In this contribution, external liquid cooling designs are described, which are avoiding two constraints. First, in the cell active area, no liquid coolant

J. Scholta; M. Messerschmidt; L. Jörissen; Ch. Hartnig

2009-01-01

373

PBI-based composite membranes for polymer fuel cells  

Microsoft Academic Search

In the present study poly(2,2-(2,6-pyridin)-5,5-bibenzimidazole) was used for the preparation of novel MEAs for high-temperature polymer fuel cells (HT-PEMFCs). We prepared hybrid materials with two types of silica fillers in order to increase the MEA performances using this polymer. The membranes were characterized in terms of their microstructure and thermal stability. Cell operation tests and Electrochemical Impedance Spectroscopy were used

V. Kurdakova; E. Quartarone; P. Mustarelli; A. Magistris; E. Caponetti; M. L. Saladino

2010-01-01

374

EPR investigation of cell membrane fluidity upon external oxidative stimulus  

Microsoft Academic Search

The perturbation of the physical state of cell membranes triggered by an external oxidative stimulus has been studied with\\u000a sperm cells which were chosen as a model system. Electron paramagnetic resonance (EPR) spectroscopy was applied and three\\u000a different nitroxides bearing a paramagnetic group on the 5th, 7th and 13th carbon of the stearic acid acyl chain were used\\u000a to probe

M. Kveder; R. Rakoš; M. Gavella; V. Lipovac; G. Pifat; S. Pe?ar; M. Schara

2004-01-01

375

Elastic Thickness Compressibilty of the Red Cell Membrane  

Microsoft Academic Search

We have used an ultrasensitive force probe and optical interferometry to examine the thickness compressibility of the red cell membrane in situ. Pushed into the centers of washed-white red cell ghosts lying on a coverglass, the height of the microsphere-probe tip relative to its closest approach on the adjacent glass surface revealed the apparent material thickness, which began at ?90nm

Volkmar Heinrich; Ken Ritchie; Narla Mohandas; Evan Evans

2001-01-01

376

Influence of water and membrane microstructure on the transport properties of proton exchange membrane fuel cells  

NASA Astrophysics Data System (ADS)

Proton transport in proton exchange membranes (PEMs) depends on interaction between water and acid groups covalently bound to the polymer. Although the presence of water is important in maintaining the PEM's functions, a thorough understanding of this topic is still lacking. The objective of this work is to provide a better understanding of how the nature water, confined to ionic domains of the polymer, influences the membrane's ability to transport protons, methanol and water. Understanding this topic will facilitate development of new materials with favorable transport properties for fuel cells use. Five classes of polymer membranes were used in this work: polyacrylonitrile-graft-poly(styrenesulfonic) acid (PAN-g-macPSSA); poly(vinylidene difluoride) irradiation-graft-poly(styrenesulfonic) acid (PVDF-g-PSSA); poly(ethylenetetrafluoroethylene) irradiation-graft-poly(styrenesulfonic) acid (ETFE-gPSSA); PVDF-g-PSSA with hydroxyethylmethacrylate (HEMA); and perfluorosulfonic acid membrane (Nafion). The nature of water within the polymers (freezable versus non-freezable states) was measured by systematically freezing samples, and observing the temperature at which water freezes and the amount of heat released in the process. Freezing water-swollen membranes resulted in a 4-fold decrease in the proton conductivity of the PEM. Activation energies of proton transport before and after freezing were ˜ 0.15 eV and 0.5 eV, consistent with proton transport through liquid water and bound water, respectively. Reducing the content of water in membrane samples decreased the amount of freezable and non-freezable water. Calorimetric measurements of membranes in various degrees of hydration showed that water molecules became non-freezable when lambda, (water molecules per sulfonic acid group) was less than ˜14. Proton conduction through membranes containing only non-freezable water was demonstrated to be feasible. Diffusion experiments showed that the permeability of methanol decreased when the content of free water in the membranes decreased. Variation in permeability trends observed for the different polymer classes of the same content of free water was explained on the basis of tortuosity and interaction of methanol within the ionic network. Finally, a novel set of polymers containing non-ionic hydrophilic segments were examined for enhanced water transport in order to see if such domains might offset the flux of water due to electro-osmosis.

Siu, Ana Rosa

377

Distinct Apical and Basolateral Membrane Requirements for Stretch-induced Membrane Traffic at the Apical Surface of Bladder Umbrella Cells  

Microsoft Academic Search

Epithelial cells respond to mechanical stimuli by increasing exocytosis, endocytosis, and ion transport, but how these processes are initiated and coordinated and the mechanotransduction pathways involved are not well understood. We observed that in response to a dynamic mechanical environment, increased apical membrane tension, but not pressure, stimulated apical membrane exocytosis and ion transport in bladder umbrella cells. The exocytic

Weiqun Yu; Puneet Khandelwal; Gerard Apodaca

2009-01-01

378

Stabilized composite membranes and membrane electrode assemblies for high temperature\\/low relative humidity polymer electrolyte fuel cell operation  

