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Interfacial interactions between natural RBC membranes and synthetic polymeric nanoparticles  

NASA Astrophysics Data System (ADS)

The unique structural features and stealth properties of a recently developed red blood cell membrane-cloaked nanoparticle (RBC-NP) platform raise curiosity over the interfacial interactions between natural cellular membranes and polymeric nanoparticle substrates. Herein, several interfacial aspects of the RBC-NPs are examined, including completeness of membrane coverage, membrane sidedness upon coating, and the effects of polymeric particles' surface charge and surface curvature on the membrane cloaking process. The study shows that RBC membranes completely cover negatively charged polymeric nanoparticles in a right-side-out manner and enhance the particles' colloidal stability. The membrane cloaking process is applicable to particle substrates with a diameter ranging from 65 to 340 nm. Additionally, the study reveals that both surface glycans on RBC membranes and the substrate properties play a significant role in driving and directing the membrane-particle assembly. These findings further the understanding of the dynamics between cellular membranes and nanoscale substrates and provide valuable information toward future development and characterization of cellular membrane-cloaked nanodevices.The unique structural features and stealth properties of a recently developed red blood cell membrane-cloaked nanoparticle (RBC-NP) platform raise curiosity over the interfacial interactions between natural cellular membranes and polymeric nanoparticle substrates. Herein, several interfacial aspects of the RBC-NPs are examined, including completeness of membrane coverage, membrane sidedness upon coating, and the effects of polymeric particles' surface charge and surface curvature on the membrane cloaking process. The study shows that RBC membranes completely cover negatively charged polymeric nanoparticles in a right-side-out manner and enhance the particles' colloidal stability. The membrane cloaking process is applicable to particle substrates with a diameter ranging from 65 to 340 nm. Additionally, the study reveals that both surface glycans on RBC membranes and the substrate properties play a significant role in driving and directing the membrane-particle assembly. These findings further the understanding of the dynamics between cellular membranes and nanoscale substrates and provide valuable information toward future development and characterization of cellular membrane-cloaked nanodevices. Electronic supplementary information (ESI) available: Theoretical calculations and supporting figures. See DOI: 10.1039/c3nr06371b

Luk, Brian T.; Jack Hu, Che-Ming; Fang, Ronnie H.; Dehaini, Diana; Carpenter, Cody; Gao, Weiwei; Zhang, Liangfang



Canine RBC osmotic tolerance and membrane permeability.  


The objective of this study was to determine the cryobiological characteristics of canine red blood cells (RBC). These included the hydraulic conductivity (L(p)), the permeability coefficients (P(s)) of common cryoprotectant agents (CPAs), the associated reflection coefficient (sigma), the activation energies (E(a)) of L(p) and P(s) and the osmotic tolerance limits. By using a stopped-flow apparatus, the changes of fluorescence intensity emitted by intracellularly entrapped 5-carboxyfluorescein diacetate (CFDA) were recorded when cells were experiencing osmotic volume changes. After the determination of the relationship between fluorescence intensity and cell volume, cell volume changes were calculated. These volume changes were used in three-parameter fitting calculations to determine the values of L(p), P(s), and sigma for common CPAs. These volume measurements and data analyses were repeated at three different temperatures (22, 14, 7 degrees C). Using the Arrhenius equation, the activation energies of L(p) and P(s) in the presence of CPAs were determined. The osmotic tolerance limits for canine RBC were determined by measuring the percentage of free hemoglobin in NaCl solutions with various osmolalities compared to that released by RBC incubated in double distilled water. The upper and lower osmotic tolerance limits were found to be 150mOsm (1.67V(iso)) and 1200mOsm (0.45V(iso)), respectively. These parameters were then used to calculate the amount of non-permeating solute needed to keep cell volume excursions within the osmotic tolerance limits during CPA addition and removal. PMID:12237091

Liu, J; Christian, J A; Critser, J K



Less is more: removing membrane attachments stiffens the RBC cytoskeleton  

NASA Astrophysics Data System (ADS)

The polymerized network of the cytoskeleton of the red-blood cell (RBC) contains different protein components that maintain its overall integrity and attachment to the lipid bilayer. One of these key components is the band 3 ankyrin complex that attaches the spectrin filaments to the fluid bilayer. Defects in this particular component result in the shape transformation called spherocytosis, through the shedding of membrane nano-vesicles. We show here that this transition and membrane shedding can be explained through the increased stiffness of the network when the band 3 ankyrin complexes are removed. ATP-induced transient dissociations lead to network softening, which offsets the stiffening to some extent, and causes increased fragility of these mutant cells, as is observed.

Gov, Nir S.



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



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


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



Comparative analysis of RBC membrane lipids in thalassemia, and iron deficiency anemia in relation to hypochromia and oxidant injury  

Microsoft Academic Search

The effect of an intrinsic defect in the red cell and pronounced hypochromia on oxidative damage to RBC membrane lipids was\\u000a compared in beta-thalassemia and iron deficiency anemia (IDA), which have a varied etiology but equivalent low hemogiobin\\u000a content. The study was planned to correlate the etiology of the disorders to the severity of lipid imbalance and RBC hemolysis\\u000a in

S. P. Sanghani; V. A. Haldankar; K. K. Shalia; S. K. Bichlle



RBC count  


... renal cell carcinoma) Low blood oxygen levels (hypoxia) Pulmonary fibrosis Polycythemia vera Your RBC count will increase for several weeks when you move to a higher altitude. Drugs that can increase the RBC count include: Gentamicin ...


Spectrin Folding versus Unfolding Reactions and RBC Membrane Stiffness  

PubMed Central

Spectrin (Sp), a key component of the erythrocyte membrane, is routinely stretched to near its fully folded contour length during cell deformations. Such dynamic loading may induce domain unfolding as suggested by recent experiments. Herein we develop a model to describe the folding/unfolding of spectrin during equilibrium or nonequilibrium extensions. In both cases, our model indicates that there exists a critical extension beyond which unfolding occurs. We further deploy this model, together with a three-dimensional model of the junctional complex in the erythrocyte membrane, to explore the effect of Sp unfolding on the membrane's mechanical properties, and on the thermal fluctuation of membrane-attached beads. At large deformations our results show a distinctive strain-induced unstiffening behavior, manifested in the slow decrease of the shear modulus, and accompanied by an increase in bead fluctuation. Bead fluctuation is also found to be influenced by mode switching, a phenomenon predicted by our three-dimensional model. The amount of stiffness reduction, however, is modest compared with that reported in experiments. A possible explanation for the discrepancy is the occurrence of spectrin head-to-head disassociation which is also included within our modeling framework and used to analyze bead motion as observed via experiment.

Zhu, Qiang; Asaro, Robert J.



Comparative analysis of RBC membrane lipids in thalassemia, and iron deficiency anemia in relation to hypochromia and oxidant injury.  


The effect of an intrinsic defect in the red cell and pronounced hypochromia on oxidative damage to RBC membrane lipids was compared in beta-thalassemia and iron deficiency anemia (IDA), which have a varied etiology but equivalent low hemogiobin content. The study was planned to correlate the etiology of the disorders to the severity of lipid imbalance and RBC hemolysis in membranes of both the conditions. Results indicated a fall of lysophosphatidylcholine(LPC), phosphatidylethanolamine(PE) and the unsaturated to saturated fatty acid ratio in both conditions, while phosphatidylcholine(PC) increased only in thalassemia. However, irrespective of the disease, sphingomyelin(SM), total cholesterol and phospholipid levels elevated and the hydrogen peroxide stress test indicated increased susceptibility of both pathologic RBCs to peroxidation. Present findings indicate that IDA and thalassemla, allow for considerable amounts of oxidative damage to membrane lipids, irrespective of their etiologles, and thus point hypochromia as an important contributor for inducing lipid imbalance and RBC hemolysis. PMID:23105304

Sanghani, S P; Haldankar, V A; Shalia, K K; Bichlle, S K



Increased anticoagulant osmolality improves separation of leukocytes from red blood cells (RBC)  

Microsoft Academic Search

Background: The bottom-and-top (BAT) procedure separates the buffy coat (BC) from plasma and red blood cells (RBC). The contents of mononuclear cells (MNC) remaining in the RBC are about 1×106 cells\\/unit, whereas the granulocytes are removed less effectively, 500–800×106 or more remaining in the RBC unit. The aim was to improve the separation efficacy by collecting the blood in an

F Knutson; H Lööf; C. F Högman



Theoretical model and experimental study of red blood cell (RBC) deformation in microchannels.  


The motion and deformation of red blood cells (RBCs) flowing in a microchannel were studied using a theoretical model and a novel automated rheoscope. The theoretical model was developed to predict the cells deformation under shear as a function of the cells geometry and mechanical properties. Fluid dynamics and membrane mechanics are incorporated, calculating the traction and deformation in an iterative manner. The model was utilized to evaluate the effect of different biophysical parameters, such as: inner cell viscosity, membrane shear modulus and surface to volume ratio on deformation measurements. The experimental system enables the measurement of individual RBCs velocity and their deformation at defined planes within the microchannel. Good agreement was observed between the simulation results, the rheoscope measurements and published ektacytometry results. The theoretical model results imply that such deformability measuring techniques are weakly influenced by changes in the inner viscosity of the cell or the ambient fluid viscosity. However, these measurements are highly sensitive to RBC shear modulus. The shear modulus, estimated by the model and the rheoscope measurements, falls between the values obtained by micropipette aspiration and laser trapping. The study demonstrates the integration of a theoretical model with a microfabricated device in order to achieve a better understanding of RBC mechanics and their measurement using microfluidic shear assays. The system and the model have the potential of serving as quantitative clinical tools for diagnosing deformability disorders in RBCs. PMID:17188279

Korin, Natanel; Bransky, Avishay; Dinnar, Uri



RBC nuclear scan  


An RBC nuclear scan uses small amounts of radioactive material to mark (tag) red blood cells (RBCs). Your body is then ... radiation -- it does not give off radiation. Most nuclear scans (including an RBC scan) are not recommended ...


Treatment of whole blood (WB) and red blood cells (RBC) with S-303 inactivates pathogens and retains in vitro quality of stored RBC.  


A pathogen inactivation (PI) process has been developed using the frangible anchor linker effector (FRALE) compound S-303. A series of experiments were performed in whole blood (WB) to measure the level of viral and bacterial inactivation. The results showed that 0.2mM S-303 and 2mM glutathione (GSH) inactivated >6.5 logs of HIV, >5.7 logs of Bluetongue virus, >7.0 logs of Yersinia enterocolitica, 4.2 logs of Serratia marcescens, and 7.5 logs of Staphylococcus epidermidis. Recent development for S-303 is focused on optimization of the PI process for red blood cell concentrates (RBC). A series of studies in RBC showed that 0.2mM S-303 and 20mM GSH inactivated approximately 5 logs or greater of Y. enterocolitica, E. coli, S. marcescens, S. aureus, HIV, bovine viral diarrhoea virus, bluetongue virus and human adenovirus 5. In both applications of the S-303 process, in vitro parameters of RBC function and physiology were retained compared to conventional RBC. Results from these studies indicate that S-303 can be applicable for PI of RBC and WB. PMID:19995680

Mufti, N A; Erickson, A C; North, A K; Hanson, D; Sawyer, L; Corash, L M; Lin, L



Measurement of RBC agglutination with microscopic cell image analysis in a microchannel chip.  


Since Landsteiner's discovery of ABO blood groups, RBC agglutination has been one of the most important immunohematologic techniques for ABO and RhD blood groupings. The conventional RBC agglutination grading system for RhD blood typings relies on macroscopic reading, followed by the assignment of a grade ranging from (-) to (4+) to the degree of red blood cells clumping. However, with the new scoring method introduced in this report, microscopically captured cell images of agglutinated RBCs, placed in a microchannel chip, are used for analysis. Indeed, the cell images' pixel number first allows the differentiation of agglutinated and non-agglutinated red blood cells. Finally, the ratio of agglutinated RBCs per total RBC counts (CRAT) from 90 captured images is then calculated. During the trial, it was observed that the agglutinated group's CRAT was significantly higher (3.77-0.003) than that of the normal control (0). Based on these facts, it was established that the microchannel method was more suitable for the discrimination between agglutinated RBCs and non-agglutinated RhD negative, and thus more reliable for the grading of RBCs agglutination than the conventional method. PMID:23364023

Cho, Chi Hyun; Kim, Ju Yeon; Nyeck, Agnes E; Lim, Chae Seung; Hur, Dae Sung; Chung, Chanil; Chang, Jun Keun; An, Seong Soo A; Shin, Sehyun



Podocalyxin Is a Glycoprotein Ligand of the Human Pluripotent Stem Cell-Specific Probe rBC2LCN  

PubMed Central

In comprehensive glycome analysis with a high-density lectin microarray, we have previously shown that the recombinant N-terminal domain of the lectin BC2L-C from Burkholderia cenocepacia (rBC2LCN) binds exclusively to undifferentiated human induced pluripotent stem (iPS) cells and embryonic stem (ES) cells but not to differentiated somatic cells. Here we demonstrate that podocalyxin, a heavily glycosylated type 1 transmembrane protein, is a glycoprotein ligand of rBC2LCN on human iPS cells and ES cells. When analyzed by DNA microarray, podocalyxin was found to be highly expressed in both iPS cells and ES cells. Western and lectin blotting revealed that rBC2LCN binds to podocalyxin with a high molecular weight of more than 240 kDa in undifferentiated iPS cells of six different origins and four ES cell lines, but no binding was observed in either differentiated mouse feeder cells or somatic cells. The specific binding of rBC2LCN to podocalyxin prepared from a large set of iPS cells (138 types) and ES cells (15 types) was also confirmed using a high-throughput antibody-overlay lectin microarray. Alkaline digestion greatly reduced the binding of rBC2LCN to podocalyxin, indicating that the major glycan ligands of rBC2LCN are presented on O-glycans. Furthermore, rBC2LCN was found to exhibit significant affinity to a branched O-glycan comprising an H type 3 structure (Ka, 2.5 × 104 M?1) prepared from human 201B7 iPS cells, indicating that H type 3 is a most probable potential pluripotency marker. We conclude that podocalyxin is a glycoprotein ligand of rBC2LCN on human iPS cells and ES cells.

Tateno, Hiroaki; Matsushima, Asako; Hiemori, Keiko; Onuma, Yasuko; Ito, Yuzuru; Hasehira, Kayo; Nishimura, Ken; Ohtaka, Manami; Takayasu, Satoko; Nakanishi, Mahito; Ikehara, Yuzuru; Nakanishi, Mio; Ohnuma, Kiyoshi; Chan, Techuan; Toyoda, Masashi; Akutsu, Hidenori; Umezawa, Akihiro; Asashima, Makoto



Detection and characterization of red blood cell (RBC) aggregation with photoacoustics  

NASA Astrophysics Data System (ADS)

Red blood cells (RBCs) aggregate in the presence of increased plasma fibrinogen and low shear forces during blood flow. RBC aggregation has been observed in deep vein thrombosis, sepsis and diabetes. We propose using photoacoustics (PA) as a non-invasive imaging modality to detect RBC aggregation. The theoretical and experimental feasibility of PA for detecting and characterizing aggregation was assessed. A simulation study was performed to generate PA signals from non-aggregated and aggregated RBCs using a frequency domain approach and to study the PA signals' dependence on hematocrit and aggregate size. The effect of the finite bandwidth nature of transducers on the PA power spectra was also investigated. Experimental confirmation of theoretical results was conducted using porcine RBC samples exposed to 1064 nm optical wavelength using the Imagio Small Animal PA imaging system (Seno Medical Instruments, Inc., San Antonio, TX). Aggregation was induced with Dextran-70 (Sigma-Aldrich, St. Louis, MO) and the effect of hematocrit and aggregation level was investigated. The theoretical and experimental PA signal amplitude increased linearly with increasing hematocrit. The theoretical dominant frequency content of PA signals shifted towards lower frequencies (<30 MHz) and 9 dB enhancements in spectral power were observed as the size of aggregates increased compared to non-aggregating RBCs. Calibration of the PA spectra with the transducer response obtained from a 200 nm gold film was performed to remove system dependencies. Analysis of the spectral parameters from the calibrated spectra suggested that PA can assess the degree of aggregation at multiple hematocrit and aggregation levels.

Hysi, Eno; Saha, Ratan K.; Rui, Min; Kolios, Michael C.



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



RBC urine test  


Red blood cells in urine; Hematuria test; Urine - red blood cells ... A normal result is 4 RBC/HPF (red blood cells per high power field) or less when the sample is examined under a microscope. The example above is a common measurement ...


Accurate measurement of peripheral blood mononuclear cell concentration using image cytometry to eliminate RBC-induced counting error.  


Peripheral blood mononuclear cells (PBMCs) have been widely researched in the fields of immunology, infectious disease, oncology, transplantation, hematological malignancy, and vaccine development. Specifically, in immunology research, PBMCs have been utilized to monitor concentration, viability, proliferation, and cytokine production from immune cells, which are critical for both clinical trials and biomedical research. The viability and concentration of isolated PBMCs are traditionally measured by manual counting with trypan blue (TB) using a hemacytometer. One of the common issues of PBMC isolation is red blood cell (RBC) contamination. The RBC contamination can be dependent on the donor sample and/or technical skill level of the operator. RBC contamination in a PBMC sample can introduce error to the measured concentration, which can pass down to future experimental assays performed on these cells. To resolve this issue, RBC lysing protocol can be used to eliminate potential error caused by RBC contamination. In the recent years, a rapid fluorescence-based image cytometry system has been utilized for bright-field and fluorescence imaging analysis of cellular characteristics (Nexcelom Bioscience LLC, Lawrence, MA). The Cellometer image cytometry system has demonstrated the capability of automated concentration and viability detection in disposable counting chambers of unpurified mouse splenocytes and PBMCs stained with acridine orange (AO) and propidium iodide (PI) under fluorescence detection. In this work, we demonstrate the ability of Cellometer image cytometry system to accurately measure PBMC concentration, despite RBC contamination, by comparison of five different total PBMC counting methods: (1) manual counting of trypan blue-stained PBMCs in hemacytometer, (2) manual counting of PBMCs in bright-field images, (3) manual counting of acetic acid lysing of RBCs with TB-stained PBMCs, (4) automated counting of acetic acid lysing of RBCs with PI-stained PBMCs, and (5) AO/PI dual staining method. The results show comparable total PBMC counting among all five methods, which validate the AO/PI staining method for PBMC measurement in the image cytometry method. PMID:23201386

Chan, Leo Li-Ying; Laverty, Daniel J; Smith, Tim; Nejad, Parham; Hei, Hillary; Gandhi, Roopali; Kuksin, Dmitry; Qiu, Jean



Differential expression of HSP90? and heme oxygenase in cord blood RBC during preeclampsia.  


Preeclampsia is a multisystem disorder with profound implications on both mother and fetus. Analysis of umbilical cord blood red blood cell (RBC) changes shall depict the fetal response to pregnancy-specific complications like preeclampsia. This study aims to analyze the regulation relationship between HSP90? and heme oxygenase-2 (HO-2) in cord blood RBC during preeclampsia. The lipid hydroperoxide (LHP) and 3-nitrotyrosine (3-NT) levels were measured as stress markers in cord blood RBC of both subjects. The impact of stress on RBC was assessed by measuring the level of membrane bound enzymes and assessing the changes in cord blood RBC. The expression of HSP90? and HO-2 were analyzed by enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry analysis, respectively. There was significant increase in the level of LHP (p < 0.01), 3-NT (p < 0.05), HSP90? (p < 0.01) with decrease in the expression of HO-2 (p < 0.05) in cord blood RBC of preeclamptic subjects compared to normotensive subjects. Similarly, the membrane damage in preeclamptic RBC was assessed by spectrophotometrically and found to be increased by 41.7%, along with increase in number of nucleated RBC. The antiproliferative effect of carbon monoxide under stress might decrease the expression of HO-2 under conditions when there is an increasing need for RBC. The role of HSP90? level in cord blood RBC is discussed with reference to nitrative stress in preeclampsia. This study concludes the increased expression of nucleated RBC, HSP90? and corresponding decreased expression of HO-2 in such hypoxic condition may play a protective role; to prevent cord blood RBC against stress induced damage during preeclampsia. PMID:22935040

Ekambaram, Padmini; Jayachandran, Tharani; Dhakshinamoorthy, Lavanya



RBC indices  


... corpuscular hemoglobin concentration (MCHC); Mean corpuscular volume (MCV); Red blood cell indices ... and hemoglobin. The MCV reflects the size of red blood cells. The MCH and MCHC reflect the ...


Kit-labeled technetium-99m red blood cells (Tc99mRBC's) for clinical cardiac chamber imaging  

Microsoft Academic Search

We have studied 223 consecutive patients by R-wave synchronized cardiac imaging using Tc-99m-RBC's labeled by the Brookhaven kit method. Preparation of the Tc-99m-RBC's is simple and can be accomplished in less than 20 min per patient. The average percent RBC labeling was 96.89%. Only 5 of 223 patients (2.2%) had yields less than 95%. Fifty patients were randomly selected from

Steven M. Larson; Glen W. Hamilton; Powell Richards; James L. Ritchie



Comparative analysis of RBC membrane fatty acids, proteins and glycophorin in patients with heterozygous beta thalassemia and iron deficiency anemia  

Microsoft Academic Search

Membrane lipid and protein composition was compared in erythrocytes from iron deficiency anemia (IDA) and heterozygous beta\\u000a thalassemia patients. The study was planned to correlate the influence of iron deficiency with the intrinsic defect of the\\u000a heterozygous condition on the membrane structural integrity as well as to investigate whether there are differences in membrane\\u000a changes between the two conditions. Results

S. P. Sanghani; V. A. Haldankar



Palmitoylation of MPP1 (Membrane-palmitoylated Protein 1)/p55 Is Crucial for Lateral Membrane Organization in Erythroid Cells*  

PubMed Central

S-Acylation of proteins is a ubiquitous post-translational modification and a common signal for membrane association. The major palmitoylated protein in erythrocytes is MPP1, a member of the MAGUK family and an important component of the ternary complex that attaches the spectrin-based skeleton to the plasma membrane. Here we show that DHHC17 is the only acyltransferase present in red blood cells (RBC). Moreover, we give evidence that protein palmitoylation is essential for membrane organization and is crucial for proper RBC morphology, and that the effect is specific for MPP1. Our observations are based on the clinical cases of two related patients whose RBC had no palmitoylation activity, caused by a lack of DHHC17 in the membrane, which resulted in a strong decrease of the amount of detergent-resistant membrane (DRM) material. We confirmed that this loss of detergent-resistant membrane was due to the lack of palmitoylation by treatment of healthy RBC with 2-bromopalmitic acid (2-BrP, common palmitoylation inhibitor). Concomitantly, fluorescence lifetime imaging microscopy (FLIM) analyses of an order-sensing dye revealed a reduction of membrane order after chemical inhibition of palmitoylation in erythrocytes. These data point to a pathophysiological relationship between the loss of MPP1-directed palmitoylation activity and perturbed lateral membrane organization.

Lach, Agnieszka; Grzybek, Michal; Heger, Elzbieta; Korycka, Justyna; Wolny, Marcin; Kubiak, Jakub; Kolondra, Adam; Boguslawska, Dzamila M.; Augoff, Katarzyna; Majkowski, Michal; Podkalicka, Joanna; Kaczor, Jakub; Stefanko, Adam; Kuliczkowski, Kazimierz; Sikorski, Aleksander F.



