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Sample records for membrane prevents activation

  1. Constitutive activation of a plasma membrane H+-ATPase prevents abscisic acid-mediated stomatal closure

    Merlot, Sylvain; Leonhardt, Nathalie; Fenzi, Francesca; Valon, Christiane; Costa, Miguel; Piette, Laurie; Vavasseur, Alain; Genty, Bernard; Boivin, Karine; Müller, Axel; Giraudat, Jérôme; Leung, Jeffrey

    2007-01-01

    Light activates proton (H+)-ATPases in guard cells, to drive hyperpolarization of the plasma membrane to initiate stomatal opening, allowing diffusion of ambient CO2 to photosynthetic tissues. Light to darkness transition, high CO2 levels and the stress hormone abscisic acid (ABA) promote stomatal closing. The overall H+-ATPase activity is diminished by ABA treatments, but the significance of this phenomenon in relationship to stomatal closure is still debated. We report two dominant mutations in the OPEN STOMATA2 (OST2) locus of Arabidopsis that completely abolish stomatal response to ABA, but importantly, to a much lesser extent the responses to CO2 and darkness. The OST2 gene encodes the major plasma membrane H+-ATPase AHA1, and both mutations cause constitutive activity of this pump, leading to necrotic lesions. H+-ATPases have been traditionally assumed to be general endpoints of all signaling pathways affecting membrane polarization and transport. Our results provide evidence that AHA1 is a distinct component of an ABA-directed signaling pathway, and that dynamic downregulation of this pump during drought is an essential step in membrane depolarization to initiate stomatal closure. PMID:17557075

  2. Plasma membrane disruption: repair, prevention, adaptation

    McNeil, Paul L.; Steinhardt, Richard A.

    2003-01-01

    Many metazoan cells inhabit mechanically stressful environments and, consequently, their plasma membranes are frequently disrupted. Survival requires that the cell rapidly repair or reseal the disruption. Rapid resealing is an active and complex structural modification that employs endomembrane as its primary building block, and cytoskeletal and membrane fusion proteins as its catalysts. Endomembrane is delivered to the damaged plasma membrane through exocytosis, a ubiquitous Ca2+-triggered response to disruption. Tissue and cell level architecture prevent disruptions from occurring, either by shielding cells from damaging levels of force, or, when this is not possible, by promoting safe force transmission through the plasma membrane via protein-based cables and linkages. Prevention of disruption also can be a dynamic cell or tissue level adaptation triggered when a damaging level of mechanical stress is imposed. Disease results from failure of either the preventive or resealing mechanisms.

  3. Anaplasma phagocytophilum inhibits human neutrophil apoptosis via upregulation of bfl-1, maintenance of mitochondrial membrane potential and prevention of caspase 3 activation.

    Ge, Yan; Yoshiie, Kiyotaka; Kuribayashi, Futoshi; Lin, Mingqun; Rikihisa, Yasuko

    2005-01-01

    The inhibition of neutrophil apoptosis plays a central role in human granulocytic anaplasmosis. Intracellular signalling pathways through which the obligatory intracellular bacterium Anaplasma phagocytophilum inhibits the spontaneous apoptosis of human peripheral blood neutrophils were investigated. bfl-1 mRNA levels in uninfected neutrophils after 12 h in culture were reduced to approximately 5-25% of 0 h levels, but remained high in infected neutrophils. The eukaryotic RNA synthesis inhibitor, actinomycin D, prevented the maintenance of bfl-1 mRNA levels by A. phagocytophilum. Differences in mcl-1, bax, bcl-w, bad or bak mRNA levels in infected versus uninfected neutrophils were not remarkable. By using mitochondrial fluorescent dyes, Mitotracker Red and JC-1, it was found that most uninfected neutrophils lost mitochondrial membrane potential after 10-12 h incubation, whereas A. phagocytophilum-infected neutrophils maintained high membrane potential. Caspase 3 activity and the degree of apoptosis were lower in dose-dependent manner in A. phagocytophilum-infected neutrophils at 16 h post infection, as compared to uninfected neutrophils. Anti-active caspase 3 antibody labelling showed less positively stained population in infected neutrophils compared to those in uninfected neutrophils after 12 h incubation. These results suggest that A. phagocytophilum inhibits human neutrophil apoptosis via transcriptional upregulation of bfl-1 and inhibition of mitochondria-mediated activation of caspase 3.

  4. Modeling electrically active viscoelastic membranes.

    Sitikantha Roy

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

  5. Fuel-Cell Structure Prevents Membrane Drying

    Mcelroy, J.

    1986-01-01

    Embossed plates direct flows of reactants and coolant. Membrane-type fuel-cell battery has improved reactant flow and heat removal. Compact, lightweight battery produces high current and power without drying of membranes.

  6. Activation of interfacial enzymes at membrane surfaces

    Mouritsen, Ole G.; Andresen, Thomas Lars; Halperin, Avi

    2006-01-01

    A host of water-soluble enzymes are active at membrane surfaces and in association with membranes. Some of these enzymes are involved in signalling and in modification and remodelling of the membranes. A special class of enzymes, the phospholipases, and in particular secretory phospholipase A2 (s...

  7. Functional implications of plasma membrane condensation for T cell activation.

    Carles Rentero

    2008-05-01

    Full Text Available The T lymphocyte plasma membrane condenses at the site of activation but the functional significance of this receptor-mediated membrane reorganization is not yet known. Here we demonstrate that membrane condensation at the T cell activation sites can be inhibited by incorporation of the oxysterol 7-ketocholesterol (7KC, which is known to prevent the formation of raft-like liquid-ordered domains in model membranes. We enriched T cells with 7KC, or cholesterol as control, to assess the importance of membrane condensation for T cell activation. Upon 7KC treatment, T cell antigen receptor (TCR triggered calcium fluxes and early tyrosine phosphorylation events appear unaltered. However, signaling complexes form less efficiently on the cell surface, fewer phosphorylated signaling proteins are retained in the plasma membrane and actin restructuring at activation sites is impaired in 7KC-enriched cells resulting in compromised downstream activation responses. Our data emphasizes lipids as an important medium for the organization at T cell activation sites and strongly indicates that membrane condensation is an important element of the T cell activation process.

  8. Effective tension and fluctuations in active membranes.

    Loubet, Bastien; Seifert, Udo; Lomholt, Michael Andersen

    2012-03-01

    We calculate the fluctuation spectrum of the shape of a lipid vesicle or cell exposed to a nonthermal source of noise. In particular, we take constraints on the membrane area and the volume of fluid that it encapsulates into account when obtaining expressions for the dependency of the membrane tension on the noise. We then investigate three possible origins of the nonthermal noise taken from the literature: A direct force, which models an external medium pushing on the membrane, a curvature force, which models a fluctuating spontaneous curvature, and a permeation force coming from an active transport of fluid through the membrane. For the direct force and curvature force cases, we compare our results to existing experiments on active membranes.

  9. Use of rhamnolipid biosurfactant for membrane biofouling prevention and cleaning.

    Kim, Lan Hee; Jung, Yongmoon; Kim, Sung-Jo; Kim, Chang-Min; Yu, Hye-Weon; Park, Hee-Deung; Kim, In S

    2015-01-01

    Rhamnolipids were evaluated as biofouling reducing agents in this study. The permeability of the bacterial outer membrane was increased by rhamnolipids while the growth rate of Pseudomonas aeruginosa was not affected. The surface hydrophobicity was increased through the release of lipopolysaccharides and extracellular polymeric substances from the outer cell membrane. Rhamnolipids were evaluated as agents for the prevention and cleaning of biofilms. A high degree of biofilm detachment was observed when the rhamnolipids were used as a cleaning agent. In addition, effective biofilm reduction occurred when rhamnolipids were applied to various species of Gram-negative bacteria isolated from seawater samples. Biofilm reduction using rhamnolipids was comparable to commercially available surfactants. In addition, 20% of the water flux was increased after rhamnolipid treatment (300 μg ml(-1), 6 h exposure time) in a dead-end filtration system. Rhamnolipids appear to have promise as biological agents for reducing membrane biofouling.

  10. Internal Limiting Membrane Peeling to Prevent Post-vitrectomy Epiretinal Membrane Development in Retinal Detachment.

    Akiyama, Kunihiko; Fujinami, Kaoru; Watanabe, Ken; Tsunoda, Kazushige; Noda, Toru

    2016-11-01

    To determine the efficacy of internal limiting membrane (ILM) peeling during vitrectomy for rhegmatogenous retinal detachment (RRD) regarding post-vitrectomy epiretinal membrane (ERM) development and visual outcomes. Retrospective, interventional, comparative case series. Setting: Institutional. One hundred and two consecutive eyes with RRD treated with vitrectomy and followed for at least 6 months. ILM was peeled without using dye such as indocyanine green (ICG). Observational Procedures: Patients were divided into 2 groups based on postoperative ERM development: Group 1, 81 eyes without ERM formation; Group 2, 21 eyes with ERM development. Patients also were divided into 2 subgroups: those with and without ILM peeling (58 and 44 eyes, respectively). Statistical analyses were performed between the 2 groups with/without ERM formation and between the 2 subgroups with/without ILM peeling for 5 preoperative factors including foveal involvement of the RRD, 4 intraoperative factors including ILM peeling, baseline best-corrected visual acuity (BCVA), and final BCVA. An association of ILM peeling with ERM prevention and the influence of ILM peeling on visual outcomes. ILM peeling was significantly (P peeling. ILM peeling without ICG staining during the initial vitrectomy for RRDs may prevent postoperative ERM formation with favorable visual outcomes. Copyright © 2016 Elsevier Inc. All rights reserved.

  11. Mechanics of nonplanar membranes with force-dipole activity

    Lomholt, Michael Andersen

    2006-01-01

    A study is made of how active membrane proteins can modify the long wavelength mechanics of fluid membranes. The activity of the proteins is modelled as disturbing the protein surroundings through nonlocal force distributions of which a force-dipole distribution is the simplest example. An analytic...... contributions to mechanical properties such as tension and bending moments become apparent. It is also explained how the activity can induce a hydrodynamic attraction between the active proteins in the membrane....

  12. DIDS prevents ischemic membrane degradation in cultured hippocampal neurons by inhibiting matrix metalloproteinase release.

    Matthew E Pamenter

    Full Text Available During stroke, cells in the infarct core exhibit rapid failure of their permeability barriers, which releases ions and inflammatory molecules that are deleterious to nearby tissue (the penumbra. Plasma membrane degradation is key to penumbral spread and is mediated by matrix metalloproteinases (MMPs, which are released via vesicular exocytosis into the extracellular fluid in response to stress. DIDS (4,4'-diisothiocyanatostilbene-2,2'-disulphonic acid preserves membrane integrity in neurons challenged with an in vitro ischemic penumbral mimic (ischemic solution: IS and we asked whether this action was mediated via inhibition of MMP activity. In cultured murine hippocampal neurons challenged with IS, intracellular proMMP-2 and -9 expression increased 4-10 fold and extracellular latent and active MMP isoform expression increased 2-22 fold. MMP-mediated extracellular gelatinolytic activity increased ∼20-50 fold, causing detachment of 32.1±4.5% of cells from the matrix and extensive plasma membrane degradation (>60% of cells took up vital dyes and >60% of plasma membranes were fragmented or blebbed. DIDS abolished cellular detachment and membrane degradation in neurons and the pathology-induced extracellular expression of latent and active MMPs. DIDS similarly inhibited extracellular MMP expression and cellular detachment induced by the pro-apoptotic agent staurosporine or the general proteinase agonist 4-aminophenylmercuric acetate (APMA. Conversely, DIDS-treatment did not impair stress-induced intracellular proMMP production, nor the intracellular cleavage of proMMP-2 to the active form, suggesting DIDS interferes with the vesicular extrusion of MMPs rather than directly inhibiting proteinase expression or activation. In support of this hypothesis, an antagonist of the V-type vesicular ATPase also inhibited extracellular MMP expression to a similar degree as DIDS. In addition, in a proteinase-independent model of vesicular exocytosis, DIDS

  13. Stabilization of mitochondrial membrane potential prevents doxorubicin-induced cardiotoxicity in isolated rat heart

    Montaigne, David; Marechal, Xavier; Baccouch, Riadh; Modine, Thomas; Preau, Sebastien; Zannis, Konstantinos; Marchetti, Philippe; Lancel, Steve; Neviere, Remi

    2010-01-01

    The present study was undertaken to examine the effects of doxorubicin on left ventricular function and cellular energy state in intact isolated hearts, and, to test whether inhibition of mitochondrial membrane potential dissipation would prevent doxorubicin-induced mitochondrial and myocardial dysfunction. Myocardial contractile performance and mitochondrial respiration were evaluated by left ventricular tension and its first derivatives and cardiac fiber respirometry, respectively. NADH levels, mitochondrial membrane potential and glucose uptake were monitored non-invasively via epicardial imaging of the left ventricular wall of Langendorff-perfused rat hearts. Heart performance was reduced in a time-dependent manner in isolated rat hearts perfused with Krebs-Henseleit solution containing 1 μM doxorubicin. Compared with controls, doxorubicin induced acute myocardial dysfunction (dF/dt max of 105 ± 8 mN/s in control hearts vs. 49 ± 7 mN/s in doxorubicin-treated hearts; *p < 0.05). In cardiac fibers prepared from perfused hearts, doxorubicin induced depression of mitochondrial respiration (respiratory control ratio of 4.0 ± 0.2 in control hearts vs. 2.2 ± 0.2 in doxorubicin-treated hearts; *p < 0.05) and cytochrome c oxidase kinetic activity (24 ± 1 μM cytochrome c/min/mg in control hearts vs. 14 ± 3 μM cytochrome c/min/mg in doxorubicin-treated hearts; *p < 0.05). Acute cardiotoxicity induced by doxorubicin was accompanied by NADH redox state, mitochondrial membrane potential, and glucose uptake reduction. Inhibition of mitochondrial permeability transition pore opening by cyclosporine A largely prevented mitochondrial membrane potential dissipation, cardiac energy state and dysfunction. These results suggest that in intact hearts an impairment of mitochondrial metabolism is involved in the development of doxorubicin cardiotoxicity.

  14. Membrane Stabilizing Activity And Phytochemistry Of Hibiscus rosa ...

    The human erythrocyte membrane stabilizing activity of saline extract of Hibiscus rosa-sinensis leaves was investigated as part of efforts at validating its use as anti-arthritic and anti-inflammatory agent. The results of the membrane stabilizing activity of the extract, when compared to two non-steroidal anti-inflammatory drugs ...

  15. Glial membranes at the node of Ranvier prevent neurite outgrowth

    Huang, Jeffrey K; Phillips, Greg R; Roth, Alejandro D

    2005-01-01

    of neurite outgrowth, including the oligodendrocyte myelin glycoprotein (OMgp). In rat spinal cord, OMgp was not localized to compact myelin, as previously thought, but to oligodendroglia-like cells, whose processes converge to form a ring that completely encircles the nodes. In OMgp-null mice, CNS nodes......Nodes of Ranvier are regularly placed, nonmyelinated axon segments along myelinated nerves. Here we show that nodal membranes isolated from the central nervous system (CNS) of mammals restricted neurite outgrowth of cultured neurons. Proteomic analysis of these membranes revealed several inhibitors...

  16. Effective tension and fluctuations in active membranes

    Loubet, Bastien; Seifert, Udo; Lomholt, Michael Andersen

    2011-01-01

    We calculate the fluctuation spectrum of the shape of a lipid vesicle or cell exposed to a nonthermal source of noise. In particular we take into account constraints on the membrane area and the volume of fluid that it encapsulates when obtaining expressions for the dependency of the membrane tension on the noise. We then investigate three possible origins of the non-thermal noise taken from the literature: A direct force, which models an external medium pushing on the membrane. A curvature f...

  17. Ascorbic acid prevents nonreceptor specific binding of [3H]-5-hydroxytryptamine to bovine cerebral cortex membranes

    Hamblin, M.W.; Adriaenssens, P.I.; Ariani, K.; Cawthon, R.M.; Stratford, C.A.; Tan, G.L.; Ciaranello, R.D.

    1987-01-01

    [ 3 H]-5-Hydroxytryptamine ([ 3 H]-5-HT) decomposes rapidly when exposed to air in solution at physiological pH if antioxidants are not present. The decomposition products appear to bind to two saturable sites on brain membranes (apparent Kd values = 1-2 and 100-1000 nM). This binding mimics ''specific'' ligand/receptor binding in that it is inhibited by 10 microM unlabeled 5-HT. This inhibition is not competitive, but rather is due to the prevention of [ 3 H]-5-HT breakdown by excess unlabeled 5-HT. Unlike genuine ligand/receptor binding, the binding of [ 3 H]-5-HT breakdown products is essentially irreversible and does not display a tissue distribution consistent with binding to authentic 5-HT receptors. [ 3 H]-5-HT decomposition can be eliminated by the inclusion of 0.05 to 5 mM ascorbic acid. At these concentrations ascorbic acid is not deleterious to reversible [ 3 H]-5-HT binding. When [ 3 H] 5-HT exposure to air occurs in the presence of brain membranes, the apparent antioxidant activity of brain membranes themselves affords protection against [ 3 H]-5-HT degradation equal to ascorbic acid. This protection is effective below final [ 3 H]-5-HT concentrations of 10 nM. Above 10 nM [ 3 H]-5-HT, addition of ascorbic acid or other antioxidants is necessary to avoid the occurrence of additional low affinity (apparent Kd = 15-2000 nM) binding sites that are specific but nonetheless irreversible. When care is taken to limit [ 3 H]-5-HT oxidation, the only reversible and saturable specific binding sites observed are of the 5-HT1 high affinity (Kd = 1-2 nM) type. Radioligand oxidation artifacts may be involved in previous reports of low affinity (Kd = 15-250 nM) [ 3 H]-5-HT binding sites in brain membrane preparations

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

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

    2013-06-04

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

  19. Cell volume and membrane stretch independently control K+ channel activity

    Bomholtz, Sofia Hammami; Willumsen, Niels J; Olsen, Hervør L

    2009-01-01

    A number of potassium channels including members of the KCNQ family and the Ca(2+) activated IK and SK, but not BK, are strongly and reversibly regulated by small changes in cell volume. It has been argued that this general regulation is mediated through sensitivity to changes in membrane stretch...... was not affected by membrane stretch. The results indicate that (1) activation of BK channels by local membrane stretch is not mimicked by membrane stress induced by cell swelling, and (2) activation of KCNQ1 channels by cell volume increase is not mediated by local tension in the cell membrane. We conclude....... To test this hypothesis we have studied the regulation of KCNQ1 and BK channels after expression in Xenopus oocytes. Results from cell-attached patch clamp studies (approximately 50 microm(2) macropatches) in oocytes expressing BK channels demonstrate that the macroscopic volume-insensitive BK current...

  20. Turing Incompleteness of Asynchronous P Systems with Active Membranes

    Leporati, Alberto; Manzoni, Luca; Porreca, Antonio E.

    2013-01-01

    We prove that asynchronous P systems with active membranes without divi- sion rules can be simulated by place/transition Petri nets, and hence are computationally weaker than Turing machines. This result holds even if the synchronisation mechanisms provided by electrical charges and membrane dissolution are exploited.

  1. Physical activity - preventive medicine (image)

    Physical activity contributes to health by reducing the heart rate, decreasing the risk for cardiovascular disease, and reducing ... loss that is associated with age and osteoporosis. Physical activity also helps the body use calories more efficiently, ...

  2. Silver-enhanced block copolymer membranes with biocidal activity

    Madhavan, Poornima

    2014-11-12

    Silver nanoparticles were deposited on the surface and pore walls of block copolymer membranes with highly ordered pore structure. Pyridine blocks constitute the pore surfaces, complexing silver ions and promoting a homogeneous distribution. Nanoparticles were then formed by reduction with sodium borohydride. The morphology varied with the preparation conditions (pH and silver ion concentration), as confirmed by field emission scanning and transmission electron microscopy. Silver has a strong biocide activity, which for membranes can bring the advantage of minimizing the growth of bacteria and formation of biofilm. The membranes with nanoparticles prepared under different pH values and ion concentrations were incubated with Pseudomonas aeruginosa and compared with the control. The strongest biocidal activity was achieved with membranes containing membranes prepared under pH 9. Under these conditions, the best distribution with small particle size was observed by microscopy.

  3. Silver-enhanced block copolymer membranes with biocidal activity

    Madhavan, Poornima; Hong, Pei-Ying; Sougrat, Rachid; Nunes, Suzana Pereira

    2014-01-01

    Silver nanoparticles were deposited on the surface and pore walls of block copolymer membranes with highly ordered pore structure. Pyridine blocks constitute the pore surfaces, complexing silver ions and promoting a homogeneous distribution. Nanoparticles were then formed by reduction with sodium borohydride. The morphology varied with the preparation conditions (pH and silver ion concentration), as confirmed by field emission scanning and transmission electron microscopy. Silver has a strong biocide activity, which for membranes can bring the advantage of minimizing the growth of bacteria and formation of biofilm. The membranes with nanoparticles prepared under different pH values and ion concentrations were incubated with Pseudomonas aeruginosa and compared with the control. The strongest biocidal activity was achieved with membranes containing membranes prepared under pH 9. Under these conditions, the best distribution with small particle size was observed by microscopy.

  4. New insights into membrane-active action in plasma membrane of fungal hyphae by the lipopeptide antibiotic bacillomycin L.

    Zhang, Bao; Dong, Chunjuan; Shang, Qingmao; Han, Yuzhu; Li, Pinglan

    2013-09-01

    Bacillomycin L, a natural iturinic lipopeptide produced by Bacillus amyloliquefaciens, is characterized by strong antifungal activities against a variety of agronomically important filamentous fungi including Rhizoctonia solani Kühn. Prior to this study, the role of membrane permeabilization in the antimicrobial activity of bacillomycin L against plant pathogenic fungi had not been investigated. To shed light on the mechanism of this antifungal activity, the permeabilization of R. solani hyphae by bacillomycin L was investigated and compared with that by amphotericin B, a polyene antibiotic which is thought to act primarily through membrane disruption. Our results derived from electron microscopy, various fluorescent techniques and gel retardation experiments revealed that the antifungal activity of bacillomycin L may be not solely a consequence of fungal membrane permeabilization, but related to the interaction of it with intracellular targets. Our findings provide more insights into the mode of action of bacillomycin L and other iturins, which could in turn help to develop new or improved antifungal formulations or result in novel strategies to prevent fungal spoilage. Crown Copyright © 2013. Published by Elsevier B.V. All rights reserved.

  5. Structural aspects of the antioxidant activity of lutein in a model of photoreceptor membranes

    Wisniewska-Becker, Anna; Nawrocki, Grzegorz; Duda, Mariusz; Subczynski, Witold K.

    2014-01-01

    It was shown that in membranes containing raft domains, the macular xanthophylls lutein and zeaxanthin are not distributed uniformly, but are excluded from saturated raft domains and about ten times more concentrated in unsaturated bulk lipids. The selective accumulation of lutein and zeaxanthin in direct proximity to unsaturated lipids, which are especially susceptible to lipid peroxidation, could be very important as far as their antioxidant activity is concerned. Therefore, the protective role of lutein against lipid peroxidation was investigated in membranes made of raft-forming mixtures and in models of photoreceptor outer segment membranes and compared with their antioxidant activity in homogeneous membranes composed of unsaturated lipids. Lipid peroxidation was induced by photosensitized reactions using rose Bengal and monitored by an MDA-TBA test, an iodometric assay, and oxygen consumption (using EPR spectroscopy and the mHCTPO spin label as an oxygen probe). The results show that lutein protects unsaturated lipids more effectively in membranes made of raft-forming mixtures than in homogeneous membranes. This suggests that the selective accumulation of macular xanthophylls in the most vulnerable regions of photoreceptor membranes may play an important role in enhancing their antioxidant properties and ability to prevent age-related macular diseases (such as age-related macular degeneration [AMD]). PMID:22428148

  6. Ovalbumin with Glycated Carboxyl Groups Shows Membrane-Damaging Activity

    Ching-Chia Tang

    2017-02-01

    Full Text Available The aim of the present study was to investigate whether glycated ovalbumin (OVA showed novel activity at the lipid-water interface. Mannosylated OVA (Man-OVA was prepared by modification of the carboxyl groups with p-aminophenyl α-dextro (d-mannopyranoside. An increase in the number of modified carboxyl groups increased the membrane-damaging activity of Man-OVA on cell membrane-mimicking vesicles, whereas OVA did not induce membrane permeability in the tested phospholipid vesicles. The glycation of carboxyl groups caused a notable change in the gross conformation of OVA. Moreover, owing to their spatial positions, the Trp residues in Man-OVA were more exposed, unlike those in OVA. Fluorescence quenching studies suggested that the Trp residues in Man-OVA were located on the interface binds with the lipid vesicles, and their microenvironment was abundant in positively charged residues. Although OVA and Man-OVA showed a similar binding affinity for lipid vesicles, the lipid-interacting feature of Man-OVA was distinct from that of OVA. Chemical modification studies revealed that Lys and Arg residues, but not Trp residues, played a crucial role in the membrane-damaging activity of Man-OVA. Taken together, our data suggest that glycation of carboxyl groups causes changes in the structural properties and membrane-interacting features of OVA, generating OVA with membrane-perturbing activities at the lipid-water interface.