Microsoft Academic Search

Polymer electrolyte membrane fuel cells (PEMFCs) have a variety of applications in the stationary power, mobile power and automotive power sectors. Existing membrane technology presently permits fuel cell operation at temperatures less than 100°C under fully saturated conditions. However, several advantages such as easier heat rejection rates and improved impurities tolerance by the anode electrocatalyst result by operating a PEMFC

Vijay Krishna Ramani

2004-01-01

379

Alternative Sources of Adult Stem Cells: Human Amniotic Membrane  

NASA Astrophysics Data System (ADS)

Human amniotic membrane is a highly promising cell source for tissue engineering. The cells thereof, human amniotic epithelial cells (hAEC) and human amniotic mesenchymal stromal cells (hAMSC), may be immunoprivileged, they represent an early developmental status, and their application is ethically uncontroversial. Cell banking strategies may use freshly isolated cells or involve in vitro expansion to increase cell numbers. Therefore, we have thoroughly characterized the effect of in vitro cultivation on both phenotype and differentiation potential of hAEC. Moreover, we present different strategies to improve expansion including replacement of animal-derived supplements by human platelet products or the introduction of the catalytic subunit of human telomerase to extend the in vitro lifespan of amniotic cells. Characterization of the resulting cultures includes phenotype, growth characteristics, and differentiation potential, as well as immunogenic and immunomodulatory properties.

Wolbank, Susanne; van Griensven, Martijn; Grillari-Voglauer, Regina; Peterbauer-Scherb, Anja

380

Glass sealed silicon membrane solar cell  

Microsoft Academic Search

An improved silicon back surface field solar cell is described comprising a high quality very thin silicon single crystal wafer base 0.0005 to about 0.0004 inch thick. The base contains a back surface field region of the same type, P or N, as the base. The back surface field region has a thickness of about 1 m and a dopant

Mandelkorn

1987-01-01

381

Nanosecond pulsed electric field induced cytoskeleton, nuclear membrane and telomere damage adversely impact cell survival  

Microsoft Academic Search

We investigated the effects of nanosecond pulsed electric fields (nsPEF) on three human cell lines and demonstrated cell shrinkage, breakdown of the cytoskeleton, nuclear membrane and chromosomal telomere damage. There was a differential response between cell types coinciding with cell survival. Jurkat cells showed cytoskeleton, nuclear membrane and telomere damage that severely impacted cell survival compared to two adherent cell

M. Stacey; P. Fox; S. Buescher; J. Kolb

2011-01-01

382

Continuous flow magnetic cell fractionation based on antigen expression level  

Microsoft Academic Search

Cell separation is important in medical and biological research and plays an increasingly important role in clinical therapy and diagnostics, such as rare cancer cell detection in blood. The immunomagnetic labeling of cells with antibodies conjugated to magnetic nanospheres gives rise to a proportional relationship between the number of magnetic nanospheres attached to the cell and the cell surface marker

Thomas Schneider; Lee R. Moore; Ying Jing; Seungjoo Haam; P. Stephen Williams; Aaron J. Fleischman; Shuvo Roy; Jeffrey J. Chalmers; Maciej Zborowski

2006-01-01

383

T cell proliferative responses to molecular fractions of periodontopathic bacteria.  

PubMed Central

Soluble antigenic preparations of Veillonella parvula and Bacteroides gingivalis were separated by SDS-PAGE and used after electroblotting and solubilization for in vitro lymphocyte stimulation in 13 patients with severe periodontitis and 12 controls. The cellular responses of controls and patients to V. parvula antigens were represented by four main proliferation-inducing fractions with 74-66, 52-46, 22-19 and 12 kD mol. wt. These fractions induced slightly enhanced DNA synthesis in lymphocytes from eight patients who failed to respond to whole antigenic extract. Lymphocyte samples from Veillonella whole extract unresponsive patients were also examined for in vitro proliferation by B. gingivalis fractions. Almost all stimulatory activities could be classified into five regions of 84-74, 35-31, 28-25, 17-15 and 12 kD.

Ivanyi, L; Newman, H N; Marsh, P D

1991-01-01

384

Schistosomula of Schistosoma mansoni use lysophosphatidylcholine to lyse adherent human red blood cells and immobilize red cell membrane components  