Red cell membrane and cation deficiency in Rh null syndrome.  


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



Comparative analysis of RBC membrane fatty acids, proteins and glycophorin in patients with heterozygous beta thalassemia and iron deficiency anemia.  


Membrane lipid and protein composition was compared in erythrocytes from iron deficiency anemia (IDA) and heterozygous beta thalassemia patients. The study was planned to correlate the influence of iron deficiency with the intrinsic defect of the heterozygous condition on the membrane structural integrity as well as to investigate whether there are differences in membrane changes between the two conditions. Results indicate high levels of saturated fatty acids and low unsaturated fatty acids in both disorders although arachidonic acid and the unsaturation index were lower in heterozygous thalassemia than IDA. Nevertheless, neither of the conditions provoked any alterations in membrane protein or glycophorin suggesting alterations in the lipid moiety only. Present findings indicate that irrespective to the etiology, both, iron deficiency and the heterozygous condition show a common pattern of lipid derangement, which may in turn result in increased membrane rigidity and decreased cellular deformability. PMID:23105609

Sanghani, S P; Haldankar, V A



Evaluation of Membrane Systems for Washing/Deglycerolizing Packed Red Blood Cells.  

National Technical Information Service (NTIS)

The objective of this contract is to develop a membrane-based process for deglycerolizing previously frozen, packed red blood cells (RBCs) prior to transfusion. This membrane-based process should reduce the glycerol concentration in the RBC solution in 35...

J. M. Radovich K. R. Pearson R. J. Wedel



Composite fuel cell membranes  


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.



Composite fuel cell membranes  


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



Red blood cell membrane viscoelasticity, agglutination and zeta potential measurements with double optical tweezers  

Microsoft Academic Search

The red blood cell (RBC) viscoelastic membrane contains proteins and glycolproteins embedded in, or attached, to a fluid lipid bilayer and are negatively charged, which creates a repulsive electric (zeta) potential between the cells and prevents their aggregation in the blood stream. There are techniques, however, to decrease the zeta potential to allow cell agglutination which are the basis of

Adriana Fontes; Heloise P. Fernandes; Maria L. Barjas-Castro; André A. de Thomaz; Liliana de Ysasa Pozzo; Luiz C. Barbosa; Carlos L. Cesar



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.



Antigens protected functional red blood cells by the membrane grafting of compact hyperbranched polyglycerols.  


Red blood cell (RBC) transfusion is vital for the treatment of a number of acute and chronic medical problems such as thalassemia major and sickle cell anemia. Due to the presence of multitude of antigens on the RBC surface (~308 known antigens), patients in the chronic blood transfusion therapy develop alloantibodies due to the miss match of minor antigens on transfused RBCs. Grafting of hydrophilic polymers such as polyethylene glycol (PEG) and hyperbranched polyglycerol (HPG) forms an exclusion layer on RBC membrane that prevents the interaction of antibodies with surface antigens without affecting the passage of small molecules such as oxygen, glucose, and ions. At present no method is available for the generation of universal red blood donor cells in part because of the daunting challenge presented by the presence of large number of antigens (protein and carbohydrate based) on the RBC surface and the development of such methods will significantly improve transfusion safety, and dramatically improve the availability and use of RBCs. In this report, the experiments that are used to develop antigen protected functional RBCs by the membrane grafting of HPG and their characterization are presented. HPGs are highly biocompatible compact polymers, and are expected to be located within the cell glycocalyx that surrounds the lipid membrane and mask RBC surface antigens. PMID:23328980

Chapanian, Rafi; Constantinescu, Iren; Brooks, Donald E; Scott, Mark D; Kizhakkedathu, Jayachandran



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



Analysis of nanostructure of red blood cells membranes by space Fourier transform of AFM images.  


Atomic force microscopy (AFM) allows a researcher to obtain images of red blood cells (RBC) and their membranes. Various effects on blood lead to surface alterations of cell membranes. Such alterations are estimated by a corrugation of membrane surface. This problem is complicated for statistical analysis because the membrane is the ensemble of structures with different sizes. In the present work we used the space Fourier transform to decompose the complex AFM image of the surface into three simpler ones. The parameters of spectral windows were selected according to the natural structures of RBC membranes. This method allowed us to obtain high resolution images for the corresponding spectral windows, to establish specificity of alterations from each effect, to estimate quantitatively the membrane nanostructures at different space scales and to compare their sizes statistically after actions of different agents. The blood intoxication was modeled by adding hemin, furosemide, chlorpromazine and zinc ions into blood, in vitro. PMID:22854216

Kozlova, Elena K; Chernysh, Alexander M; Moroz, Victor V; Kuzovlev, Artem N



Affinity driven molecular transfer from erythrocyte membrane to target cells.  


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



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.



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.



Enhancing Effect of Radioresistant Spleen Cells on the Primary Immune Response Against Sheep Rbc by Mouse Spleen Cells in Vitro.  

National Technical Information Service (NTIS)

Irradiated spleen cells cultured for 3 days caused a stimulation of the primary in vitro immune response by normal spleen cells. These irradiated spleen cells were fractionated by velocity sedimentation and the fractions were tested for their stimulating ...

F. H. Lubbe O. B. Zaalberg



Two-dimensional strain-hardening membrane model for large deformation behavior of multiple red blood cells in high shear conditions  

PubMed Central

Background Computational modeling of Red Blood Cell (RBC) flow contributes to the fundamental understanding of microhemodynamics and microcirculation. In order to construct theoretical RBC models, experimental studies on single RBC mechanics have presented a material description for RBC membranes based on their membrane shear, bending and area moduli. These properties have been directly employed in 3D continuum models of RBCs but practical flow analysis with 3D models have been limited by their computationally expensive nature. As such, various researchers have employed 2D models to efficiently and qualitatively study microvessel flows. Currently, the representation of RBC dynamics using 2D models is a limited methodology that breaks down at high shear rates due to excessive and unrealistic stretching. Methods We propose a localized scaling of the 2D elastic moduli such that it increases with RBC local membrane strain, thereby accounting for effects such as the Poisson effect and membrane local area incompressibility lost in the 2D simplification. Validation of our 2D Large Deformation (2D-LD) RBC model was achieved by comparing the predicted RBC deformation against the 3D model from literature for the case of a single RBC in simple shear flow under various shear rates (dimensionless shear rate G?=?0.05, 0.1, 0.2, 0.5). The multi-cell flow of RBCs (38% Hematocrit) in a 20 ?m width microchannel under varying shear rates (50, 150, 150 s-1) was then simulated with our proposed model and the popularly-employed 2D neo-Hookean model in order to evaluate the efficacy of our proposed 2D-LD model. Results The validation set indicated similar RBC deformation for both the 2D-LD and the 3D models across the studied shear rates, highlighting the robustness of our model. The multi-cell simulation indicated that the 2D neo-Hookean model predicts noodle-like RBC shapes at high shear rates (G?=?0.5) whereas our 2D-LD model maintains sensible RBC deformations. Conclusion The ability of the 2D-LD model to limit RBC strain even at high shear rates enables this proposed model to be employed in practical simulations of high shear rate microfluidic flows such as blood separation channels.



Light scattering of human red blood cells during metabolic remodeling of the membrane  

NASA Astrophysics Data System (ADS)

We present the light scattering properties of individual human red blood cells (RBCs). We show that both the RBC static and dynamic scattering signals are altered by adenosine 5'-triphosphate (ATP)-driven membrane metabolic remodeling. To measure the light scattering signal from individual RBCs, we use diffraction phase microscopy together with a Fourier transform light scattering technique. RBC cytosolic ATPs are both chemically and metabolically depleted, and the corresponding scattering signals are compared with the light scattering signal of normal RBCs having physiologic levels of ATP.

Park, Yongkeun; Best-Popescu, Catherine A.; Dasari, Ramachandra R.; Popescu, Gabriel



Role of enzyme-treated cells in RBC antibody screening using the gel test: a study of anti-RH1, -RH2, and -RH3 antibodies.  


The role of enzyme-treated cells (ETCs) in red blood cell (RBC) antibody screening has been the subject of controversy, and its place in the clinical routine remains to be determined. In this work, plasma samples containing anti-RH1 (anti-D; N = 10), anti-RH2 (anti-C; N = 10), or anti-RH3 (anti-E; N = 10) antibodies were studied. The samples were diluted in nonbuffered or buffered normal saline, as well as in a pool of AB plasma samples. Titers and scores were determined by means of the gel test, using the indirect antiglobulin test (IAT) as well as ETCs, with R(0)r, r'r, or r''r test cells. Our results showed that compared to the IAT, ETCs allowed a clearer detection of anti-RH2 and anti-RH3, but not of anti-RH1 antibodies. Based on our study, it is not clear whether the ETC phase of the gel test should be maintained for RBC antibody screening. PMID:17385674

Conne, Jocelyne; Schneider, Philippe; Tissot, Jean-Daniel



Cyclosporin A-Associated Changes in Red Blood Cell Membrane Composition, Deformability, Blood and Plasma Viscosity in Rats  

Microsoft Academic Search

Most of the studies concerning the effects of cyclosporin A (Cs A) on red blood cell (RBC) rheology were carried out in human transplant recipients who may still have residual insufficiency and concomitant administration of other immunosuppressive and antihypertensive drugs. The aim of this study is to evaluate the effects of Cs A on red cell rheology and membrane composition

Evin Ademoglu; Sule Tamer; Isil Albeniz; Cahide Gokkusu; Sevda Tanrikulu



The Effect of Alcohols on Red Blood Cell Mechanical Properties and Membrane Fluidity Depends on Their Molecular Size  

PubMed Central

The role of membrane fluidity in determining red blood cell (RBC) deformability has been suggested by a number of studies. The present investigation evaluated alterations of RBC membrane fluidity, deformability and stability in the presence of four linear alcohols (methanol, ethanol, propanol and butanol) using ektacytometry and electron paramagnetic resonance (EPR) spectroscopy. All alcohols had a biphasic effect on deformability such that it increased then decreased with increasing concentration; the critical concentration for reversal was an inverse function of molecular size. EPR results showed biphasic changes of near-surface fluidity (i.e., increase then decrease) and a decreased fluidity of the lipid core; rank order of effectiveness was butanol > propanol > ethanol > methanol, with a significant correlation between near-surface fluidity and deformability (r = 0.697; p<0.01). The presence of alcohol enhanced the impairment of RBC deformability caused by subjecting cells to 100 Pa shear stress for 300 s, with significant differences from control being observed at higher concentrations of all four alcohols. The level of hemolysis was dependent on molecular size and concentration, whereas echinocytic shape transformation (i.e., biconcave disc to crenated morphology) was observed only for ethanol and propanol. These results are in accordance with available data obtained on model membranes. They document the presence of mechanical links between RBC deformability and near-surface membrane fluidity, chain length-dependence of the ability of alcohols to alter RBC mechanical behavior, and the biphasic response of RBC deformability and near-surface membrane fluidity to increasing alcohol concentrations.

Sonmez, Melda; Ince, Huseyin Yavuz; Yalcin, Ozlem; Ajdzanovic, Vladimir; Spasojevic, Ivan; Meiselman, Herbert J.; Baskurt, Oguz K.



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.



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.



Evaluation of Red Cell Membrane Cytoskeletal Disorders Using a Flow Cytometric Method in South Iran  

PubMed Central

Objective: The diagnosis of hereditary red blood cell (RBC) membrane disorders, and in particular hereditary spherocytosis (HS) and Southeast Asian ovalocytosis (SAO), is based on clinical history, RBC morphology, and other conventional tests such as osmotic fragility. However, there are some milder cases of these disorders that are difficult to diagnose. The application of eosin-5’-maleimide (EMA) was evaluated for screening of RBC membrane defects along with some other anemias. We used EMA dye, which binds mostly to band 3 protein and to a lesser extent some other membrane proteins, for screening of some membrane defects such as HS. Materials and Methods: Fresh RBCs from hematologically normal controls and patients with HS, SAO, hereditary elliptocytosis, hereditary spherocytosis with pincered cells, severe iron deficiency, thalassemia minor, and autoimmune hemolytic anemia were stained with EMA dye and analyzed for mean fluorescent intensity (MFI) using a flow cytometer. Results: RBCs from patients with HS and iron deficiency showed a significant reduction in MFI compared to those from normal controls (p<0.0001 and p<0.001, respectively), while macrocytic RBCs showed a significant increase in MFI (p<0.01). A significant correlation was shown between mean corpuscular volume and MFI, with the exceptions of HS and thalassemia minor. Conclusion: Our results showed that the flow cytometric method could be a reliable diagnostic method for screening and confirmation, with higher sensitivity and specificity (95% and 93%, respectively) than conventional routine tests for HS patients prior to further specific membrane protein molecular tests.

Golafshan, Habib Alah; Ranjbaran, Reza; Kalantari, Tahereh; Moezzi, Leili; Karimi, Mehran; Behzad- Behbahani, Abbas; Aboualizadeh, Farzaneh; Sharifzadeh, Sedigheh



Elastic properties of the red blood cell membrane that determine echinocyte deformability.  


The natural biconcave shape of red blood cells (RBC) may be altered by injury or environmental conditions into a spiculated form (echinocyte). An analysis is presented of the effect of such a transformation on the resistance of RBC to entry into capillary sized cylindrical tubes. The analysis accounts for the elasticity of the membrane skeleton in dilation and shear, and the local and nonlocal resistance of the bilayer to bending, the latter corresponding to different area strains in the two leaflets of the bilayer. The shape transformation is assumed to be driven by the equilibrium area difference (delta A(0), the difference between the equilibrium areas of the bilayer leaflets), which also affects the energy of deformation. The cell shape is approximated by a parametric model. Shape parameters, skeleton shear deformation, and the skeleton density of deformed membrane relative to the skeleton density of undeformed membrane are obtained by minimization of the corresponding thermodynamic potential. Experimentally, delta A(0) is modified and the corresponding discocyte-echinocyte shape transition obtained by high-pressure aspiration into a narrow pipette, and the deformability of the resulting echinocyte is examined by whole cell aspiration into a larger pipette. We conclude that the deformability of the echinocyte can be accounted for by the mechanical behavior of the normal RBC membrane, where the equilibrium area difference delta A(0) is modified. PMID:13680208

Kuzman, D; Svetina, S; Waugh, R E; Zeks, B



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)



Comparative efficacy of blood cell immunocamouflage by membrane grafting of methoxypoly(ethylene glycol) and polyethyloxazoline.  


The grafting of low-immunogenic polymers to cells dramatically reduces antigenic recognition and immunogenicity of allogeneic donor cells consequent to steric and charge camouflage (i.e., immunocamouflage). While methoxypoly(ethylene glycol) [mPEG] has historically been utilized for the immunocamouflage of cells, other low-immunogenic polymers such as polyethyloxazoline propionic acid (PEOZ) may also be capable of conferring immunoprotection. Moreover, PEOZ may have attributes that could have enhanced pharmacological and biological utility relative to mPEG. To evaluate the immunocamouflage efficacy of PEOZ relative to mPEG, human red blood cells (RBC) and leukocytes were modified with mPEG or PEOZ. The differential effects of mPEG and PEOZ was assessed via grafting efficacy, cell morphology and viability, immunocamouflage of surface antigens, and the prevention of in vitro immune recognition (RhD and HLA). Although membrane grafting of mPEG and PEOZ were similar, mPEG demonstrated superior immunocamouflage efficacy as measured by antibody binding and phagocytosis of opsonized RBC while PEOZ showed improved RBC morphology. While mPEG appears to be superior to PEOZ in the immunocamouflage of cells, PEOZ may still be a valuable addition to our repertoire of immunomodulatory polymers. Moreover, our results demonstrate the importance of indirect immunocamouflage of antigens found in membrane protein complexes. PMID:24074839

Kyluik-Price, Dana L; Li, Li; Scott, Mark D




Microsoft Academic Search

There is a hypothesis that lack of n-3 polyunsaturated fatty acids (n-3 PUFA) is of etiological importance in de- pression. Docosahexaenoic acid (DHA), a member of the n-3 PUFA family, is a crucial component of synaptic cell membranes. Objective: The aim of this study was to measure red blood cell (RBC) membrane n-3 fatty acids in a group of depressed



Marathon Running Fails to Influence RBC Survival Rates in Iron-Replete Women.  

ERIC Educational Resources Information Center

This study used radiolabeling to measure red blood cell (RBC) survival rates in six iron-replete female marathon runners, and urinary tests were conducted to search for secondary evidence of RBC damage. The hypothesized RBC fragmentation was not disclosed. (Author/MT)

Steenkamp, Irene; And Others



Metabolic remodeling of the human red blood cell membrane measured by quantitative phase microscopy  

NASA Astrophysics Data System (ADS)

We have quantitatively and systemically measured the morphologies and dynamics of fluctuations in human RBC membranes using a full-field laser interferometry technique that accurately measures dynamic membrane fluctuations. We present conclusive evidence that the presence of adenosine 5'-triphosphate (ATP) facilitates nonequilibrium dynamic fluctuations in the RBC membrane and that these fluctuations are highly correlated with specific regions in the biconcave shape of RBCs. Spatial analysis reveals that these nonequilibrium membrane fluctuations are enhanced at the scale of the spectrin mesh size. Our results indicate the presence of dynamic remodeling in the RBC membrane cortex powered by ATP, which results in nonequilibrium membrane fluctuations.

Park, Yongkeun; Best, Catherine; Auth, Thorsten; Gov, Nir S.; Safran, Samuel; Popescu, Gabriel



Importance of spectrin network reorganization in computer simulations of RBC shapes  

NASA Astrophysics Data System (ADS)

The shape of red blood cells (RBCs) has been the subject of intensive investigations in both experiments and theoretical models. Various computational models for RBCs have also been developed. However, a rigorous quantitative comparison of the observed shapes is still lacking. We have developed a flexible model that allows to study the influence of the various contributions to the membrane stress and their relevance for RBC shape. Our model reveals that a pure curvature model does not fully explain the experimentally observed discocyte shapes. We demonstrate that the in-plane stresses of the spectrin network have a crucial effect on the cell shapes and their transitions, and that the dynamic relaxation of the stresses due to spectrin reorganization is important. We present an extended model that incorporates the effects of dynamic spectrin remodeling and study their role on the dynamics of RBC shapes.

Schiller, Ulf; Ladd, Tony



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.



Effects of age-dependent membrane transport changes on the homeostasis of senescent human red blood cells.  


Little is known about age-related changes in red blood cell (RBC) membrane transport and homeostasis. We investigated first whether the known large variation in plasma membrane Ca(2+) (PMCA) pump activity was correlated with RBC age. Glycated hemoglobin, Hb A1c, was used as a reliable age marker for normal RBCs. We found an inverse correlation between PMCA strength and Hb A1c content, indicating that PMCA activity declines monotonically with RBC age. The previously described subpopulation of high-Na(+), low-density RBCs had the highest Hb A1c levels, suggesting it represents a late homeostatic condition of senescent RBCs. Thus, the normal densification process of RBCs with age must undergo late reversal, requiring a membrane permeability increase with net NaCl gain exceeding KCl loss. Activation of a nonselective cation channel, Pcat, was considered the key link in this density reversal. Investigation of Pcat properties showed that its most powerful activator was increased intracellular Ca(2+). Pcat was comparably selective to Na(+), K(+), choline, and N-methyl-D-glucamine, indicating a fairly large, poorly selective cation permeability pathway. Based on these observations, a working hypothesis is proposed to explain the mechanism of progressive RBC densification with age and of the late reversal to a low-density condition with altered ionic gradients. PMID:17456724

Lew, Virgilio L; Daw, Nuala; Etzion, Zipora; Tiffert, Teresa; Muoma, Adaeze; Vanagas, Laura; Bookchin, Robert M



Spatially-Resolved Eigenmode Decomposition of Red Blood Cells Membrane Fluctuations Questions the Role of ATP in Flickering  

PubMed Central

Red blood cells (RBCs) present unique reversible shape deformability, essential for both function and survival, resulting notably in cell membrane fluctuations (CMF). These CMF have been subject of many studies in order to obtain a better understanding of these remarkable biomechanical membrane properties altered in some pathological states including blood diseases. In particular the discussion over the thermal or metabolic origin of the CMF has led in the past to a large number of investigations and modeling. However, the origin of the CMF is still debated. In this article, we present an analysis of the CMF of RBCs by combining digital holographic microscopy (DHM) with an orthogonal subspace decomposition of the imaging data. These subspace components can be reliably identified and quantified as the eigenmode basis of CMF that minimizes the deformation energy of the RBC structure. By fitting the observed fluctuation modes with a theoretical dynamic model, we find that the CMF are mainly governed by the bending elasticity of the membrane and that shear and tension elasticities have only a marginal influence on the membrane fluctations of the discocyte RBC. Further, our experiments show that the role of ATP as a driving force of CMF is questionable. ATP, however, seems to be required to maintain the unique biomechanical properties of the RBC membrane that lead to thermally excited CMF.

Boss, Daniel; Hoffmann, Annick; Rappaz, Benjamin; Depeursinge, Christian; Magistretti, Pierre J.; Van de Ville, Dimitri; Marquet, Pierre



Protective effects of tea polyphenols against oxidative damage to red blood cells  

Microsoft Academic Search

Tea polyphenols (TPP) from black and green teas were evaluated for their antioxidant effects on normal red blood cells (RBC) and ?-thalassemic RBC membranes challenged with exogenous oxidants in vitro. The TPP of both types protected RBC against primaquine-induced lysis; they also protected the whole cells and the membranes against H2O2-induced lipid peroxidation so that about 80% protection was reached

Leonid N. Grinberg; Harold Newmark; Nahum Kitrossky; Ezra Rahamim; Mordechai Chevion; Eliezer A. Rachmilewitz



The effect of curvature on the undulation spectrum of Red Blood Cell membranes  

NASA Astrophysics Data System (ADS)

The human red blood cell (RBC) membrane has a composite structure of a fluid lipid bilayer tethered to an elastic 2D spectrin network. The study of the mechanical properties of RBCs is crucial to our understanding of their ability withstand large amplitude deformations during their passage through the microvasculature. The linear mechanical response of this composite membrane can be measured by observing its undulatory dynamics in thermal equilibrium, i.e. microrheology. Previous models of these dynamics postulated an effective surface tension. In this talk, we show that surface tension is not necessary. Rather, the coupling of membrane bending to spectrin network compression by curvature can account for the observed dynamics. We use a simplified theoretical model to describe the undulatory dynamics of RBCs, measured experimentally by the Popescu group.ootnotetextG. Popescu et al. ``Imaging red blood cell dynamics by quantitative phase microscopy, Blood Cells, Molecules, and Diseases, (2008), in print'' Analyzing their data using our model, we observe dramatic changes in RBC membrane elasticity associated with cells' morphological transition from discocytes to echinocyte to spherocyte.

Kuriabova, Tatiana; Henle, Mark L.; Levine, Alex J.



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



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



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



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.