  7. Modulation of hyaluronan synthase activity in cellular membrane fractions.

    Vigetti, Davide; Genasetti, Anna; Karousou, Evgenia; Viola, Manuela; Clerici, Moira; Bartolini, Barbara; Moretto, Paola; De Luca, Giancarlo; Hascall, Vincent C; Passi, Alberto

    2009-10-30

    Hyaluronan (HA), the only non-sulfated glycosaminoglycan, is involved in morphogenesis, wound healing, inflammation, angiogenesis, and cancer. In mammals, HA is synthesized by three homologous HA synthases, HAS1, HAS2, and HAS3, that polymerize the HA chain using UDP-glucuronic acid and UDP-N-acetylglucosamine as precursors. Since the amount of HA is critical in several pathophysiological conditions, we developed a non-radioactive assay for measuring the activity of HA synthases (HASs) in eukaryotic cells and addressed the question of HAS activity during intracellular protein trafficking. We prepared three cellular fractions: plasma membrane, cytosol (containing membrane proteins mainly from the endoplasmic reticulum and Golgi), and nuclei. After incubation with UDP-sugar precursors, newly synthesized HA was quantified by polyacrylamide gel electrophoresis of fluorophore-labeled saccharides and high performance liquid chromatography. This new method measured HAS activity not only in the plasma membrane fraction but also in the cytosolic membranes. This new technique was used to evaluate the effects of 4-methylumbeliferone, phorbol 12-myristate 13-acetate, interleukin 1beta, platelet-derived growth factor BB, and tunicamycin on HAS activities. We found that HAS activity can be modulated by post-translational modification, such as phosphorylation and N-glycosylation. Interestingly, we detected a significant increase in HAS activity in the cytosolic membrane fraction after tunicamycin treatment. Since this compound is known to induce HA cable structures, this result links HAS activity alteration with the capability of the cell to promote HA cable formation.

  8. Physical Activity and the Prevention of Hypertension

    Diaz, Keith M.; Shimbo, Daichi

    2013-01-01

    As the worldwide prevalence of hypertension continues to increase, the primary prevention of hypertension has become an important global public health initiative. Physical activity is commonly recommended as an important lifestyle modification that may aid in the prevention of hypertension. Recent epidemiologic evidence has demonstrated a consistent, temporal, and dose-dependent relationship between physical activity and the development of hypertension. Experimental evidence from interventional studies have further confirmed a relationship between physical activity and hypertension as the favorable effects of exercise on blood pressure reduction have been well characterized in recent years. Despite the available evidence strongly supporting a role for physical activity in the prevention of hypertension, many unanswered questions regarding the protective benefits of physical activity in high-risk individuals, the factors that may moderate the relationship between physical activity and hypertension, and the optimal prescription for hypertension prevention remain. We review the most recent evidence for the role of physical activity in the prevention of hypertension and discuss recent studies that have sought to address these unanswered questions. PMID:24052212

  9. A study of monoamine oxidase activity in fetal membranes.

    Sekizawa, A; Ishikawa, H; Morimoto, T; Hirose, K; Suzuki, A; Saito, H; Yanaihara, T; Arai, Y; Oguchi, K

    1996-05-01

    To study the role of decidual monoamine oxidase (MAO)-A and -B activities before delivery, the relationship between MAO activity in fetal membranes and catecholamine (CA) concentration in amniotic fluid (AF) was determined. Fetal membranes and AF were obtained at the time of elective Cesarean section (CS group, n = 11) and Cesarean section due to fetal distress without labor pains (FD group, n = 5). MAO-A and -B activities were radiometrically measured using 14C-5-hydroxytriptamine for MAO-A substrate and 14C-benzylamine for MAO-B substrate. CA concentrations in AF were measured by high performance liquid chromatograph with an electro-chemical detector. Both MAO-A and -B activities in decidua obtained from CS were significantly lower than those obtained from FD. Both norepinephrine (NE) and epinephrine (EP) concentrations were significantly lower in the CS group than the FD group. A significant positive correlation between decidual MAO-A activity and NE concentration in AF was observed. No significant correlation was observed between MAO-B activity and the concentration of NE in AF. There was no correlation between EP concentrations and MAO activities. These results suggest that CA concentration in AF may be related to the activity of MAO in fetal membranes, determined by certain physiological processes during pregnancy. It has been suggested that metabolism of monoamines in fetal membranes also plays an important role in reducing monoamine influx into maternal myometrium from the AF.

  10. Role of the membrane skeleton in preventing the shedding of procoagulant-rich microvesicles from the platelet plasma membrane

    1990-01-01

    The platelet plasma membrane is lined by a membrane skeleton that appears to contain short actin filaments cross-linked by actin-binding protein. Actin-binding protein is in turn associated with specific plasma membrane glycoproteins. The aim of this study was to determine whether the membrane skeleton regulates properties of the plasma membrane. Platelets were incubated with agents that disrupted the association of the membrane skeleton with membrane glycoproteins. The consequences of this c...

  11. Aquatic biofouling prevention by electrically charged nanocomposite polymer thin film membranes.

    de Lannoy, Charles-François; Jassby, David; Gloe, Katie; Gordon, Alexander D; Wiesner, Mark R

    2013-03-19

    Electrically conductive polymer-nanocomposite (ECPNC) tight nanofiltration (NF) thin film membranes were demonstrated to have biofilm-preventing capabilities under extreme bacteria and organic material loadings. A simple route to the creation and application of these polyamide-carbon nanotube thin films is also reported. These thin films were characterized with SEM and TEM as well as FTIR to demonstrate that the carbon nanotubes are embedded within the polyamide and form ester bonds with trimesoyl chloride, one of the monomers of polyamide. These polymer nanocomposite thin film materials boast high electrical conductivity (∼400 S/m), good NaCl rejection (>95%), and high water permeability. To demonstrate these membranes' biofouling capabilities, we designed a cross-flow water filtration vessel with insulated electrical leads connecting the ECPNC membranes to an arbitrary waveform generator. In all experiments, conducted in highly bacterially contaminated LB media, flux tests were run until fluxes decreased by 45 ± 3% over initial flux. Biofilm-induced, nonreversible flux decline was observed in all control experiments and a cross-flow rinse with the feed solution failed to induce flux recovery. In contrast, flux decrease for the ECPNC membranes with an electric potential applied to their surface was only caused by deposition of bacteria rather than bacterial attachment, and flux was fully recoverable following a short rinse with the feed solution and no added cleaning agents. The prevention of biofilm formation on the ECPNC membranes was a long-term effect, did not decrease with use, and was highly reproducible.

  12. [Ketoconazole and clindamycin efficacy vs oral clindamycin in premature membranes rupture prevention].

    Castillo Huerta, Eugenio; Garibay Valencia, Miguel; Mirabent-González, Felio

    2008-07-01

    Vulvovaginitis is one of the main causes of premature membrane rupture. To evaluate effectiveness of a combination of ketoconazole (400 mg) and clindamycin (100 mg) in vaginal tablets, compared with clindamicyn alone (600 mg/daily) orally, for six days, to prevent premature membrane rupture in patients with vulvovaginitis. Longitudinal, prospective, comparative, randomized, double-blind, double-dummy study in patients older than 18 years, during them third trimester of normoevolutive pregnancy with symptomatic vulvovaginitis. Patients were monitored as out patient. Genital secretion culture and fresh studies were made. Signs and symptoms were evaluated in regular intervals: 4, 7 and 11 days. Pregnancy control was performed every three weeks, until childbirth or premature membrane rupture. 105 patients were included: 53 in the group of ketoconazole and clindamicyn (1), and 52 in the group of clindamycin alone (2). Symptoms were similar in both groups of treatment, without statistically significant differences. A case of group 2 has premature membrane rupture (p = 0.495). C. albicans was cultured in 35% of group 2 and in 11% of group 2. No adverse events with treatments were reported. The combination of ketoconazole and cindamycin was effective to prevent premature membrane rupture in patients with vulvovaginitis.

  13. PLGA nanofiber membranes loaded with epigallocatechin-3-O-gallate are beneficial to prevention of postsurgical adhesions

    Shin YC

    2014-08-01

    both scavenging reactive oxygen species and extending activated partial thromboplastin time. Macroscopic observation after 1 week of surgical treatment revealed that the antiadhesion efficacy of E-PLGA nanofiber membranes was significantly superior to those of untreated controls and pure PLGA equivalents, which was comparable to that of a commercial tissue-adhesion barrier. In conclusion, the E-PLGA hybrid nanofiber can be exploited to craft strategies for the prevention of postsurgical adhesions. Keywords: nanofiber membrane, poly(lactic-co-glycolic acid, epigallocatechin-3-O-gallate, antiadhesion, tissue-adhesion barrier

  14. Large-Aperture Membrane Active Phased-Array Antennas

    Karasik, Boris; McGrath, William; Leduc, Henry

    2009-01-01

    Large-aperture phased-array microwave antennas supported by membranes are being developed for use in spaceborne interferometric synthetic aperture radar systems. There may also be terrestrial uses for such antennas supported on stationary membranes, large balloons, and blimps. These antennas are expected to have areal mass densities of about 2 kg/sq m, satisfying a need for lightweight alternatives to conventional rigid phased-array antennas, which have typical areal mass densities between 8 and 15 kg/sq m. The differences in areal mass densities translate to substantial differences in total mass in contemplated applications involving aperture areas as large as 400 sq m. A membrane phased-array antenna includes patch antenna elements in a repeating pattern. All previously reported membrane antennas were passive antennas; this is the first active membrane antenna that includes transmitting/receiving (T/R) electronic circuits as integral parts. Other integral parts of the antenna include a network of radio-frequency (RF) feed lines (more specifically, a corporate feed network) and of bias and control lines, all in the form of flexible copper strip conductors on flexible polymeric membranes. Each unit cell of a prototype antenna (see Figure 1) contains a patch antenna element and a compact T/R module that is compatible with flexible membrane circuitry. There are two membrane layers separated by a 12.7-mm air gap. Each membrane layer is made from a commercially available flexible circuit material that, as supplied, comprises a 127-micron-thick polyimide dielectric layer clad on both sides with 17.5-micron-thick copper layers. The copper layers are patterned into RF, bias, and control conductors. The T/R module is located on the back side of the ground plane and is RF-coupled to the patch element via a slot. The T/R module is a hybrid multilayer module assembled and packaged independently and attached to the membrane array. At the time of reporting the information for

  15. Conditions of activation of yeast plasma membrane ATPase.

    Sychrová, H; Kotyk, A

    1985-04-08

    The in vivo activation of the H+-ATPase of baker's yeast plasma membrane found by Serrano in 1983 was demonstrated with D-glucose aerobically and anaerobically (as well as in a respiration-deficient mutant) and, after suitable induction, with maltose, trehalose, and galactose. The activated but not the control ATPase was sensitive to oligomycin. No activation was possible in a cell-free extract with added glucose. The ATPase was not activated in yeast protoplasts which may account for the absence of glucose-stimulated secondary active transports in these wall-less cells and provide support for a microscopic coupling between ATPase activity and these transports in yeast cells.

  16. Activated sludge filterability and full-scale membrane bioreactor operation

    Krzeminski, P.

    2013-01-01

    Despite continuous developments in the field of MBR technology, membrane fouling together with the associated energy demand and related costs issues remain major challenges. The efficiency of the filtration process in an MBR is governed by the activated sludge filterability, which is still limitedly

  17. Anti-glycophorin C induces mitochondrial membrane depolarization and a loss of extracellular regulated kinase 1/2 protein kinase activity that is prevented by pretreatment with cytochalasin D: implications for hemolytic disease of the fetus and newborn caused by anti-Ge3.

    Micieli, Jonathan A; Wang, Duncheng; Denomme, Gregory A

    2010-08-01

    Anti-glycophorin C (GPC), blood group antibodies of which cause hemolytic disease of the fetus and newborn (HDFN), is a potent inhibitor of erythroid progenitor cell growth. The cellular mechanism for growth inhibition has not been characterized. K562 cells were incubated in the presence of either anti-GPC, an immunoglobulin G isotype control, an inhibitor of actin polymerization called cytochalasin D with anti-GPC, or cytochalasin D alone. The JC-1 cationic dye was used to detect mitochondrial depolarization and the activity of the mitogen-activated protein kinases was assessed by Western blotting. Anti-GPC inhibits the activity of extracellular regulated kinase (ERK)1/2 within 10 minutes but does not alter the activity of p38 or c-Jun N-terminal kinase. After 24 hours there was a significant loss of mitochondrial membrane potential compared to isotype control–treated cells. Both the ERK1/2 inhibition and the loss of mitochondrial potential were prevented by pretreatment with cytochalasin D. A cell surface antibody can cause anemia by altering the signaling pathways in erythroid cells by promoting depolarization of mitochondria via cytoskeletal rearrangement. The observation that neonates with anti-GPC HDFN are unresponsive to erythropoietin can be explained by the antibody inhibiting a protein kinase through which this hematopoietic growth factor achieves its effects.

  18. DETRIMENTAL EFFECTS OF ACTIVE INTERNAL LIMITING MEMBRANE PEELING DURING EPIRETINAL MEMBRANE SURGERY: Microperimetric Analysis.

    Deltour, Jean-Baptiste; Grimbert, Pierre; Masse, Helene; Lebreton, Olivier; Weber, Michel

    2017-03-01

    The aim of the study was to assess the microperimetric consequences of active internal limiting membrane (ILM) peeling during idiopathic epimacular membrane (IEMM) surgery. This retrospective monocentric study included 32 eyes of 31 consecutive patients who underwent IEMM surgery. Internal limiting membrane integrity was assessed by ILM Blue staining after IEMM removal: peeling was spontaneous (Group S) or active (Group A). Preprocedure and postprocedure (1 and 6 months) examinations were performed using visual acuity determination, spectral domain optical coherence tomography and microperimetry. Twenty-two eyes had an "active ILM peeling" and 10 a "spontaneous ILM peeling." Both groups had comparable and significant improvements in visual acuity 6 months after surgery (+1.82 lines [+9 letters] [Group A] and +1.51 lines [+8 letters] [Group S], P peeling has progressively become generalized in IEMM surgery to reduce recurrences. This additional procedure does not change the postoperative visual acuity but increases the development of deeper microscotomas. The real impact on the quality of vision remains unclear. Active ILM peeling in IEMM surgery may be responsible for visual impairment related to its microtraumatic effects.

  19. The cAMP-activated GTP exchange factor, Epac1 Upregulates Plasma Membrane and Nuclear Akt Kinase Activities in 8-CPT-2-O-Me-cAMP-Stimulated Macrophages: Gene silencing of the cAMP-activated GTP exchange Epac1 prevents 8-CPT-2-O-Me-cAMP activation of Akt activity in macrophages*

    Misra, Uma K.; Kaczowka, Steven; Pizzo, Salvatore V.

    2008-01-01

    cAMP regulates a wide range of processes through its downstream effectors including PKA, and the family of guanine nucleotide exchange factors. Depending on the cell type, cAMP inhibits or stimulates growth and proliferation in a PKA-dependent or independent manner. PKA-independent effects are mediated by PI 3-kinases-Akt signaling and EPAC1 (exchange protein directly activated by cAMP) activation. Recently, we reported PKA-independent activation of the protein kinase Akt as well co-immunopre...

  20. Antimycotic activity of fengycin C biosurfactant and its interaction with phosphatidylcholine model membranes.

    González-Jaramillo, Lina María; Aranda, Francisco José; Teruel, José Antonio; Villegas-Escobar, Valeska; Ortiz, Antonio

    2017-08-01

    Lipopeptide biosurfactants constitute one of the most promising groups of compounds for the treatment and prevention of fungal diseases in plants. Bacillus subtilis strain EA-CB0015 produces iturin A, fengycin C and surfactin and it has been proven useful for the treatment of black Sigatoka disease in banana plants, an important pathology caused by the fungus Mycosphaerella fijiensis (Morelet). We have found that B. subtilis EA-CB0015 cell free supernatants and purified fractions inhibit M. fijiensis cellular growth. The effect of the purified lipopeptides mentioned above on fungal growth has been also evaluated, observing that iturin A and fengycin C inhibit mycelial growth and ascospore germination, whereas surfactin is not effective. On the hypothesis that the antifungal action of the lipopeptides is associated to their incorporation into biological membranes, ultimately leading to membrane permeabilization, a detailed biophysical study on the interaction of a new isoform of fengycin C with model dipalmitoyphosphatidylcholine (DPPC) membranes has been carried out. Differential scanning calorimetry shows that fengycin C alters the thermotropic phase transitions of DPPC, and is laterally segregated in the fluid bilayer forming domains. Fluorescent probe polarization measurements show that fengycin C does not affect the hydrophobic interior of the membrane. This latter perturbation is concomitant with a strong dehydration of the polar region of DPPC, as shown by FTIR. Fengycin-rich domains, where the surrounding DPPC molecules are highly dehydrated, may well constitute sites of membrane permeabilization leading to a leaky target membrane. These results are a solid support to explain the membrane perturbing action of fengycin, which has been related to its antifungal activity. Copyright © 2017 Elsevier B.V. All rights reserved.

  1. Preparation of Cu{sub 2}O nanowire-blended polysulfone ultrafiltration membrane with improved stability and antimicrobial activity

    Xu, Zehai; Ye, Shuaiju; Fan, Zheng; Ren, Fanghua; Gao, Congjie [Zhejiang University of Technology, Institute of Oceanic and Environmental Chemical Engineering, College of Chemical Engineering and Material Science and College of Ocean, and State Key Lab Breeding Base of Green Chemical Synthesis Technology and Zhejiang Collaborative Innovation Center of Membrane Separation and Water Treatment (China); Li, Qingbiao; Li, Guoqing [Quanzhou Normal University, College of Chemistry and Life Science (China); Zhang, Guoliang, E-mail: membrane86571@163.com, E-mail: guoliangz@zjut.edu.cn [Zhejiang University of Technology, Institute of Oceanic and Environmental Chemical Engineering, College of Chemical Engineering and Material Science and College of Ocean, and State Key Lab Breeding Base of Green Chemical Synthesis Technology and Zhejiang Collaborative Innovation Center of Membrane Separation and Water Treatment (China)

    2015-10-15

    Polysulfone (PSF) membranes have been widely applied in water and wastewater treatment, food-processing and biomedical fields. In this study, we report the preparation of modified PSF membranes by blending PSF with Cu{sub 2}O nanowires (NWs) to improve their stability and antifouling activity. Synthesis of novel Cu{sub 2}O NWs/PSF-blended ultrafiltration membrane was achieved via phase inversion method by dispersing one-dimensional Cu{sub 2}O nanowires in PSF casting solutions. Various techniques such as XRD, SEM, TEM, and EDS were applied to characterize and investigate the properties of nanowires and membranes. The introduced Cu{sub 2}O nanowires can firmly be restricted into micropores of PSF membranes, and therefore, they can effectively prevent the serious leaking problem of inorganic substances in separation process. The blended PSF membranes also provided enhanced antimicrobial activity and superior permeation property compared to pure PSF membrane. The overall work can not only provide a new way for preparation of novel blended membranes with multidimensional nanomaterials, but can also be beneficial to solve the annoying problem of biofouling.

  2. Crime prevention through sports and physical activity

    Dimovski Darko

    2015-01-01

    Full Text Available Starting from the definition of sport, the author has presented the possibilities its application in the prevention of crime and delinquency. In that context, the author analyzes the rate of juvenile delinquency in specific countries, such as Canada, and underlines the fact that the classical criminal measures do not give adequate results. The author points out that it is, therefore, necessary to apply some other preventive measures, which embody the application of sports and physical activity. The author provides examples of good practice in the states which has achieved the best results in the development of such programs. Finally, in view of the increasing number of reported criminal offences committed by both juveniles and adults, the author highlights the need for developing such programs in the Republic of Serbia.

  3. Modification of Salmonella Lipopolysaccharides Prevents the Outer Membrane Penetration of Novobiocin

    Nobre, Thatyane M.; Martynowycz, Michael W.; Andreev, Konstantin; Kuzmenko, Ivan; Nikaido, Hiroshi; Gidalevitz, David

    2015-12-01

    Small hydrophilic antibiotics traverse the outer membrane of Gram-negative bacteria through porin channels. Large lipophilic agents traverse the outer membrane through its bilayer, containing a majority of lipopolysaccharides in its outer leaflet. Genes controlled by the two-component regulatory system PhoPQ modify lipopolysaccharides. We isolate lipopolysaccharides from isogenic mutants of Salmonella sp., one lacking the modification, the other fully modified. These lipopolysaccharides were reconstituted asmonolayers at the air-water interface, and their properties, aswell as their interaction with a large lipophilic drug, novobiocin, was studied. X-ray reflectivity showed that the drug penetrated the monolayer of the unmodified lipopolysaccharides reaching the hydrophobic region,butwas prevented fromthis penetration intothemodified lipopolysaccharides.Results correlatewith behavior of bacterial cells, which become resistant to antibiotics after PhoPQ-regulated modifications. Grazing incidence x-ray diffraction showed that novobiocin produced a striking increase in crystalline coherence length, and the size of the near-crystalline domains.

  4. Photosynthetic solar cell using nanostructured proton exchange membrane for microbial biofilm prevention.

    Lee, Dong Hyun; Oh, Hwa Jin; Bai, Seoung Jae; Song, Young Seok

    2014-06-24

    Unwanted biofilm formation has a detrimental effect on bioelectrical energy harvesting in microbial cells. This issue still needs to be solved for higher power and longer durability and could be resolved with the help of nanoengineering in designing and manufacturing. Here, we demonstrate a photosynthetic solar cell (PSC) that contains a nanostructure to prevent the formation of biofilm by micro-organisms. Nanostructures were fabricated using nanoimprint lithography, where a film heater array system was introduced to precisely control the local wall temperature. To understand the heat and mass transfer phenomena behind the manufacturing and energy harvesting processes of PSC, we carried out a numerical simulation and experimental measurements. It revealed that the nanostructures developed on the proton exchange membrane enable PSC to produce enhanced output power due to the retarded microbial attachment on the Nafion membrane. We anticipate that this strategy can provide a pathway where PSC can ensure more renewable, sustainable, and efficient energy harvesting performance.

  5. Erythrocyte membrane stabilization effect and antioxidant activity of methyl methacrylate

    Popov, B.

    2004-01-01

    Methyl methacrylate (MMK) is a synthetic product with mild impact on human health that is not well studied on cellular basis. Here, human erythrocytes were used to investigate the effects MMK exerts on acid and heat-induced hemolysis. Biphasic effect of MMK was observed for acid-induced hemolysis; i.e., protection at low (0 - 0.05% v/v) and stimulation at higher (0.1- 0.4% v/v) concentrations. The maximal protective effect was produced at 0.03% (v/v). At this concentration MMK increased the temperatures of heat denaturation of erythrocyte membrane proteins, spectrin and integral proteins, by about 2 0 C and inhibited the heat-induced hemolysis by 20 %. This membrane stabilization effect of MMK is similar to that produced by some anti-inflammatory and antirheumatic drugs. The increased acid resistance possibly indicated anti-oxidant properties of MMK. The nonenzymatic antioxidant activity test evidenced that MMK has no superoxide dismutase-like activity but demonstrates strong catalase-like activity (about 900 kU/mmol at 0.05-0.1 mmol/l concentration). The results indicate that at low concentration MMK exerts benign effect on cellular membrane that could find therapeutic usage. (author)

  6. Photosynthesis Activates Plasma Membrane H+-ATPase via Sugar Accumulation.

    Okumura, Masaki; Inoue, Shin-Ichiro; Kuwata, Keiko; Kinoshita, Toshinori

    2016-05-01

    Plant plasma membrane H(+)-ATPase acts as a primary transporter via proton pumping and regulates diverse physiological responses by controlling secondary solute transport, pH homeostasis, and membrane potential. Phosphorylation of the penultimate threonine and the subsequent binding of 14-3-3 proteins in the carboxyl terminus of the enzyme are required for H(+)-ATPase activation. We showed previously that photosynthesis induces phosphorylation of the penultimate threonine in the nonvascular bryophyte Marchantia polymorpha However, (1) whether this response is conserved in vascular plants and (2) the process by which photosynthesis regulates H(+)-ATPase phosphorylation at the plasma membrane remain unresolved issues. Here, we report that photosynthesis induced the phosphorylation and activation of H(+)-ATPase in Arabidopsis (Arabidopsis thaliana) leaves via sugar accumulation. Light reversibly phosphorylated leaf H(+)-ATPase, and this process was inhibited by pharmacological and genetic suppression of photosynthesis. Immunohistochemical and biochemical analyses indicated that light-induced phosphorylation of H(+)-ATPase occurred autonomously in mesophyll cells. We also show that the phosphorylation status of H(+)-ATPase and photosynthetic sugar accumulation in leaves were positively correlated and that sugar treatment promoted phosphorylation. Furthermore, light-induced phosphorylation of H(+)-ATPase was strongly suppressed in a double mutant defective in ADP-glucose pyrophosphorylase and triose phosphate/phosphate translocator (adg1-1 tpt-2); these mutations strongly inhibited endogenous sugar accumulation. Overall, we show that photosynthesis activated H(+)-ATPase via sugar production in the mesophyll cells of vascular plants. Our work provides new insight into signaling from chloroplasts to the plasma membrane ion transport mechanism. © 2016 American Society of Plant Biologists. All Rights Reserved.