PubMed Central

Human red blood cells (RBCs) adhere to and are lysed by schistosomula of Schistosoma mansoni. We have investigated the mechanism of RBC lysis by comparing the dynamic properties of transmembrane protein and lipid probes in adherent ghost membranes with those in control RBCs and in RBCs treated with various membrane perturbants. Fluorescence photobleaching recovery was used to measure the lateral mobility of two integral membrane proteins, glycophorin and band 3, and two lipid analogues, fluorescein phosphatidylethanolamine (Fl-PE) and carbocyanine dyes, in RBCs and ghosts adherent to schistosomula. Adherent ghosts manifested 95-100% immobilization of both membrane proteins and 45-55% immobilization of both lipid probes. In separate experiments, diamide-induced cross-linking of RBC cytoskeletal proteins slowed transmembrane protein diffusion by 30-40%, without affecting either transmembrane protein fractional mobility or lipid probe lateral mobility. Wheat germ agglutinin- and polylysine-induced cross-linking of glycophorin at the extracellular surface caused 80-95% immobilization of the transmembrane proteins, without affecting the fractional mobility of the lipid probe. Egg lysophosphatidylcholine (lysoPC) induced both lysis of RBCs and a concentration-dependent decrease in the lateral mobility of glycophorin, band 3, and Fl-PE in ghost membranes. At a concentration of 8.4 micrograms/ml, lysoPC caused a pattern of protein and lipid immobilization in RBC ghosts identical to that in ghosts adherent to schistosomula. Schistosomula incubated with labeled palmitate released lysoPC into the culture medium at a rate of 1.5 fmol/h per 10(3) organisms. These data suggest that lysoPC is transferred from schistosomula to adherent RBCs, causing their lysis.

1986-01-01

385

Phosphoinositide phosphorylation and hydrolysis in pancreatic islet cell membrane  

SciTech Connect

Membranes were isolated from dispersed rat pancreatic islet cells by attachment to Sephadex beads. When these membranes were exposed to (gamma-32P)ATP, formation of 32P-labeled phosphatidate, phosphatidylinositol 4-phosphate, and phosphatidylinositol 4,5-bisphosphate was observed. Carbamylcholine, added 10 s prior to lipid extraction, caused a dose-related fall in 32P-labeled phospholipids. The effect of the cholinergic agent was suppressed by atropine, ethylene glycol bis(beta-aminoethyl ether)-N,N'-tetraacetic acid, and verapamil, and simulated, in part, by an increase in Ca2+ concentration. When the membranes were derived from islet cells prelabeled with (U-14C)arachidonate, carbamylcholine stimulation, in addition to decreasing labeled polyphosphoinositides, was accompanied by an increased production of labeled diacylglycerol, without a concomitant increase in labeled phosphatidylinositol. These results indicate that activation of a plasma membrane-associated phospholipase C directed against polyphosphoinositides represents a primary event in the functional response of the pancreatic beta cell to cholinergic agents.

Dunlop, M.E.; Malaisse, W.J.

1986-02-01

386

A Novel Unitized Regenerative Proton Exchange Membrane Fuel Cell  

NASA Technical Reports Server (NTRS)

A difficulty encountered in designing a unitized regenerative proton exchange membrane (PEM) fuel cell lies in the incompatibility of electrode structures and electrocatalyst materials optimized for either of the two functions (fuel cell or electrolyzer) with the needs of the other function. This difficulty is compounded in previous regenerative fuel cell designs by the fact that water, which is needed for proton conduction in the PEM during both modes of operation, is the reactant supplied to the anode in the electrolyzer mode of operation and the product formed at the cathode in the fuel cell mode. Drawbacks associated with existing regenerative fuel cells have been addressed. In a first innovation, electrodes function either as oxidation electrodes (hydrogen ionization or oxygen evolution) or as reduction electrodes (oxygen reduction or hydrogen evolution) in the fuel cell and electrolyzer modes, respectively. Control of liquid water within the regenerative fuel cell has been brought about by a second innovation. A novel PEM has been developed with internal channels that permit the direct access of water along the length of the membrane. Lateral diffusion of water along the polymer chains of the PEM provides the water needed at electrode/PEM interfaces. Fabrication of the novel single cell unitized regenerative fuel cell and results obtained on testing it are presented.

Murphy, O. J.; Cisar, A. J.; Gonzalez-Martin, A.; Salinas, C. E.; Simpson, S. F.

1996-01-01

387

Membrane electrode assemblies for unitised regenerative polymer electrolyte fuel cells  

NASA Astrophysics Data System (ADS)

Membrane electrode assemblies for regenerative polymer electrolyte fuel cells were made by hot pressing and sputtering. The different MEAs are examined in fuel cell and water electrolysis mode at different pressure and temperature conditions. Polarisation curves and ac impedance spectra are used to investigate the influence of the changes in coating technique. The hydrogen gas permeation through the membrane is determined by analysing the produced oxygen in electrolysis mode. The analysis shows, that better performances in both process directions can be achieved with an additional layer of sputtered platinum on the oxygen electrode. Thus, the electrochemical round-trip efficiency can be improved by more than 4%. Treating the oxygen electrode with PTFE solution shows better performance in fuel cell and less performance in electrolysis mode. The increase of the round-trip efficiency is negligible. A layer sputtered directly on the membrane shows good impermeability, and hence results in high voltages at low current densities. The mass transportation is apparently constricted. The gas diffusion layer on the oxygen electrode, in this case a titanium foam, leads to flooding of the cell in fuel cell mode. Stable operation is achieved after pretreatment of the GDL with a PTFE solution.

Wittstadt, U.; Wagner, E.; Jungmann, T.

388