Defected red blood cell membranes and direct correlation with the uraemic milieu: the connection with the decreased red blood cell lifespan observed in haemodialysis patients  

NASA Astrophysics Data System (ADS)

Together with impaired production of erythropoietin and iron deficiency, the decreased lifespan of red blood cells (RBCs) is a main factor contributing to the chronic anaemia observed in haemodialysis (HD) patients. Atomic force microscopy is employed in this work to thoroughly survey the membrane of intact RBCs (iRBCs) of HD patients in comparison to those of healthy donors, aiming to obtain direct information on the structural status of RBCs that can be related to their decreased lifespan. We observed that the iRBC membrane of the HD patients is overpopulated with extended circular defects, termed ‘orifices’, that have typical dimension ranging between 0.2 and 1.0 ?m. The ‘orifice’ index—that is, the mean population of ‘orifices’ per top membrane surface—exhibits a pronounced relative increase of order 54 ± 12% for the HD patients as compared to healthy donors. Interestingly, for the HD patients, the ‘orifice’ index, which relates to the structural status of the RBC membrane, correlates strongly with urea concentration, which is a basic index of the uraemic milieu. Thus, these results indicate that the uraemic milieu downgrades the structural status of the RBC membrane, possibly triggering biochemical processes that result in their premature elimination from the circulation. This process could decrease the lifespan of RBCs, as observed in HD patients.

Stamopoulos, D.; Grapsa, E.; Manios, E.; Gogola, V.; Bakirtzi, N.



Following-up changes in red blood cell deformability and membrane stability in the presence of PTFE graft implanted into the femoral artery in a canine model  

NASA Astrophysics Data System (ADS)

It is known that a moderate mechanical stress can even improve the red blood cells' (RBC) micro-rheological characteristics, however, a more significant stress causes deterioration in the deformability. In this study, we aimed to investigate the effect of the presence of artificial graft on the RBC deformability and membrane stability in beagles. In the Control group only anesthesia was induced and in the postoperative (p.o.) period blood samplings were carried out. In the Grafted group under general anesthesia, the left femoral artery was isolated, from which a 3.5 cm segment was resected and a PTFE graft (O.D.: 3 mm) of equal in length was implanted into the gap. On the 1st, 3rd, 5th, 7th and 14th p.o. days blood was collected the cephalic veins and RBC deformability was determined ektacytometry (LoRRca MaxSis Osmoscan). Membrane stability test consisted of two deformability measurements before and after the cells were being exposed to mechanical stress (60 or 100 Pa for 300 seconds). Compared to the Control group and the baseline values the red blood cell deformability showed significant deterioration on the 3rd, 5th and mainly on the 7th postoperative day after the graft implantation. The membrane stability of erythrocyte revealed marked inter-group difference on the 3rd, 5th and 7th day: in the Grafted group the deformability decreased and during the membrane stability test smaller difference was observed between the states before and after shearing. We concluded that the presence of a PTFE graft in the femoral artery may cause changes in RBC deformability in the first p.o. week. RBC membrane stability investigation shows a lower elongation index profile for the grafted group and a narrowed alteration in the deformability curves due to mechanical stress.

Toth, Csaba; Kiss, Ferenc; Klarik, Zoltan; Gergely, Eszter; Toth, Eniko; Peto, Katalin; Vanyolos, Erzsebet; Miko, Iren; Nemeth, Norbert



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.



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



Specific binding of Thiobacillus ferrooxidans RbcR to the intergenic sequence between the rbc operon and the rbcR gene.  

PubMed Central

The presence of two sets (rbcL1-rbcS1 and rbcL2-rbcS2) of rbc operons has been demonstrated in Thiobacillus ferrooxidans Fe1 (T. Kusano, T. Takeshima, C. Inoue, and K. Sugawara, J. Bacteriol. 173:7313-7323, 1991). A possible regulatory gene, rbcR, 930 bp long and possibly translated into a 309-amino-acid protein, was found upstream from the rbcL1 gene as a single copy. The gene is located divergently to rbcL1 with a 144-bp intergenic sequence. As in the cases of the Chromatium vinosum RbcR and Alcaligenes eutrophus CfxR, T. ferrooxidans RbcR is thought to be a new member of the LysR family, and these proteins share 46.5 and 42.8% identity, respectively. Gel mobility shift assays showed that T. ferrooxidans RbcR, produced in Escherichia coli, binds specifically to the intergenic sequence between rbcL1 and rbcR. Footprinting and site-directed mutagenesis experiments further demonstrated that RbcR binds to overlapping promoter elements of the rbcR and rbcL1 genes. The above data strongly support the participation of RbcR in regulation of the rbcL1-rbcS1 operon and the rbcR gene in T. ferrooxidans. Images

Kusano, T; Sugawara, K



Elastic behavior of a red blood cell with the membrane's nonuniform natural state: equilibrium shape, motion transition under shear flow, and elongation during tank-treading motion.  


Direct numerical simulations of the mechanics of a single red blood cell (RBC) were performed by considering the nonuniform natural state of the elastic membrane. A RBC was modeled as an incompressible viscous fluid encapsulated by an elastic membrane. The in-plane shear and area dilatation deformations of the membrane were modeled by Skalak constitutive equation, while out-of-plane bending deformation was formulated by the spring model. The natural state of the membrane with respect to in-plane shear deformation was modeled as a sphere ([Formula: see text]), biconcave disk shape ([Formula: see text]) and their intermediate shapes ([Formula: see text]) with the nonuniformity parameter [Formula: see text], while the natural state with respect to out-of-plane bending deformation was modeled as a flat plane. According to the numerical simulations, at an experimentally measured in-plane shear modulus of [Formula: see text] and an out-of-plane bending rigidity of [Formula: see text] of the cell membrane, the following results were obtained. (i) The RBC shape at equilibrium was biconcave discoid for [Formula: see text] and cupped otherwise; (ii) the experimentally measured fluid shear stress at the transition between tumbling and tank-treading motions under shear flow was reproduced for [Formula: see text]; (iii) the elongation deformation of the RBC during tank-treading motion from the simulation was consistent with that from in vitro experiments, irrespective of the [Formula: see text] value. Based on our RBC modeling, the three phenomena (i), (ii), and (iii) were mechanically consistent for [Formula: see text]. The condition [Formula: see text] precludes a biconcave discoid shape at equilibrium (i); however, it gives appropriate fluid shear stress at the motion transition under shear flow (ii), suggesting that a combined effect of [Formula: see text] and the natural state with respect to out-of-plane bending deformation is necessary for understanding details of the RBC mechanics at equilibrium. Our numerical results demonstrate that moderate nonuniformity in a membrane's natural state with respect to in-plane shear deformation plays a key role in RBC mechanics. PMID:24104211

Tsubota, Ken-Ichi; Wada, Shigeo; Liu, Hao



Competition between Na + and Li + for Unsealed and Cytoskeleton-Depleted Human Red Blood Cell Membrane: A 23Na Multiple Quantum Filtered and 7Li NMR Relaxation Study  

NASA Astrophysics Data System (ADS)

Evidence for competition between Li + and Na + for binding sites of human unsealed and cytoskeleton-depleted human red blood cell (csdRBC) membranes was obtained from the effect of added Li + upon the 23Na double quantum filtered (DQF) and triple quantum filtered (TQF) NMR signals of Na +-containing red blood cell (RBC) membrane suspensions. We found that, at low ionic strength, the observed quenching effect of Li + on the 23Na TQF and DQF signal intensity probed Li +/Na + competition for isotropic binding sites only. Membrane cytoskeleton depletion significantly decreased the isotropic signal intensity, strongly affecting the binding of Na + to isotropic membrane sites, but had no effect on Li +/Na + competition for those sites. Through the observed 23Na DQF NMR spectra, which allow probing of both isotropic and anisotropic Na + motion, we found anisotropic membrane binding sites for Na + when the total ionic strength was higher than 40 mM. This is a consequence of ionic strength effects on the conformation of the cytoskeleton, in particular on the dimer-tetramer equilibrium of spectrin. The determinant involvement of the cytoskeleton in the anisotropy of Na + motion at the membrane surface was demonstrated by the isotropy of the DQF spectra of csdRBC membranes even at high ionic strength. Li + addition initially quenched the isotropic signal the most, indicating preferential Li +/Na + competition for the isotropic membrane sites. High ionic strength also increased the intensity of the anisotropic signal, due to its effect on the restructuring of the membrane cytoskeleton. Further Li + addition competed with Na + for those sites, quenching the anisotropic signal. 7Li T1 relaxation data for Li +-containing suspensions of unsealed and csdRBC membranes, in the absence and presence of Na + at low ionic strength, showed that cytoskeleton depletion does not affect the affinity of Na + for the RBC membrane, but increases the affinity of Li + by 50%. This clearly indicates that cytoskeleton depletion favors Li + relative to Na + binding, and thus Li +/Na + competition for its isotropic sites. Thus, this relaxation technique proves to be very sensitive to alkali metal binding to the membrane, detecting a more pronounced steric hindrance effect of the cytoskeleton network to binding of the larger hydrated Li + ion to the membrane phosphate groups.

Srinivasan, Chandra; Minadeo, Nicole; Toon, Jason; Graham, Daniel; Mota de Freitas, Duarte; Geraldes, Carlos F. G. C.



Changes in the properties of normal human red blood cells during in vivo aging  

PubMed Central

The changes in red blood cells (RBC) as they age and the mechanisms for their eventual removal have been of interest for many years. Proposed age-related changes include dehydration with increased density and decreased size, increased membrane IgG, loss of membrane phospholipid asymmetry, and decreased activity of KCl cotransport. The biotin RBC label allows unambiguous identification of older cells and exploration of their properties as they age. Autologous normal human RBC were labeled ex vivo and, after reinfusion, compared with unlabeled RBC throughout their lifespan. RBC density increased with age, with most of the change in the first weeks. Near the end of their lifespan, RBC had increased surface IgG. However, there was no evidence for elevated external phosphatidylserine (PS) even though older RBC had significantly lower activity of aminophospholipid translocase (APLT). KCl cotransport activity persisted well past the reticulocyte stage, but eventually decreased as the RBC became older. These studies place limitations on the use of density fractionation for the study of older human RBC, and do not support loss of phospholipid asymmetry as a mechanism for human RBC senescence. However, increased levels of IgG were associated with older RBC, and may contribute to their removal from the circulation.

Franco, Robert S.; Puchulu-Campanella, M. Estela; Barber, Latorya A.; Palascak, Mary B.; Joiner, Clinton H.; Low, Philip S.; Cohen, Robert M.



Computational analysis on the mechanical interaction between a thrombus and red blood cells: possible causes of membrane damage of red blood cells at microvessels.  


Previous studies investigating thrombus formation have not focused on the physical interaction between red blood cells (RBCs) and thrombus, although they have been speculated that some pathological conditions such as microangiopathic hemolytic anemia (MAHA) stem from interactions between RBCs and thrombi. In this study, we investigated the mechanical influence of RBCs on primary thrombi during hemostasis. We also explored the mechanics and aggravating factors of intravascular hemolysis. Computer simulations of primary thrombogenesis in the presence and the absence of RBCs demonstrated that RBCs are unlikely to affect the thrombus height and coverage, although their presence may change microvessel hemodynamics and platelet transportation to the injured wall. Our results suggest that intravascular hemolysis owing to RBC membrane damage would be promoted by three hemodynamic factors: (1) dispersibility of platelet thrombi, because more frequent spatial thrombus formation decreases the time available for an RBC to recover its shape and enforces more severe deformation; (2) platelet thrombus stiffness, because a stiffer thrombus increases the degree of RBC deformation upon collision; and (3) vessel size and hemocyte density, because a smaller vessel diameter and higher hemocyte density decrease the room for RBCs to escape as they come closer to a thrombus, thereby enhancing thrombus-RBC interactions. PMID:22356820

Kamada, Hiroki; Imai, Yohsuke; Nakamura, Masanori; Ishikawa, Takuji; Yamaguchi, Takami



Red blood cell in simple shear flow  

NASA Astrophysics Data System (ADS)

The dynamics of red blood cells (RBC) in blood flow is critical for oxygen transport, and it also influences inflammation (white blood cells), thrombosis (platelets), and circulatory tumor migration. The physical properties of a RBC can be captured by modeling RBC as lipid membrane linked to a cytoskeletal spectrin network that encapsulates cytoplasm rich in hemoglobin, with bi-concave equilibrium shape. Depending on the shear force, RBC elasticity, membrane viscosity, and cytoplasm viscosity, RBC can undergo tumbling, tank-treading, or oscillatory motion. We investigate the dynamic state diagram of RBC in shear and pressure-driven flow using a combined immersed boundary-lattice Boltzmann method with a multi-scale RBC model that accurately captures the experimentally established RBC force-deformation relation. It is found that the tumbling (TU) to tank-treading (TT) transition occurs as shear rate increases for cytoplasm/outer fluid viscosity ratio smaller than 0.67. The TU frequency is found to be half of the TT frequency, in agreement with experiment observations. Larger viscosity ratios lead to the disappearance of stable TT phase and unstable complex dynamics, including the oscillation of the symmetry axis of the bi-concave shape perpendicular to the flow direction. The dependence on RBC bending rigidity, shear modulus, the order of membrane spectrin network and fluid field in the unstable region will also be discussed.

Chien, Wei; Hew, Yayu; Chen, Yeng-Long



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.



Anion exchanger 1 (band 3) is required to prevent erythrocyte membrane surface loss but not to form the membrane skeleton.  


The red blood cell (RBC) membrane protein AE1 provides high affinity binding sites for the membrane skeleton, a structure critical to RBC integrity. AE1 biosynthesis is postulated to be required for terminal erythropoiesis and membrane skeleton assembly. We used targeted mutagenesis to assess AE1 function in vivo. RBCs lacking AE1 spontaneously shed membrane vesicles and tubules, leading to severe spherocytosis and hemolysis, but the levels of the major skeleton components, the synthesis of spectrin in mutant erythroblasts, and skeletal architecture are normal or nearly normal. The results indicate that AE1 does not regulate RBC membrane skeleton assembly in vivo but is essential for membrane stability. We postulate that stabilization is achieved through AE1-lipid interactions and that loss of these interactions is a key pathogenic event in hereditary spherocytosis. PMID:8808627

Peters, L L; Shivdasani, R A; Liu, S C; Hanspal, M; John, K M; Gonzalez, J M; Brugnara, C; Gwynn, B; Mohandas, N; Alper, S L; Orkin, S H; Lux, S E



Determination of red blood cell membrane viscosity from rheoscopic observations of tank-treading motion.  

PubMed Central

Measurements of the dimensions and membrane rotational frequency of individual erythrocytes steadily tank-treading in a rheoscope are used to deduce the surface shear viscosity of the membrane. The method is based on an integral energy principle which says that the power supplied to the tank-treading cell by the suspending fluid is equal to the rate at which energy is dissipated by viscous action in the membrane and cytoplasm. The integrals involved are formulated with the aid of an idealized mathematical model of the tank-treading red blood cell (RBC) (Keller and Skalak, 1982, J. Fluid Mech., 120:24-27) and evaluated numerically. The outcome is a surface-averaged value of membrane viscosity which is representative of a finite interval of membrane shear rate. The numerical values computed show a clear shear-thinning characteristic as well as a significant augmentation of viscosity with cell age and tend toward agreement with those determined for the rapid phase of shape recovery in micropipettes (Chien, S., K.-L. P. Sung, R. Skalak, S. Usami, and A. Tozeren, 1978, Biophys. J., 24:463-487). The computations also indicate that the rate of energy dissipation in the membrane is always substantially greater than that in the cytoplasm. Images FIGURE 2

Tran-Son-Tay, R; Sutera, S P; Rao, P R



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



Toxic effects of Litsea elliptica Blume essential oil on red blood cells of Sprague-Dawley rats*  

PubMed Central

Litsea elliptica Blume leaves have been traditionally used as medicinal herbs because of its antimutagenicity, chemopreventative and insecticidal properties. In this study, the toxic effects of L. elliptica essential oil against Sprague-Dawley rat’s red blood cells (RBCs) were evaluated. L. elliptica essential oil was given by oral gavage 5 times per week for 3 treated groups in the doses of 125, 250, and 500 mg/(kg body weight), respectively, and the control group received distilled water. Full blood count, RBC osmotic fragility, RBC morphological changes, and RBC membrane lipid were analyzed 28 d after the treatment. Although L. elliptica essential oil administration had significantly different effects on hemoglobin (Hb), mean cell hemoglobin concentration (MCHC), mean cell volume (MCV), and mean cell hemoglobin (MCH) in the experimental groups as compared to the control group (P<0.05), the values were still within the normal range. L. elliptica induced morphological changes of RBC into the form of echinocyte. The percentage of echinocyte increased significantly among the treated groups in a dose-response manner (P<0.001). The concentrations of RBC membrane phospholipids and cholesterol of all treated groups were significantly lower than those of control group (P<0.001). However, the RBC membrane osmotic fragility and total proteins of RBC membrane findings did not differ significantly between control and treated groups (P>0.05). It is concluded that structural changes in the RBC membrane due to L. elliptica essential oil administration did not cause severe membrane damage.

Taib, Izatus Shima; Budin, Siti Balkis; Siti Nor Ain, Seri Maseran; Mohamed, Jamaludin; Louis, Santhana Raj; Das, Srijit; Sallehudin, Sulaiman; Rajab, Nor Fadilah; Hidayatulfathi, Othman



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)



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)



Hereditary xerocytosis: a report of six unrelated Spanish families with leaky red cell syndrome and increased heat stability of the erythrocyte membrane.  


Hereditary xerocytosis (HX) is a rare haemolytic disease due to dehydrated red blood cells (RBCs). A unique feature of this syndrome is that affected members often show normal or near normal haemoglobin levels despite clinical and laboratory evidence of mild to moderate haemolysis. The diagnostic clue is the association of markedly increased RBC Na+ + K+ fluxes with low total cation (Na+ + K+) content. 11 patients of six unrelated families of Spanish origin with HX have been studied from clinical, genetical and biological points of view. In addition, we have investigated the sensitivity of RBC membrane to heat at three different incubation times (15, 30 and 60 min) and two different temperature values (46 degrees C and 49 degrees C). Under these conditions control RBCs (50 normal subjects) exhibited at 49 degrees C and 30 min a maximum of 30% fragmented RBCs. This value increased to 80% after 60 min of incubation. In contrast, patients with HX showed significantly lower percentages of fragmented RBCs at both 30 and 60 min of incubation (maximum 10% and 30%, respectively). In an attempt to determine if increased heat stability was unique to HX RBCs, several other congenital membranopathies with haemolytic anaemia were also studied. The degree of fragmentation, except in one case of HPP (which was strongly increased), did not differ from the control group. Electrophoretic studies of membrane proteins performed in RBCs of all the patients with HX did not explain any qualitative nor quantitative abnormality. In addition to its physiopathological interest, study of RBC heat stability, together with other haematological parameters (increased MCHC and decreased RBC osmotic fragility), may be useful for HX diagnosis, especially in laboratories which are not equipped to evaluate RBC membrane permeability. PMID:7669659

Vives Corrons, J L; Besson, I; Aymerich, M; Ayala, S; Alloisio, N; Delaunay, J; Gonzalez, I; Manrubia, E



Suppression of fluid membrane fluctuations by a periodic pinning potential: Applications to red blood cells.  

NASA Astrophysics Data System (ADS)

The membrane of the red blood cell (RBC) is tethered to a two- dimensional triangular network of semi-flexible elastic spectrin filaments. This network allows the cell to maintain its structural integrity during the large shape deformations that occur as it circulates through the microvasculature. The lipid membrane is anchored to the spectrin filaments at the nodes of the network. Consequently, these attachments impose a two-dimensional periodic pinning potential upon the membrane. In this talk, we investigate the effect of this pinning potential on the thermal bending fluctuations of the membrane. We show that there is an exact mapping of this system onto the classic problem of non-interacting electrons subject to a periodic potential; we exploit this mapping to obtain an exact analytic solution for a defect-free triangular array of harmonic pinning sites. The pinning potential affects both the local and global structure of the bending fluctuations. To investigate the local structure we consider the bending correlations between two nearby points in the membrane, while for the global structure we consider the total area stored in the fluctuations. We also investigate the effective area modulus of the membrane/spectrin composite structure.

Henle, Mark L.; Levine, Alex J.



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.



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



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



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



Imaging of membrane systems and membrane traffic in living cells.  


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



Drag reducing polymers improve tissue perfusion via modification of the RBC traffic in microvessels  

PubMed Central

This paper reports a novel, physiologically significant, microfluidic phenomenon generated by nanomolar concentrations of drag-reducing polymers (DRP) dissolved in flowing blood, which may explain previously demonstrated beneficial effects of DRP on tissue perfusion. In microfluidic systems used in this study, DRP additives were found to significantly modify traffic of red blood cells (RBC) into microchannel branches as well as reduce the near-wall cell-free layer, which normally is found in microvessels with a diameter smaller than 0.3 mm. The reduction in plasma layer size led to attenuation of the so-called “plasma skimming” effect at microchannel bifurcations, increasing the number of RBC entering branches. In vivo, these changes in RBC traffic may facilitate gas transport by increasing the near vessel wall concentration of RBC and capillary hematocrit. In addition, an increase in near-wall viscosity due to the redirection of RBC in this region may potentially decrease vascular resistance as a result of increased wall shear stress, which promotes endothelium mediated vasodilation. These microcirculatory phenomena may explain the previously reported beneficial effects of DRP on hemodynamics in vivo observed in many animal studies. We also report here our finding that DRP additives reduce flow separations at microchannel expansions, deflecting RBC closer to the wall and eliminating the plasma recirculation zone. Although the exact mechanism of the DRP effects on RBC traffic in microchannels is yet to be elucidated, these findings may further DRP progress toward clinical use.

Marhefka, J.N.; Zhao, R.; Wu, Z.; Velankar, S.S.; Antaki, J.F.; Kameneva, M.V.



Urinary proteins and red blood cell membrane negative charges in diabetes mellitus.  