  7. Exploring the biophysical properties of phytosterols in the plasma membrane for novel cancer prevention strategies.

    Fakih, Omar; Sanver, Didem; Kane, David; Thorne, James L

    2018-05-03

    Cancer is a global problem with no sign that incidences are reducing. The great costs associated with curing cancer, through developing novel treatments and applying patented therapies, is an increasing burden to developed and developing nations alike. These financial and societal problems will be alleviated by research efforts into prevention, or treatments that utilise off-patent or repurposed agents. Phytosterols are natural components of the diet found in an array of seeds, nuts and vegetables and have been added to several consumer food products for the management of cardio-vascular disease through their ability to lower LDL-cholesterol levels. In this review, we provide a connected view between the fields of structural biophysics and cellular and molecular biology to evaluate the growing evidence that phytosterols impair oncogenic pathways in a range of cancer types. The current state of understanding of how phytosterols alter the biophysical properties of plasma membrane is described, and the potential for phytosterols to be repurposed from cardio-vascular to oncology therapeutics. Through an overview of the types of biophysical and molecular biology experiments that have been performed to date, this review informs the reader of the molecular and biophysical mechanisms through which phytosterols could have anti-cancer properties via their interactions with the plasma cell membrane. We also outline emerging and under-explored areas such as computational modelling, improved biomimetic membranes and ex vivo tissue evaluation. Focus of future research in these areas should improve understanding, not just of phytosterols in cancer cell biology but also to give insights into the interaction between the plasma membrane and the genome. These fields are increasingly providing meaningful biological and clinical data but iterative experiments between molecular biology assays, biosynthetic membrane studies and computational membrane modelling improve and refine our

  8. Thin Film Polyamide Membranes with Photoresponsive Antibacterial Activity

    Duong, Phuoc H. H.

    2017-08-09

    Membranes containing a photosensitizer molecule as part of the selective layer are proposed with demonstrated anti-biofouling activity. For the membrane preparation, mixtures of an amine-functionalized photosensitizer molecule, (5,10,15,20-(tetra-4-aminophenyl)porphyrin) and m-phenylene diamine (MPD) reacted with trimesoyl chloride (TMC) by interfacial polymerization to form thin polyamide films on top of an asymmetric porous support. A highly permeable membrane (35.4 Lm−2h−1bar−1) with 99 % rejection of Brilliant Blue R (826 g/mol) was obtained using 0.25 wt% porphyrin and 0.75 wt% MPD as amine monomers. Under visible light exposure, singlet oxygen (1O2) is generated in the porphyrin containing-polyamide film, reaching the bacteria in the feed by diffusion and enhancing the biofouling resistance and anti-microbial activity. Anti-biofouling and anti-microbial photoactivity in solution are demonstrated on Staphylococcus aureus at different porphyrin concentrations and light exposure time.

  9. Thin Film Polyamide Membranes with Photoresponsive Antibacterial Activity

    Duong, Phuoc H. H.; Hong, Pei-Ying; Musteata, Valentina-Elena; Peinemann, Klaus-Viktor; Nunes, Suzana Pereira

    2017-01-01

    Membranes containing a photosensitizer molecule as part of the selective layer are proposed with demonstrated anti-biofouling activity. For the membrane preparation, mixtures of an amine-functionalized photosensitizer molecule, (5,10,15,20-(tetra-4-aminophenyl)porphyrin) and m-phenylene diamine (MPD) reacted with trimesoyl chloride (TMC) by interfacial polymerization to form thin polyamide films on top of an asymmetric porous support. A highly permeable membrane (35.4 Lm−2h−1bar−1) with 99 % rejection of Brilliant Blue R (826 g/mol) was obtained using 0.25 wt% porphyrin and 0.75 wt% MPD as amine monomers. Under visible light exposure, singlet oxygen (1O2) is generated in the porphyrin containing-polyamide film, reaching the bacteria in the feed by diffusion and enhancing the biofouling resistance and anti-microbial activity. Anti-biofouling and anti-microbial photoactivity in solution are demonstrated on Staphylococcus aureus at different porphyrin concentrations and light exposure time.

  10. Modulation of Erythrocyte Plasma Membrane Redox System Activity by Curcumin

    Prabhakar Singh

    2016-01-01

    Full Text Available Plasma membrane redox system (PMRS is an electron transport chain system ubiquitously present throughout all cell types. It transfers electron from intracellular substrates to extracellular acceptors for regulation of redox status. Curcumin, isolated from Curcuma longa, has modulatory effects on cellular physiology due to its membrane interaction ability and antioxidant potential. The present study investigates the effect of curcumin on PMRS activity of erythrocytes isolated from Wistar rats in vitro and in vivo and validated through an in silico docking simulation study using Molegro Virtual Docker (MVD. Effects of curcumin were also evaluated on level of glutathione (GSH and the oxidant potential of plasma measured in terms of plasma ferric equivalent oxidative potentials (PFEOP. Results show that curcumin significantly (p<0.01 downregulated the PMRS activity in a dose-dependent manner. Molecular docking results suggest that curcumin interacts with amino acids at the active site cavity of cytochrome b5 reductase, a key constituent of PMRS. Curcumin also increased the GSH level in erythrocytes and plasma while simultaneously decreasing the oxidant potential (PFEOP of plasma. Altered PMRS activity and redox status are associated with the pathophysiology of several health complications including aging and diabetes; hence, the above finding may explain part of the role of curcumin in health beneficial effects.

  11. AN-69 membrane reactions are pH-dependent and preventable.

    Brophy, P D; Mottes, T A; Kudelka, T L; McBryde, K D; Gardner, J J; Maxvold, N J; Bunchman, T E

    2001-07-01

    We report two pediatric patients who required blood priming for continuous venovenous hemodiafiltration. Both of these patients developed a significant hypotensive episode with initiation of continuous venovenous hemodiafiltration with immediate resolution on discontinuation. The most notable common characteristics of these patients were the use of the Multi-flo 60 (AN-69) dialyzer membrane and blood priming. No similar episodes were encountered when patients were primed with saline or albumin. The AN-69 membrane is exquisitely pH sensitive. The lower the pH concentration of the blood passing by the membrane, the greater the activation of bradykinin, a known hypotensive-inducing agent, by the dialyzer. On review of blood available from our blood bank, the following parameters became apparent. The pH of standard blood available from our blood bank ranged from 6.1 to 6.4. The blood obtained from our blood bank had significant hyperkalemia, hyponatremia, and hypocalcemia. No reactions were noted when patients were primed with normal saline, which has a pH of around 5.9. We speculate that the presence of endogenous blood substances, such as bradykinin, may have induced the hypotensive episodes. We describe two techniques we developed that should allow for the increased safe and effective use of the AN-69 membranes in continuous venovenous hemodiafiltration circuits. These observations indicate the requirement for careful and close attention to detail when delivering renal replacement therapy to anyone, but especially patients weighing less than 10 kg.

  12. Detecting Extracellular Carbonic Anhydrase Activity Using Membrane Inlet Mass Spectrometry

    Delacruz, Joannalyn; Mikulski, Rose; Tu, Chingkuang; Li, Ying; Wang, Hai; Shiverick, Kathleen T.; Frost, Susan C.; Horenstein, Nicole A.; Silverman, David N.

    2010-01-01

    Current research into the function of carbonic anhydrases in cell physiology emphasizes the role of membrane-bound carbonic anhydrases, such as carbonic anhydrase IX that has been identified in malignant tumors and is associated with extracellular acidification as a response to hypoxia. We present here a mass spectrometric method to determine the extent to which total carbonic anhydrase activity is due to extracellular carbonic anhydrase in whole cell preparations. The method is based on the biphasic rate of depletion of 18O from CO2 measured by membrane inlet mass spectrometry. The slopes of the biphasic depletion are a sensitive measure of the presence of carbonic anhydrase outside and inside of the cells. This property is demonstrated here using suspensions of human red cells in which external carbonic anhydrase was added to the suspending solution. It is also applied to breast and prostate cancer cells which both express exofacial carbonic anhydrase IX. Inhibition of external carbonic anhydrase is achieved by use of a membrane impermeant inhibitor that was synthesized for this purpose, p-aminomethylbenzenesulfonamide attached to a polyethyleneglycol polymer. PMID:20417171

  13. Surface Functionalization of Polyethersulfone Membrane with Quaternary Ammonium Salts for Contact-Active Antibacterial and Anti-Biofouling Properties

    Xiao Hu

    2016-05-01

    Full Text Available Biofilm is a significant cause for membrane fouling. Antibacterial-coated surfaces can inhibit biofilm formation by killing bacteria. In this study, polyethersulfone (PES microfiltration membrane was photografted by four antibiotic quaternary ammonium compounds (QACs separately, which were synthesized from dimethylaminoethyl methacrylate (DMAEMA by quaternization with butyl bromide (BB, octyl bromide (OB, dodecyl bromide (DB, or hexadecyl bromide (HB. XPS, ATR-FTIR, and SEM were used to confirm the surfaces’ composition and morphology. After modification, the pores on PES-g-DMAEMA-BB and PES-g-DMAEMA-OB were blocked, while PES-g-DMAEMA-DB and PES-g-DMAEMA-HB were retained. We supposed that DMAEMA-BB and DMAEMA-OB aggregated on the membrane surface due to the activities of intermolecular or intramolecular hydrogen bonds. Bacteria testing found the antibacterial activities of the membranes increased with the length of the substituted alkyl chain. Correspondingly, little bacteria were observed on PES-g-DMAEMA-DB and PES-g-DMAEMA-HB by SEM. The antifouling properties were investigated by filtration of a solution of Escherichia coli. Compared with the initial membrane, PES-g-DMAEMA-DB and PES-g-DMAEMA-HB showed excellent anti-biofouling performance with higher relative flux recovery (RFR of 88.3% and 92.7%, respectively. Thus, surface functionalization of the PES membrane with QACs can prevent bacteria adhesion and improve the anti-biofouling activity by the contact-active antibacterial property.

  14. Membrane curvature stress and antibacterial activity of lactoferricin derivatives.

    Zweytick, Dagmar; Tumer, Sabine; Blondelle, Sylvie E; Lohner, Karl

    2008-05-02

    We have studied correlation of non-lamellar phase formation and antimicrobial activity of two cationic amphipathic peptides, termed VS1-13 and VS1-24 derived from a fragment (LF11) of human lactoferricin on Escherichia coli total lipid extracts. Compared to LF11, VS1-13 exhibits minor, but VS1-24 significantly higher antimicrobial activity. X-ray experiments demonstrated that only VS1-24 decreased the onset of cubic phase formation of dispersions of E. coli lipid extracts, significantly, down to physiological relevant temperatures. Cubic structures were identified to belong to the space groups Pn3m and Im3m. Formation of latter is enhanced in the presence of VS1-24. Additionally, the presence of this peptide caused membrane thinning in the fluid phase, which may promote cubic phase formation. VS1-24 containing a larger hydrophobic volume at the N-terminus than its less active counterpart VS1-13 seems to increase curvature stress in the bilayer and alter the behaviour of the membrane significantly enhancing disruption.

  15. Protein kinase C prevents oligodendrocyte differentiation : Modulation of actin cytoskeleton and cognate polarized membrane traffic

    Baron, W; de Vries, EJ; de Vries, H; Hoekstra, D

    1999-01-01

    In a previous study, we showed that activation of protein kinase C (PKC) prevents oligodendrocyte differentiation at the pro-oligodendrocyte stage. The present study was undertaken to identify downstream targets of PKC action in oligodendrocyte progenitor cells. Activation of PKC induced the

  16. Lateralized Difference in Tympanic Membrane Temperature: Emotion and Hemispheric Activity

    Ruth E Propper

    2013-03-01

    Full Text Available We review literature examining relationships between tympanic membrane temperature (TMT, affective/motivational orientation, and hemispheric activity. Lateralized differences in TMT might enable real-time monitoring of hemispheric activity in real-world conditions, and could serve as a corroborating marker of mental illnesses associated with specific affective dysregulation. We support the proposal that TMT holds potential for broadly indexing lateralized brain physiology during tasks demanding the processing and representation of emotional and/or motivational states, and for predicting trait-related affective/motivational orientations. The precise nature of the relationship between TMT and brain physiology, however, remains elusive. Indeed the limited extant research has sampled different participant populations and employed largely different procedures and measures, making for seemingly discrepant findings and implications. We propose, however, that many of these discrepancies can be resolved by considering how emotional states map onto motivational systems, and further examining how validated methods for inducing lateralized brain activity might affect TMT.

  17. Membrane-associated IL 1-like activity on rat dendritic cells

    Nagelkerken, L.M.; van Breda Vriesman, P.J.C.

    1986-01-01

    The secretion of interleukin 1 (IL 1) by rat dendritic cells (DC) was studied in relation to their ability to induce the production interleukin 2 (IL 2 ) and to induce IL 2 responsiveness. IL 1 (or IL 1-like activity) was measured by its capacity to enhance IL 2 production by EL4 cells. In contrast to peritoneal exudate cells (PEC) or splenic adherent cells, DC from thoracic duct lymph (TD-DC) or from spleen did not secrete detectable amounts of IL 1 on stimulation with LPS/Silica. However, TD-DC and splenic DC were able to enhance IL 2 production by EL4 cells directly, and were only two times less effective than PEC. By preventing cell-to-cell contact between stimulator cells and EL4 cells, it was demonstrated that most of the IL 2-inducing activity of TD-DC and PEC was associated with the cell membrane. Treatment with 1% paraformaldehyde (PFA) to abolish metabolic activity resulted in a 50% decrease (or inactivation) of IL 2-inducing activity of TD-DC in the EL4 assay. Moreover, UVB-irradiation (300 mJ/cm 2 ) of TD-DC, which has been described to inhibit the release of IL 1 by macrophages, caused a 70% decrease in IL 2-inducing activity. These results suggest that membrane-associated structures, that are identical to or mimic Il 1, are involved in the activation of T cells by DC

  18. Bilateral PLA/alginate membranes for the prevention of postsurgical adhesions.

    Kessler, Martina; Esser, Eva; Groll, Jürgen; Tessmar, Jörg

    2016-11-01

    A bilateral barrier membrane for the prevention of postsurgical adhesions was developed. Thereby, a smooth PLA side was supposed to keep the affected tissues glidingly separated, while a mucoadhesive side made of alginate was meant to keep the barrier resident on the site of injury so that suturing becomes redundant or at least the membrane stays long enough to facilitate surgical handling. Because hydrophilic alginate and lipophilic PLA films show only low cohesion, solution electrospun meshes of PLA and PLA-PEG-PLA triblock copolymers with varying poly(ethylene glycol) [PEG] content were investigated as cohesion promoter to avoid an easy separation of the functionally different layers. Using direct electrospinning onto the PLA film, a modified contact surface of the mesh was created, which allowed the tested alginate solutions (3%, 5%) to infiltrate to different extents. Thereby, an increasing content of hydrophilic PEG within the mesh copolymer and a lower alginate concentration facilitated the infiltration. As a result, the PLA film with a PLA35k-PEG10k-PLA35k (racemic PLA chains) mesh and an alginate layer cast from a 3% alginate solution appeared to be the most effective combination as examined by means of a t peel test, a mucoadhesion test, a tensile test and optical evaluations. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 104B: 1563-1570, 2016. © 2015 Wiley Periodicals, Inc.

  19. The hydroxyflavone, fisetin, suppresses mast cell activation induced by interaction with activated T cell membranes

    Nagai, K; Takahashi, Y; Mikami, I; Fukusima, T; Oike, H; Kobori, M

    2009-01-01

    Background and purpose: Cell-to-cell interactions between mast cells and activated T cells are increasingly recognized as a possible mechanism in the aetiology of allergic or non-allergic inflammatory disorders. To determine the anti-allergic effect of fisetin, we examined the ability of fisetin to suppress activation of the human mast cell line, HMC-1, induced by activated Jurkat T cell membranes. Experimental approach: HMC-1 cells were incubated with or without fisetin for 15 min and then co-cultured with Jurkat T cell membranes activated by phorbol-12-myristate 13-acetate for 16 h. We determined gene expression in activated HMC-1 cells by DNA microarray and quantitative reverse transcription (RT)-PCR analysis. We also examined activation of the transcription factor NF-κB and MAP kinases (MAPKs) in activated HMC-1 cells. Key results: Fisetin suppresses cell spreading and gene expression in HMC-1 cells stimulated by activated T cell membranes. Additionally, we show that these stimulated HMC-1 cells expressed granzyme B. The stimulatory interaction also induced activation of NF-κB and MAPKs; these activations were suppressed by fisetin. Fisetin also reduced the amount of cell surface antigen CD40 and intercellular adhesion molecule-1 (ICAM-1) on activated HMC-1 cells. Conclusions and implications: Fisetin suppressed activation of HMC-1 cells by activated T cell membranes by interfering with cell-to-cell interaction and inhibiting the activity of NF-κB and MAPKs and thereby suppressing gene expression. Fisetin may protect against the progression of inflammatory diseases by limiting interactions between mast cells and activated T cells. PMID:19702784

  20. Amniotic membrane grafts for the prevention of esophageal stricture after circumferential endoscopic submucosal dissection.

    Barret, Maximilien; Pratico, Carlos Alberto; Camus, Marine; Beuvon, Frédéric; Jarraya, Mohamed; Nicco, Carole; Mangialavori, Luigi; Chaussade, Stanislas; Batteux, Frédéric; Prat, Frédéric

    2014-01-01

    The prevention of esophageal strictures following circumferential mucosal resection remains a major clinical challenge. Human amniotic membrane (AM) is an easily available material, which is widely used in ophthalmology due to its wound healing, anti-inflammatory and anti-fibrotic properties. We studied the effect of AM grafts in the prevention of esophageal stricture after endoscopic submucosal dissection (ESD) in a swine model. In this prospective, randomized controlled trial, 20 swine underwent a 5 cm-long circumferential ESD of the lower esophagus. In the AM Group (n = 10), amniotic membrane grafts were placed on esophageal stents; a subgroup of 5 swine (AM 1 group) was sacrificed on day 14, whereas the other 5 animals (AM 2 group) were kept alive. The esophageal stent (ES) group (n = 5) had ES placement alone after ESD. Another 5 animals served as a control group with only ESD. The prevalence of symptomatic strictures at day 14 was significantly reduced in the AM group and ES groups vs. the control group (33%, 40% and 100%, respectively, p = 0.03); mean esophageal diameter was 5.8±3.6 mm, 6.8±3.3 mm, and 2.6±1.7 mm for AM, ES, and control groups, respectively. Median (range) esophageal fibrosis thickness was 0.87 mm (0.78-1.72), 1.19 mm (0.28-1.95), and 1.65 mm (0.7-1.79) for AM 1, ES, and control groups, respectively. All animals had developed esophageal strictures by day 35. The anti-fibrotic effect of AM on esophageal wound healing after ESD delayed the development of esophageal stricture in our model. However, this benefit was of limited duration in the conditions of our study.

  1. Amniotic membrane grafts for the prevention of esophageal stricture after circumferential endoscopic submucosal dissection.

    Maximilien Barret

    Full Text Available The prevention of esophageal strictures following circumferential mucosal resection remains a major clinical challenge. Human amniotic membrane (AM is an easily available material, which is widely used in ophthalmology due to its wound healing, anti-inflammatory and anti-fibrotic properties. We studied the effect of AM grafts in the prevention of esophageal stricture after endoscopic submucosal dissection (ESD in a swine model.In this prospective, randomized controlled trial, 20 swine underwent a 5 cm-long circumferential ESD of the lower esophagus. In the AM Group (n = 10, amniotic membrane grafts were placed on esophageal stents; a subgroup of 5 swine (AM 1 group was sacrificed on day 14, whereas the other 5 animals (AM 2 group were kept alive. The esophageal stent (ES group (n = 5 had ES placement alone after ESD. Another 5 animals served as a control group with only ESD.The prevalence of symptomatic strictures at day 14 was significantly reduced in the AM group and ES groups vs. the control group (33%, 40% and 100%, respectively, p = 0.03; mean esophageal diameter was 5.8±3.6 mm, 6.8±3.3 mm, and 2.6±1.7 mm for AM, ES, and control groups, respectively. Median (range esophageal fibrosis thickness was 0.87 mm (0.78-1.72, 1.19 mm (0.28-1.95, and 1.65 mm (0.7-1.79 for AM 1, ES, and control groups, respectively. All animals had developed esophageal strictures by day 35.The anti-fibrotic effect of AM on esophageal wound healing after ESD delayed the development of esophageal stricture in our model. However, this benefit was of limited duration in the conditions of our study.

  2. Avidin/PSS membrane microcapsules with biotin-binding activity.

    Endo, Yoshihiro; Sato, Katsuhiko; Sugimoto, Kentaro; Anzai, Jun-ichi

    2011-08-15

    Polyelectrolyte microcapsules with avidin-poly(styrene sulfonate) (PSS) membrane were prepared by a layer-by-layer deposition technique. The uptake and release of biotin-labeled fluorescein (b-FITC) as well as immobilization of biotin-labeled glucose oxidase (b-GOx) to the microcapsule were studied. The polyelectrolyte microcapsules were prepared by coating the surface of calcium carbonate (CaCO(3)) microparticles with an avidin/PSS multilayer membrane, followed by dissolution of CaCO(3) core in an ethylenediaminetetraacetic acid solution. Inner and outer poly(allylamine)/PSS films were required to isolate the microcapsules, whereas microcapsules could not be formed without the support. The uptake of b-FITC into the microcapsule was highly enhanced through a strong binding of b-FITC to avidin as compared with the uptake of biotin-free FITC. Release of b-FITC from the microcapsule was accelerated upon addition of biotin due to a competitive binding of the added biotin to the binding site of avidin. Similarly, the surface of microcapsule was modified with b-GOx with retaining its catalytic activity. Copyright © 2011 Elsevier Inc. All rights reserved.

  3. Blood-group-Ii-active gangliosides of human erythrocyte membranes

    Feizi, T.; Childs, R.A.; Hakomori, S.-I.; Powell, M.E.

    1978-01-01

    More than ten new types of gangliosides, in addition to haematoside and sialosylparagloboside, were isolated from human erythrocyte membranes. These were separated by successive chromatographies on DAEA-Sephadex, on porous silica-gel columns and on thin-layer silica gel as acetylated compounds. Highly potent blood-group-Ii and moderate blood-group-H activities were demonstrated in some of the ganglioside fractions. The gangliosides incorporated into chlolesterol/phosphatidylcholine liposomes stoicheiometrically inhibited binding of anti-(blood-group-I and i) antibodies to a radioiodinated blood-group-Ii-active glycoprotein. The fraction with the highest blood-group-I activity, I(g) fraction, behaved like sialosyl-deca- to dodeca-glycosylceramides on t.l.c. Certain blood-group-I and most of the i-determinants were in partially or completely cryptic form and could be unmasked by sialidase treatment. Thus the I and i antigens, which are known to occur on internal structures of blood-group-ABH-active glycoproteins in secretions, also occur in the interior of the carbohydrate chains of erythrocyte gangliosides. (author)

  4. Broad spectrum antibacterial and antifungal polymeric paint materials: synthesis, structure-activity relationship, and membrane-active mode of action.

    Hoque, Jiaul; Akkapeddi, Padma; Yadav, Vikas; Manjunath, Goutham B; Uppu, Divakara S S M; Konai, Mohini M; Yarlagadda, Venkateswarlu; Sanyal, Kaustuv; Haldar, Jayanta

    2015-01-28

    Microbial attachment and subsequent colonization onto surfaces lead to the spread of deadly community-acquired and hospital-acquired (nosocomial) infections. Noncovalent immobilization of water insoluble and organo-soluble cationic polymers onto a surface is a facile approach to prevent microbial contamination. In the present study, we described the synthesis of water insoluble and organo-soluble polymeric materials and demonstrated their structure-activity relationship against various human pathogenic bacteria including drug-resistant strains such as methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant enterococci (VRE), and beta lactam-resistant Klebsiella pneumoniae as well as pathogenic fungi such as Candida spp. and Cryptococcus spp. The polymer coated surfaces completely inactivated both bacteria and fungi upon contact (5 log reduction with respect to control). Linear polymers were more active and found to have a higher killing rate than the branched polymers. The polymer coated surfaces also exhibited significant activity in various complex mammalian fluids such as serum, plasma, and blood and showed negligible hemolysis at an amount much higher than minimum inhibitory amounts (MIAs). These polymers were found to have excellent compatibility with other medically relevant polymers (polylactic acid, PLA) and commercial paint. The cationic hydrophobic polymer coatings disrupted the lipid membrane of both bacteria and fungi and thus showed a membrane-active mode of action. Further, bacteria did not develop resistance against these membrane-active polymers in sharp contrast to conventional antibiotics and lipopeptides, thus the polymers hold great promise to be used as coating materials for developing permanent antimicrobial paint.

  5. Is there evidence that barrier membranes prevent bone resorption in autologous bone grafts during the healing period? A systematic review

    Gielkens, Pepijn F. M.; Bos, Ruud R. M.; Raghoebar, Gerry M.; Stegenga, Boudewijn

    2007-01-01

    Introduction: Autologous bone is considered the "reference standard" for bone-grafting procedures. A barrier membrane covering an autologous bone graft (guided bone regeneration [GBR]) is expected to prevent graft resorption. Good clinical results have been reported for GBR, although potential

  6. Spermine modulates fungal morphogenesis and activates plasma membrane H+-ATPase during yeast to hyphae transition

    Antônio Jesus Dorighetto Cogo

    2018-02-01

    Full Text Available Polyamines play a regulatory role in eukaryotic cell growth and morphogenesis. Despite many molecular advances, the underlying mechanism of action remains unclear. Here, we investigate a mechanism by which spermine affects the morphogenesis of a dimorphic fungal model of emerging relevance in plant interactions, Yarrowia lipolytica, through the recruitment of a phytohormone-like pathway involving activation of the plasma membrane P-type H+-ATPase. Morphological transition was followed microscopically, and the H+-ATPase activity was analyzed in isolated membrane vesicles. Proton flux and acidification were directly probed at living cell surfaces by a non-invasive selective ion electrode technique. Spermine and indol-3-acetic acid (IAA induced the yeast-hypha transition, influencing the colony architecture. Spermine induced H+-ATPase activity and H+ efflux in living cells correlating with yeast-hypha dynamics. Pharmacological inhibition of spermine and IAA pathways prevented the physio-morphological responses, and indicated that spermine could act upstream of the IAA pathway. This study provides the first compelling evidence on the fungal morphogenesis and colony development as modulated by a spermine-induced acid growth mechanism analogous to that previously postulated for the multicellular growth regulation of plants.

  7. Two Outer Membrane Proteins Contribute to Caulobacter crescentus Cellular Fitness by Preventing Intracellular S-Layer Protein Accumulation

    Overton, K. Wesley; Park, Dan M.; Yung, Mimi C.; Dohnalkova, Alice; Smit, John; Jiao, Yongqin

    2016-09-23

    ABSTRACT

    Surface layers, or S-layers, are two-dimensional protein arrays that form the outermost layer of many bacteria and archaea. They serve several functions, including physical protection of the cell from environmental threats. The high abundance of S-layer proteins necessitates a highly efficient export mechanism to transport the S-layer protein from the cytoplasm to the cell exterior.Caulobacter crescentusis unique in that it has two homologous, seemingly redundant outer membrane proteins, RsaFaand RsaFb, which together with other components form a type I protein translocation pathway for S-layer export. These proteins have homology toEscherichia coliTolC, the outer membrane channel of multidrug efflux pumps. Here we provide evidence that, unlike TolC, RsaFaand RsaFbare not involved in either the maintenance of membrane stability or the active export of antimicrobial compounds. Rather, RsaFaand RsaFbare required to prevent intracellular accumulation and aggregation of the S-layer protein RsaA; deletion of RsaFaand RsaFbled to a general growth defect and lowered cellular fitness. Using Western blotting, transmission electron microscopy, and transcriptome sequencing (RNA-seq), we show that loss of both RsaFaand RsaFbled to accumulation of insoluble RsaA in the cytoplasm, which in turn caused upregulation of a number of genes involved in protein misfolding and degradation pathways. These findings provide new insight into the requirement for RsaFaand RsaFbin cellular fitness and tolerance to antimicrobial agents and further our understanding of the S-layer export mechanism on both the transcriptional and translational levels in

  8. Activation of TRPV1 channels inhibits mechanosensitive Piezo channel activity by depleting membrane phosphoinositides

    Borbiro, Istvan; Badheka, Doreen; Rohacs, Tibor

    2015-01-01

    Capsaicin is an activator of the heat-sensitive TRPV1 (transient receptor potential vanilloid 1) ion channels and has been used as a local analgesic. We found that activation of TRPV1 channels with capsaicin either in dorsal root ganglion neurons or in a heterologous expression system inhibited the mechanosensitive Piezo1 and Piezo2 channels by depleting phosphatidylinositol 4,5-bisphosphate [PI(4,5)P2] and its precursor PI(4)P from the plasma membrane through Ca2+-induced phospholipase Cδ (PLCδ) activation. Experiments with chemically inducible phosphoinositide phosphatases and receptor-induced activation of PLCβ indicated that inhibition of Piezo channels required depletion of both PI(4)P and PI(4,5)P2. The mechanically activated current amplitudes decreased substantially in the excised inside-out configuration, where the membrane patch containing Piezo1 channels is removed from the cell. PI(4,5)P2 and PI(4)P applied to these excised patches inhibited this decrease. Thus, we concluded that Piezo channel activity requires the presence of phosphoinositides, and the combined depletion of PI(4,5)P2 or PI(4)P reduces channel activity. In addition to revealing a role for distinct membrane lipids in mechanosensitive ion channel regulation, these data suggest that inhibition of Piezo2 channels may contribute to the analgesic effect of capsaicin. PMID:25670203

  9. Hydro-osmotic Instabilities in Active Membrane Tubes

    Al-Izzi, Sami C.; Rowlands, George; Sens, Pierre; Turner, Matthew S.

    2018-03-01

    We study a membrane tube with unidirectional ion pumps driving an osmotic pressure difference. A pressure-driven peristaltic instability is identified, qualitatively distinct from similar tension-driven Rayleigh-type instabilities on membrane tubes. We discuss how this instability could be related to the function and biogenesis of membrane bound organelles, in particular, the contractile vacuole complex. The unusually long natural wavelength of this instability is in agreement with that observed in cells.

  10. Removal of Zn (II) and Ga (III) from waste waters using activated composite membranes

    Melita, L.; Meghea, A.; Munoz Tapia, M.; Gives, J. de

    2001-01-01

    The present study refers to the preparation of activated composite membrane (ACM) containing Aliquat 336 as a carrier, and testing their properties towards the selective transport of Ga and Zn cations. A new type of liquid membrane was prepared, named Activated Composite Membrane (ACM). The stability of these membrane increases, referring to other common membranes used before. These membranes have also good characteristics to separate metals. We cast membranes in two steps, first we used non-woven fabric (Hollytex 3329, France) as a support to manufacture reinforced polysulfone (PS) membrane which was obtained by the phase inversion technique, and second, a thin top layer of polyamide containing Aliquat 336 of two different concentrations (0.5 and 1 M) was obtained by interfacial polymerisation. The membrane thus prepared is composed of polyamide and polysulfone layers containing carrier. The surface texture of the membrane under study was examined by scanning electron microscopy (SEM) using a JSM-6300 scanning electron microscope. The chemical elemental analysis of freshly prepared membranes was performed, by X-ray diffraction measuring the energy distribution of the X-ray signal generated by a focused electron beam. A correlation between the carrier content in the membrane and the concentration of metal separated was obtained from the results of the membrane analysis by using the inductively coupled plasma (ICP) technique. The competition between gallium and zinc in the membrane surface is presented by the retaining membrane capacity. This type of membrane is relatively new for metal removal (Ga and Zn) from waste waters and the best cation retention was obtained for Zn. (authors)

  11. Maintenance of membrane organization in the aging mouse brain as the determining factor for preventing receptor dysfunction and for improving response to anti-Alzheimer treatments.

    Colin, Julie; Thomas, Mélanie H; Gregory-Pauron, Lynn; Pinçon, Anthony; Lanhers, Marie-Claire; Corbier, Catherine; Claudepierre, Thomas; Yen, Frances T; Oster, Thierry; Malaplate-Armand, Catherine

    2017-06-01

    Although a major risk factor for Alzheimer's disease (AD), the "aging" parameter is not systematically considered in preclinical validation of anti-AD drugs. To explore how aging affects neuronal reactivity to anti-AD agents, the ciliary neurotrophic factor (CNTF)-associated pathway was chosen as a model. Comparison of the neuroprotective properties of CNTF in 6- and 18-month old mice revealed that CNTF resistance in the older animals is associated with the exclusion of the CNTF-receptor subunits from rafts and their subsequent dispersion to non-raft cortical membrane domains. This age-dependent membrane remodeling prevented both the formation of active CNTF-receptor complexes and the activation of prosurvival STAT3 and ERK1/2 pathways, demonstrating that age-altered membranes impaired the reactivity of potential therapeutic targets. CNTF-receptor distribution and CNTF signaling responses were improved in older mice receiving dietary docosahexaenoic acid, with CNTF-receptor functionality being similar to those of younger mice, pointing toward dietary intervention as a promising adjuvant strategy to maintain functional neuronal membranes, thus allowing the associated receptors to respond appropriately to anti-AD agents. Copyright © 2017 Elsevier Inc. All rights reserved.

  12. Sch proteins are localized on endoplasmic reticulum membranes and are redistributed after tyrosine kinase receptor activation

    Lotti, L V; Lanfrancone, L; Migliaccio, E

    1996-01-01

    area of the cell and mostly associated with the cytosolic side of rough endoplasmic reticulum membranes. Upon epidermal growth factor treatment and receptor tyrosine kinase activation, the immunolabeling became peripheral and was found to be associated with the cytosolic surface of the plasma membrane....... The rough endoplasmic reticulum localization of Shc proteins in unstimulated cells and their massive recruitment to the plasma membrane, endocytic structures, and peripheral cytosol following receptor tyrosine kinase activation could account for multiple putative functions of the adaptor protein....

  13. Soluble products of Escherichia coli induce mitochondrial dysfunction-related sperm membrane lipid peroxidation which is prevented by lactobacilli.

    Arcangelo Barbonetti

    Full Text Available Unidentified soluble factors secreted by E. coli, a frequently isolated microorganism in genitourinary infections, have been reported to inhibit mitochondrial membrane potential (ΔΨm, motility and vitality of human spermatozoa. Here we explore the mechanisms involved in the adverse impact of E. coli on sperm motility, focusing mainly on sperm mitochondrial function and possible membrane damage induced by mitochondrial-generated reactive oxygen species (ROS. Furthermore, as lactobacilli, which dominate the vaginal ecosystem of healthy women, have been shown to exert anti-oxidant protective effects on spermatozoa, we also evaluated whether soluble products from these microorganisms could protect spermatozoa against the effects of E. coli. We assessed motility (by computer-aided semen analysis, ΔΨm (with JC-1 dye by flow cytometry, mitochondrial ROS generation (with MitoSOX red dye by flow cytometry and membrane lipid-peroxidation (with the fluorophore BODIPY C11 by flow cytometry of sperm suspensions exposed to E. coli in the presence and in the absence of a combination of 3 selected strains of lactobacilli (L. brevis, L. salivarius, L. plantarum. A Transwell system was used to avoid direct contact between spermatozoa and microorganisms. Soluble products of E. coli induced ΔΨm loss, mitochondrial generation of ROS and membrane lipid-peroxidation, resulting in motility loss. Soluble factors of lactobacilli prevented membrane lipid-peroxidation of E. coli-exposed spermatozoa, thus preserving their motility. In conclusion, sperm motility loss by soluble products of E. coli reflects a mitochondrial dysfunction-related membrane lipid-peroxidation. Lactobacilli could protect spermatozoa in the presence of vaginal disorders, by preventing ROS-induced membrane damage.

  14. Fouling on ion-exchange membranes: Classification, characterization and strategies of prevention and control.

    Mikhaylin, Sergey; Bazinet, Laurent

    2016-03-01

    The environmentally friendly ion-exchange membrane (IEM) processes find more and more applications in the modern industries in order to demineralize, concentrate and modify products. Moreover, these processes may be applied for the energy conversion and storage. However, the main drawback of the IEM processes is a formation of fouling, which significantly decreases the process efficiency and increases the process cost. The present review is dedicated to the problematic of IEM fouling phenomena. Firstly, the major types of IEM fouling such as colloidal fouling, organic fouling, scaling and biofouling are discussed along with consideration of the main factors affecting fouling formation and development. Secondly, the review of the possible methods of IEM fouling characterization is provided. This section includes the methods of fouling visualization and characterization as well as methods allowing investigations of characteristics of the fouled IEMs. Eventually, the reader will find the conventional and modern strategies of prevention and control of different fouling types. Copyright © 2015 Elsevier B.V. All rights reserved.

  15. Active calcium transport in plasma membrane vesicles from developing cotyledons of common bean

    Huang Jianzhong; Chen Ziyuan

    1995-01-01

    Plasma membrane vesicles were prepared from the developing cotyledons of common bean (Phaseolus vulgaris L cv Diyundou) by aqueous two-phase partitioning and characterized as to their purity by assaying marker enzymes for other membranes. The putative plasma membrane fraction was minimally contaminated by membranes other than plasma membrane and hence was of high purity. It exhibited a Ca 2+ -dependent ATPase activity, which was inhibited by 1 μmol/L EB and promoted by calcium ionophore A23187. Such an activity was responsible for the observed ATP-dependent 45 Ca 2+ uptake into inside-out plasma membrane vesicles. This process was stimulated by 0.6 μmol/L CaM and 20 μmol/L IAA but inhibited by 2 μmol/L ABA and abolished by A23187. Possible role of cytoplasmic Ca 2+ in mediating phytohormones activity is discussed

  16. Oxygen activation at the plasma membrane: relation between superoxide and hydroxyl radical production by isolated membranes.

    Heyno, Eiri; Mary, Véronique; Schopfer, Peter; Krieger-Liszkay, Anja

    2011-07-01

    Production of reactive oxygen species (hydroxyl radicals, superoxide radicals and hydrogen peroxide) was studied using EPR spin-trapping techniques and specific dyes in isolated plasma membranes from the growing and the non-growing zones of hypocotyls and roots of etiolated soybean seedlings as well as coleoptiles and roots of etiolated maize seedlings. NAD(P)H mediated the production of superoxide in all plasma membrane samples. Hydroxyl radicals were only produced by the membranes of the hypocotyl growing zone when a Fenton catalyst (FeEDTA) was present. By contrast, in membranes from other parts of the seedlings a low rate of spontaneous hydroxyl radical formation was observed due to the presence of small amounts of tightly bound peroxidase. It is concluded that apoplastic hydroxyl radical generation depends fully, or for the most part, on peroxidase localized in the cell wall. In soybean plasma membranes from the growing zone of the hypocotyl pharmacological tests showed that the superoxide production could potentially be attributed to the action of at least two enzymes, an NADPH oxidase and, in the presence of menadione, a quinone reductase.

  17. Barodiffusion phenomena at active transport of na+ and K+ ions through the cell membrane

    Khrapijchuk, G.V.; Chalyi, A.V.; Nurishchenko, N.Je.

    2010-01-01

    The influence of ultrasound as the significant motive force of barodiffusion phenomena at the processes of active transport of Na + and K + ions through the cell membrane is considered. The dependence of membrane potential is theoretically estimated at active transport of natrium and potassium ions on the ultrasound intensity and pressure overfall between external and internal medium of the cell.

  18. A Loss in the Plasma Membrane ATPase Activity and Its Recovery Coincides with Incipient Freeze-Thaw Injury and Postthaw Recovery in Onion Bulb Scale Tissue 1

    Arora, Rajeev; Palta, Jiwan P.

    1991-01-01

    Plasma membrane ATPase has been proposed to be functionally altered during early stages of injury caused by a freeze-thaw stress. Complete recovery from freezing injury in onion cells during the postthaw period provided evidence in support of this proposal. During recovery, a simultaneous decrease in ion leakage and disappearance of water soaking (symptoms of freeze-thaw injury) has been noted. Since reabsorption of ions during recovery must be an active process, recovery of plasma membrane ATPase (active transport system) functions has been implicated. In the present study, onion (Allium cepa L. cv Downing Yellow Globe) bulbs were subjected to a freeze-thaw stress which resulted in a reversible (recoverable) injury. Plasma membrane ATPase activity in the microsomes (isolated from the bulb scales) and ion leakage rate (efflux/hour) from the same scale tissue were measured immediately following thawing and after complete recovery. In injured tissue (30-40% water soaking), plasma membrane ATPase activity was reduced by about 30% and this was paralleled by about 25% higher ion leakage rate. As water soaking disappeared during recovery, the plasma membrane ATPase activity and the ion leakage rate returned to about the same level as the respective controls. Treatment of freeze-thaw injured tissue with vanadate, a specific inhibitor of plasma membrane ATPase, during postthaw prevented the recovery process. These results indicate that recovery of freeze-injured tissue depends on the functional activity of plasma membrane ATPase. PMID:16668063

  19. Study on application of safety checklist in preventive maintenance activities

    Shi Jin; Chen Song; Liu Jingquan

    2013-01-01

    The paper describes the principles and the characteristics of safety checklist as a risk evaluation method. Examples of application of safety checklists to preventive maintenance activities such as criteria comparison and checkup items in place in nuclear power plants are illustrated in details with issues appeared in the checklist establishment. Checklist has a good application in the RCM analysis or in the actual preventive maintenance program for Chashma Nuclear Power Plant indicated by concrete instances. In the light of safety checklist which is used to sustain preventive maintenance as a simple and applicable risk analysis approach, we can get deep knowledge of risks of nuclear power plant to perfect preventive maintenance activities. (authors)

  20. Membrane Heterogeneity in Akt Activation in Prostate Cancer

    Hager, Martin H

    2008-01-01

    This project focuses on the novel finding from our group that the serine-threonine kinase Akt1 partitions into specialized membrane microdomains, termed lipid rafts, and that this localization event...

  1. Chitosan/Carboxymethylcellulose/Ionic Liquid/Ag(0) Nanoparticles Form a Membrane with Antimicrobial Activity

    Quadros, C.; Faria, V.W.; Scheeren, C.W.; Klein, M.P.; Hertz, P.F.

    2013-01-01

    Silver metal nanoparticles were immobilized in chitosan/carboxymethylcellulose/BMI.BF4(1-n-butyl-3-methylimidazolium tetrafluoroborate ionic liquid) (CS/CMC/IL) to form polymeric membrane with 20 μm thickness. The CS/CMC/IL polymeric membrane was prepared using a simple solution blending method. Irregularly shaped Ag(0) nanoparticles with monomodal size distributions of nm Ag(0) were immobilized in the membrane. The presence of small Ag(0) nanoparticles induced an augmentation in the CS/CMC/IL film surface areas. The CS/CMC/IL membrane containing Ag(0) showed increase antimicrobial activity the Ag(0) concentration increased up to saturation at 10 mg. CS/CMC/IL membrane that contains Ag(0) nanoparticles has enhanced durability of the membrane and exhibited stronger antimicrobial activity against Escherichia coli and Staphylococcus aureus.

  2. One-dimensional statistical model of active transfer of substances in membranes

    Melkikh, A.V.; Seleznev, V.D.

    1995-01-01

    A matrix of kinetic coefficients that describe the processes of particle and heat transfer in a double membrane is obtained. The kinetic coefficients are considered as functions of the membrane parameters. Conditions are found under which the energy expended on transfer of one atom through the membrane is minimum. The obtained theoretical value of this energy is compared with the corresponding value for active transport of ions in biomembranes

  3. FAM21 directs SNX27–retromer cargoes to the plasma membrane by preventing transport to the Golgi apparatus

    Lee, Seongju; Chang, Jaerak; Blackstone, Craig

    2016-01-01

    The endosomal network maintains cellular homeostasis by sorting, recycling and degrading endocytosed cargoes. Retromer organizes the endosomal sorting pathway in conjunction with various sorting nexin (SNX) proteins. The SNX27–retromer complex has recently been identified as a major endosomal hub that regulates endosome-to-plasma membrane recycling by preventing lysosomal entry of cargoes. Here, we show that SNX27 directly interacts with FAM21, which also binds retromer, within the Wiskott–Aldrich syndrome protein and SCAR homologue (WASH) complex. This interaction is required for the precise localization of SNX27 at an endosomal subdomain as well as for recycling of SNX27-retromer cargoes. Furthermore, FAM21 prevents cargo transport to the Golgi apparatus by controlling levels of phosphatidylinositol 4-phosphate, which facilitates cargo dissociation at the Golgi. Together, our results demonstrate that the SNX27–retromer–WASH complex directs cargoes to the plasma membrane by blocking their transport to lysosomes and the Golgi. PMID:26956659

  4. Zinc can increase the activity of protein kinase C and contributes to its binding to plasma membranes in T lymphocytes

    Csermely, P.; Szamel, M.; Resch, K.; Somogyi, J.

    1988-01-01

    In the primary structure of protein kinase C, the presence of a putative metal-binding site has been suggested. In the present report, the authors demonstrate that the most abundant intracellular heavy metal, zinc, can increase the activity of cytosolic protein kinase C. Zinc reversibly binds the enzyme to plasma membranes,and it may contribute to the calcium-induced binding as well. The intracellular heavy metal chelator N,N,N',N'-tetrakis(2-pyridylmethyl) ethylenediamine prevents the phorbol ester- and antigen-induced translocation of protein kinase C. This effect can be totally reversed by the concomitant addition of Zn 2+ , while Fe 2+ and Mn 2+ are only partially counteractive. The results suggest that zinc can activate protein kinase C and contributes to its binding to plasma membranes in T lymphocytes induced by Ca 2+ , phorbol ester, or antigen

  5. Zinc can increase the activity of protein kinase C and contributes to its binding to plasma membranes in T lymphocytes

    Csermely, P.; Szamel, M.; Resch, K.; Somogyi, J.

    1988-05-15

    In the primary structure of protein kinase C, the presence of a putative metal-binding site has been suggested. In the present report, the authors demonstrate that the most abundant intracellular heavy metal, zinc, can increase the activity of cytosolic protein kinase C. Zinc reversibly binds the enzyme to plasma membranes,and it may contribute to the calcium-induced binding as well. The intracellular heavy metal chelator N,N,N',N'-tetrakis(2-pyridylmethyl) ethylenediamine prevents the phorbol ester- and antigen-induced translocation of protein kinase C. This effect can be totally reversed by the concomitant addition of Zn/sup 2 +/, while Fe/sup 2 +/ and Mn/sup 2 +/ are only partially counteractive. The results suggest that zinc can activate protein kinase C and contributes to its binding to plasma membranes in T lymphocytes induced by Ca/sup 2 +/, phorbol ester, or antigen.

  6. Microbial products activate monocytic cells through detergent-resistant membrane microdomains.

    Epelman, Slava; Berenger, Byron; Stack, Danuta; Neely, Graham G; Ma, Ling Ling; Mody, Christopher H

    2008-12-01

    Patients with cystic fibrosis suffer recurrent pulmonary infections that are characterized by an overactive yet ineffective and destructive inflammatory response that is associated with respiratory infections by Pseudomonas aeruginosa, a pathogen that produces a number of phlogistic molecules. To better understand this process, we used exoenzyme S (ExoS), one of the key P. aeruginosa-secreted exoproducts, which is known to stimulate cells via the Toll-like receptor (TLR) pathway. We found that ExoS induced proinflammatory cytokine production via the NF-kappaB, Erk1/2, and Src kinase pathways. Because Src kinases are concentrated within cholesterol-containing, detergent-resistant membrane microdomains (DRM) (also called lipid rafts) and DRM act as signaling platforms and amplifiers on the surface of cells, we addressed the role of DRM in ExoS signaling. ExoS bound directly to a subset of DRM and induced the phosphorylation of multiple proteins within DRM, including Src kinases. Disruption of DRM by cholesterol extraction prevented NF-kappaB and Erk 1/2 activation and TNF-alpha production in response to ExoS. Activation of monocytic cells by other TLR and Nod-like receptor agonists, such as lipoteichoic acid, lipopolysaccharide, and peptidoglycan, were also dependent on DRM, and disruption prevented TNF-alpha production. Disruption of DRM did not prevent ExoS binding but did release the Src kinase, Lyn, from the DRM fraction into the detergent-soluble fraction, a site in which Src kinases are not active. These studies show that ExoS, a TLR agonist, requires direct binding to DRM for optimal signaling, which suggests that DRM are possible therapeutic targets in cystic fibrosis.

  7. HIV/AIDS Prevention Activities of Faith-Based Christian ...

    Based on these findings, it was recommended that the faith-based Christian organizations should collaborate with relevant agencies, community based workers and non-formal education providers to improve HIV/AIDS prevention service delivery in the study area. Keywords: Prevention, Activities, HIV/AIDS, Empowerment, ...