The nature and origin of proteinuria in diabetes mellitus have been investigated by measuring the urinary excretion of seven specific proteins of low (beta 2-microglobin, retinol-binding protein) or high molecular weight (albumin, transferrin, hemopexin and IgG). Using the Alcian Blue binding test, we also measured negative charges on red blood cell (RBC) membrane which according to recent studies might mirror the glomerular polyanion charge. A group of 190 diabetics was examined, including 90 patients with type I diabetes, 23 type II diabetics treated with diet and/or hypoglycaemic agents and 77 longstanding type II diabetics requiring insulin therapy. With the exception of beta 2-microglobulin all proteins measured were excreted in the urine of diabetics in significantly higher amounts than in controls. The assay of transferrin proved the most sensitive (58% positive) followed by albumin (49%), IgG (34%), hemopexin (28%) and retinol-binding protein (26%). Practically the same ranking was obtained when only type I diabetics were considered. RBC membrane negative charges were diminished in diabetics and negatively correlated with the urinary excretion of albumin (r = -0.61, n = 190). RBC charges were also negatively correlated with other urinary proteins of high molecular mass (r between - 0.5 and - 0.2) but presented no relation with urinary beta 2-microglobulin or retinol-binding protein. The loss of RBC charges in diabetics most likely reflects the concomitant depletion of the glomerular polyanion responsible for the increased glomerular leakage of high molecular mass plasma proteins. The preferential increase in transferrin excretion together with the progressive rise in the urinary excretion of IgG lead us to postulate that the loss of glomerular polyanion in diabetes is accompanied, from the early stage, by a progressive decrease in the size-selectivity of the glomerular filter. The urinary excretion of retinol-binding protein was weakly correlated with albuminuria (r = 0.26, n = 186). Eight % of diabetics showed an elevation of urinary retinol-binding protein without evidence of microalbuminuria, which clearly demonstrates that a proximal tubular impairment can occur independently of the glomerular alterations in the course of diabetic nephropathy. PMID:2253403

Bernard, A; Amor, A O; Goemare-Vanneste, J; Antoine, J L; Lauwerys, R; Colin, I; Vandeleene, B; Lambert, A



Protective effects of stem bark of Harungana madgascariensis on the red blood cell membrane  

PubMed Central

Background Anemia is a condition that has multiple origins. One such origin is the destruction of red blood cells’ (RBCs) membrane induced by free radicals. Treatment of anemia could therefore be enhanced by the use of free radicals’ scavengers potentially found in some medicinal plants. In this study, the protective effect of Harungana madagascariensis on the RBCs’ membrane physiology was investigated in vitro and in vivo. Methods In vitro hemolytic anemia was induced by incubation of fresh human RBCs with carbontetrachloride (CCl4) in Olive oil (Oo). Relaxation times of protons excited at 20 MHz (Carr-Purcell-Meiboom-Gill pulse sequence) in the absence or presence of paramagnetic Mn2+ ions (T2i for “extracellular” water and T2a for “intracellular” water, respectively) were determined at several temperatures (25–37°C) via Nuclear Magnetic Resonance (NMR) on a Bruker Minispec spectrometer. Water exchange times (Te) were consequently calculated using the Conlon-Outhred equation: 1/Te = (1/T2a) – (1/T2i). Morphological characteristics (mean cell volume, V, and cell surface area, A) were determined by photonic microscopy and the RBCs’ diffusional water permeability (Pd) was calculated as Pd = (1/Te)*(Va/A), where Va is the aqueous volume in the RBC and is about 0.7 of the cell volume (V). The activation energy of the diffusional process (Ea) for the respective temperature range was estimated using the Arrhenius modified equation k = A(T/T0)n*e-Ea/RT. Inhibition of the water diffusion induced by incubation with para-chloro-mercuribenzoic acid (PCMB) at 25, 30 and 37°C was calculated as I(%) = [(Pd control – Pd sample)/Pd control]*100. To investigate the protective influence of the extract on the RBC membrane, inhibition of the water permeability was evaluated on membranes pre-incubated with the Harungana madagascariensis extract. Male rats were used in in vivo investigations. Malondialdehyde (MDA) and cholesterol in the RBC membrane were estimated by induction of lipid peroxidation while the antioxidant properties of catalase (CAT) and superoxide dismutase (SOD) on the membrane were evaluated in regard to their antioxidant properties on the membrane. Results T2a significantly decreased at each temperature. Te results were higher in both RBCs and RBCs + extract groups incubated with PCMB compared to non-incubated controls, but differences were not statistically significant. A high percentage (73.81 ± 7.22) of RBCs pre-incubated with the extract presented the regular biconcave shape. Inhibition by PCMB of the RBCs’ membrane water permeability was increased at 30°C and decreased in the presence of extract (25°C and 37°C), while Ea decreased from 30.52 ± 1.3 KJ/mol to 25.49 ± 1.84 KJ/mol. Presence of the Harungana madagascariensis extract normalized the SOD and CAT activities as well as the MDA and membrane cholesterol concentrations altered by the CCl4-induced oxidative stress. Conclusion Harungana madagascariensis could protect the RBCs’ membrane through its antioxidative properties.



Membrane elastic properties and cell function.  


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



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



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.



Ligation of complement receptor 1 increases erythrocyte membrane deformability  

PubMed Central

Microbes as well as immune complexes and other continuously generated inflammatory particles are efficiently removed from the human circulation by red blood cells (RBCs) through a process called immune-adherence clearance. During this process, RBCs use complement receptor 1 (CR1, CD35) to bind circulating complement-opsonized particles and transfer them to resident macrophages in the liver and spleen for removal. We here show that ligation of RBC CR1 by antibody and complement-opsonized particles induces a transient Ca++ influx that is proportional to the RBC CR1 levels and is inhibited by T1E3 pAb, a specific inhibitor of TRPC1 channels. The CR1-elicited RBC Ca++ influx is accompanied by an increase in RBC membrane deformability that positively correlates with the number of preexisting CR1 molecules on RBC membranes. Biochemically, ligation of RBC CR1 causes a significant increase in phosphorylation levels of ?-spectrin that is inhibited by preincubation of RBCs with DMAT, a specific casein kinase II inhibitor. We hypothesize that the CR1-dependent increase in membrane deformability could be relevant for facilitating the transfer of CR1-bound particles from the RBCs to the hepatic and splenic phagocytes.

Glodek, Aleksandra M.; Mirchev, Rossen; Golan, David E.; Khoory, Joseph A.; Burns, Jennie M.; Shevkoplyas, Sergey S.; Nicholson-Weller, Anne



Determination of spontaneous curvature and internal pressure of vesicles and red cells by their shape and the flexoelectric effect of membrane  

SciTech Connect

Three geometric relations for vesicle equilibrium within the Helfrich elasticity model have been derived. The relations can serve to determine the spontaneous curvature c[sub o] and cell internal pressure -[Delta]p=p[sup in]-p[sup ext] from the vesicle shape. In analogy with the theory of the flexoelectric effect of nematic liquid crystals we have also obtained a relation between c[sub o] and the membrane potential. By applying these predictions to the human red blood cell (RBC) shape measured by Evans and Fung we found good agreement of the calculated potential with measured values previously given by other biologists by direct impaling RBC with a microelectrode. In this paper, the calculated value of the internal pressure is discussed by comparison with previous

Ou-Yang, Z.C.; Hu, J.G.; Liu, J.X. (Inst. of Theoretical Physics, P.O. Box 2735, Beijing-100080 (China))



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



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



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


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



Direct measurement of the area expansion and shear moduli of the human red blood cell membrane skeleton.  

PubMed Central

The area expansion and the shear moduli of the free spectrin skeleton, freshly extracted from the membrane of a human red blood cell (RBC), are measured by using optical tweezers micromanipulation. An RBC is trapped by three silica beads bound to its membrane. After extraction, the skeleton is deformed by applying calibrated forces to the beads. The area expansion modulus K(C) and shear modulus mu(C) of the two-dimensional spectrin network are inferred from the deformations measured as functions of the applied stress. In low hypotonic buffer (25 mOsm/kg), one finds K(C) = 4.8 +/- 2.7 microN/m, mu(C) = 2.4 +/- 0.7 microN/m, and K(C)/mu(C) = 1.9 +/- 1.0. In isotonic buffer, one measures higher values for K(C), mu(C), and K(C)/mu(C), partly because the skeleton collapses in a high-ionic-strength environment. Some data concerning the time evolution of the mechanical properties of the skeleton after extraction and the influence of ATP are also reported. In the Discussion, it is shown that the measured values are consistent with estimates deduced from experiments carried out on the intact membrane and agree with theoretical and numerical predictions concerning two-dimensional networks of entropic springs.

Lenormand, G; Henon, S; Richert, A; Simeon, J; Gallet, F



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.



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



Thermoelasticity of red blood cell membrane.  


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



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.




EPA Science Inventory

This paper presents an alternative method for designing rotating biological contactors (RBC) for use as a secondary treatment operation. The method uses a combination of chemical kinetics, good engineering practice, operational simplicity, and cost effectiveness to design a RBC s...


Computational analysis of dynamic interaction of two red blood cells in a capillary.  


The dynamic interaction of two red blood cells (RBCs) in a capillary is investigated computationally by the two-fluid model, including their deformable motion and interaction. For characterization of the deformation, the RBC membrane is treated as a curved two-dimensional shell with finite thickness by the shell model, and allowed to undergo the stretching strain and bending deformation. Moreover, a Morse potential is adopted to model the intercellular interaction for the aggregation behavior, which is characterized as the weak attraction at far distance and strong repulsion at near distance. For validation of the present technique, the dynamic interaction of two RBCs in static blood plasma is simulated firstly, where the RBCs aggregate slowly until a balanced configuration is achieved between the deformation and aggregation forces. The balanced configuration is in good agreement with the results reported previously. Three important effects on the dynamic behavior of RBCs are then analyzed, and they are the initial RBC shape, RBC deformability, and the intercellular interaction strength. It is found that the RBC is less deformed into a well-known parachute shape when the initial RBC shape is larger. Similarly, if the elastic shear modulus and bending stiffness of RBC membrane increase, the RBC resistance to deformation becomes higher, such that the RBC is less deformed. The simulation results also demonstrate that the RBC deformability strongly depends on the intercellular interaction strength. The RBCs deform more easily as the intercellular interaction strength increases. PMID:24590262

Li, Hua; Ye, Ting; Lam, K Y



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


An inherited RBC characteristic, NOR, resulting in erythrocyte polyagglutination.  


This report describes a new inheritable red blood cell characteristic, termed NOR, that is associated with polyagglutination. The NOR factor was shown to be inherited in an apparent dominant manner by 5 members of a two-generation family. NOR was clearly demonstrated to be a unique form of polyagglutination distinguishable from previously reported forms by serological tests using seed lectins, snail agglutinins, enzymes and absorption techniques. The NOR factor was characterized by the lack of reactivity with various seed and snail lectins, enhanced serum agglutinability after RBC proteolytic enzyme treatment, and the ability of hydatid cyst fluid to inhibit the serum hemagglutination reaction. PMID:7072192

Harris, P A; Roman, G K; Moulds, J J; Bird, G W; Shah, N G



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



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



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)



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



Alterations of the Erythrocyte Membrane during Sepsis  

PubMed Central

Erythrocytes have been long considered as “dead” cells with transport of oxygen (O2) as their only function. However, the ability of red blood cells (RBCs) to modulate the microcirculation is now recognized as an important additional function. This capacity is regulated by a key element in the rheologic process: the RBC membrane. This membrane is a complex unit with multiple interactions between the extracellular and intracellular compartments: blood stream, endothelium, and other blood cells on the one hand, and the intracytoplasmic compartment with possible rapid adaptation of erythrocyte metabolism on the other. In this paper, we review the alterations in the erythrocyte membrane observed in critically ill patients and the influence of these alterations on the microcirculatory abnormalities observed in such patients. An understanding of the mechanisms of RBC rheologic alterations in sepsis and their effects on blood flow and on oxygen transport may be important to help reduce morbidity and mortality from severe sepsis.

Serroukh, Yasmina; Djebara, Sarah; Lelubre, Christophe; Zouaoui Boudjeltia, Karim; Biston, Patrick; Piagnerelli, Michael



Alterations of the Erythrocyte Membrane during Sepsis.  


Erythrocytes have been long considered as "dead" cells with transport of oxygen (O(2)) as their only function. However, the ability of red blood cells (RBCs) to modulate the microcirculation is now recognized as an important additional function. This capacity is regulated by a key element in the rheologic process: the RBC membrane. This membrane is a complex unit with multiple interactions between the extracellular and intracellular compartments: blood stream, endothelium, and other blood cells on the one hand, and the intracytoplasmic compartment with possible rapid adaptation of erythrocyte metabolism on the other. In this paper, we review the alterations in the erythrocyte membrane observed in critically ill patients and the influence of these alterations on the microcirculatory abnormalities observed in such patients. An understanding of the mechanisms of RBC rheologic alterations in sepsis and their effects on blood flow and on oxygen transport may be important to help reduce morbidity and mortality from severe sepsis. PMID:22675622

Serroukh, Yasmina; Djebara, Sarah; Lelubre, Christophe; Zouaoui Boudjeltia, Karim; Biston, Patrick; Piagnerelli, Michael



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



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.



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.



Catalytic membranes for fuel cells  


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)



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)



Different activities of the reovirus FAST proteins and influenza hemagglutinin in cell-cell fusion assays and in response to membrane curvature agents  

SciTech Connect

The reovirus fusion-associated small transmembrane (FAST) proteins evolved to induce cell-cell, rather than virus-cell, membrane fusion. It is unclear whether the FAST protein fusion reaction proceeds in the same manner as the enveloped virus fusion proteins. We now show that fluorescence-based cell-cell and cell-RBC hemifusion assays are unsuited for detecting lipid mixing in the absence of content mixing during FAST protein-mediated membrane fusion. Furthermore, membrane curvature agents that inhibit hemifusion or promote pore formation mediated by influenza hemagglutinin had no effect on p14-induced cell-cell fusion, even under conditions of limiting p14 concentrations. Standard assays used to detect fusion intermediates induced by enveloped virus fusion proteins are therefore not applicable to the FAST proteins. These results suggest the possibility that the nature of the fusion intermediates or the mechanisms used to transit through the various stages of the fusion reaction may differ between these distinct classes of viral fusogens.

Clancy, Eileen K.; Barry, Chris; Ciechonska, Marta [Department of Microbiology and Immunology, Dalhousie University, Halifax, Nova Scotia, B3H 1X5 (Canada); Duncan, Roy, E-mail: roy.duncan@dal.c [Department of Microbiology and Immunology, Dalhousie University, Halifax, Nova Scotia, B3H 1X5 (Canada); Department of Pediatrics, Dalhousie University, Halifax, Nova Scotia, B3H 1X5 (Canada)



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.



Monte Carlo simulation of OCT signals from aggregating and sedimenting RBC suspension  

NASA Astrophysics Data System (ADS)

In the present paper, Monte Carlo simulation is used to obtain model OCT signals from horizontally oriented 1.1-mm thick blood layer at different stages of red blood cell (RBC) aggregation and sedimentation processes. The parameters for aggregating and sedimenting blood cells were chosen basing on the data available from literature and our earlier studies. We consider two different cases: a suspension of washed RBC in blood plasma, which provides necessary conditions for aggregation. Nice correspondence of the obtained simulation results with the available experimental results shows that chosen optical parameters are reasonable.

Kirillin, M. Y.; Priezzhev, A. V.; Tuchin, V. V.; Myllyla, R.



Challenges for red blood cell biomarker discovery through proteomics.  


Red blood cells are rather unique body cells, since they have lost all organelles when mature, which results in lack of potential to replace proteins that have lost their function. They maintain only a few pathways for obtaining energy and reducing power for the key functions they need to fulfill. This makes RBCs highly sensitive to any aberration. If so, these RBCs are quickly removed from circulation, but if the RBC levels reduce extremely fast, this results in hemolytic anemia. Several causes of HA exist, and proteome analysis is the most straightforward way to obtain deeper insight into RBC functioning under the stress of disease. This should result in discovery of biomarkers, typical for each source of anemia. In this review, several challenges to generate in-depth RBC proteomes are described, like to obtain pure RBCs, to overcome the wide dynamic range in protein expression, and to establish which of the identified/quantified proteins are active in RBCs. The final challenge is to acquire and validate suited biomarkers unique for the changes that occur for each of the clinical questions; in red blood cell aging (also important for transfusion medicine), for thalassemias or sickle cell disease. Biomarkers for other hemolytic anemias that are caused by dysfunction of RBC membrane proteins (the RBC membrane defects) or RBC cytosolic proteins (the enzymopathies) are sometimes even harder to discover, in particular for the patients with RBC rare diseases with unknown cause. This article is part of a Special Issue entitled: Biomarkers: A Proteomic Challenge. PMID:24129076

Barasa, Benjamin; Slijper, Monique



Cholesterol promotes hemifusion and pore widening in membrane fusion induced by influenza hemagglutinin.  


Cholesterol-specific interactions that affect membrane fusion were tested for using insect cells; cells that have naturally low cholesterol levels (< 4 mol %). Sf9 cells were engineered (HAS cells) to express the hemagglutinin (HA) of the influenza virus X-31 strain. Enrichment of HAS cells with cholesterol reduced the delay between triggering and lipid dye transfer between HAS cells and human red blood cells (RBC), indicating that cholesterol facilitates membrane lipid mixing prior to fusion pore opening. Increased cholesterol also increased aqueous content transfer between HAS cells and RBC over a broad range of HA expression levels, suggesting that cholesterol also favors fusion pore expansion. This interpretation was tested using both trans-cell dye diffusion and fusion pore conductivity measurements in cholesterol-enriched cells. The results of this study support the hypothesis that host cell cholesterol acts at two stages in membrane fusion: (1) early, prior to fusion pore opening, and (2) late, during fusion pore expansion. PMID:18443361

Biswas, Subrata; Yin, Shu-Rong; Blank, Paul S; Zimmerberg, Joshua



Cholesterol Promotes Hemifusion and Pore Widening in Membrane Fusion Induced by Influenza Hemagglutinin  

PubMed Central

Cholesterol-specific interactions that affect membrane fusion were tested for using insect cells; cells that have naturally low cholesterol levels (<4 mol %). Sf9 cells were engineered (HAS cells) to express the hemagglutinin (HA) of the influenza virus X-31 strain. Enrichment of HAS cells with cholesterol reduced the delay between triggering and lipid dye transfer between HAS cells and human red blood cells (RBC), indicating that cholesterol facilitates membrane lipid mixing prior to fusion pore opening. Increased cholesterol also increased aqueous content transfer between HAS cells and RBC over a broad range of HA expression levels, suggesting that cholesterol also favors fusion pore expansion. This interpretation was tested using both trans-cell dye diffusion and fusion pore conductivity measurements in cholesterol-enriched cells. The results of this study support the hypothesis that host cell cholesterol acts at two stages in membrane fusion: (1) early, prior to fusion pore opening, and (2) late, during fusion pore expansion.

Biswas, Subrata; Yin, Shu-Rong; Blank, Paul S.; Zimmerberg, Joshua



Colloidal RBC-Shaped, Hydrophilic, and Hollow Mesoporous Carbon Nanocapsules for Highly Efficient Biomedical Engineering.  


A facile and versatile synthetic route (templated framework pyrolysis) toward the construction of a new colloidal carbon nanosystem based on red blood cell (RBC)-shaped, hydrophilic, and hollow mesoporous carbon nanocapsules (HMCNs) is developed for highly efficient biomedical engineering, including stimuli-responsive drug release, inhibiting the metastasis, and circumventing the multidrug resistance of cancer cells. PMID:24687452

Chen, Yu; Xu, Pengfei; Wu, Meiying; Meng, Qingshuo; Chen, Hangrong; Shu, Zhu; Wang, Jin; Zhang, Lingxia; Li, Yaping; Shi, Jianlin



Fuel cell membranes and crossover prevention  


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)



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



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



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



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



Red blood cell populations and membrane levels of peroxiredoxin 2 as candidate biomarkers to reveal blood doping  

PubMed Central

Background Blood doping represents one main trend in doping strategies. Blood doping refers to the practice of boosting the number of red blood cells (RBCs) in the bloodstream in order to enhance athletic performance, by means of blood transfusions, administration of erythropoiesis-stimulating substances, blood substitutes, natural or artificial altitude facilities, and innovative gene therapies. While detection of recombinant EPO and homologous transfusion is already feasible through electrophoretic, mass spectrometry or flow cytometry-based approaches, no method is currently available to tackle doping strategies relying on autologous transfusions. Materials and methods. We exploited an in vitro model of autologous transfusion through a 1:10 dilution of concentrated RBCs after 30 days of storage upon appropriate dilution in freshly withdrawn RBCs from the same donor. Western blot towards membrane Prdx2 and Percoll density gradients were exploited to assess their suitability as biomarkers of transfusion. Results Membrane Prdx2 was visible in day 30 samples albeit not in day 0, while it was still visible in the 1:10 dilution of day 30 in day 0 RBCs. Cell gradients also highlighted changes in the profile of the RBC subpopulations upon dilution of stored RBCs in the fresh ones. Discussion. From this preliminary in vitro investigation it emerges that Prdx2 and RBC populations might be further tested as candidate biomarkers of blood doping through autologous transfusion, though it is yet to be assessed whether the kinetics in vivo of Prdx2 exposure in the membrane of transfused RBCs will endow a sufficient time-window to allow reliable anti-doping testing.

Marrocco, Cristina; Pallotta, Valeria; D'Alessandro, Angelo; Alves, Gilda; Zolla, Lello



Changes in red blood cell membrane structure in type 2 diabetes: a scanning electron and atomic force microscopy study  

PubMed Central

Red blood cells (RBCs) are highly deformable and possess a robust membrane that can withstand shear force. Previous research showed that in diabetic patients, there is a changed RBC ultrastructure, where these cells are elongated and twist around spontaneously formed fibrin fibers. These changes may impact erythrocyte function. Ultrastructural analysis of RBCs in inflammatory and degenerative diseases can no longer be ignored and should form a fundamental research tool in clinical studies. Consequently, we investigated the membrane roughness and ultrastructural changes in type 2 diabetes. Atomic force microscopy (AFM) was used to study membrane roughness and we correlate this with scanning electron microscopy (SEM) to compare results of both the techniques with the RBCs of healthy individuals. We show that the combined AFM and SEM analyses of RBCs give valuable information about the disease status of patients with diabetes. Effectiveness of treatment regimes on the integrity, cell shape and roughness of RBCs may be tracked, as this cell’s health status is crucial to the overall wellness of the diabetic patient.



Comparison of in vitro RBC labeling with the UltraTag RBC kit versus in vivo labeling  

SciTech Connect

This study compared cardiac-gated equilibrium blood-pool imaging studies using in vitro technetium-99m- (99mTc) labeled red blood cells (RBCs) prepared with the UltraTag RBC kit to in vivo labeling with stannous (pyro- and trimeta-) phosphates. The in vitro labeling procedure takes approximately 25 min and does not require centrifugation to separate free from bound 99mTc. Imaging studies were performed in 30 patients using the in vitro labeling procedure and in 30 patients with in vivo labeling. Regions of interest were placed over the center of the left ventricle, inferior and lateral to the left ventricle (background), and over the right midlung. The mean +/- s.e. in vitro RBC labeling efficiency was 98.5 +/- 0.2%. The heart-to-background ratios were significantly higher with in vitro labeling. The heart-to-background ratios, averaged among two blinded reviewers, were 4.6 and 3.4 for the in vitro and in vivo methods, respectively. The heart-to-lung ratio was generally higher with the in vitro procedure (3.6) than that observed with the in vivo method (3.2) but failed to attain statistical significance (p = 0.059). These results demonstrate the superiority of the in vitro labeling procedure over in vivo labeling for gated equilibrium blood-pool imaging.

Patrick, S.T.; Glowniak, J.V.; Turner, F.E.; Robbins, M.S.; Wolfangel, R.G. (VA Medical Center, Portland, OR (USA))



Sialoglycosylation of RBC in Visceral Leishmaniasis Leads to Enhanced Oxidative Stress, Calpain-Induced Fragmentation of Spectrin and Hemolysis  

PubMed Central

Visceral leishmaniasis (VL) caused by the intracellular parasite Leishmania donovani accounts for an estimated 12 million cases of human infection. It is almost always associated with anemia, which severely complicates the disease course. However, the pathological processes leading to anemia in VL have thus far not been adequately characterized to date. In studying the glycosylation patterns of peripheral blood cells we found that the red blood cells (RBC) of VL patients (RBCVL) express eight 9-O-acetylated sialoglycoproteins (9-O-AcSGPs) that are not detected in the RBC of healthy individuals (RBCN). At the same time, the patients had high titers of anti-9-O-AcSGP IgG antibodies in their sera. These two conditions appear to be linked and related to the anemic state of the patients, as exposure of RBCVL but not RBCN to anti-9-O-AcSGPs antibodies purified from patient sera triggered a series of responses. These included calcium influx via the P/Q-type but not L-type channels, activation of calpain I, proteolysis of spectrin, enhanced oxidative stress, lipid peroxidation, externalization of phosphatidyl serine with enhanced erythrophagocytosis, enhanced membrane fragility and, finally, hemolysis. Taken together, this study suggests that the enhanced hemolysis is linked to an impairment of membrane integrity in RBCVL which is mediated by ligand-specific interaction of surface 9-O-AcSGPs. This affords a potential explanation for the structural and functional features of RBCVL which are involved in the hemolysis related to the anemia which develops in VL patients.