  8. XF-73, a novel antistaphylococcal membrane-active agent with rapid bactericidal activity.

    Ooi, Nicola; Miller, Keith; Hobbs, Joanne; Rhys-Williams, William; Love, William; Chopra, Ian

    2009-10-01

    XF-73 is a novel porphyrin antibacterial agent previously reported to inhibit a range of gram-positive bacterial species, including Staphylococcus aureus. Its mode of action is unknown. Using S. aureus as a model organism we sought to examine the basis of its antibacterial activity. The effects of XF-73 on the growth and survival of S. aureus SH1000 were investigated by viable count and culture absorbance techniques. Inhibition of macromolecular synthesis and disruption of membrane integrity after exposure to XF-73 were examined by radiolabelling experiments, the BacLight fluorescent dye assay and measurement of K(+) and ATP leakage from the cell. The effect of XF-73 on a staphylococcal coupled transcription-translation system was also investigated. XF-73 was rapidly bactericidal against S. aureus SH1000 and demonstrated more rapid killing kinetics than all other comparator agents when tested at an equivalent multiple (4x) of the MIC. Exposure of S. aureus to XF-73 for 10 min completely inhibited DNA, RNA and protein synthesis. XF-73 had no effect on transcription and translation in vitro. Cells exposed to XF-73 gave a positive response in the BacLight assay, which detects membrane damage. The drug also caused substantial loss of K(+) and ATP from the cell, but did not promote bacterial lysis. XF-73 exhibited rapid membrane-perturbing activity, which is likely to be responsible for inhibition of macromolecular synthesis and the death of staphylococci exposed to the drug.

  9. [Pollution prevention and control of aqueous extract of astragali radix processed with ZrO2 inorganic ceramic membrane micro-filtration].

    Pan, Lin-Men; Huang, Min-Yan; Guo, Li-Wei

    2012-11-01

    To study the measures for preventing and controlling the pollution of aqueous extract of Astragali Radix proceeded with inorganic ceramic membrane micro-filtration, in order to find effective measures for preventing and controlling the membrane pollution. The resistance distribution, polymer removal and changes in physical and chemical parameters of the zirconium oxide film of different pore diameters were determined to analyze the state or location of pollutants as well as the regularity of formation. Meanwhile, recoil and ultrasonic physical measures were adopted to strengthen the membrane process, in order to explore the methods for preventing and controlling the membrane pollution. When 0.2 microm of ZrO2 micro-filtrated aqueous extract of Astragali Radix, the rate of pollution was as high as 44.9%. The hole blocking resistance and the concentration polarization resistance were the main filtration resistances, while the surface deposit resistance decreased with the increase in the membrane's hold diameter; after micro-filtration, the liquid turbidity significantly reduced, with slight changes in both pH and viscosity. The 0.2 microm ZrO2 micro-filtration membrane performed better than the 0.05 microm pore size membrane in terms of conductivity. The 0. 2 microm and 0.05 microm pore diameter membranes showed better performance in the removal of pectin. The ultrasonic measure to strengthen membranes is more suitable to this system, with a flux rate up by 41.7%. The membrane optimization process adopts appropriate measures for preventing and controlling the membrane pollution, in order to reduce the membrane pollution, recover membrane performance and increase filtration efficiency.

  10. Forced-flow bioreactor for sucrose inversion using ceramic membrane activated by silanization.

    Nakajima, M; Watanabe, A; Jimbo, N; Nishizawa, K; Nakao, S

    1989-02-20

    A forced-flow enzyme membrane reactor system for sucrose inversion was investigated using three ceramic membranes having different pore sizes. Invertase was immobilized chemically to the inner surface of a ceramic membrane activated by a silane-glutaraldehyde technique. With the cross-flow filtration of sucrose solution, the reaction rate was a function of the permeate flux, easily controlled by pressure. Using 0.5 microm support pore size of membrane, the volumetric productivity obtained was 10 times higher than that in a reported immobilized enzyme column reactor, with a short residence time of 5 s and 100% conversion of the sucrose inversion.

  11. Improved PVDF membrane performance by doping extracellular polymeric substances of activated sludge.

    Guan, Yan-Fang; Huang, Bao-Cheng; Qian, Chen; Wang, Long-Fei; Yu, Han-Qing

    2017-04-15

    Polyvinylidene fluoride (PVDF) membrane has been widely applied in water and wastewater treatment because of its high mechanical strength, thermal stability and chemical resistance. However, the hydrophobic nature of PVDF membrane makes it readily fouled, substantially reducing water flux and overall membrane rejection ability. In this work, an in-situ blending modifier, i.e., extracellular polymeric substances (EPS) from activated sludge, was used to enhance the anti-fouling ability of PVDF membrane. Results indicate that the pure water flux of the membrane and its anti-fouling performance were substantially improved by blending 8% EPS into the membrane. By introducing EPS, the membrane hydrophilicity was increased and the cross section morphology was changed when it interacted with polyvinl pyrrolidone, resulting in the formation of large cavities below the finger-like pores. In addition, the fraction of pores with a size of 100-500 nm increased, which was also beneficial to improving membrane performance. Surface thermodynamic calculations indicate the EPS-functionalized membrane had a higher cohesion free energy, implying its good pollutant rejection and anti-fouling ability. This work provides a simple, efficient and cost-effective method to improve membrane performance and also extends the applications of EPS. Copyright © 2017 Elsevier Ltd. All rights reserved.

  12. A Catalytically Active Membrane Reactor for Fast, Highly Exothermic, Heterogeneous Gas Reactions. A Pilot Plant Study

    Veldsink, Jan W.; Versteeg, Geert F.; Swaaij, Wim P.M. van

    1995-01-01

    Membrane reactors have been frequently studied because of their ability to combine chemical activity and separation properties into one device. Due to their thermal stability and mechanical strength, ceramic membranes are preferred over polymeric ones, but small transmembrane fluxes obstruct a

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

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

    2016-09-01

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

  14. The influence of activation of heterogeneous ion-exchange membranes on their electrochemical properties

    Brožová, Libuše; Křivčík, J.; Neděla, D.; Kysela, V.; Žitka, Jan

    2015-01-01

    Roč. 56, č. 12 (2015), s. 3228-3232 ISSN 1944-3994. [International Conference on Membrane and Electromembrane Processes - MELPRO 2014. Prague, 18.05.2014-21.05.2014] Institutional support: RVO:61389013 Keywords : heterogeneous ion-exchange membranes * electrochemical properties * activation Subject RIV: JP - Industrial Processing Impact factor: 1.272, year: 2015

  15. Preventing Anabolic Steroid Use: Guidelines and Activities.

    Nutter, June; Rauhe, Betty

    1997-01-01

    Information about anabolic steroids should be included in the school health curriculum as early as possible. The paper presents suggestions for planning education programs and offers a variety of activities and strategies appropriate for many age groups, including case studies, story completion, posters, demonstrations, projects, creative writing,…

  16. Preventing Running Injuries through Barefoot Activity

    Hart, Priscilla M.; Smith, Darla R.

    2008-01-01

    Running has become a very popular lifetime physical activity even though there are numerous reports of running injuries. Although common theories have pointed to impact forces and overpronation as the main contributors to chronic running injuries, the increased use of cushioning and orthotics has done little to decrease running injuries. A new…

  17. Mental disorder prevention and physical activity in Iranian elderly

    Seyede Salehe Mortazavi

    2012-01-01

    Conclusions: Physical activity significantly prevents mental disorder in older adults. Although it has effects on anxiety, social dysfunction, and depression, the greatest influence is on improving the somatization symptoms.

  18. Preparation, Characterization and Performance Studies of Active PVDF Ultrafiltration-Surfactants Membranes Containing PVP as Additive

    Nur Izzah Md Fadilah; Abdul Rahman Hassan

    2016-01-01

    The role of surfactants in the formation of active Poly(vinylidene fluoride) (PVDF) ultrafiltration (AUF) membranes was studied. The effect combination of surfactants that are Sodium dodecyl sulfate (SDS)/ Tween 80 and Tween 80/ Triton X-100 formulations on performance and morphological structures were investigated for the first time. The influence of surfactants blends on the membrane pores was also examined. Experimental data showed that combination of Tween 80/ Triton X-100 give the highest BSA permeation flux with a value of 285.51 Lm -2 h -1 . With combination of SDS/ Tween 80, the AUF membrane showed the highest protein rejection up to 93 % and 79 % for Bovine Serum Albumin (BSA) and Egg Albumin (EA), respectively. Moreover, membranes characterization demonstrated that the addition of SDS/ Tween 80 and Tween 80/ Triton X-100 were found to affect the performance, surface morphologies and membrane pores of AUF PVDF membranes. (author)

  19. Prevention Research Matters-Communities Working to Improve Physical Activity

    2018-02-15

    We know that children who are physically active every day are less likely to develop chronic diseases as adults, including obesity. Dr. Sandy Slater, a researcher with the University of Illinois, Chicago Prevention Research Center, discusses how a park improvement project in Chicago helped engage communities to improve areas for play and activity.  Created: 2/15/2018 by National Center for Chronic Disease Prevention and Health Promotion (NCCDPHP).   Date Released: 2/15/2018.

  20. G-protein activity in Percoll-purified plasma membranes, bulk plasma membranes, and low-density plasma membranes isolated from rat cerebral cortex

    Bouřová, Lenka; Stöhr, Jiří; Lisý, Václav; Rudajev, Vladimír; Novotný, Jiří; Svoboda, Petr

    2009-01-01

    Roč. 15, č. 4 (2009), BR111-BR122 ISSN 1234-1010 R&D Projects: GA MŠk(CZ) LC554; GA MŠk(CZ) LC06063; GA ČR(CZ) GA309/06/0121; GA AV ČR(CZ) IAA500110606 Institutional research plan: CEZ:AV0Z50110509 Keywords : rat cerebral cortex * plasma membrane * G-protein activity Subject RIV: CE - Biochemistry Impact factor: 1.543, year: 2009

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

    Shunsuke Yagi

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

  2. Cell activation and cellular-cellular interactions during hemodialysis: effect of dialyzer membrane.

    Sirolli, V; Ballone, E; Di Stante, S; Amoroso, L; Bonomini, M

    2002-06-01

    During hemodialysis (HD), circulating blood cells can be activated and also engage in dynamic interplay. These phenomena may be important factors behind dialysis membrane bio(in)compatibility. In the present prospective cross-over study, we have used flow cytometry to evaluate the influence of different dialysis membranes on the activation of circulating blood cells (leukocytes, platelets) and their dynamic interactions (formation of circulating platelet-leukocyte and platelet-erythrocyte aggregates) during in vivo HD. Each patient (n = 10) was treated with dialyzers containing membranes of cellulose diacetate, polysulfone and ethylenevinylalcohol (EVAL) in a randomized order. Upregulation of adhesion receptor expression (CD15s, CD11b/CD18) occurred mainly with the cellulosic membrane, though an increase in CD11b/CD18 circulating on neutrophils was also found with both synthetic membranes. Circulating activated platelets (P-selectin/CD63-positive platelets) increased during HD sessions with cellulose diacetate and polysulfone. An increased formation of platelet-neutrophil aggregates was found at 15 and 30 min during dialysis with cellulose diacetate and polysulfone but not with EVAL. Platelet-erythrocyte aggregates also increased with cellulose diacetate and at 15 min with polysulfone as well. Generally in concomitance with the increase in platelet-neutrophil coaggregates, there was an increased hydrogen peroxide production by neutrophils. The results of this study indicate that cellular mechanisms can be activated during HD largely depending on the membrane material, EVAL causing less reactivity than the other two membranes. It appears that each dialysis membrane has multiple and different characteristics that may contribute to interactions with blood components. Our results also indicate that derivatizing cellulose (cellulose diacetate) may be a useful way to improve the biocompatibility of the cellulose polymer and that there may be great variability in the

  3. Cellulose microfibril deposition: coordinated activity at the plant plasma membrane

    Lindeboom, J.J.; Mulder, B.; Vos, J.W.; Ketelaar, M.J.; Emons, A.M.C.

    2008-01-01

    Plant cell wall production is a membrane-bound process. Cell walls are composed of cellulose microfibrils, embedded inside a matrix of other polysaccharides and glycoproteins. The cell wall matrix is extruded into the existing cell wall by exocytosis. This same process also inserts the cellulose

  4. The roles of membrane microdomains (rafts) in T cell activation

    Hořejší, Václav

    2003-01-01

    Roč. 191, - (2003), s. 148-164 ISSN 0105-2896 R&D Projects: GA MŠk LN00A026 Grant - others:Wellcome Trust(GB) J1116W24Z Institutional research plan: CEZ:AV0Z5052915 Keywords : membrane microdomain * raft * T cell Subject RIV: EC - Immunology Impact factor: 7.052, year: 2003

  5. In vitro antioxidant and membrane stabilization activities of the fruit ...

    Arthritis is an inflammation of one or more joints. There is considerable experimental evidence linking lysosomal enzymes with tissue damage in arthritis. This study investigated anti-arthritic properties of Tetrapleura tetraptera (TT) using membrane stabilization assay (MSA). Powdered TT fruit sample was extracted by ...

  6. Plasma Membrane ATPase Activity following Reversible and Irreversible Freezing Injury 1

    Iswari, S.; Palta, Jiwan P.

    1989-01-01

    Plasma membrane ATPase has been proposed as a site of functional alteration during early stages of freezing injury. To test this, plasma membrane was purified from Solanum leaflets by a single step partitioning of microsomes in a dextran-polyethylene glycol two phase system. Addition of lysolecithin in the ATPase assay produced up to 10-fold increase in ATPase activity. ATPase activity was specific for ATP with a Km around 0.4 millimolar. Presence of the ATPase enzyme was identified by immunoblotting with oat ATPase antibodies. Using the phase partitioning method, plasma membrane was isolated from Solanum commersonii leaflets which had four different degrees of freezing damage, namely, slight (reversible), partial (partially reversible), substantial and total (irreversible). With slight (reversible) damage the plasma membrane ATPase specific activity increased 1.5- to 2-fold and its Km was decreased by about 3-fold, whereas the specific activity of cytochrome c reductase and cytochrome c oxidase in the microsomes were not different from the control. However, with substantial (lethal, irreversible) damage, there was a loss of membrane protein, decrease in plasma membrane ATPase specific activity and decrease in Km, while cytochrome c oxidase and cytochrome c reductase were unaffected. These results support the hypothesis that plasma membrane ATPase is altered by slight freeze-thaw stress. Images Figure 1 Figure 2 PMID:16666856

  7. Development and Characterization a Single-Active-Chamber Piezoelectric Membrane Pump with Multiple Passive Check Valves.

    Zhang, Ronghui; You, Feng; Lv, Zhihan; He, Zhaocheng; Wang, Haiwei; Huang, Ling

    2016-12-12

    In order to prevent the backward flow of piezoelectric pumps, this paper presents a single-active-chamber piezoelectric membrane pump with multiple passive check valves. Under the condition of a fixed total number of passive check valves, by means of changing the inlet valves and outlet valves' configuration, the pumping characteristics in terms of flow rate and backpressure are experimentally investigated. Like the maximum flow rate and backpressure, the testing results show that the optimal frequencies are significantly affected by changes in the number inlet valves and outlet valves. The variation ratios of the maximum flow rate and the maximum backpressure are up to 66% and less than 20%, respectively. Furthermore, the piezoelectric pump generally demonstrates very similar flow rate and backpressure characteristics when the number of inlet valves in one kind of configuration is the same as that of outlet valves in another configuration. The comparison indicates that the backflow from the pumping chamber to inlet is basically the same as the backflow from the outlet to the pumping chamber. No matter whether the number of inlet valves or the number of outlet valves is increased, the backflow can be effectively reduced. In addition, the backpressure fluctuation can be significantly suppressed with an increase of either inlet valves or outlet valves. It also means that the pump can prevent the backflow more effectively at the cost of power consumption. The pump is very suitable for conditions where more accurate flow rates are needed and wear and fatigue of check valves often occur.

  8. The Sur7 protein regulates plasma membrane organization and prevents intracellular cell wall growth in Candida albicans.

    Alvarez, Francisco J; Douglas, Lois M; Rosebrock, Adam; Konopka, James B

    2008-12-01

    The Candida albicans plasma membrane plays important roles in cell growth and as a target for antifungal drugs. Analysis of Ca-Sur7 showed that this four transmembrane domain protein localized to stable punctate patches, similar to the plasma membrane subdomains known as eisosomes or MCC that were discovered in S. cerevisiae. The localization of Ca-Sur7 depended on sphingolipid synthesis. In contrast to S. cerevisiae, a C. albicans sur7Delta mutant displayed defects in endocytosis and morphogenesis. Septins and actin were mislocalized, and cell wall synthesis was very abnormal, including long projections of cell wall into the cytoplasm. Several phenotypes of the sur7Delta mutant are similar to the effects of inhibiting beta-glucan synthase, suggesting that the abnormal cell wall synthesis is related to activation of chitin synthase activity seen under stress conditions. These results expand the roles of eisosomes by demonstrating that Sur7 is needed for proper plasma membrane organization and cell wall synthesis. A conserved Cys motif in the first extracellular loop of fungal Sur7 proteins is similar to a characteristic motif of the claudin proteins that form tight junctions in animal cells, suggesting a common role for these tetraspanning membrane proteins in forming specialized plasma membrane domains.

  9. The Sur7 Protein Regulates Plasma Membrane Organization and Prevents Intracellular Cell Wall Growth in Candida albicans

    Alvarez, Francisco J.; Douglas, Lois M.; Rosebrock, Adam

    2008-01-01

    The Candida albicans plasma membrane plays important roles in cell growth and as a target for antifungal drugs. Analysis of Ca-Sur7 showed that this four transmembrane domain protein localized to stable punctate patches, similar to the plasma membrane subdomains known as eisosomes or MCC that were discovered in S. cerevisiae. The localization of Ca-Sur7 depended on sphingolipid synthesis. In contrast to S. cerevisiae, a C. albicans sur7Δ mutant displayed defects in endocytosis and morphogenesis. Septins and actin were mislocalized, and cell wall synthesis was very abnormal, including long projections of cell wall into the cytoplasm. Several phenotypes of the sur7Δ mutant are similar to the effects of inhibiting β-glucan synthase, suggesting that the abnormal cell wall synthesis is related to activation of chitin synthase activity seen under stress conditions. These results expand the roles of eisosomes by demonstrating that Sur7 is needed for proper plasma membrane organization and cell wall synthesis. A conserved Cys motif in the first extracellular loop of fungal Sur7 proteins is similar to a characteristic motif of the claudin proteins that form tight junctions in animal cells, suggesting a common role for these tetraspanning membrane proteins in forming specialized plasma membrane domains. PMID:18799621

  10. Type-1 cannabinoid receptors reduce membrane fluidity of capacitated boar sperm by impairing their activation by bicarbonate.

    Barbara Barboni

    Full Text Available BACKGROUND: Mammalian spermatozoa acquire their full fertilizing ability (so called capacitation within the female genital tract, where they are progressively exposed to inverse gradients of inhibiting and stimulating molecules. METHODOLOGY/PRINCIPAL FINDINGS: In the present research, the effect on this process of anandamide, an endocannabinoid that can either activate or inhibit cannabinoid receptors depending on its concentration, and bicarbonate, an oviductal activatory molecule, was assessed, in order to study the role exerted by the type 1 cannabinoid receptor (CB1R in the process of lipid membrane remodeling crucial to complete capacitation. To this aim, boar sperm were incubated in vitro under capacitating conditions (stimulated by bicarbonate in the presence or in the absence of methanandamide (Met-AEA, a non-hydrolysable analogue of anandamide. The CB1R involvement was studied by using the specific inhibitor (SR141716 or mimicking its activation by adding a permeable cAMP analogue (8Br-cAMP. By an immunocytochemistry approach it was shown that the Met-AEA inhibits the bicarbonate-dependent translocation of CB1R from the post-equatorial to equatorial region of sperm head. In addition it was found that Met-AEA is able to prevent the bicarbonate-induced increase in membrane disorder and the cholesterol extraction, both preliminary to capacitation, acting through a CB1R-cAMP mediated pathway, as indicated by MC540 and filipin staining, EPR spectroscopy and biochemical analysis on whole membranes (CB1R activity and on membrane enriched fraction (C/P content and anisotropy. CONCLUSIONS/SIGNIFICANCE: Altogether, these data demonstrate that the endocannabinoid system strongly inhibits the process of sperm capacitation, acting as membrane stabilizing agent, thus increasing the basic knowledge on capacitation-related signaling and potentially opening new perspectives in diagnostics and therapeutics of male infertility.

  11. Cancer-preventive activities of tocopherols and tocotrienols.

    Ju, Jihyeung; Picinich, Sonia C; Yang, Zhihong; Zhao, Yang; Suh, Nanjoo; Kong, Ah-Ng; Yang, Chung S

    2010-04-01

    The cancer-preventive activity of vitamin E has been studied. Whereas some epidemiological studies have suggested a protective effect of vitamin E against cancer formation, many large-scale intervention studies with alpha-tocopherol (usually large doses) have not demonstrated a cancer-preventive effect. Studies on alpha-tocopherol in animal models also have not demonstrated robust cancer prevention effects. One possible explanation for the lack of demonstrable cancer-preventive effects is that high doses of alpha-tocopherol decrease the blood and tissue levels of delta-tocopherols. It has been suggested that gamma-tocopherol, due to its strong anti-inflammatory and other activities, may be the more effective form of vitamin E in cancer prevention. Our recent results have demonstrated that a gamma-tocopherol-rich mixture of tocopherols inhibits colon, prostate, mammary and lung tumorigenesis in animal models, suggesting that this mixture may have a high potential for applications in the prevention of human cancer. In this review, we discuss biochemical properties of tocopherols, results of possible cancer-preventive effects in humans and animal models and possible mechanisms involved in the inhibition of carcinogenesis. Based on this information, we propose that a gamma-tocopherol-rich mixture of tocopherols is a very promising cancer-preventive agent and warrants extensive future research.

  12. Enhancement of antibacterial activity in nanofillers incorporated PSF/PVP membranes

    Pramila, P.; Gopalakrishnan, N.

    2018-04-01

    An attempt has been made to investigate the nanofillers incorporated polysulfone (PSF) and polyvinylpyrrolidone (PVP) polymer membranes prepared by phase inversion method. Initially, the nanofillers, viz, Zinc Oxide (ZnO) nanoparticle, Graphene Oxide-Zinc Oxide (GO-ZnO) nanocomposite were synthesized and then directly incorporated into PSF/PVP blend during the preparation of membranes. The prepared membranes have been subjected to FE-SEM, AFM, BET, contact angle, tensile test and anti-bacterial studies. Significant membrane morphologies and nanoporous properties have been observed by FE-SEM and BET, respectively. It has been observed that hydrophilicity, mechanical strength and water permeability of the ZnO and GO-ZnO incorporated membranes were enhanced than bare membrane. Antibacterial activity was assessed by measuring the inhibition zones formed around the membrane by disc-diffusion method using Escherichia coli (gram-negative) as a model bacterium. Again, it has been observed that nanofillers incorporated membrane exhibits high antibacterial performance compared to bare membrane.

  13. Modification of trout sperm membranes associated with activation and cryopreservation. Implications for fertilizing potential

    Abstract We investigated the effects of two trout sperm activation solutions on sperm physiology and membrane organization prior to and following cryopreservation using flow cytometry and investigated their impact on in vitro fertility. Cryopreservation caused greater phospholipid disorder (high pl...

  14. The Tower: Modelling, Analysis and Construction of Bending Active Tensile Membrane Hybrid Structures

    Holden Deleuran, Anders; Schmeck, Michel; Charles Quinn, Gregory

    2015-01-01

    The project is the result of an interdisciplinary research collaboration between CITA, KET and Fibrenamics exploring the design of integrated hybrid structures employing bending active elements and tensile membranes with bespoke material properties and detailing. Hybrid structures are defined her...

  15. Role of Membrane Cholesterol Levels in Activation of Lyn upon Cell Detachment

    Takao Morinaga

    2018-06-01

    Full Text Available Cholesterol, a major component of the plasma membrane, determines the physicalproperties of biological membranes and plays a critical role in the assembly of membranemicrodomains. Enrichment or deprivation of membrane cholesterol affects the activities of manysignaling molecules at the plasma membrane. Cell detachment changes the structure of the plasmamembrane and influences the localizations of lipids, including cholesterol. Recent studies showedthat cell detachment changes the activities of a variety of signaling molecules. We previously reportedthat the localization and the function of the Src-family kinase Lyn are critically regulated by itsmembrane anchorage through lipid modifications. More recently, we found that the localization andthe activity of Lyn were changed upon cell detachment, although the manners of which vary betweencell types. In this review, we highlight the changes in the localization of Lyn and a role of cholesterolin the regulation of Lyn’s activation following cell detachment.

  16. Machine learning-enabled discovery and design of membrane-active peptides.

    Lee, Ernest Y; Wong, Gerard C L; Ferguson, Andrew L

    2017-07-08

    Antimicrobial peptides are a class of membrane-active peptides that form a critical component of innate host immunity and possess a diversity of sequence and structure. Machine learning approaches have been profitably employed to efficiently screen sequence space and guide experiment towards promising candidates with high putative activity. In this mini-review, we provide an introduction to antimicrobial peptides and summarize recent advances in machine learning-enabled antimicrobial peptide discovery and design with a focus on a recent work Lee et al. Proc. Natl. Acad. Sci. USA 2016;113(48):13588-13593. This study reports the development of a support vector machine classifier to aid in the design of membrane active peptides. We use this model to discover membrane activity as a multiplexed function in diverse peptide families and provide interpretable understanding of the physicochemical properties and mechanisms governing membrane activity. Experimental validation of the classifier reveals it to have learned membrane activity as a unifying signature of antimicrobial peptides with diverse modes of action. Some of the discriminating rules by which it performs classification are in line with existing "human learned" understanding, but it also unveils new previously unknown determinants and multidimensional couplings governing membrane activity. Integrating machine learning with targeted experimentation can guide both antimicrobial peptide discovery and design and new understanding of the properties and mechanisms underpinning their modes of action. Copyright © 2017 Elsevier Ltd. All rights reserved.

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

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

    2017-09-08

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

  18. The Effects of Altered Membrane Cholesterol Levels on Sodium Pump Activity in Subclinical Hypothyroidism

    Suparna Roy

    2017-02-01

    Full Text Available BackgroundMetabolic dysfunctions characteristic of overt hypothyroidism (OH start at the early stage of subclinical hypothyroidism (SCH. Na+/K+-ATPase (the sodium pump is a transmembrane enzyme that plays a vital role in cellular activities in combination with membrane lipids. We evaluated the effects of early changes in thyroid hormone and membrane cholesterol on sodium pump activity in SCH and OH patients.MethodsIn 32 SCH patients, 35 OH patients, and 34 euthyroid patients, sodium pump activity and cholesterol levels in red blood cell membranes were measured. Serum thyroxine (T4 and thyroid stimulating hormone (TSH levels were measured using enzyme-linked immunosorbent assays. Differences in their mean values were analysed using post hoc analysis of variance. We assessed the dependence of the sodium pump on other metabolites by multiple regression analysis.ResultsSodium pump activity and membrane cholesterol were lower in both hypothyroid groups than in control group, OH group exhibiting lower values than SCH group. In SCH group, sodium pump activity showed a significant direct dependence on membrane cholesterol with an inverse relationship with serum TSH levels. In OH group, sodium pump activity depended directly on membrane cholesterol and serum T4 levels. No dependence on serum cholesterol was observed in either case.ConclusionDespite the presence of elevated serum cholesterol in hypothyroidism, membrane cholesterol contributed significantly to maintain sodium pump activity in the cells. A critical reduction in membrane cholesterol levels heralds compromised enzyme activity, even in the early stage of hypothyroidism, and this can be predicted by elevated TSH levels alone, without any evident clinical manifestations.

  19. Designing CNC Knit for Hybrid Membrane And Bending Active Structures

    Tamke, Martin; Holden Deleuran, Anders; Gengnagel, Christoph

    2015-01-01

    specific properties and detailing. CNC knitting with high tenacity yarn enables this practice and offers an alternative to current woven membranes. The design and fabrication of an 8m high fabric tower through an interdisciplinary team of architects, structural and textile engineers, allowed to investigate...... means to design, specify, make and test CNC knit as material for hybrid structures in architectural scale. This paper shares the developed process, identifies challenges, potentials and future work...

  20. High-pressure stainless steel active membrane microvalves

    Sharma, G; Svensson, S; Ogden, S; Klintberg, L; Hjort, K

    2011-01-01

    In this work, high-pressure membrane microvalves have been designed, manufactured and evaluated. The valves were able to withstand back-pressures of 200 bar with a response time of less than 0.6 s. These stainless steel valves, manufactured with back-end batch production, utilize the large volume expansion coupled to the solid–liquid phase transition in paraffin wax. When membrane materials were evaluated, parylene coated stainless steel was found to be the best choice as compared to polydimethylsiloxane and polyimide. Also, the influence of the orifice placement and diameter is included in this work. If the orifice is placed too close to the rim of the membrane, the valve can stay sealed even after turning the power off, and the valve will not open until the pressure in the system is released. The developed steel valves, evaluated for both water and air, provide excellent properties in terms of mechanical stability, ease of fabrication, and low cost. Possible applications include sampling at high pressures, chemical microreactors, high performance liquid chromatography, pneumatics, and hydraulics

  1. LH-RH binding to purified pituitary plasma membranes: absence of adenylate cyclase activation.

    Clayton, R N; Shakespear, R A; Marshall, J C

    1978-06-01

    Purified bovine pituitary plasma membranes possess two specific LH-RH binding sites. The high affinity site (2.5 X 10(9) l/mol) has low capacity (9 X 10(-15) mol/mg membrane protein) while the low affinity site 6.1 X 10(5) l/mol) has a much higher capacity (1.1 X 10(-10) mol/mg). Specific LH-RH binding to plasma membranes is increased 8.5-fold during purification from homogenate whilst adenylate cyclase activity is enriched 7--8-fold. Distribution of specific LH-RH binding to sucrose density gradient interface fractions parallels that of adenylate cyclase activity. Mg2+ and Ca2+ inhibit specific [125I]LH-RH binding at micromolar concentrations. Synthetic LH-RH, up to 250 microgram/ml, failed to stimulate adenylase cyclase activity of the purified bovine membranes. Using a crude 10,800 g rat pituitary membrane preparation, LH-RH similarly failed to activate adenylate cyclase even in the presence of guanyl nucleotides. These data confirm the presence of LH-RH receptor sites on pituitary plasma membranes and suggest that LH-RH-induced gonadotrophin release may be mediated by mechanisms other than activation of adenylate cyclase.

  2. Recognition of acidic phospholipase A2 activity in plasma membranes of resident peritoneal macrophages

    Shibata, Y.; Abiko, Y.; Ohno, H.; Araki, T.; Takiguchi, H.

    1988-01-01

    Phospholipase (PLase) activities in the plasma membrane of guinea pig peritoneal macrophages were studied, as these enzymes having such activity may be candidates for the release of arachidonic acid (AA) from phosphatidylcholine (PC). An AA release system operating at acidic pH was identified in the macrophage plasma membrane and characterized. This membrane-bound acidic PLase A 2 had an optimum pH at 4.5, and enzyme activation was observed in Ca ++ -free medium; but the maximum activity was found at 0.5 mM Ca ++ concentration. The Km value for PC of acidic PLase A 2 was 4.2 μM, and a Michaelis-Menten relationship was evident. Calcium might act as a cofactor at some intermediate step during the activation of acidic PLase A 2 in light of the uncompetitive manner of Ca ++ action. Furthermore, the release of [ 3 H]-AA from preradiolabelled macrophage plasma membranes occurred with the addition of Ca ++ at pH 4.5. These data suggest that the acid PLase A 2 is a component of the plasma membrane and is not due to lysosomal contamination since membrane-bound acidic PLase A 2 properties are opposite to those found for lysosomal PLase A 2

  3. Estimating the magnitude of near-membrane PDE4 activity in living cells.

    Xin, Wenkuan; Feinstein, Wei P; Britain, Andrea L; Ochoa, Cristhiaan D; Zhu, Bing; Richter, Wito; Leavesley, Silas J; Rich, Thomas C

    2015-09-15

    Recent studies have demonstrated that functionally discrete pools of phosphodiesterase (PDE) activity regulate distinct cellular functions. While the importance of localized pools of enzyme activity has become apparent, few studies have estimated enzyme activity within discrete subcellular compartments. Here we present an approach to estimate near-membrane PDE activity. First, total PDE activity is measured using traditional PDE activity assays. Second, known cAMP concentrations are dialyzed into single cells and the spatial spread of cAMP is monitored using cyclic nucleotide-gated channels. Third, mathematical models are used to estimate the spatial distribution of PDE activity within cells. Using this three-tiered approach, we observed two pharmacologically distinct pools of PDE activity, a rolipram-sensitive pool and an 8-methoxymethyl IBMX (8MM-IBMX)-sensitive pool. We observed that the rolipram-sensitive PDE (PDE4) was primarily responsible for cAMP hydrolysis near the plasma membrane. Finally, we observed that PDE4 was capable of blunting cAMP levels near the plasma membrane even when 100 μM cAMP were introduced into the cell via a patch pipette. Two compartment models predict that PDE activity near the plasma membrane, near cyclic nucleotide-gated channels, was significantly lower than total cellular PDE activity and that a slow spatial spread of cAMP allowed PDE activity to effectively hydrolyze near-membrane cAMP. These results imply that cAMP levels near the plasma membrane are distinct from those in other subcellular compartments; PDE activity is not uniform within cells; and localized pools of AC and PDE activities are responsible for controlling cAMP levels within distinct subcellular compartments. Copyright © 2015 the American Physiological Society.

  4. Analysis of functional damages in membrane receptor caused by radiation exposure and development of damage-preventing techniques by modification of membrane lipids

    Amano, Fumio; Tanaka, Yasuhito; Asano, Toshihiko; Ogura, Atsuro; Suzuki, Osamu; Matsuda, Junichiro

    1998-01-01

    This study was made to assess the physiological effects of radiation exposure using ligand-binding ability of the receptor on cell membrane. The effects of exposure to low-dose radiation were investigated in macrophage lines, J774.1 and RAW264.7 and human monocyte cell line, U937 using γ-radiation apparatus. In macrophages, the production of active oxygen (O 2 - ) was dose-dependently increased by γ-ray radiation, suggesting that the production might occur through the activation of proteinkinase C. Arachidonic acid bending BSA (AA-BSA) and eicosapentadienic acid binding BSA (EP-BSA) were used to modify the fatty acids in J774.1 cell and both BSA-treated cells exhibited significantly higher colony-forming abilities after γ-ray radiation. The modification of membrane lipids by AA-BSA or EPA-BSA resulted to reduce the inhibition of cell growth and change the cell adhesion ability and the fine structure of cell surface. Furthermore, the effects of radiation on behavior were investigated in PC mouse and the circadian rhythm of locomotive activity was found to be affected by radiation at 5-10 Gy. (M.N.)

  5. Calmodulin and CaMKII modulate ENaC activity by regulating the association of MARCKS and the cytoskeleton with the apical membrane.

    Alli, Abdel A; Bao, Hui-Fang; Liu, Bing-Chen; Yu, Ling; Aldrugh, Summer; Montgomery, Darrice S; Ma, He-Ping; Eaton, Douglas C

    2015-09-01

    Phosphatidylinositol bisphosphate (PIP2) regulates epithelial sodium channel (ENaC) open probability. In turn, myristoylated alanine-rich C kinase substrate (MARCKS) protein or MARCKS-like protein 1 (MLP-1) at the plasma membrane regulates the delivery of PIP2 to ENaC. MARCKS and MLP-1 are regulated by changes in cytosolic calcium; increasing calcium promotes dissociation of MARCKS from the membrane, but the calcium-regulatory mechanisms are unclear. However, it is known that increased intracellular calcium can activate calmodulin and we show that inhibition of calmodulin with calmidazolium increases ENaC activity presumably by regulating MARCKS and MLP-1. Activated calmodulin can regulate MARCKS and MLP-1 in two ways. Calmodulin can bind to the effector domain of MARCKS or MLP-1, inactivating both proteins by causing their dissociation from the membrane. Mutations in MARCKS that prevent calmodulin association prevent dissociation of MARCKS from the membrane. Calmodulin also activates CaM kinase II (CaMKII). An inhibitor of CaMKII (KN93) increases ENaC activity, MARCKS association with ENaC, and promotes MARCKS movement to a membrane fraction. CaMKII phosphorylates filamin. Filamin is an essential component of the cytoskeleton and promotes association of ENaC, MARCKS, and MLP-1. Disruption of the cytoskeleton with cytochalasin E reduces ENaC activity. CaMKII phosphorylation of filamin disrupts the cytoskeleton and the association of MARCKS, MLP-1, and ENaC, thereby reducing ENaC open probability. Taken together, these findings suggest calmodulin and CaMKII modulate ENaC activity by destabilizing the association between the actin cytoskeleton, ENaC, and MARCKS, or MLP-1 at the apical membrane. Copyright © 2015 the American Physiological Society.

  6. Differential effects of inhibitors and detergents on the Ca2+-ATPase and Mg2+-ATPase activities of the plasma membrane of a human oat cell carcinoma

    Knowles, A.F.; Lawrence, C.M.

    1986-01-01

    Plasma membranes of human oat cell carcinoma possess Mg 2+ - and Ca 2+ -dependent ATPase activities of similar magnitude. These activities exhibit the unusual characteristic of being inactiviated by prolonged incubation of the membrane with 1-2 mM dithiothreitol (DTT). Inactivation by DTT was prevented by lowering the incubation temperature, elevation of the membrane protein concentration, and addition of ATP. Fluorosulfonylbenzoyl adenosine (FSBA), an affinity ATP analog, also inactivates these activities. The Ca 2+ -ATPase activity appears to be more sensitive to both DTT and FSBA. The Ca 2+ -ATPase activity is more easily inactivated by Triton X-100, while the Mg 2+ -ATPase is preferentially activated by digitonin. These differential effects of inhibitors and detergents suggest that the Ca 2+ -ATPase and Mg 2+ -ATPase are separate enzymes. Incubation of oat cell carcinoma plasma membrane with [ 3 H]FSBA resulted in the labeling of several proteins. A labelled 35,000 dalton protein corresponds to the molecular weight of the oat cell carcinoma plasma membrane Ca 2+ -ATPase previously purified in this laboratory. The identity of one or more of the other labelled proteins with the Mg 2+ -ATPase has not been demonstrated, but is presently under investigation

  7. The higher level of complexity of K-Ras4B activation at the membrane

    Jang, Hyunbum; Banerjee, Avik; Chavan, Tanmay S.; Lu, Shaoyong; Zhang, Jian; Gaponenko, Vadim; Nussinov, Ruth

    2016-01-01

    Is nucleotide exchange sufficient to activate K-Ras4B? To signal, oncogenic rat sarcoma (Ras) anchors in the membrane and recruits effectors by exposing its effector lobe. With the use of NMR and molecular dynamics (MD) simulations, we observed that in solution, farnesylated guanosine 5′-diphosphate (GDP)-bound K-Ras4B is predominantly autoinhibited by its hypervariable region (HVR), whereas the GTP-bound state favors an activated, HVR-released state. On the anionic membrane, the catalytic domain adopts multiple orientations, including parallel (∼180°) and perpendicular (∼90°) alignments of the allosteric helices, with respect to the membrane surface direction. In the autoinhibited state, the HVR is sandwiched between the effector lobe and the membrane; in the active state, with membrane-anchored farnesyl and unrestrained HVR, the catalytic domain fluctuates reinlessly, exposing its effector-binding site. Dimerization and clustering can reduce the fluctuations. This achieves preorganized, productive conformations. Notably, we also observe HVR-autoinhibited K-Ras4B-GTP states, with GDP-bound-like orientations of the helices. Thus, we propose that the GDP/GTP exchange may not be sufficient for activation; instead, our results suggest that the GDP/GTP exchange, HVR sequestration, farnesyl insertion, and orientation/localization of the catalytic domain at the membrane conjointly determine the active or inactive state of K-Ras4B. Importantly, K-Ras4B-GTP can exist in active and inactive states; on its own, GTP binding may not compel K-Ras4B activation.—Jang, H., Banerjee, A., Chavan, T. S, Lu, S., Zhang, J., Gaponenko, V., Nussinov, R. The higher level of complexity of K-Ras4B activation at the membrane. PMID:26718888

  8. The higher level of complexity of K-Ras4B activation at the membrane.

    Jang, Hyunbum; Banerjee, Avik; Chavan, Tanmay S; Lu, Shaoyong; Zhang, Jian; Gaponenko, Vadim; Nussinov, Ruth

    2016-04-01

    Is nucleotide exchange sufficient to activate K-Ras4B? To signal, oncogenic rat sarcoma (Ras) anchors in the membrane and recruits effectors by exposing its effector lobe. With the use of NMR and molecular dynamics (MD) simulations, we observed that in solution, farnesylated guanosine 5'-diphosphate (GDP)-bound K-Ras4B is predominantly autoinhibited by its hypervariable region (HVR), whereas the GTP-bound state favors an activated, HVR-released state. On the anionic membrane, the catalytic domain adopts multiple orientations, including parallel (∼180°) and perpendicular (∼90°) alignments of the allosteric helices, with respect to the membrane surface direction. In the autoinhibited state, the HVR is sandwiched between the effector lobe and the membrane; in the active state, with membrane-anchored farnesyl and unrestrained HVR, the catalytic domain fluctuates reinlessly, exposing its effector-binding site. Dimerization and clustering can reduce the fluctuations. This achieves preorganized, productive conformations. Notably, we also observe HVR-autoinhibited K-Ras4B-GTP states, with GDP-bound-like orientations of the helices. Thus, we propose that the GDP/GTP exchange may not be sufficient for activation; instead, our results suggest that the GDP/GTP exchange, HVR sequestration, farnesyl insertion, and orientation/localization of the catalytic domain at the membrane conjointly determine the active or inactive state of K-Ras4B. Importantly, K-Ras4B-GTP can exist in active and inactive states; on its own, GTP binding may not compel K-Ras4B activation.-Jang, H., Banerjee, A., Chavan, T. S, Lu, S., Zhang, J., Gaponenko, V., Nussinov, R. The higher level of complexity of K-Ras4B activation at the membrane. © FASEB.

  9. Estradiol-mediated internalisation of the non-activated estrogen receptor from the goat uterine plasma membrane: identification of the proteins involved.

    Sreeja, S; Thampan, Raghava Varman

    2004-04-01

    An indirect approach has been made to study the molecular details associated with the estradiol-induced internalisation of the non-activated estrogen receptor (naER) from the goat uterine plasma membrane. The internalisation of naER appears to be an energy dependent process. Exposure of the plasma membrane to estradiol results in the activation of a Mg2+ dependent ATPase associated with the membrane fraction. Presence of quercetin in the medium prevented the activation of the Mg2+ ATPase as well as the dissociation of naER from the plasma membrane. Using isolated plasma membrane preparations it has been possible to identify the proteins which interact with naER during various stages of its internalisation. The main proteins identified are: (1) a 58 kDa protein, p58, which apparently recognizes the nuclear localization signals on the naER and transports it to the nucleus: (2) hsp70: (3) hsp90, the functional roles of which remain unknown at this stage; (4) a 50 kDa protein associated with the clathrin coated vesicles, presumed to be involved in recognizing the tyrosine based internalisation signals on the naER; (5) actin which mediates the plasma membrane-to-nucleus movement of the naER-p58 complex.

  10. Ion channel activity of membrane vesicles released from sea urchin sperm during the acrosome reaction

    Schulz, Joseph R.; Vega-Beltran, Jose L. de la; Beltran, Carmen; Vacquier, Victor D.; Darszon, Alberto

    2004-01-01

    The sperm acrosome reaction (AR) involves ion channel activation. In sea urchin sperm, the AR requires Ca 2+ and Na + influx and K + and H + efflux. During the AR, the plasma membrane fuses with the acrosomal vesicle membrane forming hybrid membrane vesicles that are released from sperm into the medium. This paper reports the isolation and preliminary characterization of these acrosome reaction vesicles (ARVs), using synaptosome-associated protein of 25 kDa (SNAP-25) as a marker. Isolated ARVs have a unique protein composition. The exocytosis regulatory proteins vesicle-associated membrane protein and SNAP-25 are inside ARVs, as judged by protease protection experiments, and membrane associated based on Triton X-114 partitioning. ARVs fused with planar bilayers display three main types of single channel activity. The most frequently recorded channel is cationic, weakly voltage dependent and has a low open probability that increases with negative potentials. This channel is activated by cAMP, blocked by Ba 2+ , and has a PK + /PNa + selectivity of 4.5. ARVs represent a novel membrane preparation suitable to deepen our understanding of ion channel activity in the AR and during fertilization

  11. Activation of Endothelial Nitric Oxide (eNOS Occurs through Different Membrane Domains in Endothelial Cells.

    Jason Tran

    Full Text Available Endothelial cells respond to a large range of stimuli including circulating lipoproteins, growth factors and changes in haemodynamic mechanical forces to regulate the activity of endothelial nitric oxide synthase (eNOS and maintain blood pressure. While many signalling pathways have been mapped, the identities of membrane domains through which these signals are transmitted are less well characterized. Here, we manipulated bovine aortic endothelial cells (BAEC with cholesterol and the oxysterol 7-ketocholesterol (7KC. Using a range of microscopy techniques including confocal, 2-photon, super-resolution and electron microscopy, we found that sterol enrichment had differential effects on eNOS and caveolin-1 (Cav1 colocalisation, membrane order of the plasma membrane, caveolae numbers and Cav1 clustering. We found a correlation between cholesterol-induced condensation of the plasma membrane and enhanced high density lipoprotein (HDL-induced eNOS activity and phosphorylation suggesting that cholesterol domains, but not individual caveolae, mediate HDL stimulation of eNOS. Vascular endothelial growth factor (VEGF-induced and shear stress-induced eNOS activity was relatively independent of membrane order and may be predominantly controlled by the number of caveolae on the cell surface. Taken together, our data suggest that signals that activate and phosphorylate eNOS are transmitted through distinct membrane domains in endothelial cells.

  12. Nonlinear Dielectric Spectroscopy as an Indirect Probe of Metabolic Activity in Thylakoid Membrane

    John H. Miller

    2011-01-01

    Full Text Available Nonlinear dielectric spectroscopy (NDS is a non-invasive probe of cellular metabolic activity with potential application in the development of whole-cell biosensors. However, the mechanism of NDS interaction with metabolic membrane proteins is poorly understood, partly due to the inherent complexity of single cell organisms. Here we use the light-activated electron transport chain of spinach thylakoid membrane as a model system to study how NDS interacts with metabolic activity. We find protein modification, as opposed to membrane pump activity, to be the dominant source of NDS signal change in this system. Potential mechanisms for such protein modifications include reactive oxygen species generation and light-activated phosphorylation.

  13. Caffeine inhibits erythrocyte membrane derangement by antioxidant activity and by blocking caspase 3 activation.

    Tellone, Ester; Ficarra, Silvana; Russo, Annamaria; Bellocco, Ersilia; Barreca, Davide; Laganà, Giuseppina; Leuzzi, Ugo; Pirolli, Davide; De Rosa, Maria Cristina; Giardina, Bruno; Galtieri, Antonio

    2012-02-01

    The aim of this research was to investigate the effect of caffeine on band 3 (the anion exchanger protein), haemoglobin function, caspase 3 activation and glucose-6-phosphate metabolism during the oxygenation-deoxygenation cycle in human red blood cells. A particular attention has been given to the antioxidant activity by using in vitro antioxidant models. Caffeine crosses the erythrocyte membrane and interacts with the two extreme conformational states of haemoglobin (the T and the R-state within the framework of the simple two states allosteric model) with different binding affinities. By promoting the high affinity state (R-state), the caffeine-haemoglobin interaction does enhance the pentose phosphate pathway. This is of benefit for red blood cells since it leads to an increase of NADPH availability. Moreover, caffeine effect on band 3, mediated by haemoglobin, results in an extreme increase of the anion exchange, particularly in oxygenated erythrocytes. This enhances the transport of the endogenously produced CO(2) thereby avoiding the production of dangerous secondary radicals (carbonate and nitrogen dioxide) which are harmful to the cellular membrane. Furthermore caffeine destabilizes the haeme-protein interactions within the haemoglobin molecule and triggers the production of superoxide and met-haemoglobin. However this damaging effect is almost balanced by the surprising scavenger action of the alkaloid with respect to the hydroxyl radical. These experimental findings are supported by in silico docking and molecular dynamics studies and by what we may call the "caspase silence"; in fact, there is no evidence of any caspase 3 activity enhancement; this is likely due to the promotion of positive metabolic conditions which result in an increase of the cellular reducing power. Copyright © 2011 Elsevier Masson SAS. All rights reserved.

  14. Tuning Liposome Membrane Permeability by Competitive Peptide Dimerization and Partitioning-Folding Interactions Regulated by Proteolytic Activity

    Lim, Seng Koon; Sandén, Camilla; Selegård, Robert; Liedberg, Bo; Aili, Daniel

    2016-02-01

    Membrane active peptides are of large interest for development of drug delivery vehicles and therapeutics for treatment of multiple drug resistant infections. Lack of specificity can be detrimental and finding routes to tune specificity and activity of membrane active peptides is vital for improving their therapeutic efficacy and minimize harmful side effects. We describe a de novo designed membrane active peptide that partition into lipid membranes only when specifically and covalently anchored to the membrane, resulting in pore-formation. Dimerization with a complementary peptide efficiently inhibits formation of pores. The effect can be regulated by proteolytic digestion of the inhibitory peptide by the matrix metalloproteinase MMP-7, an enzyme upregulated in many malignant tumors. This system thus provides a precise and specific route for tuning the permeability of lipid membranes and a novel strategy for development of recognition based membrane active peptides and indirect enzymatically controlled release of liposomal cargo.