Samanta, Sajal; Ghoshal, Angana; Bhattacharya, Kaushik; Saha, Bibhuti; Walden, Peter; Mandal, Chitra



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



Fuel cell membrane hydration and fluid metering  


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)



Fuel cell membrane hydration and fluid metering  


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)



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



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



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.


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


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



Membrane rafts of the human red blood cell.  


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



Measurement of red blood cell mechanics during morphological changes  

NASA Astrophysics Data System (ADS)

The human red blood cell (RBC) membrane, a fluid lipid bilayer tethered to an elastic 2D spectrin network, provides the principal control of the cell's morphology and mechanics. These properties, in turn, influence the ability of RBCs to transport oxygen in circulation. Current mechanical measurements of RBCs rely on external loads. Here we apply a Noncontact optical interferometric technique to quantify the thermal fluctuations of RBC membranes with 3 nm accuracy over a broad range of spatial and temporal frequencies. Combining this technique with a new mathematical model describing RBC membrane undulations, we measure the mechanical changes of RBCs as they undergo a transition from the normal discoid shape to the abnormal echinocyte and spherical shapes. These measurements indicate that, coincident with this morphological transition, there is a significant increase in the membrane's shear and bending moduli. This mechanical transition can alter cell circulation and impede oxygen delivery.

Popescu, Gabriel; Park, Yongkeun; Best, Catherine; Dasari, Ramachandra; Feld, Michael; Kuriabova, Tatiana; Henle, Mark; Levine, Alex



Theoretical analysis of membrane tension in moving cells.  


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



Use of the Bo-RBC-SCID Mouse Model for Isolation of a Babesia Parasite from Grazing Calves in Japan  

Microsoft Academic Search

SCID mice with circulating bovine red blood cells (Bo-RBC-SCID mice) were used to isolate Babesia parasites from grazing calves in Japan. Although the initial blood samples obtained from the calves contained both Babesia and Theileria parasites, we were able to isolate only the Babesia parasites by repeated blood passages in Bo-RBC-SCID mice, taking advantage of the much more rapid growth

M. Tsuji; Y. Terada; S. Arai; H. Okada; C. Ishihara



Magnetically responsive microflaps reveal cell membrane boundaries from multiple angles.  


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



Temperature transitions of protein properties in human red blood cells.  

PubMed Central

Human red blood cells (RBC) undergo a sudden change from blocking to passing through 1.3 +/- 0.2-micrometer micropipettes at a transition temperature (Tc) of 36.4 degrees C. For resealed RBC ghosts this transition occurs at 28.3 degrees C (Tg). These findings are attributed to an elastomeric transition of hemoglobin from being gel-like to a fluid and to an elastomeric transition of membrane proteins such as spectrin. Spectrin shows a uniform distribution along the aspirated RBC tongue above Tg in contrast to the linear gradient below Tg.

Artmann, G M; Kelemen, C; Porst, D; Buldt, G; Chien, S



Erythrocyte NADPH oxidase activity modulated by Rac GTPases, PKC, and plasma cytokines contributes to oxidative stress in sickle cell disease.  


Chronic inflammation has emerged as an important pathogenic mechanism in sickle cell disease (SCD). One component of this inflammatory response is oxidant stress mediated by reactive oxygen species (ROS) generated by leukocytes, endothelial cells, plasma enzymes, and sickle red blood cells (RBC). Sickle RBC ROS generation has been attributed to sickle hemoglobin auto-oxidation and Fenton chemistry reactions catalyzed by denatured heme moieties bound to the RBC membrane. In this study, we demonstrate that a significant part of ROS production in sickle cells is mediated enzymatically by NADPH oxidase, which is regulated by protein kinase C, Rac GTPase, and intracellular Ca(2+) signaling within the sickle RBC. Moreover, plasma from patients with SCD and isolated cytokines, such as transforming growth factor ?1 and endothelin-1, enhance RBC NADPH oxidase activity and increase ROS generation. ROS-mediated damage to RBC membrane components is known to contribute to erythrocyte rigidity and fragility in SCD. Erythrocyte ROS generation, hemolysis, vaso-occlusion, and the inflammatory response to tissue damage may therefore act in a positive-feedback loop to drive the pathophysiology of sickle cell disease. These findings suggest a novel pathogenic mechanism in SCD and may offer new therapeutic targets to counteract inflammation and RBC rigidity and fragility in SCD. PMID:23349388

George, Alex; Pushkaran, Suvarnamala; Konstantinidis, Diamantis G; Koochaki, Sebastian; Malik, Punam; Mohandas, Narla; Zheng, Yi; Joiner, Clinton H; Kalfa, Theodosia A



Predicting dynamics and rheology of blood flow: A comparative study of multiscale and low-dimensional models of red blood cells  

SciTech Connect

In this work we compare the predictive capability of two mathematical models for red blood cells (RBCs) focusing on blood flow in capillaries and arterioles. Both RBC models as well as their corresponding blood flows are based on the dissipative particle dynamics (DPD) method, a coarse-grained molecular dynamics approach. The first model employs a multiscale description of the RBC (MS-RBC), with its membrane represented by hundreds or even thousands of DPD-particles connected by springs into a triangular network in combination with out-of-plane elastic bending resistance. Extra dissipation within the network accounts for membrane viscosity, while the characteristic biconcave RBC shape is achieved by imposition of constraints for constant membrane area and constant cell volume. The second model is based on a low-dimensional description (LD-RBC) constructed as a closed torus-like ring of only 10 large DPD colloidal particles. They are connected into a ring by worm-like chain (WLC) springs combined with bending resistance. The LD-RBC model can be fitted to represent the entire range of nonlinear elastic deformations as measured by optical-tweezers for healthy and for infected RBCs in malaria. MS-RBCs suspensions model the dynamics and rheology of blood flow accurately for any size vessel but this approach is computationally expensive above 100 microns. Surprisingly, the much more economical suspensions of LD-RBCs also capture the blood flow dynamics and rheology accurately except for vessels with sizes comparable to RBC diameter. In particular, the LD-RBC suspensions are shown to properly capture the experimental data for the apparent viscosity of blood and its cell-free layer (CFL) in tube flow. Taken together, these findings suggest a hierarchical approach in modeling blood flow in the arterial tree, whereby the MS-RBC model should be employed for capillaries and arterioles below 100 microns, the LD-RBC model for arterioles, and the continuum description for arteries.

Pan, Wenxiao; Fedosov, Dmitry A.; Caswell, Bruce; Karniadakis, George E.



Membrane disorder and phospholipid scrambling in electropermeabilized and viable cells.  


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



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



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


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



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



Anticancer effects of human amniotic membrane and its epithelial cells.  


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



Simulated Red Blood Cell Motion in Microvessel Bifurcations: Effects of Cell-Cell Interactions on Cell Partitioning  

PubMed Central

Partitioning of red blood cell (RBC) fluxes between the branches of a diverging microvessel bifurcation is generally not proportional to the flow rates, as RBCs preferentially enter the higher-flow branch. A two-dimensional model for RBC motion and deformation is used to investigate the effects of cell-cell mechanical interactions on RBC partitioning in bifurcations. The RBC membrane and cytoplasm are represented by sets of viscoelastic elements immersed in a low Reynolds number flow. Several types of two-cell interactions that can affect partitioning are found. In the most frequent interactions, a `trade-off' occurs, in which a cell entering one branch causes a following cell to enter the other branch. Other types of interactions include `herding,' where the leading cell is caused to enter the same branch as the following cell, and `following,' where the trailing cell is caused to enter the same branch as the leading cell. The combined effect of these cell-cell interactions is a tendency towards more uniform partitioning, which results from the trade-off effect but is reduced by the herding and following effects. With increasing hematocrit, the frequency of interactions increases, and more uniform partitioning results. This prediction is consistent with experimental observations on how hematocrit affects RBC partitioning.

Barber, Jared O.; Restrepo, Juan M.; Secomb, Timothy W.



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



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



Isolation of plasma membranes from purified mouse spermatogenic cells.  


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. 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



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.



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.



Phosphorylation and Activation of the Plasma Membrane Na+/H+ Exchanger (NHE1) during Osmotic Cell Shrinkage  

PubMed Central

The Na+/H+ Exchanger isoform 1 (NHE1) is a highly versatile, broadly distributed and precisely controlled transport protein that mediates volume and pH regulation in most cell types. NHE1 phosphorylation contributes to Na+/H+ exchange activity in response to phorbol esters, growth factors or protein phosphatase inhibitors, but has not been observed during activation by osmotic cell shrinkage (OCS). We examined the role of NHE1 phosphorylation during activation by OCS, using an ideal model system, the Amphiuma tridactylum red blood cell (atRBC). Na+/H+ exchange in atRBCs is mediated by an NHE1 homolog (atNHE1) that is 79% identical to human NHE1 at the amino acid level. NHE1 activity in atRBCs is exceptionally robust in that transport activity can increase more than 2 orders of magnitude from rest to full activation. Michaelis-Menten transport kinetics indicates that either OCS or treatment with the phosphatase inhibitor calyculin-A (CLA) increase Na+ transport capacity without affecting transport affinity (Km?=?44 mM) in atRBCs. CLA and OCS act non-additively to activate atNHE1, indicating convergent, phosphorylation-dependent signaling in atNHE1 activation. In situ 32P labeling and immunoprecipitation demonstrates that the net phosphorylation of atNHE1 is increased 4-fold during OCS coinciding with a more than 2-order increase in Na+ transport activity. This is the first reported evidence of increased NHE1 phosphorylation during OCS in any vertebrate cell type. Finally, liquid chromatography and mass spectrometry (LC-MS/MS) analysis of atNHE1 immunoprecipitated from atRBC membranes reveals 9 phosphorylated serine/threonine residues, suggesting that activation of atNHE1 involves multiple phosphorylation and/or dephosphorylation events.

Rigor, Robert R.; Damoc, Catalina; Phinney, Brett S.; Cala, Peter M.



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



A Multiscale Red Blood Cell Model with Accurate Mechanics, Rheology, and Dynamics  

PubMed Central

Abstract Red blood cells (RBCs) have highly deformable viscoelastic membranes exhibiting complex rheological response and rich hydrodynamic behavior governed by special elastic and bending properties and by the external/internal fluid and membrane viscosities. We present a multiscale RBC model that is able to predict RBC mechanics, rheology, and dynamics in agreement with experiments. Based on an analytic theory, the modeled membrane properties can be uniquely related to the experimentally established RBC macroscopic properties without any adjustment of parameters. The RBC linear and nonlinear elastic deformations match those obtained in optical-tweezers experiments. The rheological properties of the membrane are compared with those obtained in optical magnetic twisting cytometry, membrane thermal fluctuations, and creep followed by cell recovery. The dynamics of RBCs in shear and Poiseuille flows is tested against experiments and theoretical predictions, and the applicability of the latter is discussed. Our findings clearly indicate that a purely elastic model for the membrane cannot accurately represent the RBC's rheological properties and its dynamics, and therefore accurate modeling of a viscoelastic membrane is necessary.

Fedosov, Dmitry A.; Caswell, Bruce; Karniadakis, George Em



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.



Effect of Irradiation and/or Leucocyte Filtration on RBC Storage Lesions  

PubMed Central

Red blood cell (RBC) storage lesions have been shown to be associated with some adverse reactions; numerous studies have focused on the lesions caused by storage, and few data on the RBC storage lesions caused by prestorage treatments of leucocyte filtration and irradiation. In this study, we examined the changes related with the RBC storage lesions, including 2,3-diphosphatidylglyceric acid (2,3-DPG), pH, free hemoglobin (Hb), supernatant free K+ and Na+ concentration, mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH). Along with the increasing storage time, decreases in 2, 3-DPG levels, pH and Na+ concentration, increases in K+ and free Hb concentrations, and significant morphological changes in RBC in all groups were found. The changes in the groups of irradiation, leucocyte filtration and the combined irradiation and leucocyte filtration were more significant than those in the untreated group. Meanwhile, the MCV levels of the three treated groups were significantly lower than those in the untreated group, while the MCH variations were significantly higher. Our results suggest that irradiation and leucocyte filtration before storage may aggravate blood storage lesions.

Xiao, Yanni; Zhao, Jiang; Ye, Xingde; Li, Zhongjun



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.



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



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


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)



Hemoglobin s polymerization and red cell membrane changes.  


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



Spring-network-based model of a red blood cell for simulating mesoscopic blood flow.  


We developed a mechanical model of a red blood cell (RBC) that is capable of expressing its characteristic behaviors in shear flows. The RBC was modeled as a closed shell membrane consisting of spring networks in the framework of the energy minimum concept. The fluid forces acting on RBCs were modeled from Newton's viscosity law and the conservation of momentum. In a steady shear flow, the RBC model exhibited various behaviors, depending on the shear rate; it tumbled, tank-treaded, or both. The transition from tumbling to tank-treading occurred at a shear rate of 20 s(?-?1). The simulation of an RBC in steady and unsteady parallel shear flows (Couette flows) showed that the deformation parameters of the RBC were consistent with experimental results. The RBC in Poiseuille flow migrated radially towards the central axis of the flow channel. Axial migration became faster with an increase in the viscosity of the media, qualitatively consistent with experimental results. These results demonstrate that the proposed model satisfies the essential conditions for simulating RBC behavior in blood flow. Finally, a large-scale RBC flow simulation was implemented to show the capability of the proposed model for analyzing the mesoscopic nature of blood flow. PMID:23293072

Nakamura, Masanori; Bessho, Sadao; Wada, Shigeo



Cytolytic peptides induce biphasic permeability changes in mammalian cell membranes.  


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



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



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



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.



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




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




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



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



The human erythrocyte plasma membrane: a Rosetta Stone for decoding membrane-cytoskeleton structure.  


The mammalian erythrocyte, or red blood cell (RBC), is a unique experiment of nature: a cell with no intracellular organelles, nucleus or transcellular cytoskeleton, and a plasma membrane with uniform structure across its entire surface. By virtue of these specialized properties, the RBC membrane has provided a template for discovery of the fundamental actin filament network machine of the membrane skeleton, now known to confer mechanical resilience, anchor membrane proteins, and organize membrane domains in all cells. This chapter provides a historical perspective and critical analysis of the biochemistry, structure, and physiological functions of this actin filament network in RBCs. The core units of this network are nodes of ~35-37 nm-long actin filaments, interconnected by long strands of (?1?1)?-spectrin tetramers, forming a 2D isotropic lattice with quasi-hexagonal symmetry. Actin filament length and stability is critical for network formation, relying upon filament capping at both ends: tropomodulin-1 at pointed ends and ??-adducin at barbed ends. Tropomodulin-1 capping is essential for precise filament lengths, and is enhanced by tropomyosin, which binds along the short actin filaments. ??-adducin capping recruits spectrins to sites near barbed ends, promoting network formation. Accessory proteins, 4.1R and dematin, also promote spectrin binding to actin and, with ??-adducin, link to membrane proteins, targeting actin nodes to the membrane. Dissection of the molecular organization within the RBC membrane skeleton is one of the paramount achievements of cell biological research in the past century. Future studies will reveal the structure and dynamics of actin filament capping, mechanisms of precise length regulation, and spectrin-actin lattice symmetry. PMID:24210427

Fowler, Velia M



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



Glucose transport in human red cell membranes. Dependence of age, ATP, and insulin.  


Glucose self-exchange flux (Jex) and net efflux (Jnet) in human red cells and ghosts were studied at 25 degrees C and pH 7.2 by means of the combined use of the Millipore-Swinnex filtering method and the continuous flow tube method to show the dependence of time of storage after aspiration, ATP and insulin. In fresh cells (RBC), ghosts (G), ghosts with 2 mM ATP (G +), and cells stored at 4 degrees C greater than 60 days (OC) both Jex and Jnet follow simple Michaelis-Menten kinetics where J = Jmax X Ci X (K1/2 + Ci)-1. Jmaxex and Jmaxnet (nmol X cm-2 X s-1), respectively, was: (RBC) 0.27 and 0.19, (G) 0.24 and 0.27, (G +) 0.23 and 0.24, (OC) 0.23 and 0.20. K1/2,ex and K1/2,net (mM), respectively, was: (RBC) 7.5 and 1.3, (G) 4.8 and 14.2, (G +) 11.6 and 6.8, (OC) 3.8 and 9.0. In ghosts, the ATP-dependent fraction of the permeability shows a hyperbolic dependence on glucose concentrations lower than 80 mM. Insulin up to 1 microM had effect on neither Jex nor Jnet in RBC. Based on reported values of cytochalasin B binding sites the turnover rate per site in RBC appears to be as high as in maximally insulin-stimulated fat cells. Our results suggest that the number of transport sites remains constant, independent of age, ATP and insulin. PMID:3297147

Jensen, M R; Brahm, J



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


Numerical analysis of a red blood cell flowing through a thin micropore  

NASA Astrophysics Data System (ADS)

Red blood cell (RBC) deformability plays a key role in microcirculation, especially in vessels that have diameters even smaller than the nominal cell size. In this study, we numerically investigate the dynamics of an RBC in a thin micropore. The RBC is modeled as a capsule with a thin hyperelastic membrane. In a numerical simulation, we employ a boundary element method for fluid mechanics and a finite element method for membrane mechanics. The resulting RBC deformation towards the flow direction is suppressed considerably by increased cytoplasm viscosity, whereas the gap between the cell membrane and solid wall becomes smaller with higher cytoplasm viscosity. We also measure the transit time of the RBC and find that nondimensional transit time increases nonlinearly with respect to the viscosity ratio, whereas it is invariant to the capillary number. In conclusion, cytoplasmic viscosity plays a key role in the dynamics of an RBC in a thin pore. The results of this study will be useful for designing a microfluidic device to measure cytoplasmic viscosity.

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



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



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




EPA Science Inventory

The major objective of this study was to operate a full scale rotating biological contactor (RBC) to determine if it could produce an effluent that would meet the definition of a secondary effluent (BOD ...


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



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



Hydrodynamic extrusion of membrane nanotubes: the role of the cytoskeleton  

NASA Astrophysics Data System (ADS)

We have investigated membrane-cytoskeleton adhesion properties by extrusion of tubes from tethered vesicles and cells using hydrodynamic flows. Our experimental results show that impermeable membranes (giant vesicles) act as entropic springs, i.e. the extruded tubes reach a stationary length, whereas porous membranes (vesicles decorated with pores) lead to tubes, which extrude at constant velocity without reaching a stationary length. On the other hand, experiments on red blood cells (RBC) suggest that the dynamics of extruded tubes is dominated by the detachment of the membrane from the cytoskeleton and the flow of lipids through the binding membrane proteins. We have estimated the membrane-cytoskeleton binding energy and the viscosity of the membrane for RBC-s. Tube extrusion from other cell types (S180, MDCK, BON) show phenomena such as healing time for the membrane-cytoskeleton rebinding, and cell aging (breakage of the tube after a few consecutive extrusions). We will discuss how these phenomena depend on the properties of the cytoskeleton and on the presence of cell adhesion molecules.

Guevorkian, Karine; Borghi, Nicolas; Kremer, Séastien; Buguin, Axel; Brochard, Françise



Novel epinephrine and cyclic AMP-mediated activation of BCAM/Lu-dependent sickle (SS) RBC adhesion.  


The vasoocclusive crisis is the major clinical feature of sickle cell anemia, which is believed to be initiated or sustained by sickle (SS) red blood cell (RBC) adhesion to the vascular wall. SS RBCs, but not unaffected (AA) RBCs, adhere avidly to multiple components of the vascular wall, including laminin. Here we report a novel role for epinephrine and cyclic adenosine monophosphate (cAMP) in the regulation of human SS RBC adhesiveness via the laminin receptor, basal cell adhesion molecule/Lutheran (BCAM/Lu). Our data demonstrate that peripheral SS RBCs contain greater than 4-fold more cAMP than AA RBCs under basal conditions. Forskolin or the stress mediator epinephrine further elevates cAMP in SS RBCs and increases adhesion of SS RBCs to laminin in a protein kinase A (PKA)-dependent manner, with the low-density population being the most responsive. Epinephrine-stimulated adhesion to laminin, mediated primarily via the beta 2-adrenergic receptor, occurred in SS RBC samples from 46% of patients and was blocked by recombinant, soluble BCAM/Lu, implicating this receptor as a target of cAMP signaling. Thus, these studies demonstrate a novel, rapid regulation of SS RBC adhesion by a cAMP-dependent pathway and suggest that components of this pathway, particularly PKA, the beta 2-adrenergic receptor, and BCAM/Lu, should be further explored as potential therapeutic targets to inhibit SS RBC adhesion. PMID:12506027

Hines, Patrick C; Zen, Qin; Burney, Sharran N; Shea, Deborah A; Ataga, Kenneth I; Orringer, Eugene P; Telen, Marilyn J; Parise, Leslie V



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



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



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.



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



Erythrocyte membrane modification in malignant diseases of myeloid and lymphoreticular tissues. I. Tn-polyagglutination in acute myelocytic leukaemia.  


Tn polyagglutination (persistent mixed-field polyagglutination) occurring in a patient with an acute myelocytic leukemia is described. The association of this RBC membrane change, thought to be due to somatic mutation at stem cell level, with a myeloproliferative disorder, has not been previously reported. PMID:1063594

Bird, G W; Wingham, J; Pippard, M J; Hoult, J G; Melikian, V



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.



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



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



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



Human hepatocytes and endothelial cells in organotypic membrane systems.  


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



Membrane fusion in cells: molecular machinery and mechanisms.  


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



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



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


Effect of hydroperoxides on red blood cell membrane mechanical properties.  


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



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.



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.



From artificial red blood cells, oxygen carriers, and oxygen therapeutics to artificial cells, nanomedicine, and beyond.  


The first experimental artificial red blood cells have all three major functions of red blood cells (rbc). However, the first practical one is a simple polyhemoglobin (PolyHb) that only has an oxygen-carrying function. This is now in routine clinical use in South Africa and Russia. An oxygen carrier with antioxidant functions, PolyHb-catalase-superoxide dismutase, can fulfill two of the three functions of rbc. Even more complete is one with all three functions of rbc in the form of PolyHb-catalase-superoxide dismutase-carbonic anhydrase. The most advanced ones are nanodimension artificial rbc with either PEG-lipid membrane or PEG-PLA polymer membrane. Extensions into oxygen therapeutics include a PolyHb-tyrosinase that suppresses the growth of melanoma in a mice model. Another is a PolyHb-fibrinogen that is an oxygen carrier with platelet-like function. Research has now extended well beyond the original research on artificial rbc into many areas of artificial cells. These include nanoparticles, nanotubules, lipid vesicles, liposomes, polymer-tethered lipid vesicles, polymersomes, microcapsules, bioencapsulation, nanocapules, macroencapsulation, synthetic cells, and others. These are being used in nanotechnology, nanomedicine, regenerative medicine, enzyme/gene therapy, cell/stem cell therapy, biotechnology, drug delivery, hemoperfusion, nanosensers, and even by some groups in agriculture, industry, aquatic culture, nanocomputers, and nanorobotics. PMID:22409281

Chang, Thomas M S



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



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.