  15. Cisplatin impairs rat liver mitochondrial functions by inducing changes on membrane ion permeability: Prevention by thiol group protecting agents

    Custodio, Jose B.A.; Cardoso, Carla M.P.; Santos, Maria S.; Almeida, Leonor M.; Vicente, Joaquim A.F.; Fernandes, Maria A.S.

    2009-01-01

    Cisplatin (CisPt) is the most important platinum anticancer drug widely used in the treatment of head, neck, ovarian and testicular cancers. However, the mechanisms by which CisPt induces cytotoxicity, namely hepatotoxicity, are not completely understood. The goal of this study was to investigate the influence of CisPt on rat liver mitochondrial functions (Ca 2+ -induced mitochondrial permeability transition (MPT), mitochondrial bioenergetics, and mitochondrial oxidative stress) to better understand the mechanism underlying its hepatotoxicity. The effect of thiol group protecting agents and some antioxidants against CisPt-induced mitochondrial damage was also investigated. Treatment of rat liver mitochondria with CisPt (20 nmol/mg protein) induced Ca 2+ -dependent mitochondrial swelling, depolarization of membrane potential (ΔΨ), Ca 2+ release, and NAD(P)H fluorescence intensity decay. These effects were prevented by cyclosporine A (CyA), a potent and specific inhibitor of the MPT. In the concentration range of up to 40 nmol/mg protein, CisPt slightly inhibited state 3 and stimulated state 2 and state 4 respiration rates using succinate as respiratory substrate. The respiratory indexes, respiratory control ratio (RCR) and ADP/O ratios, the ΔΨ, and the ADP phosphorylation rate were also depressed. CisPt induced mitochondrial inner membrane permeabilization to protons (proton leak) but did not induce significant changes on mitochondrial H 2 O 2 generation. All the effects induced by CisPt on rat liver mitochondria were prevented by thiol group protecting agents namely, glutathione (GSH), dithiothreitol (DTT), N-acetyl-L-cysteine (NAC) and cysteine (CYS), whereas superoxide-dismutase (SOD), catalase (CAT) and ascorbate (ASC) were without effect. In conclusion, the anticancer drug CisPt: (1) increases the sensitivity of mitochondria to Ca 2+ -induced MPT; (2) interferes with mitochondrial bioenergetics by increasing mitochondrial inner membrane permeabilization to

  16. [Physical activity in basic and primary prevention of cardiovascular disease].

    Sobieszczańska, Małgorzata; Kałka, Dariusz; Pilecki, Witold; Adamus, Jerzy

    2009-06-01

    On account of the frequency of appearing and character of atherosclerosis cardiac vascular disease, one of the most crucial elements of effective fight against it is preparation of complex preventive programs including as vast number of population as possible. Consequently, Benjamin and Smitch suggested attaching the notion of basic prevention to the standard division into primary and secondary one. The basic prevention, carrying out in the general population, should concern genetic predisposition, psychosocial factors, keeping up proper body weight, healthy eating and physical activity. Especially high hopes are connected with high efficiency, simplicity and low money-consumption of preventive activities associated with physical activity modification, which has a crucial influence on reducing negative impact of atherosclerosis hazard. The results of numerous scientific research, carried out in many countries and on various, large groups, proved undoubtedly that at the healthy adult people of both sex the systematic physical activity of moderate intensification plays an essential part in preventing CVD and decreasing the death risk because of that reason as well. Moreover, systematic physical exercises show many other health-oriented actions, thanks to which they have an influence on decreasing premature and total death rate. The risk of incidence of civilization-related diseases such as diabetes type II, hypertension, obesity, osteoporosis, tumors (of large intestine, breast, prostatic gland) and depression has decreased significantly. Unequivocally positive influence has been proved at many observations dedicated to health recreational physical activity and physical activity connected with professional work based on aerobe effort. The positive effects have been also observed at children population and senior population which is more and more numerous and the most at risk. The beneficial action of physical activity is connected with direct effect on organism

  17. Forming a Learning Culture to Promote Fracture Prevention Activities

    Hjalmarson, Helene V.; Strandmark, Margaretha

    2012-01-01

    Purpose: The purpose of this paper is to explore interprofessional experiences of incorporating fracture prevention activities in clinical practice inspired by an empowerment approach. Design/methodology/approach: Data collection consisted primarily of focus groups interviews, systematized and analyzed by the grounded theory method. The study took…

  18. Characterizing preventive activity in Camagüey province

    Hernández, María

    2012-01-01

    The article describes the characteristic features of preventive activities within the System of Education at Camaguey province. This information herein gathers results from a research investigation project and might be of interest for teachers, principals and investigators as a professional tool.

  19. Radionuclide assay of membrane Na+, K+-ATPase activity of peserved red blood cells

    Trusov, V.V.; Zelenin, A.A.; Marizin, S.A.

    1986-01-01

    The radionuclide tests were used to investigate the influence of varying blood preservatives on erythrocylic membrane Na + , K + -ATPase activity in samples of whole blood and packed red blood cells from normal donors prepared by standard methods. The tests were performed before and after seven days of preservation under standard conditions. It was found that blood preservations lowered membrane Na + , K + -ATPase activity: its minimum reduction was recorded with citroglucopnosphate, while glugicir induced a significant drop in Na + , K + -ATPase activity of preserved red blood cells regardless of the type of the blood transfusion solution. The assay of membrane Na + , K + -ATPase activity of preserved red blood cells with the use of 86 Rb could be recommended as an evaluation test for preserved blood and its components

  20. Influence of Active Layer on Separation Potentials of Nanofiltration Membranes for Inorganic Ions.

    Wadekar, Shardul S; Vidic, Radisav D

    2017-05-16

    Active layers of two fully aromatic and two semi-aromatic nanofiltration membranes were studied along with surface charge at different electrolyte composition and effective pore size to elucidate their influence on separation mechanisms for inorganic ions by steric, charge, and dielectric exclusion. The membrane potential method used for pore size measurement is underlined as the most appropriate measurement technique for this application owing to its dependence on the diffusional potentials of inorganic ions. Crossflow rejection experiments with dilute feed composition indicate that both fully aromatic membranes achieved similar rejection despite the differences in surface charge, which suggests that rejection by these membranes is exclusively dependent on size exclusion and the contribution of charge exclusion is weak. Rejection experiments with higher ionic strength and different composition of the feed solution confirmed this hypothesis. On the other hand, increase in the ionic strength of feed solution when the charge exclusion effects are negligible due to charge screening strongly influenced ion rejection by semi-aromatic membranes. The experimental results confirmed that charge exclusion contributes significantly to the performance of semi-aromatic membranes in addition to size exclusion. The contribution of dielectric exclusion to overall ion rejection would be more significant for fully aromatic membranes.

  1. Improving the performance of membrane bioreactors by powdered activated carbon dosing with cost considerations.

    Yang, W; Paetkau, M; Cicek, N

    2010-01-01

    Effects of powdered activated carbon (PAC) dosing on the overall performance of membrane bioreactors (MBR) were investigated in two bench-scale submerged MBRs. Positive impacts of PAC dosing on membrane fouling and the removal of 17beta-estradiol (E2) and 17alpha-ethyinylestradiol (EE2) were demonstrated over a six-month stable operational period. PAC dosing in the MBR increased the removal rates of E2 and EE2 by 3.4% and 15.8%, respectively. The average soluble extracellular polymeric substances (EPS) and colloidal total organic carbon (TOC) concentrations in the PAC-MBR sludge was 60.1% and 61.8% lower than the control MBR sludge, respectively. Lower soluble EPS and colloidal TOC concentrations in the PAC-MBR sludge resulted in a slower rate of trans-membrane pressure (TMP) increase during MBRs operation, which could prolong the lifespan of membranes. Cost assessment showed that PAC dosing could reduce the operating cost for membrane cleaning and/or membrane replacement by about 25%. The operating cost for PAC dosing could be offset by the benefit from its reducing the cost for membrane maintenance.

  2. Preparation of amino-functionalized regenerated cellulose membranes with high catalytic activity.

    Wang, Wei; Bai, Qian; Liang, Tao; Bai, Huiyu; Liu, Xiaoya

    2017-09-01

    The modification of regenerated cellulose (RC) membranes was carried out by using silane coupling agents presenting primary and secondary amino-groups. The grafting of the amino groups onto the modified cellulose molecule was confirmed by X-ray photoelectron spectroscopies and 13 C nuclear magnetic resonance spectroscopic analyses. The crystallinity of the cellulose membranes (CM) decreased after chemical modification as indicated by the X-ray diffraction results. Moreover, a denser structure was observed at the surface and cross section of the modified membranes by SEM images. The contact angle measurements showed that the silane coupling treatment enhanced the hydrophobicity of the obtained materials. Then the catalytic properties of two types of modified membranes were studied in a batch process by evaluating their catalytic performance in a Knoevenagel condensation. The results indicated that the cellulose membrane grafted with many secondary amines exhibited a better catalytic activity compared to the one grafted only by primary amines. In addition, the compact structure of the modified membranes permitted their application in a pervaporation catalytic membrane reactor. Therefore, functional CM that prepared in this paper represented a promising material in the field of industrial catalysis. Copyright © 2017 Elsevier B.V. All rights reserved.

  3. Sphingomyelinase D activity in model membranes: structural effects of in situ generation of ceramide-1-phosphate.

    Roberto P Stock

    Full Text Available The toxicity of Loxosceles spider venom has been attributed to a rare enzyme, sphingomyelinase D, which transforms sphingomyelin to ceramide-1-phosphate. The bases of its inflammatory and dermonecrotic activity, however, remain unclear. In this work the effects of ceramide-1-phosphate on model membranes were studied both by in situ generation of this lipid using a recombinant sphingomyelinase D from the spider Loxosceles laeta and by pre-mixing it with sphingomyelin and cholesterol. The systems of choice were large unilamellar vesicles for bulk studies (enzyme kinetics, fluorescence spectroscopy and dynamic light scattering and giant unilamellar vesicles for fluorescence microscopy examination using a variety of fluorescent probes. The influence of membrane lateral structure on the kinetics of enzyme activity and the consequences of enzyme activity on the structure of target membranes containing sphingomyelin were examined. The findings indicate that: 1 ceramide-1-phosphate (particularly lauroyl ceramide-1-phosphate can be incorporated into sphingomyelin bilayers in a concentration-dependent manner and generates coexistence of liquid disordered/solid ordered domains, 2 the activity of sphingomyelinase D is clearly influenced by the supramolecular organization of its substrate in membranes and, 3 in situ ceramide-1-phosphate generation by enzymatic activity profoundly alters the lateral structure and morphology of the target membranes.

  4. Plasma membrane factor XIIIA transglutaminase activity regulates osteoblast matrix secretion and deposition by affecting microtubule dynamics.

    Hadil F Al-Jallad

    2011-01-01

    Full Text Available Transglutaminase activity, arising potentially from transglutaminase 2 (TG2 and Factor XIIIA (FXIIIA, has been linked to osteoblast differentiation where it is required for type I collagen and fibronectin matrix deposition. In this study we have used an irreversible TG-inhibitor to 'block -and-track' enzyme(s targeted during osteoblast differentiation. We show that the irreversible TG-inhibitor is highly potent in inhibiting osteoblast differentiation and mineralization and reduces secretion of both fibronectin and type I collagen and their release from the cell surface. Tracking of the dansyl probe by Western blotting and immunofluorescence microscopy demonstrated that the inhibitor targets plasma membrane-associated FXIIIA. TG2 appears not to contribute to crosslinking activity on the osteoblast surface. Inhibition of FXIIIA with NC9 resulted in defective secretory vesicle delivery to the plasma membrane which was attributable to a disorganized microtubule network and decreased microtubule association with the plasma membrane. NC9 inhibition of FXIIIA resulted in destabilization of microtubules as assessed by cellular Glu-tubulin levels. Furthermore, NC9 blocked modification of Glu-tubulin into 150 kDa high-molecular weight Glu-tubulin form which was specifically localized to the plasma membrane. FXIIIA enzyme and its crosslinking activity were colocalized with plasma membrane-associated tubulin, and thus, it appears that FXIIIA crosslinking activity is directed towards stabilizing the interaction of microtubules with the plasma membrane. Our work provides the first mechanistic cues as to how transglutaminase activity could affect protein secretion and matrix deposition in osteoblasts and suggests a novel function for plasma membrane FXIIIA in microtubule dynamics.

  5. Tissue Factor Coagulant Activity is Regulated by the Plasma Membrane Microenvironment.

    Yu, Yuanjie; Böing, Anita N; Hau, Chi M; Hajji, Najat; Ruf, Wolfram; Sturk, Auguste; Nieuwland, Rienk

    2018-06-01

     Tissue factor (TF) can be present in a non-coagulant and coagulant form. Whether the coagulant activity is affected by the plasma membrane microenvironment is unexplored.  This article studies the presence and coagulant activity of human TF in plasma membrane micro-domains.  Plasma membranes were isolated from human MIA PaCa2 cells, MDA-MB-231 cells and human vascular smooth muscle cells by Percoll gradient ultracentrifugation after cell disruption. Plasma membranes were fractionated by OptiPrep gradient ultracentrifugation, and the presence of TF, flotillin, caveolin, clathrin, protein disulphide isomerase (PDI), TF pathway inhibitor (TFPI) and phosphatidylserine (PS) were determined.  Plasma membranes contain two detergent-resistant membrane (DRM) compartments differing in density and biochemical composition. High-density DRMs (DRM-H) have a density ( ρ ) of 1.15 to 1.20 g/mL and contain clathrin, whereas low-density DRMs (DRM-L) have a density between 1.09 and 1.13 g/mL and do not contain clathrin. Both DRMs contain TF, flotillin and caveolin. PDI is detectable in DRM-H, TFPI is not detectable in either DMR-H or DRM-L and PS is detectable in DRM-L. The DRM-H-associated TF (> 95% of the TF antigen) lacks detectable coagulant activity, whereas the DRM-L-associated TF triggers coagulation. This coagulant activity is inhibited by lactadherin and thus PS-dependent, but seemed insensitive to 16F16, an inhibitor of PDI.  Non-coagulant and coagulant TF are present within different types of DRMs in the plasma membrane, and the composition of these DRMs may affect the TF coagulant activity. Schattauer GmbH Stuttgart.

  6. Development and Characterization a Single-Active-Chamber Piezoelectric Membrane Pump with Multiple Passive Check Valves

    Ronghui Zhang

    2016-12-01

    Full Text Available In order to prevent the backward flow of piezoelectric pumps, this paper presents a single-active-chamber piezoelectric membrane pump with multiple passive check valves. Under the condition of a fixed total number of passive check valves, by means of changing the inlet valves and outlet valves’ configuration, the pumping characteristics in terms of flow rate and backpressure are experimentally investigated. Like the maximum flow rate and backpressure, the testing results show that the optimal frequencies are significantly affected by changes in the number inlet valves and outlet valves. The variation ratios of the maximum flow rate and the maximum backpressure are up to 66% and less than 20%, respectively. Furthermore, the piezoelectric pump generally demonstrates very similar flow rate and backpressure characteristics when the number of inlet valves in one kind of configuration is the same as that of outlet valves in another configuration. The comparison indicates that the backflow from the pumping chamber to inlet is basically the same as the backflow from the outlet to the pumping chamber. No matter whether the number of inlet valves or the number of outlet valves is increased, the backflow can be effectively reduced. In addition, the backpressure fluctuation can be significantly suppressed with an increase of either inlet valves or outlet valves. It also means that the pump can prevent the backflow more effectively at the cost of power consumption. The pump is very suitable for conditions where more accurate flow rates are needed and wear and fatigue of check valves often occur.

  7. Comparison of the Modeling Approach between Membrane Bioreactor and Conventional Activated Sludge Processes

    Jiang, Tao; Sin, Gürkan; Spanjers, Henri

    2009-01-01

    Activated sludge models (ASM) have been developed and largely applied in conventional activated sludge (CAS) systems. The applicability of ASM to model membrane bioreactors (MBR) and the differences in modeling approaches have not been studied in detail. A laboratory-scale MBR was modeled using ASM...

  8. Slack sodium-activated potassium channel membrane expression requires p38 mitogen-activated protein kinase phosphorylation.

    Gururaj, Sushmitha; Fleites, John; Bhattacharjee, Arin

    2016-04-01

    p38 MAPK has long been understood as an inducible kinase under conditions of cellular stress, but there is now increasing evidence to support its role in the regulation of neuronal function. Several phosphorylation targets have been identified, an appreciable number of which are ion channels, implicating the possible involvement of p38 MAPK in neuronal excitability. The KNa channel Slack is an important protein to be studied as it is highly and ubiquitously expressed in DRG neurons and is important in the maintenance of their firing accommodation. We sought to examine if the Slack channel could be a substrate of p38 MAPK activity. First, we found that the Slack C-terminus contains two putative p38 MAPK phosphorylation sites that are highly conserved across species. Second, we show via electrophysiology experiments that KNa currents and further, Slack currents, are subject to tonic modulation by p38 MAPK. Third, biochemical approaches revealed that Slack channel regulation by p38 MAPK occurs through direct phosphorylation at the two putative sites of interaction, and mutating both sites prevented surface expression of Slack channels. Based on these results, we conclude that p38 MAPK is an obligate regulator of Slack channel function via the trafficking of channels into the membrane. The present study identifies Slack KNa channels as p38 MAPK substrates. Copyright © 2015 Elsevier Ltd. All rights reserved.

  9. Membranolytic Activity of Bile Salts: Influence of Biological Membrane Properties and Composition

    Alfred Blume

    2007-10-01

    Full Text Available The two main steps of the membranolytic activity of detergents: 1 the partitioning of detergent molecules in the membrane and 2 the solubilisation of the membrane are systematically investigated. The interactions of two bile salt molecules, sodium cholate (NaC and sodium deoxycholate (NaDC with biological phospholipid model membranes are considered. The membranolytic activity is analysed as a function of the hydrophobicity of the bile salt, ionic strength, temperature, membrane phase properties, membrane surface charge and composition of the acyl chains of the lipids. The results are derived from calorimetric measurements (ITC, isothermal titration calorimetry. A thermodynamic model is described, taking into consideration electrostatic interactions, which is used for the calculation of the partition coefficient as well as to derive the complete thermodynamic parameters describing the interaction of detergents with biological membranes (change in enthalpy, change in free energy, change in entropy etc. The solubilisation properties are described in a so-called vesicle-to-micelle phase transition diagram. The obtained results are supplemented and confirmed by data obtained from other biophysical techniques (DSC differential scanning calorimetry, DLS dynamic light scattering, SANS small angle neutron scattering.

  10. Pervaporation dehydration of ethanol by hyaluronic acid/sodium alginate two-active-layer composite membranes.

    Gao, Chengyun; Zhang, Minhua; Ding, Jianwu; Pan, Fusheng; Jiang, Zhongyi; Li, Yifan; Zhao, Jing

    2014-01-01

    The composite membranes with two-active-layer (a capping layer and an inner layer) were prepared by sequential spin-coatings of hyaluronic acid (HA) and sodium alginate (NaAlg) on the polyacrylonitrile (PAN) support layer. The SEM showed a mutilayer structure and a distinct interface between the HA layer and the NaAlg layer. The coating sequence of two-active-layer had an obvious influence on the pervaporation dehydration performance of membranes. When the operation temperature was 80 °C and water concentration in feed was 10 wt.%, the permeate fluxes of HA/Alg/PAN membrane and Alg/HA/PAN membrane were similar, whereas the separation factor were 1130 and 527, respectively. It was found that the capping layer with higher hydrophilicity and water retention capacity, and the inner layer with higher permselectivity could increase the separation performance of the composite membranes. Meanwhile, effects of operation temperature and water concentration in feed on pervaporation performance as well as membrane properties were studied. Copyright © 2013 Elsevier Ltd. All rights reserved.

  11. Receptor kinase-mediated control of primary active proton pumping at the plasma membrane

    Fuglsang, Anja Thoe; Kristensen, Astrid; Cuin, Tracey A.

    2014-01-01

    Acidification of the cell wall space outside the plasma membrane is required for plant growth and is the result of proton extrusion by the plasma membrane-localized H+-ATPases. Here we show that the major plasma membrane proton pumps in Arabidopsis, AHA1 and AHA2, interact directly in vitro...... and in planta with PSY1R, a receptor kinase of the plasma membrane that serves as a receptor for the peptide growth hormone PSY1. The intracellular protein kinase domain of PSY1R phosphorylates AHA2/AHA1 at Thr-881, situated in the autoinhibitory region I of the C-terminal domain. When expressed in a yeast...... heterologous expression system, the introduction of a negative charge at this position caused pump activation. Application of PSY1 to plant seedlings induced rapid in planta phosphorylation at Thr-881, concomitant with an instantaneous increase in proton efflux from roots. The direct interaction between AHA2...

  12. Heteronanostructure of Ag particle on titanate nanowire membrane with enhanced photocatalytic properties and bactericidal activities

    Shang Lu; Li Bingjie; Dong Wenjun; Chen Benyong; Li Chaorong; Tang Weihua; Wang Ge; Wu Jian; Ying Yibin

    2010-01-01

    A novel seed induced method has been developed for syntheses of Ag particles on titanate nanowires, and then the heteronanostructured Ag/titanate nanowires were assembled into porous, flexible membranes. These titanate nanowires were about several hundreds micrometers in length and about 80 nm in diameter. The size of the Ag particle can be tuned within 300-700 nm. The pore size and thickness of the heteronanostructured membrane were easily controlled. An Ag/titanate nanowire membrane reactor has been developed to study the photocatalytic degradation of methamidophos in aqueous solution, and 87.0% of the methamidophos can be degraded in a concurrent filtration and photocatalytic oxidation process. The antibacterial activity was also investigated on the heteronanostructured membrane with UVA light (365 nm) irradiation, and a 99.99% satisfactory antibacterial effect on Escherichia coli was achieved.

  13. Protein kinase and phosphatase activities of thylakoid membranes

    Michel, H.; Shaw, E.K.; Bennett, J.

    1987-01-01

    Dephosphorylation of the 25 and 27 kDa light-harvesting Chl a/b proteins (LHCII) of the thylakoid membranes is catalyzed by a phosphatase which differs from previously reported thylakoid-bound phosphatases in having an alkaline pH optimum (9.0) and a requirement for Mg 2+ ions. Dephosphorylation of the 8.3 kDa psb H gene product requires a Mg 2+ ion concentration more than 200 fold higher than that for dephosphorylation of LHC II. The 8.3 kDa and 27 kDa proteins appear to be phosphorylated by two distinct kinases, which differ in substrate specificity and sensitivity to inhibitors. The plastoquinone antagonist 2,5-dibromo-3-methyl-6-isopropyl-benzoquinone (DBMIB) inhibits phosphorylation of the 27 kDa LHC II much more readily than phosphorylation of the 8.3 kDa protein. A similar pattern of inhibition is seen for two synthetic oligopeptides (MRKSATTKKAVC and ATQTLESSSRC) which are analogs of the phosphorylation sites of the two proteins. Possible modes of action of DBMIB are discussed. 45 refs., 7 figs., 3 tabs

  14. Membrane-associated proteolytic activity in Escherichia coli that is stimulated by ATP

    Klemes, Y.; Voellmy, R.W.; Goldberg, A.L.

    1986-01-01

    The degradation of proteins in bacteria requires metabolism energy. One important enzyme in this process is protease La, a soluble ATP-dependent protease encoded by the lon gene. However, lon mutants that lack a functional protease La still show some ATP-dependent protein breakdown. The authors have reported an ATP-stimulated endoproteolytic activity associated with the inner membrane of E. coli. This ATP-stimulated activity is found in normal levels in membranes derived from lon mutants, including strains carrying insertions in the lon gene. The membrane-bound activity hydrolyzes 14 C-methylglobin at a linear rate for up to 3 hours. These fractions also contain appreciable proteolytic activity that is not affected by ATP. The stimulation by ATP requires the presence of Mg 2+ . Nonhydrolyzable ATP analogs (e.g. AMPPNP or ATP-γ-S) and ADP do not enhance proteolysis. Unlike protease La, the membrane-associated enzyme does not degrade the fluorometric substrate, Glt-Ala-Ala-Phe-MNA, in an ATP-stimulated fashion, and its level is not influenced by high temperature of by the gene which regulates the heat-shock response. The enzyme is inhibited by dichloroisocoumarin and certain peptide chloromethyl ketones. They conclude that E. coli contain at least two ATP-dependent proteases with distinct specificities: one is soluble and the other is membrane-associated

  15. Blueberry polyphenols prevent cardiomyocyte death by preventing calpain activation and oxidative stress.

    Louis, Xavier Lieben; Thandapilly, Sijo Joseph; Kalt, Wilhelmina; Vinqvist-Tymchuk, Melinda; Aloud, Basma Milad; Raj, Pema; Yu, Liping; Le, Hoa; Netticadan, Thomas

    2014-08-01

    The purpose of this study was to examine the efficacy of an aqueous wild blueberry extract and five wild blueberry polyphenol fractions on an in vitro model of heart disease. Adult rat cardiomyocytes were pretreated with extract and fractions, and then exposed to norepinephrine (NE). Cardiomyocyte hypertrophy, cell death, oxidative stress, apoptosis and cardiomyocyte contractile function as well as the activities of calpain, superoxide dismutase (SOD) and catalase (CAT) were measured in cardiomyocytes treated with and without NE and blueberry fraction (BF). Four of five blueberry fractions prevented cell death and cardiomyocyte hypertrophy induced by NE. Total phenolic fraction was used for all further analysis. The NE-induced increase in oxidative stress, nuclear condensation, calpain activity and lowering of SOD and CAT activities were prevented upon pretreatment with BF. Reduced contractile function was also significantly improved with BF pretreatment. Blueberry polyphenols prevent NE-induced adult cardiomyocyte hypertrophy and cell death. The protective effects of BF may be in part attributed to a reduction in calpain activity and oxidative stress.