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)



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


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



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.


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



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



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



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



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


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



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


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



Design of efficient methanol impermeable membranes for fuel cell applications.  


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



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



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.



Influence of hydrogen ion concentration on refractive index value in red blood cells of diabetes patients  

Microsoft Academic Search

Since hyperglycaemia changes fluidity of erythrocyte cell membrane and impair cell deformability, our goal was to characterize light refractive properties of haemoglobin and red blood cells (RBC) in diabetes patients. Microscopic investigation was carried out on intact and in methanol fixed RBCs of diabetes patients with long-term hyperglycaemia (glycosylated haemoglobin > 7,5%). Interference microscopy was used for refractive index (RI)

Gunta Mazarevica; Talivaldis Freivalds; Antra Jurka



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



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.



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



Membrane and actin reorganization in electropulse-induced cell fusion.  


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



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


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



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



Strategies for the cell-free expression of membrane proteins.  


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



99mTc-RBC Scintigraphy for diagnosis of intestinal stromal tumor hemorrhage: a case report  

PubMed Central

Background Gastrointestinal stromal tumors (GISTs) are rare and our understanding of the natural history and optimal treatment of GISTs are continually evolving. They are characterized by a remarkable cellular variability and their malignant potential is sometimes difficult to predict. Case presentation We report the case of intestinal stromal tumor in a 44 years old patient with a long history of anemia and recurrent hemafecia. By using 99mTc-labeled red blood cell (99mTc-RBC) scintigraphy, extensive tracer accumulation in the jejunum was detected. Immunohistochemically, the tumor strongly expresses the KIT (CD117) protein. The intestinal tumor was successfully resected with a postoperative favorable outcome. Conclusion 99mTc-RBC scintigraphy is an established technique for the identification and localization of gastrointestinal bleeding. Abdominal scintigraphy appears to be a valuable supplement to conventional diagnostic methods for the diagnosis of gastrointestinal stromal tumor hemorrhage.

Wang, Jiang; Zhao, Rong



Thrombelastographic evaluation of the influence of 2-RBC apheresis on donor's coagulation system.  


Rotation thrombelastogram (ROTEM®/TEG®) assays allow rapid global assessment of hemostatic function using whole blood. Since published data about the effects of automated red cell collection on coagulation system are scarce, we aimed to investigate the effects of 2-RBC apheresis on donor's coagulation system using ROTEM® assays. In INTEM assay, CFT was significantly shortened 24h after apheresis compared with baseline value (p<0.05) and MCF was significantly prolonged immediately after apheresis and 24h after apheresis compared with baseline value (p<0.05 and p<0.01, respectively). In EXTEM assay, CFT was significantly prolonged immediately after apheresis and 24h after apheresis compared with baseline value (p=0.001 and p<0.001, respectively) and MCF was significantly prolonged 24h after apheresis compared with baseline value (p<0,001). Our results demonstrate thromboelastographic signs of hypercoagulability in donors undergoing 2-RBC apheresis. PMID:23660469

Akay, Olga Meltem; Karagulle, Mustafa; Kus, Gokhan; Mutlu, Fezan Sah?n; Gunduz, Eren



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.


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


Shear stress-induced improvement of red blood cell deformability.  


Classically, it is known that red blood cell (RBC) deformability is determined by the geometric and material properties of these cells. Experimental evidence accumulated during the last decade has introduced the concept of active regulation of RBC deformability. This regulation is mainly related to altered associations between membrane skeletal proteins and integral proteins, with the latter serving to anchor the skeleton to the lipid matrix. It has been hypothesized that shear stress induces alterations of RBC deformability: the current study investigated the dynamics of the transient improvement in deformability induced by shear stress at physiologically-relevant levels. RBC were exposed to various levels of shear stress (SS) in a Couette type shearing system that is part of an ektacytometer, thus permitting the changes in RBC deformability during the application of SS to be monitored. Initial studies showed that there is an increase in deformability of the RBC subjected to SS in the range of 5-20 Pa, with kinetics characterized by time constants of a few seconds. Such improvement in deformability, expressed by an elongation index (EI), was faster with higher levels of SS and hence yielded shorter time constants: absolute values of EI increased by 3-8% of the starting level. Upon the removal of the shear stress, this response by RBC was reversible with a slower time course compared to the increase in EI during application of SS. Increased calcium concentration in the RBC suspending medium prevented the improvement of deformability. It is suggested that the improvement of RBC deformability by shear forces may have significant effects on blood flow dynamics, at least in tissues supplied by blood vessels with impaired vasomotor reserve, and may therefore serve as a compensating mechanism for the maintenance of adequate microcirculatory perfusion. PMID:23863281

Meram, Ece; Yilmaz, Bahar D; Bas, Ceren; Atac, Nazl?; Yalcin, O; Meiselman, Herbert J; Baskurt, Oguz K



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



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


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.



Analysis of RBC damage using laser tweezers Raman spectroscopy (LTRS) during femtosecond laser optical trapping  

NASA Astrophysics Data System (ADS)

We monitored cell viability and damage under femtosecond laser irradiation using aser weezers Raman pectroscopy (LTRS) which is becoming a powerful tool for the analysis of biological materials. Femtosecond lasers are more frequently used as a light source for optical tweezers since they enable nonlinear optical phenomena such as two-photon absorption or second harmonic generation trapping. Femtosecond laser optical trapping similar to thee CW laser optical trapping except that optical damage can be easily induced due to extremely high peak power of femtosecond pulses. We monitored the Raman signal changes as a marker for optical damage. We used red blood cell (RBC) as a target sample and first used the CW laser beams to trap the RBC from the bottom of the chamber. After the trapped RBC is moved to a desired depth, we switched the laser mode to mode-locked mode and monitored the Raman signals as a function of the laser irradiation time. It was observed that the Raman shift at 1543 cm -1 may be a good marker for optical damage both for CW and femtosecond laser trapping.

Ju, Sung-bin; Pyo, Jin-woo; Jang, Jae-young; Lee, Seungduk; Kim, Beop-Min



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


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



Efficient and Specific Analysis of Red Blood Cell Glycerophospholipid Fatty Acid Composition  

PubMed Central

Background Red blood cell (RBC) n-3 fatty acid status is related to various health outcomes. Accepted biological markers for the fatty acid status determination are RBC phospholipids, phosphatidylcholine, and phosphatidyletholamine. The analysis of these lipid fractions is demanding and time consuming and total phospholipid n-3 fatty acid levels might be affected by changes of sphingomyelin contents in the RBC membrane during n-3 supplementation. Aim We developed a method for the specific analysis of RBC glycerophospholipids. The application of the new method in a DHA supplementation trial and the comparison to established markers will determine the relevance of RBC GPL as a valid fatty acid status marker in humans. Methods Methyl esters of glycerophospholipid fatty acids are selectively generated by a two step procedure involving methanolic protein precipitation and base-catalysed methyl ester synthesis. RBC GPL solubilisation is facilitated by ultrasound treatment. Fatty acid status in RBC glycerophospholipids and other established markers were evaluated in thirteen subjects participating in a 30 days supplementation trial (510 mg DHA/d). Outcome The intra-assay CV for GPL fatty acids ranged from 1.0 to 10.5% and the inter-assay CV from 1.3 to 10.9%. Docosahexaenoic acid supplementation significantly increased the docosahexaenoic acid contents in all analysed lipid fractions. High correlations were observed for most of the mono- and polyunsaturated fatty acids, and for the omega-3 index (r?=?0.924) between RBC phospholipids and glycerophospholipids. The analysis of RBC glycerophospholipid fatty acids yields faster, easier and less costly results equivalent to the conventional analysis of RBC total phospholipids.

Klem, Sabrina; Klingler, Mario; Demmelmair, Hans; Koletzko, Berthold



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



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



Controlled permeation of cell membrane by single bubble acoustic cavitation.  


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



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


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



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



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)



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.



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



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



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



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.



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



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



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.



The protective effect of aqueous extracts of roselle (Hibiscus sabdariffa L. UKMR-2) against red blood cell membrane oxidative stress in rats with streptozotocin-induced diabetes  

PubMed Central

OBJECTIVES: The aim of this study was to investigate the protective effects of aqueous extracts of roselle (Hibiscus sabdariffa L. UKMR-2) against red blood cell (RBC) membrane oxidative stress in rats with streptozotocin-induced diabetes. METHODS: Forty male Sprague-Dawley rats weighing 230-250 g were randomly divided into four groups (n?=?10 rats each): control group (N), roselle-treated control group, diabetic group, and roselle-treated diabetic group. Roselle was administered by force-feeding with aqueous extracts of roselle (100 mg/kg body weight) for 28 days. RESULTS: The results demonstrated that the malondialdehyde levels of the red blood cell membranes in the diabetic group were significantly higher than the levels in the roselle-treated control and roselle-treated diabetic groups. The protein carbonyl level was significantly higher in the roselle-treated diabetic group than in the roselle-treated control group but lower than that in the diabetic group. A significant increase in the red blood cell membrane superoxide dismutase enzyme was found in roselle-treated diabetic rats compared with roselle-treated control rats and diabetic rats. The total protein level of the red blood cell membrane, osmotic fragility, and red blood cell morphology were maintained. CONCLUSION: The present study demonstrates that aqueous extracts of roselle possess a protective effect against red blood cell membrane oxidative stress in rats with streptozotocin-induced diabetes. These data suggest that roselle can be used as a natural antioxidative supplement in the prevention of oxidative damage in diabetic patients.

Mohamed, Jamaludin; Shing, Saw Wuan; Md Idris, Muhd Hanis; Budin, Siti Balkis; Zainalabidin, Satirah



A synbiotic containing Lactobacillus gasseri [corrected] CHO-220 and inulin improves irregularity of red blood cells.  


This randomized, double-blind, placebo-controlled, and parallel-design study was conducted to investigate the effect of a synbiotic product containing Lactobacillus gasseri [corrected] CHO-220 and inulin on the irregularity in shape of red blood cells (RBC) in hypercholesterolemic subjects. The subjects (n=32) were randomly allocated to 2 groups, a treatment group (synbiotic product) and a control group (placebo), and received 4 capsules of either synbiotic or placebo daily for 12 wk. Morphological representation via scanning electron microscopy showed that the occurrence of spur RBC was improved upon supplementation of the synbiotic. In addition, the supplementation of synbiotic reduced the cholesterol:phospholipids ratio of the RBC membrane by 47.02% over 12 wk, whereas the control showed insignificant changes. Our present study also showed that supplementation of the synbiotic reduced the concentration of saturated fatty acids (SFA), increased unsaturated fatty acids (UFA), and increased the ratio of UFA:SFA over 12 wk, whereas the control showed inconspicuous changes. The alteration of RBC membrane was assessed using fluorescence anisotropy (FAn) and fluorescence probes with different affinities for varying sections of the membrane phospholipid bilayer. A noticeable decrease in FAn of three fluorescent probes was observed in the synbiotic group compared with the control over 12 wk, indicative of increased membrane fluidity and reduced cholesterol enrichment in the RBC membrane. PMID:20854987

Ooi, L-G; Bhat, R; Rosma, A; Yuen, K-H; Liong, M-T



Fuel cell electrolyte membrane with basic polymer  


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)



Fuel cell electrolyte membrane with basic polymer  


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.



Clinical application of sodium-23 nuclear magnetic resonance for measurement of red cell sodium concentrations.  


Red cell sodium (RBC-Na+) concentrations were measured using 23Na nuclear magnetic resonance (NMR), without the destruction of erythrocyte membranes. Subjects were categorized into four groups: 20 normotensive subjects (NT group), 20 age-matched essential hypertensive patients (EHT group), 10 patients with primary aldosteronism (PA group), and 18 patients treated with digoxin (DIG group). Although RBC-Na+ concentrations were similar between the NT group (6.14 +/- 0.80 (Mean +/- SD) mmol/l) and the EHT group (5.92 +/- 0.99), they were significantly higher in both the PA group (7.55 +/- 0.88, p less than 0.001) and the DIG group (8.43 +/- 3.81, p less than 0.02). In the PA group, RBC-Na+ concentrations decreased significantly after resection of the adenoma, and there was an inverse relationship between serum potassium and RBC-Na+ concentrations (r = -0.65, p less than 0.01). In the DIG group, RBC-Na+ concentrations tended to increase in proportion to serum digoxin levels (r = 0.53, p less than 0.05). These results support the view that RBC-Na+ concentrations are determined primarily by Na+/K+-pump activity of red cell membranes. This study showed also that Na+ NMR is an useful method determining intracellular Na+ concentrations. PMID:2595244

Kojima, S; Kanashiro, M; Hayashi, F; Yoshida, K; Abe, H; Imanishi, M; Kawamura, M; Kawano, Y; Ashida, T; Kimura, G



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



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.




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



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



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.



An elastic network model based on the structure of the red blood cell membrane skeleton.  

PubMed Central

A finite element network model has been developed to predict the macroscopic elastic shear modulus and the area expansion modulus of the red blood cell (RBC) membrane skeleton on the basis of its microstructure. The topological organization of connections between spectrin molecules is represented by the edges of a random Delaunay triangulation, and the elasticity of an individual spectrin molecule is represented by the spring constant, K, for a linear spring element. The model network is subjected to deformations by prescribing nodal displacements on the boundary. The positions of internal nodes are computed by the finite element program. The average response of the network is used to compute the shear modulus (mu) and area expansion modulus (kappa) for the corresponding effective continuum. For networks with a moderate degree of randomness, this model predicts mu/K = 0.45 and kappa/K = 0.90 in small deformations. These results are consistent with previous computational models and experimental estimates of the ratio mu/kappa. This model also predicts that the elastic moduli vary by 20% or more in networks with varying degrees of randomness. In large deformations, mu increases as a cubic function of the extension ratio lambda 1, with mu/K = 0.62 when lambda 1 = 1.5. Images FIGURE 1 FIGURE 2 FIGURE 3 FIGURE 4 FIGURE 7 FIGURE 8 FIGURE 9 FIGURE 10 FIGURE 11 FIGURE 12 FIGURE 13 FIGURE 14 FIGURE 15 FIGURE 16 FIGURE 17 FIGURE 18 FIGURE 20 FIGURE A1 FIGURE A2 FIGURE A3

Hansen, J C; Skalak, R; Chien, S; Hoger, A



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



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



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.



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


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?



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



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



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



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



Probing red blood cell mechanics, rheology and dynamics with a two-component multi-scale model.  


This study is partially motivated by the validation of a new two-component multi-scale cell model we developed recently that treats the lipid bilayer and the cytoskeleton as two distinct components. Here, the whole cell model is validated and compared against several available experiments that examine red blood cell (RBC) mechanics, rheology and dynamics. First, we investigated RBC deformability in a microfluidic channel with a very small cross-sectional area and quantified the mechanical properties of the RBC membrane. Second, we simulated twisting torque cytometry and compared predicted rheological properties of the RBC membrane with experimental measurements. Finally, we modelled the tank-treading (TT) motion of a RBC in a shear flow and explored the effect of channel width variation on the TT frequency. We also investigated the effects of bilayer-cytoskeletal interactions on these experiments and our simulations clearly indicated that they play key roles in the determination of cell membrane mechanical, rheological and dynamical properties. These simulations serve as validation tests and moreover reveal the capabilities and limitations of the new whole cell model. PMID:24982252

Li, Xuejin; Peng, Zhangli; Lei, Huan; Dao, Ming; Karniadakis, George Em



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


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.



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



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



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



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



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


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



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


From artificial red blood cells, oxygen carriers, and oxygen therapeutics to artificial cells, nanomedicine, and beyond  

PubMed Central

The first experimental artificial red blood cells have all three major functions of red blood cells (rbc). However, the first practical one is a simple polyhemoglobin (PolyHb) that only has an oxygen-carrying function. This is now in routine clinical use in South Africa and Russia. An oxygen carrier with antioxidant functions, PolyHb-catalase-superoxide dismutase, can fulfill two of the three functions of rbc. Even more complete is one with all three functions of rbc in the form of PolyHb-catalase-superoxide dismutase-carbonic anhydrase. The most advanced ones are nanodimension artificial rbc with either PEG-lipid membrane or PEG-PLA polymermembrane. Extensions in to oxygen therapeutics include a PolyHb-tyrosinase that suppresses the growth of melanoma in a mice model. Another is a PolyHb-fibrinogen that is an oxygen carrier with platelet-like function. Research has now extended well beyond the original research on artificial rbc into many areas of artificial cells. These include nanoparticles, nanotubules, lipid vesicles, liposomes, polymer-tethered lipid vesicles, polymersomes, microcapsules, bioencapsulation, nanocapules, macroencapsulation, synthetic cells, and others. These are being used in nanotechnology, nanomedicine, regenerative medicine, enzyme/gene therapy, cell/stem cell therapy, biotechnology, drug delivery, hemoperfusion, nanosensers, and even by some groups in agriculture, industry, aquatic culture, nanocomputers, and nanorobotics.

Chang, Thomas M. S.



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



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.



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



Nitrite-Induced Improved Blood Circulation Associated with an Increase in a Pool of RBC-NO with NO Bioactivity  

PubMed Central

The reduction of nitrite by RBCs producing NO can play a role in regulating vascular tone. This hypothesis was investigated in rats by measuring the effect of nitrite infusion on mean arterial blood pressure, cerebral blood flow and cerebrovascular resistance in conjunction with the accumulation of RBC-NO. The nitrite infusion reversed the increase in MAP and decrease in CBF produced by L-NAME inhibition of e-NOS. At the same time there was a dramatic increase in RBC-NO. Correlations of RBC-NO for individual rats support a role for the regulating vascular tone by this pool of NO. Furthermore, data obtained prior to treatment with L-NAME or nitrite is consistent with a contribution of RBC reduced nitrite in regulating vascular tone even under normal conditions. The role of the red cell in delivering NO to the vasculature was explained by the accumulation of a pool of bioactive NO in the red cell when nitrite is reduced by deoxygenated hemoglobin chains. A comparison of R and T state hemoglobin demonstrated a potential mechanism for the release of this NO in the T-state present at reduced oxygen pressures when blood enters the microcirculation. Coupled with enhanced hemoglobin binding under theses conditions the NO can be released to the vasculature.

Rifkind, Joseph M.; Nagababu, Enika; Cao, Zeling; Barbiro-Michaely, Efrat; Mayevsky, Avraham



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



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



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



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



Proton-conductive nanochannel membrane for fuel-cell applications.  


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



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


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



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.



Photothermal microscopy for cell imaging and diagnostics  

NASA Astrophysics Data System (ADS)

Dual-laser photothermal (PT) techniques are applied for live blood cell studies. The measurement of the action of a pump pulse (400 - 600 nm, 10 ns, 1-500 (mu) J) on a single cell provides the imaging of the cell's absorbing structure/thermal field, spectroscopy and control of the cell's state. The imaging technique includes acquisition of three types of images: optical image, pulsed laser image with 5 ns temporal resolution, and PT-image with the same temporal and 0.5 micrometers spatial resolution. In PT-images of WBC the details of a cell structure have been found that cannot be obtained from optical images of these cells. Quantitative analysis of PT-images enables abnormal cell differentiation in a case of anaemia (RBC) and pulmonary diseases (neutrophyls). Analysis of cell population according to the cell's ability to survive a pump pulse is considered as the viability test. This technique was used for RBC comparative study. Significant differences in RBC populations have been found. Also the viability test was applied for monitoring of action of the drugs upon RBC membrane. A fully automated experimental system was built for PT-studies of the cell populations. Our set up is technically compatible to optical microscope. Operation rate is 1 cell/s. The required capacity of cell suspension is about 1 (mu) l.

Lapotko, Dmitry; Kuchinsky, Georgy



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.



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)



Lactic acid fermentation in cell-recycle membrane bioreactor.  


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



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



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.



Photobleaching regions of living cells to monitor membrane traffic.  


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



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



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.



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



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


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



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.



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



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



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


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



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.



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.



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



Electrospun fiber membranes enable proliferation of genetically modified cells.  


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



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


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



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



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.



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



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.



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.



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



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



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



Fibronectin-degrading proteases from the membranes of transformed cells.  


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



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



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



MG53 regulates membrane budding and exocytosis in muscle cells.  


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



Cholesterol-mediated membrane surface area dynamics in neuroendocrine cells.  


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



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



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



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



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



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


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



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



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


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




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



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.



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.



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.



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.


Dynamics of flat membranes and flickering in red blood cells  

NASA Astrophysics Data System (ADS)

A theory of the dynamics of polymerized membranes in the flat phase is presented. The dynamics of dilute membrane solutions is strongly influenced by long-ranged hydrodynamic interactions among the monomers, mediated by the intervening solvent. We discuss the renormalization of the kinetic coefficients for the undulation and phonon modes due to hydrodynamic “backflow” (Zimm behavior). The dynamics is also studied for free draining membranes (Rouse dynamics) corresponding to the Brownian dynamics method used in Monte Carlo simulations. The long time behavior of the dynamic structure factor is given by stretched exponentials with stretching exponents determined by the exponents of the elastic coefficients and the wave vector dependence of the Oseen tensor. We also study the dynamics of the thickness fluctuations in red blood cells (flicker phenomenon) taking into account the underlying polymerized spectrin skeleton. Qualitatively different dynamical behavior is predicted for spectrin skeletons isolated from heir natural lipid environment.

Frey, Erwin; Nelson, David R.



BIOCHEMISTRY: Signaling Across the Cell Membrane  

NSDL National Science Digital Library

Access to the article is free, however registration and sign-in are required. Structural and functional studies shed light on how G protein-coupled receptors sense external stimuli. G protein-coupled receptors (GPCRs)--the largest and most diverse group of tranmembrane receptors--occur in nearly every eukaryotic cell and can sense photons, cations, small molecules, peptides, and proteins (1, 2). Two research articles in this issue (4, 5) and a recent article in Nature (6) report important steps towards understanding how GPCRs operate.

Rama Ranganathan (University of Texas Southwestern Medical Center;Green Center for Systems Biology and Department of Pharmacology)



Sertoli cell plasma membrane polypeptides involved in spermatogenic cell-Sertoli cell adhesion.  


This study concerns Sertoli cell-spermatogenic cell adhesive interactions in the seminiferous tubule. Sertoli cell surface polypeptides involved in germ cell-Sertoli cell adhesion were identified by serological inhibition of an in vitro Sertoli-germ cell adhesion assay. This assay was modified from a previously reported adhesion assay, and employs a scanning laser cytometer for quantification of adherent cells. Reactivity of the polyclonal antiserum raised against rat Sertoli cells was also assessed via immunofluorescent microscopy. The addition of antiserum to the adhesion assay resulted in a 42% to 66% inhibition of cell-cell adhesion. Moreover, preincubation of antiserum with Sertoli cell monolayers resulted in a significant reduction of spermatogenic cell binding. Conversely, preincubation of antiserum with germ cells resulted in no reduction. Western blot analysis of the antiserum against purified Sertoli cell membranes indicated reactivity with four polypeptides. The data suggest that one or more of these polypeptides are directly involved in the adhesion of germ cells to Sertoli cell monolayers in vitro. PMID:1597400

Newton, S C; Millette, C F



Enhanced production of a thermostable mannanase by immobilized cells of Bacillus subtilis on various membranes  

Microsoft Academic Search

Cells of the thermophilic Bacillus subtilis WY34 were immobilized on various formaldehyde-activated polymer membranes and the immobilized cells were used for the production\\u000a of thermostable mannanase in flasks. The results showed that polyethersulfone membranes (PES) and nylon-6 membranes were the\\u000a most suitable supports for cell immobilization to produce the mannanase. Moreover, PES and nylon-6 membranes immobilized cells\\u000a provided 1.78- and

Qiaojuan Yan; Aimin Wu; Zhengqiang Jiang; Luo Tang; Shi Bo



Use the force: Membrane tension as an organizer of cell shape and motility  

PubMed Central

Many cell phenomena that involve shape changes are affected by the intrinsic deformability of the plasma membrane. Far from being a passive participant, the plasma membrane is now known to physically, as well as biochemically, influence cell processes ranging from vesicle trafficking to actin assembly. Here we review current understanding of how changes in plasma membrane tension regulate cell shape and movement as well as how cells sense plasma membrane tension.

Diz-Munoz, Alba; Fletcher, Daniel A.; Weiner, Orion D.



Creating Transient Cell Membrane Pores Using a Standard Inkjet Printer  

PubMed Central

Bioprinting has a wide range of applications and significance, including tissue engineering, direct cell application therapies, and biosensor microfabrication.1-10 Recently, thermal inkjet printing has also been used for gene transfection.8,9 The thermal inkjet printing process was shown to temporarily disrupt the cell membranes without affecting cell viability. The transient pores in the membrane can be used to introduce molecules, which would otherwise be too large to pass through the membrane, into the cell cytoplasm.8,9,11 The application being demonstrated here is the use of thermal inkjet printing for the incorporation of fluorescently labeled g-actin monomers into cells. The advantage of using thermal ink-jet printing to inject molecules into cells is that the technique is relatively benign to cells.8, 12 Cell viability after printing has been shown to be similar to standard cell plating methods1,8. In addition, inkjet printing can process thousands of cells in minutes, which is much faster than manual microinjection. The pores created by printing have been shown to close within about two hours. However, there is a limit to the size of the pore created (~10 nm) with this printing technique, which limits the technique to injecting cells with small proteins and/or particles. 8,9,11 A standard HP DeskJet 500 printer was modified to allow for cell printing.3, 5, 8 The cover of the printer was removed and the paper feed mechanism was bypassed using a mechanical lever. A stage was created to allow for placement of microscope slides and coverslips directly under the print head. Ink cartridges were opened, the ink was removed and they were cleaned prior to use with cells. The printing pattern was created using standard drawing software, which then controlled the printer through a simple print command. 3T3 fibroblasts were grown to confluence, trypsinized, and then resuspended into phosphate buffered saline with soluble fluorescently labeled g-actin monomers. The cell suspension was pipetted into the ink cartridge and lines of cells were printed onto glass microscope cover slips. The live cells were imaged using fluorescence microscopy and actin was found throughout the cytoplasm. Incorporation of fluorescent actin into the cell allows for imaging of short-time cytoskeletal dynamics and is useful for a wide range of applications.13-15

Owczarczak, Alexander B.; Shuford, Stephen O.; Wood, Scott T.; Deitch, Sandra; Dean, Delphine



Near-Critical Fluctuations and Cytoskeleton-Assisted Phase Separation Lead to Subdiffusion in Cell Membranes  

PubMed Central

We address the relationship between membrane microheterogeneity and anomalous subdiffusion in cell membranes by carrying out Monte Carlo simulations of two-component lipid membranes. We find that near-critical fluctuations in the membrane lead to transient subdiffusion, while membrane-cytoskeleton interaction strongly affects phase separation, enhances subdiffusion, and eventually leads to hop diffusion of lipids. Thus, we present a minimum realistic model for membrane rafts showing the features of both microscopic phase separation and subdiffusion.

Ehrig, Jens; Petrov, Eugene P.; Schwille, Petra



A novel unitized regenerative proton exchange membrane fuel cell  

NASA Astrophysics Data System (ADS)

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 work performed at Lynntech. 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 unitized regenerative fuel cell and results obtained on testing it will be presented.

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



Sodium channels in membrane vesicles from cultured toad bladder cells  

SciTech Connect

Electrical potential-driven {sup 22}Na{sup +} fluxes were measured in membrane vesicles prepared from TBM-18(cl23) cells (a clone of the established cell line TB-M). Fifty to seventy percent of the tracer uptake in vesicles derived from cells that were cultivated on a porous support were blocked by the diuretic amiloride. The amiloride inhibition constant was <0.1 {mu}M, indicating that this flux is mediated by the apical Na{sup +}-specific channels. Vesicles prepared from cells that were not grown on a porous support exhibited much smaller amiloride-sensitive fluxes. Two Ca{sup 2+}-dependent processes that down-regulated the channel conductance and were previously identified in native epithelia were found in the cultured cells as well. Vesicles isolated from cells that were preincubated with 5 {times} 10{sup {minus}7} M aldosterone for 16-20 h exhibited higher amiloride-sensitive conductance than vesicles derived from control, steroid-depleted cells. Thus membrane derived from TBM-18(cl23) cells can be used to characterize the epithelial Na{sup +} channel and its hormonal regulation.

Asher, C.; Moran, A.; Rossier, B.C.; Garty, H. (Weizmann Institute of Science, Rehovot (Israel) Ben Gurion Univ., Beer-Sheva (Israel) Institut de Pharmacologie de l'Universite de Lausanne (Switzerland))



The biocompatibility research of functional Schwann cells induced from bone mesenchymal cells with chitosan conduit membrane.  


To explore the adhesion and proliferation characteristics of bone marrow mesenchymal cells (MSCs) to chitosan conduit membrane, MSCs were induced by a sequential composition Beta-mercaptoethanol, retinoic acid, forskolin, basic-FGF, PDGF and heregulin. Schwann Cell markers, namely S-100 and GFAP, were used to discriminate the induced MSCs' properties by immunofluorescent staining. These results suggested that MSCs can take on Schwann cell's phenotype in vitro and the induce MSCs were gifted with good biocompatibility to biogradable chitosan conduit membrane. The results provided the possibilities to using the induced MSCs with chitosan conduit membrane in artificial peripherial nerve fields to promote nerve regeneration. PMID:16519406

Zhang, Peixun; Xu, Hailin; Zhang, Dianying; Fu, Zhongguo; Zhang, Hongbo; Jiang, Baoguo



Membrane tether formation from outer hair cells with optical tweezers.  

PubMed Central

Optical tweezers were used to characterize the mechanical properties of the outer hair cell (OHC) plasma membrane by pulling tethers with 4.5-microm polystyrene beads. Tether formation force and tether force were measured in static and dynamic conditions. A greater force was required for tether formations from OHC lateral wall (499 +/- 152 pN) than from OHC basal end (142 +/- 49 pN). The difference in the force required to pull tethers is consistent with an extensive cytoskeletal framework associated with the lateral wall known as the cortical lattice. The apparent plasma membrane stiffness, estimated under the static conditions by measuring tether force at different tether length, was 3.71 pN/microm for OHC lateral wall and 4.57 pN/microm for OHC basal end. The effective membrane viscosity was measured by pulling tethers at different rates while continuously recording the tether force, and estimated in the range of 2.39 to 5.25 pN x s/microm. The viscous force most likely results from the viscous interactions between plasma membrane lipids and the OHC cortical lattice and/or integral membrane proteins. The information these studies provide on the mechanical properties of the OHC lateral wall is important for understanding the mechanism of OHC electromotility.

Li, Zhiwei; Anvari, Bahman; Takashima, Masayoshi; Brecht, Peter; Torres, Jorge H; Brownell, William E



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))



Cytokines modulate expression of cell-membrane complement inhibitory proteins in human lung cancer cell lines.  


Human lung cancers overexpress several cell-membrane complement inhibitory proteins (CIP). These complement inhibitory proteins are membrane cofactor protein (CD46), decay-accelerating factor (DAF; CD55), and CD59 (protectin). These cell-membrane proteins have a wide normal tissue distribution, are known to protect normal host cells from homologous complement-mediated lysis, and are thought to facilitate tumor escape from immunosurveillance. To study whether proinflammatory cytokines that are involved in cancer growth can modulate cell-membrane CIP expression in lung cancer cells, we studied the effect of interleukin (IL)-1alpha, tumor necrosis factor (TNF)-alpha, and interferon (IFN)-gamma on two human lung cancer cell lines. ChaGo K-1 and NCI-H596 cell lines, undifferentiated carcinoma and lung adenosquamous carcinoma, respectively, were stimulated with different cytokines, and the effects of incubation time and cytokine concentration on cell-membrane CIP expression were studied. Cell-membrane CIP expression was evaluated using flow cytometry and cytokine effect was calculated as percent change in mean fluorescence intensity of each CIP molecule from its untreated control. We found that DAF was the lung cancer cell-membrane CIP molecule that was the most responsive to cytokine stimulation. Maximal stimulatory effect was usually noted 72 h after a cytokine was introduced. In ChaGo K-1 and NCI-H596 lung cancer cell lines, IL-1alpha and TNF-alpha increased DAF expression. IL-1alpha (100 U/ml/72 h) increased DAF expression up to a maximal mean of 45 and 48%, respectively, in comparison with untreated cells. TNF-alpha (1, 000 U/ml/72 h) increased DAF expression up to a mean of 131 and 46%, respectively. IFN-gamma (1 U/ml/72 h) increased DAF expression in NCI-H596 cells up to a mean of 100%, but had a slight inhibitory effect on DAF expression in ChaGo K-1 cells, decreasing expression by a mean of 17% in comparison with untreated cells. We conclude that cell-membrane DAF expression in the studied human lung cancer cell lines is modulated by IL-1alpha, TNF-alpha, and IFN-gamma, and speculate that cytokine-mediated modulation of cell-membrane DAF in human lung cancer cells might affect lung cancer cell biology. PMID:9730881

Varsano, S; Rashkovsky, L; Shapiro, H; Radnay, J



Role of membrane phospholipids and glycolipids in cell-to-cell fusion by VSV.  


To identify membrane components of CER cells interacting with vesicular stomatitis virus (VSV) during fusion at acidic pH (fusion from without, FFWO) two different approaches have been used, i.e. (i) treating the whole cells with enzymes and (ii) testing the ability of isolated membrane molecules to interfere with FFWO. Phospholipase A2 and C digestion of cells greatly reduced syncytia formation, pointing towards the involvement of lipid structures as target sites for VSV. Cell susceptibility to FFWO was also reduced after neuraminidase, beta-galactosidase or periodate treatment, suggesting that carbohydrate residues may participate in a complex receptor structure required for virus fusion. When membrane molecules were examined separately for their ability to inhibit viral FFWO, phosphatidylserine, phosphatidylinositol, sphingomyelin, cholesterol and GM3 ganglioside were found to be active, confirming the role of membrane lipid moiety in the cell surface structures involved in the early phases of VSV infection. PMID:1660797

Conti, C; Mastromarino, P; Orsi, N



Time-lapse imaging of membrane traffic in living cells.  


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 describes the use of the confocal laser-scanning microscope (CLSM) for time-lapse imaging of one or more fluorescent markers. PMID:22046037

Snapp, Erik Lee; Lajoie, Patrick



Counting cells in cerebrospinal fluid collected directly on membrane filters  

PubMed Central

Accurate enumeration of cells in the cerebrospinal fluid is not feasible with the current method of counting cells in the Fuchs Rosenthal or Neubauer chamber when the count is near normal since the volume of fluid examined is too small. A sample of adequate volume to permit such accurate assessment may be collected from the lumbar puncture needle into a syringe through a ruled membrane filter in a Swinney cartridge. A study of 291 samples shows that the upper limit of the normal cerebrospinal fluid count is 2000 cells/ml (2/cmm) and not 5000/ml (5/cmm) as is currently accepted. Images

Burechailo, F.; Cunningham, T. A.



Membrane phenotypic studies in B cell lymphoproliferative disorders.  

PubMed Central

A total of 398 cases of B cell lymphoproliferative disease were phenotypically characterised by membrane mouse red blood cell (MRBC) receptor, surface immunoglobulin, common acute lymphoblastic leukaemia (CALLA), and FMC7 and T1 monoclonal antibody studies. Relations between chronic lymphocytic leukaemia (CLL), prolymphocytic leukaemia (PLL), and "prolymphocytoid" CLL variants were examined with particular reference to the expression of FMC7. In addition, the reactivity of TU1 monoclonal antibody with B cell disorders was established. The results suggest that despite some heterogeneity most cases may be characterised by their phenotypic patterns and that these investigations provide a reproducible basis for classification.

Scott, C S; Limbert, H J; MacKarill, I D; Roberts, B E



Myosin-X facilitates Shigella-induced membrane protrusions and cell-to-cell spread  

PubMed Central

Summary The intracellular pathogen Shigella flexneri forms membrane protrusions to spread from cell to cell. As protrusions form, myosin-X (Myo10) localizes to Shigella. Electron micrographs of immunogold-labelled Shigella-infected HeLa cells reveal that Myo10 concentrates at the bases and along the sides of bacteria within membrane protrusions. Time-lapse video microscopy shows that a full-length Myo10 GFP-construct cycles along the sides of Shigella within the membrane protrusions as these structures progressively lengthen. RNAi knock-down of Myo10 is associated with shorter protrusions with thicker stalks, and causes a >80% decrease in confluent cell plaque formation. Myo10 also concentrates in membrane protrusions formed by another intracellular bacteria, Listeria, and knock-down of Myo10 also impairs Listeria plaque formation. In Cos7 cells (contain low concentrations of Myo10), the expression of full-length Myo10 nearly doubles Shigella-induced protrusion length, and lengthening requires the head domain, as well as the tail-PH domain, but not the FERM domain. The GFP-Myo10-HMM domain localizes to the sides of Shigella within membrane protrusions and the GFP-Myo10-PH domain localizes to host cell membranes. We conclude that Myo10 generates the force to enhance bacterial-induced protrusions by binding its head region to actin filaments and its PH tail domain to the peripheral membrane.

Li, Wei; Dhillon, Jess; Bohil, Aparna B.; Cheney, Richard E.; Hartwig, John H.; Southwick, Frederick S.



Red Blood Cell Antibody Identification  


... Antibody ID, RBC; RBC Ab ID Formal name: Red Blood Cell Antibody Identification Related tests: Direct Antiglobulin ... None The Test Sample What is being tested? Red blood cell antibodies are proteins produced by the ...


Band 3 is a target protein of spectrin's E2/E3 activity: implication for sickle cell disease and normal red blood cell aging.  


We have demonstrated that a 125 kDa red blood cell (RBC) membrane protein, being a target of spectrin's E2/E3 activity, is ubiquitinated band 3. This demonstration was based on copurification of this biotinylated-ubiquitinated protein with band 3, immunoprecipitation with band 3 antibody and analysis of proteins associated with strepavidin sepharose by micro liquid chromatography coupled to tandem mass spectrometry (microLC/MS/MS). Further, we demonstrated the presence of ubiquitinated band 3 in vivo by Western blotting of purified band 3 with a monoclonal antibody (FK2) against ubiquitin. The implications of these results for sickle cell disease and RBC aging are discussed. PMID:15095787

Chang, T L; Cubillos, F F; Kakhniashvili, D G; Goodman, S R



Tetraspanins regulate the protrusive activities of cell membrane  

SciTech Connect

Highlights: Black-Right-Pointing-Pointer Tetraspanins regulate microvillus formation. Black-Right-Pointing-Pointer Tetraspanin CD81 promotes microvillus formation. Black-Right-Pointing-Pointer Tetraspanin CD82 inhibits microvillus formation. Black-Right-Pointing-Pointer Based on this study, we extrapolated a general cellular mechanism for tetraspanins. Black-Right-Pointing-Pointer Tetraspanins engage various functions by regulating membrane protrusion morphogenesis. -- Abstract: Tetraspanins have gained increased attention due to their functional versatility. But the universal cellular mechanism that governs such versatility remains unknown. Herein we present the evidence that tetraspanins CD81 and CD82 regulate the formation and/or development of cell membrane protrusions. We analyzed the ultrastructure of the cells in which a tetraspanin is either overexpressed or ablated using transmission electron microscopy. The numbers of microvilli on the cell surface were counted, and the radii of microvillar tips and the lengths of microvilli were measured. We found that tetraspanin CD81 promotes the microvillus formation and/or extension while tetraspanin CD82 inhibits these events. In addition, CD81 enhances the outward bending of the plasma membrane while CD82 inhibits it. We also found that CD81 and CD82 proteins are localized at microvilli using immunofluorescence. CD82 regulates microvillus morphogenesis likely by altering the plasma membrane curvature and/or the cortical actin cytoskeletal organization. We predict that membrane protrusions embody a common morphological phenotype and cellular mechanism for, at least some if not all, tetraspanins. The differential effects of tetraspanins on microvilli likely lead to the functional diversification of tetraspanins and appear to correlate with their functional propensity.

Bari, Rafijul [Cancer Center and Department of Medicine, University of Tennessee, Memphis, TN (United States)] [Cancer Center and Department of Medicine, University of Tennessee, Memphis, TN (United States); Guo, Qiusha [Cancer Center and Department of Medicine, University of Tennessee, Memphis, TN (United States) [Cancer Center and Department of Medicine, University of Tennessee, Memphis, TN (United States); Zhongnan Hospital, Wuhan University, Wuhan (China); Xia, Bing [Zhongnan Hospital, Wuhan University, Wuhan (China)] [Zhongnan Hospital, Wuhan University, Wuhan (China); Zhang, Yanhui H. [Cancer Center and Department of Medicine, University of Tennessee, Memphis, TN (United States)] [Cancer Center and Department of Medicine, University of Tennessee, Memphis, TN (United States); Giesert, Eldon E. [Department of Ophthalmology, University of Tennessee, Memphis, TN (United States)] [Department of Ophthalmology, University of Tennessee, Memphis, TN (United States); Levy, Shoshana [Department of Medicine, Stanford University, Palo Alto, CA (United States)] [Department of Medicine, Stanford University, Palo Alto, CA (United States); Zheng, Jie J. [Department of Structural Biology, St. Jude Children's Research Hospital, Memphis, TN (United States)] [Department of Structural Biology, St. Jude Children's Research Hospital, Memphis, TN (United States); Zhang, Xin A., E-mail: [Cancer Center and Department of Medicine, University of Tennessee, Memphis, TN (United States)



Red blood cell membrane cation transport in normotensive psoriatics.  


Psoriasis might be a widespread membrane disorder. To verify whether under the influence of psoriasis the red cell membrane cation metabolism is altered, we compared 46 psoriatics with 23 normotensive controls. A significant increase was observed in intracellular K+ content, in the maximal velocity of the Na(+)-K+ ATPase, of the Na(+)-K(+)-Cl- outward co-transport, of the Na+/H+ exchange, as well as in the outward passive permeability for Na+. No difference was seen between the two groups in Na+ content, Li+/Na+ exchange or in the passive permeability to K+. In 8 psoriatics treated with etretinate (10-75 mg/day for 1-36 months), Na(+)-K+ ATPase, Na(+)-K(+)-Cl- co-transport and the passive permeability for Na+ were not significantly different from controls. These results suggest that the primary abnormality might be an increased Na+ influx, in part through the Na+/H+ exchange, which is compensated by increased activity of outward transports, and confirm that the red cells are a useful model for the study of membrane transport in this disease. Our results indicate also that these membrane transport abnormalities can be corrected by etretinate treatment. PMID:2609881

Mozzato, M G; Semplicini, A; Grosso, G; Ceolotto, G; Marzola, M; Pessina, A C; Peserico, A



Effects of extracellular calcium on cell membrane resealing in sonoporation  

PubMed Central

Sonoporation has been exploited as a promising strategy for intracellular drug and gene delivery. The technique uses ultrasound to generate pores on the cell membrane to allow entry of extracellular agents into the cell. Resealing of these non-specific pores is a key factor determining both the uptake and post-ultrasound cell survival. This study examined the effects of extracellular Ca2+ on membrane resealing in sonoporation, using Xenopus oocytes as a model system. The cells were exposed to tone burst ultrasound (1.06 MHz, duration 0.2 s, acoustic pressure 0.3 MPa) in the presence of 0.1% Definity® at various extracellular [Ca2+] (0–3 mM). Sonoporation inception and resealing in a single cell were monitored in real time via the transmembrane current of the cell under voltage clamp. The time-resolved measurements of transmembrane current revealed the involvement of two or more Ca2+ related processes with different rate constants and characteristics. Rapid resealing occurred immediately after ultrasound application followed by a much slower resealing process. Complete resealing required [Ca2+] above 0.54 mM. The cells resealed in 6–26 s at 1.8 mM Ca2+, but took longer at lower concentrations, up to 58 ~170 s at 0.54 mM Ca2+.

Zhou, Yun; Shi, Jingyi; Cui, Jianmin; Deng, Cheri X.



Membrane Domains Based on Ankyrin and Spectrin Associated with Cell-Cell Interactions  

PubMed Central

Nodes of Ranvier and axon initial segments of myelinated nerves, sites of cell–cell contact in early embryos and epithelial cells, and neuromuscular junctions of skeletal muscle all perform physiological functions that depend on clustering of functionally related but structurally diverse ion transporters and cell adhesion molecules within microdomains of the plasma membrane. These specialized cell surface domains appeared at different times in metazoan evolution, involve a variety of cell types, and are populated by distinct membrane-spanning proteins. Nevertheless, recent work has shown that these domains all share on their cytoplasmic surfaces a membrane skeleton comprised of members of the ankyrin and spectrin families. This review will summarize basic features of ankyrins and spectrins, and will discuss emerging evidence that these proteins are key players in a conserved mechanism responsible for assembly and maintenance of physiologically important domains on the surfaces of diverse cells.

Bennett, Vann; Healy, Jane



Free Tubulin Modulates Mitochondrial Membrane Potential in Cancer Cells  

PubMed Central

Formation of the mitochondrial membrane potential (??) depends on flux of respiratory substrates, ATP, ADP and Pi through voltage-dependent anion channels (VDAC). Since tubulin promotes single channel closure of VDAC, we hypothesized that tubulin is a dynamic regulator of ??. ?? in cultured cancer cells was assessed by confocal microscopy of the potential-indicating fluorophore, tetramethylrhodamine methylester (TMRM). Microtubule destabilizers, rotenone, colchicine and nocodazole, and the microtubule stabilizer, paclitaxel, increased and decreased cellular free tubulin, respectively, and in parallel decreased and increased ??. Protein kinase A (PKA) activation by cAMP analogs and glycogen synthase kinase 3? (GSK-3?) inhibition decreased ??, whereas PKA inhibition hyperpolarized, consistent with reports that PKA and GSK-3? decrease and increase VDAC conductance, respectively. Plasma membrane potential assessed by DiBAC4(3) was not altered by any of the treatments. We propose that inhibition of VDAC by free tubulin limits mitochondrial metabolism in cancer cells.

Maldonado, Eduardo N.; Patnaik, Jyoti; Mullins, Matthew R.; Lemasters, John J.



Evaluation of stem cell components in retrocorneal membranes.  


The purpose of this study was to elucidate the origin and cellular composition of retrocorneal membranes (RCMs) associated with chemical burns using immunohistochemical staining for primitive cell markers. Six cases of RCMs were collected during penetrating keratoplasty. We examined RCMs with hematoxylin and eosin (H&E), periodic acid-Schiff (PAS) staining and immunohistochemical analysis using monoclonal antibodies against hematopoietic stem cells (CD34, CD133, c-kit), mesenchymal stem cells (beta-1-integrin, TGF-?, vimentin, hSTRO-1), fibroblasts (FGF-?, ?-smooth muscle actin), and corneal endothelial cells (type IV collagen, CD133, VEGF, VEGFR1). Histologic analysis of RCMs revealed an organized assembly of spindle-shaped cells, pigment-laden cells, and thin collagenous matrix structures. RCMs were positive for markers of mesenchymal stem cells including beta-1-integrin, TGF-?, vimentin, and hSTRO-1. Fibroblast markers were also positive, including FGF-? and ?-smooth muscle actin (SMA). In contrast, immunohistochemical staining was negative for hematopoietic stem cell markers including CD34, CD133 and c-kit as well as corneal endothelial cell markers such as type IV collagen, CD133 except VEGF and VEGFR1. Pigment-laden cells did not stain with any antibodies. The results of this study suggest that RCMs consist of a thin collagen matrix and fibroblast-like cells and may be a possible neogenetic structure produced from a lineage of bone marrow-derived mesenchymal stem cells. PMID:24932088

Lee, Seok Hyun; Kim, Kyoung Woo; Kim, Mi Kyung; Chun, Yeoun Sook; Kim, Jae Chan



Delivery of the Non-Membrane-Permeative Antibiotic Gentamicin into Mammalian Cells by Using Shigella flexneri Membrane Vesicles  

Microsoft Academic Search

We developed a model to test whether non-membrane-permeative therapeutic agents such as gentamicin could be delivered into mammalian cells by means of bacterial membrane vesicles. Many gram-negative bacteria bleb off membrane vesicles (MVs) during normal growth, and the quantity of these vesicles can be increased by brief exposure to gentamicin (J. L. Kadurugamuwa and T. J. Beveridge, J. Bacteriol. 177:3998-




Cytoskeleton Influence on Normal and Tangent Fluctuation Modes in the Red Blood Cells  

NASA Astrophysics Data System (ADS)

We argue that the paradoxal softness of the red blood cells (RBC) in fluctuation experiments is apparent. We show that the effective surface shear modulus ?s of the RBC obtained from fluctuation data and that measured in static deformation experiments have the same order of magnitude. In the RBC model developed for this purpose the spectrin network cytoskeleton with the bulk shear modulus estimated as ??105 165Pa contributes to both normal and tangent fluctuations of the system and confines the membrane fluctuations. The calculated ratio of the mean-square amplitudes ?Xn2?/?Xt2? is 2 3 orders of magnitude smaller than it is in the free membrane with the same bending and shear moduli.

Rochal, S. B.; Lorman, V. L.



Metal/acid ion permeable membrane fuel cell  

SciTech Connect

A fuel cell is disclosed which comprises a metal anode immersed in a base electrolyte solution and connected with an electric circuit, a cathode comprising an acid solution, a carbon catalyst and electron distributor plate in that solution and connected with an electron supply and a wettable fluid impermeable membrane and disposed between the electrolyte and cathode solution and establishing an acid-base reactor interface where hydroxyl ions are generated for conduction through the electrolyte to the anode.

Struthers, R.C.



Optimal microporous layer for proton exchange membrane fuel cell  

Microsoft Academic Search

This study elucidates how fabrication processes (screen-printing and spraying) and constituent materials (carbon paper as backing, Acetylene Black (AB) carbon (42nm), XC-72R carbon (30nm) or BP2000 (15nm) as carbon powders, and 10–50% fluorinated ethylene propylene (FEP) as hydrophobic substances) for microporous layers (MPLs) affect the performance of proton exchange membrane fuel cells. The screen-printing process produces MPLs with smaller surface

Wei-Mon Yan; Dong-Kai Wu; Xiao-Dong Wang; Ai-Lien Ong; Duu-Jong Lee; Ay Su



Investigation of oxygen gain in polymer electrolyte membrane fuel cells  

NASA Astrophysics Data System (ADS)

The polymer electrolyte membrane fuel cell (PEMFC) faces an efficiency loss, so called "oxygen gain", when the cathode gas is changed from oxygen to air due to the reduced oxygen partial pressure. To reduce the oxygen gain of a PEMFC, performance and oxygen gain of the single cells were evaluated as a function of carbon support, Pt content in the catalyst, membrane electrode assembly (MEA) fabrication process and the cathode humidification temperature. Among the tested carbon supports, Black Pearl 2000 and an undisclosed carbon produced the best performance and the lowest oxygen gain with their high surface area and high pore volume. As the Pt content in the catalyst increased from 10 to 60 wt.%, Pt surface area and the electrode thickness decreased leading to decreases in active catalyst surface area, and an ohmic and mass transfer resistance of the electrode. Due to trade-off effects, 20 wt.% Pt exhibited the highest performance. Compared to the conventional MEA, the MEA prepared using catalyst-coated membrane (CCM) method showed better performance with reduced catalyst loss into the gas diffusion media (GDM). As the cathode humidification temperature increased from 55 to 85 °C, the amount of water supplied to the cathode increased, leading to an increase in ionic conductivity of the membrane and another probability of water flooding. Thus, in the low current density region, performance of the single cell was improved with cathode humidification temperature, while in the high current density region, the single cell showed the highest performance at the cathode humidification temperature of 65 °C with water flooding at 75 and 85 °C.

Prasanna, M.; Ha, H. Y.; Cho, E. A.; Hong, S.-A.; Oh, I.-H.


Power converter for proton exchange membrane fuel cell  

Microsoft Academic Search

This paper presents the modelling and simulation of proton exchange membrane (PEM) fuel cell (FC) together with a dc-dc converter and dc-ac inverter. The need for the dc-dc converter arises as a result of the output voltage generated from the FC which fluctuates with variation in load. The FC voltage is therefore, controlled using dc-dc converter to provide a regulated

S. N. Ngema; A. K. Saha; N. M. Ijumba



CFD-based modelling of proton exchange membrane fuel cells  

Microsoft Academic Search

A comprehensive non-isothermal, 3D computational model for proton exchange membrane (PEM) fuel cells has been developed, and implemented into a computational fluid dynamic (CFD) code. The model allows parallel computing, thus making it practical to perform well-resolved simulations for large computational domains. The model accounts for convective and diffusive transport and allows prediction of the concentration of species. Distributed heat

B. R. Sivertsen; N. Djilali



Polymer Electrolyte Membrane (PEM) Fuel Cells Modeling and Optimization  

NASA Astrophysics Data System (ADS)

Performance of polymer electrolyte membrane (PEM) fuel cells is dependent on operating parameters and designing parameters. Operating parameters mainly include temperature, pressure, humidity and the flow rate of the inlet reactants. Designing parameters include reactants distributor patterns and dimensions, electrodes dimensions, and electrodes properties such as porosity, permeability and so on. This work aims to investigate the effects of various designing parameters on the performance of PEM fuel cells, and the optimum values will be determined under a given operating condition.A three-dimensional steady-state electrochemical mathematical model was established where the mass, fluid and thermal transport processes are considered as well as the electrochemical reaction. A Powell multivariable optimization algorithm will be applied to investigate the optimum values of designing parameters. The objective function is defined as the maximum potential of the electrolyte fluid phase at the membrane/cathode interface at a typical value of the cell voltage. The robustness of the optimum design of the fuel cell under different cell potentials will be investigated using a statistical sensitivity analysis. By comparing with the reference case, the results obtained here provide useful tools for a better design of fuel cells.

Zhang, Zhuqian; Wang, Xia; Shi, Zhongying; Zhang, Xinxin; Yu, Fan



Simulation for water management in membranes for polymer electrolyte fuel cells  

Microsoft Academic Search

Water management in membranes for polymer electrolyte fuel cells during their operational conditions is considered theoretically. Using a linear transport equation based on the diffusion of water and the electroosmotic drag, analytical solutions for water concentration profiles in the membrane are obtained from which membrane resistance overvoltage and other characteristic values are calculated. Specific parameters of the membranes such as

Tatsuhiro Okada; Gang Xie; Morten Meeg



Propagating Cell-Membrane Waves Driven by Curved Activators of Actin Polymerization  

Microsoft Academic Search

Cells exhibit propagating membrane waves which involve the actin cytoskeleton. One type of such membranal waves are Circular Dorsal Ruffles (CDR) which are related to endocytosis and receptor internalization. Experimentally, CDRs have been associated with membrane bound activators of actin polymerization of concave shape. We present experimental evidence for the localization of convex membrane proteins in these structures, and their

Barak Peleg; Andrea Disanza; Giorgio Scita; Nir Gov; Jerome Mathe



Expansion of conjunctival epithelial progenitor cells on amniotic membrane.  


Amniotic membrane (AM) reconstructed human conjunctival surfaces recover a goblet cell density higher than normal. Cultured rabbit conjunctival epithelial cells (RCE) on AM preferentially exhibit non-goblet epithelial differentiation. It was thus wondered if conjunctival progenitor cells that might have been preserved during ex vivo expansion on AM can still differentiate into conjunctival non-goblet epithelial and goblet cells under the influence of mesenchymal cells. Fourteen day old AM cultures of RCE were subcutaneously implanted in Balb/c athymic mice for 11 days and processed for PAS staining and immunostaining with monoclonal antibodies to conjunctival goblet cell mucin (MUC5AC, AM3), glycocalyx (AMEM2), cornea specific cytokeratins K3 (AE5) and K12 (AK2) and basal cell specific cytokeratin K14. Cell cycle kinetics were measured by BrdU labelling for 1 or 7 days. The 7 day labelled RCE were chased for 14 days in the same primary culture. After subcutaneous implantation, conjunctival non-goblet epithelial cells increased stratification and formed occasional cysts. The resultant epithelial phenotype was conjunctival with many PAS-positive, MUC5AC-positive, and AM3-positive goblet cells, AMEM2-positive suprabasal and superficial cells, and K14-positive basal cells, but was not corneal (negative to AE5 and AK2 staining). Twenty four hr BrdU labelling showed a labelling index of 42.5%. A higher labelling index or 69% was noted after continuous BrdU labelling for 7 days. A large number of label retaining basal cells with a labelling index of 84% were noted following 14 days of chase. Conjunctival epithelial progenitor cells for goblet and non-goblet cell differentiation are preserved by AM in vitro as evidenced by being able to differentiate into goblet cells in a permissive stromal environment, and being slow-cycling, and label retaining. This information is useful for future ex vivo expansion of conjunctival epithelial stem cells for conjunctival surface reconstruction. PMID:12076097

Meller, Daniel; Dabul, Vanesa; Tseng, Scheffer C G



OCT assessment of aggregation and sedimentation in concentrated RBC suspension: comparison of experimental and Monte Carlo simulated data  

NASA Astrophysics Data System (ADS)

In this work, we use Monte Carlo simulation to obtain model OCT signals from a horizontally orientated blood layer at different stages of red blood cell (RBC) aggregation and sedimentation processes. The parameters for aggregating and sedimenting blood cells were chosen basing on the data available from literature and our earlier experimental studies. Two different models of simulated medium are considered: a suspension of washed RBC in physiological solution (where the aggregation does not take place) and RBC in blood plasma (which provides necessary conditions for aggregation). Good agreement of the simulation results with the available experimental data shows that the chosen optical parameters are reasonable. Dependencies of the numbers of photons contributing to the OCT signal on the number of experienced scattering events were analyzed for each simulated signal. It was shown, that maxima on these dependencies correspond to the peaks in the OCT signals related to the interfaces between the layers of blood plasma and blood cells. Their positions can be calculated from the optical thicknesses of the layers, and the absorption and scattering coefficients of the media.

Kirillin, Mikhail Y.; Priezzhev, Alexander V.; Tuchin, Valery V.; Wang, Ruikang K.; Myllyla, Risto



The Synthesis of Proton Conducting Membranes from Polystyrene Butadiene Rubber for Fuel Cell Application: The Effect of Sulphonating Agents on the Membrane Characteristics  

Microsoft Academic Search

Out of the three major components (membrane, electrocatalyst, and bipolar plates) of proton exchange membrane fuel cell, the cost of the polymer electrolyte membrane is the highest. Therefore, reduction in cost of the membrane will help in reducing the cost of the fuel cell. This work aims at investigating the effect of sulphonating agents (sulphuric acid, acetyl sulphate, and chlorosulphonic

A. S. Abdulkareem; C. A. Idibie; A. S. Afolabi; H. C. Vz Pienaar; S. E. Iyuke; L. Van Dyk



Surface-enhanced Raman imaging of red blood cell membrane with highly uniform active substrates obtained using block copolymers self-assembly  

NASA Astrophysics Data System (ADS)

In this communication, we discuss the application of ordered, ultrahigh-density templates of nano-textured Ag-particles obtained by self-assembling of inorganic-containing polystyrene-block-poly(4-vinylpyridine) copolymer (PS-b-P4VP) micelles, for the spectroscopic surface-enhanced Raman imaging in-vitro of red blood cells (RBCs) and its capability to identify the vibrational fingerprint of the plasma membrane of the cell physisorbed to the SERS substrate. Hexagonal arrays of PS-b-P4VP micelles, with selective inclusion of Ag nanoparticles (NPs) in the polar core, prepared by in situ reduction of a suitable precursor, are obtained by polymer self-assembly upon fast solvent evaporation during spin coating on the supporting substrate. UV irradiation and/or plasma oxygen treatment remove the polymer matrix leaving immobilized nano-islands of Ag-NPs. Such a kind of SERS-active substrate consists of a reproducible and uniform twodimensional hexagonal array of silver clusters with a diameter ranging from 25 to 30 nm (single particles having typically diameters of 5 nm) and nano-island gap distances of the order of 5-8 nm on silicon and 15 nm on glass , while giving rise to high enhancement factors and addressing the issue of SERS reproducibility. The basic substrate supporting the plasmonic coating used in this work is either of silicon or glass. This last allows working in back scattering configuration permitting real time monitoring, via microscopy, of the RBCs on which Raman measurements are being carried out. The template is thus applied for surface-enhanced Raman analysis of the red blood cell (RBC) membrane in confocal micro-Raman configuration demonstrating to have SERS imaging potential thanks to the uniformity of the nano-textured substrate. The first experimental evidence of SERS imaging of a red blood cell membrane in-vitro is demonstrated.

Zito, Gianluigi; Malafronte, Anna; Dochshanov, Alden; Rusciano, Giulia; Auriemma, Finizia; Pesce, Giuseppe; De Rosa, Claudio; Sasso, Antonio



Numerical Study on Flows of Red Blood Cells with Liposome-Encapsulated Hemoglobin at Microvascular Bifurcation  

NASA Astrophysics Data System (ADS)

Flow analysis at microvascular bifurcation after partial replacement of red blood cell (RBC) with liposome-encapsulated hemoglobin (LEH) was performed using the lattice Boltzmann method. A two-dimensional bifurcation model with a parent vessel and daughter branch was considered, and the distributions of the RBC, LEH, and oxygen fluxes were calculated. The immersed boundary method was employed to incorporate the fluid--membrane interaction between the flow field and deformable RBC When only RBCs flow into the daughter branches with unevenly distributed flows, plasma separation occurred and the RBC flow to the lower-flow branch was disproportionately decreased. On the other hand, when the half of RBC are replaced by LEH, the biasing of RBC flow was enhanced whereas LEH flowed favorably into the lower-flow branch, because many LEH within the parent vessel are suspended in the plasma layer, where no RBCs exist. Consequently, the branched oxygen fluxes became nearly proportional to flows. These results indicate that LEH facilitates oxygen supply to branches that are inaccessible to RBCs.

Hyakutake, Toru; Tani, Shigeki; Akagi, Yuki; Matsumoto, Takeshi; Yanase, Shinichiro



A hybrid microbial fuel cell membrane bioreactor with a conductive ultrafiltration membrane biocathode for wastewater treatment.  


A new hybrid, air-biocathode microbial fuel cell-membrane bioreactor (MFC-MBR) system was developed to achieve simultaneous wastewater treatment and ultrafiltration to produce water for direct reclamation. The combined advantages of this system were achieved by using an electrically conductive ultrafiltration membrane as both the cathode and the membrane for wastewater filtration. The MFC-MBR used an air-biocathode, and it was shown to have good performance relative to an otherwise identical cathode containing a platinum catalyst. With 0.1 mm prefiltered domestic wastewater as the feed, the maximum power density was 0.38 W/m(2) (6.8 W/m(3)) with the biocathode, compared to 0.82 W/m(2) (14.5 W/m(3)) using the platinum cathode. The permeate quality from the biocathode reactor was comparable to that of a conventional MBR, with removals of 97% of the soluble chemical oxygen demand, 97% NH3-N, and 91% of total bacteria (based on flow cytometry). The permeate turbidity was <0.1 nephelometric turbidity units. These results show that a biocathode MFC-MBR system can achieve high levels of wastewater treatment with a low energy input due to the lack of a need for wastewater aeration. PMID:24016059

Malaeb, Lilian; Katuri, Krishna P; Logan, Bruce E; Maab, Husnul; Nunes, S P; Saikaly, Pascal E



Cell membrane glycosylation mediates the adhesion, migration, and invasion of ovarian carcinoma cells  

Microsoft Academic Search

We have previously shown that ovarian carcinoma cell adhesion to mesothelial cell monolayers and migration toward fibronectin,\\u000a type IV collagen, and laminin is partially mediated by CD44, a proteoglycan known to affect the functional abilities of tumor\\u000a cells. The purpose of this study was to determine the role of cell membrane glycosylation in the metastatic abilities of ovarian\\u000a carcinoma cells.

Rachael C. Casey; Theodore R. Oegema Jr; Keith M. Skubitz; Stefan E. Pambuccian; Suzanne M. Grindle; Amy P. N. Skubitz



Better Proton-Conducting Polymers for Fuel-Cell Membranes  

NASA Technical Reports Server (NTRS)

Polyoxyphenylene triazole sulfonic acid has been proposed as a basis for development of improved proton-conducting polymeric materials for solid-electrolyte membranes in hydrogen/air fuel cells. Heretofore, the proton-conducting membrane materials of choice have been exemplified by a family of perfluorosulfonic acid-based polymers (Nafion7 or equivalent). These materials are suitable for operation in the temperature of 75 to 85 C, but in order to reduce the sizes and/or increase the energy-conversion efficiencies of fuel-cell systems, it would be desirable to increase temperatures to as high as 120 C for transportation applications, and to as high as 180 C for stationary applications. However, at 120 C and at relative humidity values below 50 percent, the loss of water from perfluorosulfonic acid-based polymer membranes results in fuel-cell power densities too low to be of practical value. Therefore, membrane electrolyte materials that have usefully high proton conductivity in the temperature range of 180 C at low relative humidity and that do not rely on water for proton conduction at 180 C would be desirable. The proposed polyoxyphenylene triazole sulfonic acid-based materials have been conjectured to have these desirable properties. These materials would be free of volatile or mobile acid constituents. The generic molecular structure of these materials is intended to exploit the fact, demonstrated in previous research, that materials that contain ionizable acid and base groups covalently attached to thermally stable polymer backbones exhibit proton conduction even in the anhydrous state.

Narayan, Sri; Reddy, Prakash



Preparation of Highly Sulfonated Ultra-Thin Proton-Exchange Polymer Membranes for Proton Exchange Membrane Fuel Cells  

NASA Astrophysics Data System (ADS)

Sulfonated ultra-thin proton-exchange polymer membrane carrying pyridine groups was made from a plasma polymerization of styrene, 2-vinylpyridine, and trifluoromethanesulfonic acid by after-glow capacitively coupled discharge technique. Pyridine groups tethered to the polymer backbone acts as a medium through the basic nitrogen for transfer of protons between the sulfonic acid groups of proton exchange membrane. It shows that the method using present technology could effectively depress the degradation of monomers during the plasma polymerization. Spectroscopic analyses reveal that the obtained membranes are highly functionalized with proton exchange groups and have higher proton conductivity. Thus, the membranes are expected to be used in direct methanol fuel cells.

Jiang, Zhongqing; Meng, Yuedong; Jiang, Zhong-Jie; Shi, Yicai


Relationship Between the Membrane Envelope of Rhizobial Bacteroids and the Plasma Membrane of the Host Cell as Demonstrated by Histochemical Localization of Adenyl Cyclase  

PubMed Central

By using adenyl cyclase as a marker enzyme, the relationship between the membrane envelope of the bacteroids of rhizobia and the plasma membrane of the host cell was demonstrated histochemically. Electron-dense deposits were found on the outer surface of the plasma membrane of the host cell and on the inner surface of the membrane envelopes of the bacteroids, but not in vacuole membranes, endoplasmic reticula, Golgi apparatus, and mitochondrial membranes. The results suggest that the membrane envelopes of the bacteroids are closely related to the host plasma membrane, and that entry of the bacteroids into the cytoplasm is in a manner similar to endocytosis. Images

Tu, J. C.



Solvation dynamics and intermittent oscillation of cell membrane: live Chinese hamster ovary cell.  


Dynamics of the exofacial thiols (i.e., cell surface thiol containing membrane proteins) of a live Chinese hamster ovary (CHO) cell is probed by time-resolved confocal microscopy. For this purpose, a fluorescent probe, 7-(diethylamino)-3-(4-maleimidophenyl)-4-methylcoumarin (CPM) is covalently attached to the exofacial thiols. The emission maximum of CPM bound exofacial thiols indicates a highly exposed and polar environment. Using CPM, we studied solvation dynamics, for the first time, at the membrane of a live cell. The thiol containing membrane proteins shows ultraslow response with average solvation time, ??s? = 475 ps. CPM labeled exofacial thiols also show spontaneous, intermittent oscillation in fluorescence intensity with a period of 0.5-1.0 s. This is ascribed to reversible, intermittent changes in the structure and conformation of the membrane proteins. PMID:24571625

Ghosh, Shirsendu; Chattoraj, Shyamtanu; Bhattacharyya, Kankan



Long-distance relationships: do membrane nanotubes regulate cell-cell communication and disease progression?  


Metazoan cells rapidly exchange signals at tight cell-cell interfaces, including synapses and gap junctions. Advances in imaging recently exposed a third mode of intercellular cross-talk mediated by thin, actin-containing membrane extensions broadly known as "membrane" or "tunneling" nanotubes. An explosion of research suggests diverse functions for nanotubular superhighways, including cell-cell electrical coupling, calcium signaling, small-molecule exchange, and, remarkably, the transfer of bulky cargoes, including organelles or pathogenic agents. Despite great enthusiasm for all things nanotubular and their potential roles in cell signaling and pathogenesis, key questions remain regarding the mechanisms by which these structures regulate directional cell-cell exchange; how these linkages are formed and between which cells and, critically, whether nanotubes are as prevale