  16. Na+/H+ Exchange Activity in the Plasma Membrane of Arabidopsis1

    Qiu, Quan-Sheng; Barkla, Bronwyn J.; Vera-Estrella, Rosario; Zhu, Jian-Kang; Schumaker, Karen S.

    2003-01-01

    In plants, Na+/H+ exchangers in the plasma membrane are critical for growth in high levels of salt, removing toxic Na+ from the cytoplasm by transport out of the cell. The molecular identity of a plasma membrane Na+/H+ exchanger in Arabidopsis (SOS1) has recently been determined. In this study, immunological analysis provided evidence that SOS1 localizes to the plasma membrane of leaves and roots. To characterize the transport activity of this protein, purified plasma membrane vesicles were isolated from leaves of Arabidopsis. Na+/H+ exchange activity, monitored as the ability of Na to dissipate an established pH gradient, was absent in plants grown without salt. However, exchange activity was induced when plants were grown in 250 mm NaCl and increased with prolonged salt exposure up to 8 d. H+-coupled exchange was specific for Na, because chloride salts of other monovalent cations did not dissipate the pH gradient. Na+/H+ exchange activity was dependent on Na (substrate) concentration, and kinetic analysis indicated that the affinity (apparent Km) of the transporter for Na+ is 22.8 mm. Data from two experimental approaches supports electroneutral exchange (one Na+ exchanged for one proton): (a) no change in membrane potential was measured during the exchange reaction, and (b) Na+/H+ exchange was unaffected by the presence or absence of a membrane potential. Results from this research provide a framework for future studies into the regulation of the plant plasma membrane Na+/H+ exchanger and its relative contribution to the maintenance of cellular Na+ homeostasis during plant growth in salt. PMID:12805632

  17. Na+/H+ exchange activity in the plasma membrane of Arabidopsis.

    Qiu, Quan-Sheng; Barkla, Bronwyn J; Vera-Estrella, Rosario; Zhu, Jian-Kang; Schumaker, Karen S

    2003-06-01

    In plants, Na+/H+ exchangers in the plasma membrane are critical for growth in high levels of salt, removing toxic Na+ from the cytoplasm by transport out of the cell. The molecular identity of a plasma membrane Na+/H+ exchanger in Arabidopsis (SOS1) has recently been determined. In this study, immunological analysis provided evidence that SOS1 localizes to the plasma membrane of leaves and roots. To characterize the transport activity of this protein, purified plasma membrane vesicles were isolated from leaves of Arabidopsis. Na+/H+ exchange activity, monitored as the ability of Na to dissipate an established pH gradient, was absent in plants grown without salt. However, exchange activity was induced when plants were grown in 250 mm NaCl and increased with prolonged salt exposure up to 8 d. H+-coupled exchange was specific for Na, because chloride salts of other monovalent cations did not dissipate the pH gradient. Na+/H+ exchange activity was dependent on Na (substrate) concentration, and kinetic analysis indicated that the affinity (apparent Km) of the transporter for Na+ is 22.8 mm. Data from two experimental approaches supports electroneutral exchange (one Na+ exchanged for one proton): (a) no change in membrane potential was measured during the exchange reaction, and (b) Na+/H+ exchange was unaffected by the presence or absence of a membrane potential. Results from this research provide a framework for future studies into the regulation of the plant plasma membrane Na+/H+ exchanger and its relative contribution to the maintenance of cellular Na+ homeostasis during plant growth in salt.

  18. Monitoring glycolipid transfer protein activity and membrane interaction with the surface plasmon resonance technique.

    Ohvo-Rekilä, Henna; Mattjus, Peter

    2011-01-01

    The glycolipid transfer protein (GLTP) is a protein capable of binding and transferring glycolipids. GLTP is cytosolic and it can interact through its FFAT-like (two phenylalanines in an acidic tract) motif with proteins localized on the surface of the endoplasmic reticulum. Previous in vitro work with GLTP has focused mainly on the complete transfer reaction of the protein, that is, binding and subsequent removal of the glycolipid from the donor membrane, transfer through the aqueous environment, and the final release of the glycolipid to an acceptor membrane. Using bilayer vesicles and surface plasmon resonance spectroscopy, we have now, for the first time, analyzed the binding and lipid removal capacity of GLTP with a completely label-free technique. This technique is focused on the initial steps in GLTP-mediated transfer and the parameters affecting these steps can be more precisely determined. We used the new approach for detailed structure-function studies of GLTP by examining the glycolipid transfer capacity of specific GLTP tryptophan mutants. Tryptophan 96 is crucial for the transfer activity of the protein and tryptophan 142 is an important part of the proteins membrane interacting domain. Further, we varied the composition of the used lipid vesicles and gained information on the effect of membrane properties on GLTP activity. GLTP prefers to interact with more tightly packed membranes, although GLTP-mediated transfer is faster from more fluid membranes. This technique is very useful for the study of membrane-protein interactions and lipid-transfer rates and it can easily be adapted to other membrane-interacting proteins. Copyright © 2010 Elsevier B.V. All rights reserved.

  19. Chitosan derivatives targeting lipid bilayers: Synthesis, biological activity and interaction with model membranes.

    Martins, Danubia Batista; Nasário, Fábio Domingues; Silva-Gonçalves, Laiz Costa; de Oliveira Tiera, Vera Aparecida; Arcisio-Miranda, Manoel; Tiera, Marcio José; Dos Santos Cabrera, Marcia Perez

    2018-02-01

    The antimicrobial activity of chitosan and derivatives to human and plant pathogens represents a high-valued prospective market. Presently, two low molecular weight derivatives, endowed with hydrophobic and cationic character at different ratios were synthesized and characterized. They exhibit antimicrobial activity and increased performance in relation to the intermediate and starting compounds. However, just the derivative with higher cationic character showed cytotoxicity towards human cervical carcinoma cells. Considering cell membranes as targets, the mode of action was investigated through the interaction with model lipid vesicles mimicking bacterial, tumoral and erythrocyte membranes. Intense lytic activity and binding are demonstrated for both derivatives in anionic bilayers. The less charged compound exhibits slightly improved selectivity towards bacterial model membranes, suggesting that balancing its hydrophobic/hydrophilic character may improve efficiency. Observing the aggregation of vesicles, we hypothesize that the "charge cluster mechanism", ascribed to some antimicrobial peptides, could be applied to these chitosan derivatives. Copyright © 2017 Elsevier Ltd. All rights reserved.

  20. Confinement of activating receptors at the plasma membrane controls natural killer cell tolerance.

    Guia, Sophie; Jaeger, Baptiste N; Piatek, Stefan; Mailfert, Sébastien; Trombik, Tomasz; Fenis, Aurore; Chevrier, Nicolas; Walzer, Thierry; Kerdiles, Yann M; Marguet, Didier; Vivier, Eric; Ugolini, Sophie

    2011-04-05

    Natural killer (NK) cell tolerance to self is partly ensured by major histocompatibility complex (MHC) class I-specific inhibitory receptors on NK cells, which dampen their reactivity when engaged. However, NK cells that do not detect self MHC class I are not autoreactive. We used dynamic fluorescence correlation spectroscopy to show that MHC class I-independent NK cell tolerance in mice was associated with the presence of hyporesponsive NK cells in which both activating and inhibitory receptors were confined in an actin meshwork at the plasma membrane. In contrast, the recognition of self MHC class I by inhibitory receptors "educated" NK cells to become fully reactive, and activating NK cell receptors became dynamically compartmentalized in membrane nanodomains. We propose that the confinement of activating receptors at the plasma membrane is pivotal to ensuring the self-tolerance of NK cells.

  1. Sphingomyelinase D activity in model membranes: structural effects of in situ generation of ceramide-1-phosphate

    Stock, Roberto; Brewer, Jonathan R.; Wagner, Kerstin

    2012-01-01

    The toxicity of Loxosceles spider venom has been attributed to a rare enzyme, sphingomyelinase D, which transforms sphingomyelin to ceramide-1-phosphate. The bases of its inflammatory and dermonecrotic activity, however, remain unclear. In this work the effects of ceramide-1-phosphate on model...... membranes were studied both by in situ generation of this lipid using a recombinant sphingomyelinase D from the spider Loxosceles laeta and by pre-mixing it with sphingomyelin and cholesterol. The systems of choice were large unilamellar vesicles for bulk studies (enzyme kinetics, fluorescence spectroscopy...... and dynamic light scattering) and giant unilamellar vesicles for fluorescence microscopy examination using a variety of fluorescent probes. The influence of membrane lateral structure on the kinetics of enzyme activity and the consequences of enzyme activity on the structure of target membranes containing...

  2. Preventive social activities with teenages from incomplete families

    Ruslan Chubuk

    2018-01-01

    Full Text Available This article foregrounds the importance of social prevention activities with adolescents from single-parent families, which aim at overcoming external social pressures and psychological discomfort as well as creating a system of mutual support and self-help for adolescents. The aim of this article is to highlight the forms and methods of work that constitute the set of traditional methods of professional influence on adolescents from single-parent families and their parents. The author proposes a work process cycle of social prevention activities (which defines the selection of apparatus, content, forms and methods of work to  186 carry out in a logical sequence of certain stages that are characteristic of work with any singleparent family. First (pre-preparatory stage consisted of a primary formulation of goal guidelines for social preventive specialists with the aim of determination of the search criterion for the conditions and social environment of single-parent families, acquaintance with the problems of the family and the adolescent. Second (organization and information stage consisted of gathering of information, its organization and analysis. In the third (determination and diagnosis stage, the methods of detailed investigation of both adolescent and family members’ problems were chosen. In the fourth (procedure and analytics stage, the methods of detailed research of single-family problems, determined by the programme, were implemented. In the fifth (correction and results stage stipulated for the socio-educational (correctional work (if necessary with members of single-parent families to create new or to restore damaged positive relationships of the adolescent with their family and enhance social responsibility of all family members for the future of the adolescent, etc. In the sixth (control and prevention stage social workers make monitor visits to the single-parent families during a certain period. Key words: stage, incomplete

  3. Ca2+ signals, cell membrane disintegration, and activation of TMEM16F during necroptosis.

    Ousingsawat, Jiraporn; Cabrita, Inês; Wanitchakool, Podchanart; Sirianant, Lalida; Krautwald, Stefan; Linkermann, Andreas; Schreiber, Rainer; Kunzelmann, Karl

    2017-01-01

    Activated receptor-interacting protein kinase 3 (RIPK3) and mixed lineage kinase domain like (MLKL) are essential components of the necroptotic pathway. Phosphorylated MLKL (pMLKL) is thought to induce membrane leakage, leading to cell swelling and disintegration of the cell membrane. However, the molecular identity of the necroptotic membrane pore remains unclear, and the role of pMLKL for membrane permeabilization is currently disputed. We observed earlier that the phospholipid scramblase and ion channel TMEM16F/anoctamin 6 cause large membrane currents, cell swelling, and cell death when activated by a strong increase in intracellular Ca 2+ . We, therefore, asked whether TMEM16F is also central to necroptotic cell death and other cellular events during necroptosis. Necroptosis was induced by TNFα, smac mimetic, and Z-VAD (TSZ) in NIH3T3 fibroblasts and the four additional cell lines HT 29 , 16HBE, H441, and L929. Time-dependent changes in intracellular Ca 2+ , cell morphology, and membrane currents were recorded. TSZ induced a small and only transient oscillatory rise in intracellular Ca 2+ , which was paralleled by the activation of outwardly rectifying Cl - currents, which were typical for TMEM16F/ANO6. Ca 2+ oscillations were due to Ca 2+ release from endoplasmic reticulum, and were independent of extracellular Ca 2+ . The initial TSZ-induced cell swelling was followed by cell shrinkage. Using typical channel blockers and siRNA-knockdown, the Cl - currents were shown to be due to the activation of ANO6. However, the knockdown of ANO6 or inhibitors of ANO6 did not inhibit necroptotic cell death. The present data demonstrate the activation of ANO6 during necroptosis, which, however, is not essential for cell death.

  4. Energy transfer in light-adapted photosynthetic membranes: from active to saturated photosynthesis.

    Fassioli, Francesca; Olaya-Castro, Alexandra; Scheuring, Simon; Sturgis, James N; Johnson, Neil F

    2009-11-04

    In bacterial photosynthesis light-harvesting complexes, LH2 and LH1 absorb sunlight energy and deliver it to reaction centers (RCs) with extraordinarily high efficiency. Submolecular resolution images have revealed that both the LH2:LH1 ratio, and the architecture of the photosynthetic membrane itself, adapt to light intensity. We investigate the functional implications of structural adaptations in the energy transfer performance in natural in vivo low- and high-light-adapted membrane architectures of Rhodospirillum photometricum. A model is presented to describe excitation migration across the full range of light intensities that cover states from active photosynthesis, where all RCs are available for charge separation, to saturated photosynthesis where all RCs are unavailable. Our study outlines three key findings. First, there is a critical light-energy density, below which the low-light adapted membrane is more efficient at absorbing photons and generating a charge separation at RCs, than the high-light-adapted membrane. Second, connectivity of core complexes is similar in both membranes, suggesting that, despite different growth conditions, a preferred transfer pathway is through core-core contacts. Third, there may be minimal subareas on the membrane which, containing the same LH2:LH1 ratio, behave as minimal functional units as far as excitation transfer efficiency is concerned.

  5. Antibiofilm activity of Bacillus pumilus SW9 against initial biofouling on microfiltration membranes.

    Zhang, Ying; Yu, Xin; Gong, Song; Ye, Chengsong; Fan, Zihong; Lin, Huirong

    2014-02-01

    Membrane biofouling, resulting from biofilm formation on the membrane, has become the main obstacle hindering wider application of membrane technology. Initial biofouling proves to be crucial which involves early stages of microbial adhesion and biofilm formation. Biological control of microbial attachment seems to be a promising strategy due to its high efficiency and eco-friendliness. The present study investigated the effects of a bacterium Bacillus pumilus SW9 on controlling the initial fouling formed by four target bacterial strains which were pioneer species responsible for biofouling in membrane bioreactors, using microfiltration membranes as the abiotic surfaces. The results suggested that strain SW9 exhibited excellent antibiofilm activity by decreasing the attached biomass of target strains. The production of extracellular polysaccharides and proteins by four target strains was also reduced. A distinct improvement of permeate flux in dead-end filtration systems was achieved when introducing strain SW9 to microfiltration experiments. Scanning electron microscopy and confocal laser scanning microscopy were performed to further ascertain significant changes of the biofouling layers. A link between biofilm inhibition and initial biofouling mitigation was thus provided, suggesting an alternatively potential way to control membrane biofouling through bacterial interactions.

  6. Cetamolol: a new cardioselective beta-adrenoceptor blocking agent without membrane-stabilizing activity.

    Beaulieu, G; Jaramillo, J; Cummings, J R

    1984-03-01

    Cetamolol, a new beta-adrenoceptor blocker with partial agonist activity and cardioselectivity, was studied in vivo to determine its membrane-stabilizing effects. Comparisons were carried out with atenolol, pindolol, practolol, propranolol, timolol, dexpropranolol, lidocaine, and procaine. The following results indicated that cetamolol lacked membrane-stabilizing activity: (i) failure to cause local anesthesia on the rabbit cornea and motor nerve of the rat tail; (ii) ineffectiveness in reversing ventricular arrhythmias induced by coronary artery litigation in dogs; (iii) failure to reduce cardiac automaticity in catecholamine-depleted dogs as determined by the rate of a subatrial rhythm during ventricular (vagal) escape; and (iv) lack of a significant increase in atrioventricular conduction time in vagotomized or atropinized dogs in contrast to the effect in normal dogs indicating a reflex effect of cetamolol. Other results include a restoration of sinus rhythm in dogs with ventricular tachycardia induced by ouabain, and a dose-related decline in the force of cardiac contraction in anesthetized dogs at doses from 3 to 15 mg/kg, which occurred after an initial increase in force owing to intrinsic sympathomimetic activity. Although the mechanisms for the latter two effects are not clear at this time, explanations other than membrane-stabilizing activity have been considered in view of the other findings. It is concluded that cetamolol lacks membrane-stabilizing activity even at inordinately high doses.

  7. Erythrocyte membrane ATPase and calcium pumping activities in porcine malignant hyperthermia

    Thatte, H.S.; Mickelson, J.R.; Addis, P.B.; Louis, C.F.

    1987-01-01

    To investigate possible abnormalities in erythrocyte membrane enzyme activities in the pharmacogenetic disorder MH, membrane ATPase activities have been examined in erythrocyte ghosts prepared from red blood cells of MHS and normal swine. While no differences were noted in Mg2+-ATPase activities, the (Na+, K+)-ATPase activity of MHS erythrocyte ghosts was less than that of normal ghosts. Ca2+-ATPase activity exhibited low- and high-affinity Ca2+-binding sites in both types of erythrocyte ghost. While the Km for Ca2+ was greater for normal than for MHS erythrocyte ghosts at the high-affinity Ca2+-binding site, the reverse was true at the low-affinity Ca2+-binding site. Irrespective of the type of calcium binding site occupied, the Vmax for normal erythrocyte ghost Ca2+-ATPase activity was greater than that for MHS ghosts. In the presence of calmodulin, there was now no difference between MHS and normal erythrocyte ghosts in either the Km for Ca2+ or the Vmax of the Ca2+-ATPase activity. To determine if the calcium pumping activity of intact MHS and normal pig erythrocytes differed, calcium efflux from the 45 Ca-loaded erythrocytes was determined; this activity was significantly greater for MHS than for normal erythrocytes. Thus, the present study confirms that there are abnormalities in the membranes of MHS pig red blood cells. However, we conclude that these abnormalities are unlikely to result in an impaired ability of MHS erythrocytes to regulate their cytosolic Ca2+ concentration

  8. Progesterone increases nitric oxide synthesis in human vascular endothelial cells through activation of membrane progesterone receptor-α.

    Pang, Yefei; Dong, Jing; Thomas, Peter

    2015-05-15

    Progesterone exerts beneficial effects on the human cardiovascular system by inducing rapid increases in nitric oxide (NO) production in vascular endothelial cells, but the receptors mediating these nongenomic progesterone actions remain unclear. Using human umbilical vein endothelial cells (HUVECs) as a model, we show that progesterone binds to plasma membranes of HUVECs with the characteristics of membrane progesterone receptors (mPRs). The selective mPR agonist Org OD 02-0 had high binding affinity for the progesterone receptor on HUVEC membranes, whereas nuclear PR (nPR) agonists R5020 and medroxyprogesterone acetate displayed low binding affinities. Immunocytochemical and Western blot analyses confirmed that mPRs are expressed in HUVECs and are localized on their plasma membranes. NO levels increased rapidly after treatment with 20 nM progesterone, Org OD 02-0, and a progesterone-BSA conjugate but not with R5020, suggesting that this progesterone action is at the cell surface and initiated through mPRs. Progesterone and Org OD 02-0 (20 nM) also significantly increased endothelial nitric oxide synthase (eNOS) activity and eNOS phosphorylation. Knockdown of mPRα expression by treatment with small-interfering RNA (siRNA) blocked the stimulatory effects of 20 nM progesterone on NO production and eNOS phosphorylation, whereas knockdown of nPR was ineffective. Treatment with PI3K/Akt and MAP kinase inhibitors blocked the stimulatory effects of progesterone, Org OD 02-0, and progesterone-BSA on NO production and eNOS phosphorylation and also prevented progesterone- and Org OD 02-0-induced increases in Akt and ERK phosphorylation. The results suggest that progesterone stimulation of NO production in HUVECs is mediated by mPRα and involves signaling through PI3K/Akt and MAP kinase pathways. Copyright © 2015 the American Physiological Society.

  9. Increased Granulocyte Heparanase Activity in Neutrophils from Patients with Lupus Nephritis and Idiopathic Membranous Nephropathy.

    Szymczak, Maciej; Kuźniar, Jakub; Kopeć, Wacław; Żabińska, Marcelina; Marchewka, Zofia; Kościelska-Kasprzak, Katarzyna; Klinger, Marian

    2017-02-01

    Heparanase is a β-glucuronidase that cleaves sugar chains of heparan sulfate proteoglycans. It is believed that heparanase may be involved in the pathogenesis of proteinuria. The aim of this study was to assess the significance of heparanase in the pathogenesis of particular glomerulonephritis types. The evaluation of heparanase activity in serum, urine, and granulocytes and superoxide dismutase (SOD) activity in granulocytes of patients with lupus nephritis (n = 17), membranous nephropathy (n = 11), IgA nephropathy (n = 12), focal and segmental glomerulosclerosis (n = 18), mesangiocapillary glomerulonephritis (n = 12) and in 19 healthy volunteers were performed. The heparanase activity in granulocytes of patients with lupus nephritis and membranous nephropathy was higher than heparanase activity in granulocytes in the control group (p = 0.02 in both cases). This is the first observation of this phenomenon. There was no difference between SOD activity in granulocytes of patients with all assessed types of glomerulonephritis and the control group. A positive correlation between heparanase activity in urine and double-strain DNA antibodies (r = 0.51; p = 0.04), and reverse correlations between heparanase in urine and hemolytic activity of the complement (r = -0.57; p = 0.03) in the lupus nephritis group, and between heparanase activity in granulocytes and serum total protein level (r = -0.69; p = 0.02) in membranous nephropathy were observed. Increase in heparanase activity without changes in superoxide dismutase activity in the granulocytes from patients with lupus nephritis and membranous nephropathy was observed. It may be used as one of the markers of these disease activities.

  10. Atmospheric-pressure plasma activation and surface characterization on polyethylene membrane separator

    Tseng, Yu-Chien; Li, Hsiao-Ling; Huang, Chun

    2017-01-01

    The surface hydrophilic activation of a polyethylene membrane separator was achieved using an atmospheric-pressure plasma jet. The surface of the atmospheric-pressure-plasma-treated membrane separator was found to be highly hydrophilic realized by adjusting the plasma power input. The variations in membrane separator chemical structure were confirmed by Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy. Chemical analysis showed newly formed carbonyl-containing groups and high surface concentrations of oxygen-containing species on the atmospheric-pressure-plasma-treated polymeric separator surface. It also showed that surface hydrophilicity primarily increased from the polar component after atmospheric-pressure plasma treatment. The surface and pore structures of the polyethylene membrane separator were examined by scanning electron microscopy, revealing a slight alteration in the pore structure. As a result of the incorporation of polar functionalities by atmospheric-pressure plasma activation, the electrolyte uptake and electrochemical impedance of the atmospheric-pressure-plasma-treated membrane separator improved. The investigational results show that the separator surface can be controlled by atmospheric-pressure plasma surface treatment to tailor the hydrophilicity and enhance the electrochemical performance of lithium ion batteries.

  11. Antibacterial activity on electrospun poly(lactide-co-glycolide) based membranes via Magainin II grafting

    Yüksel, Emre; Karakeçili, Ayşe, E-mail: akarakecili@eng.ankara.edu.tr

    2014-12-01

    An antimicrobial peptide (AMP), Magainin II (Mag II) was covalently immobilized on poly(lactide-co-glycolide) (PLGA) and PLGA/gelatin electrospun fibrous membranes. The surface immobilization was characterized by X-ray Photoelectron Spectroscopy (XPS). Scanning Electron Microscopy (SEM) and Atomic Force Microscopy studies showed that the surface morphology of the fibers at micron scale was not affected by the immobilization process. The antibacterial activity of the bound Mag II was tested against Gram-negative Escherichia coli and Gram-positive Staphylococcus aureus. Bacterial adhesion tests, SEM and confocal analyses revealed that the attachment and survival of bacteria were inhibited on Mag II functionalized membranes. AMP immobilization strategy was introduced as a new perspective for the modulation of antibacterial properties on PLGA based materials prepared by electrospinning. - Highlights: • PLGA and PLGA/gelatin fibrous membranes were prepared by electrospinning. • Antimicrobial peptide Mag II was successfully immobilized on PLGA based membranes. • The antibacterial activity was tested against E. coli and S. aureus. • Bacterial adhesion was inhibited on Mag II functionalized membranes.

  12. Group B streptococcus activates transcriptomic pathways related to premature birth in human extraplacental membranes in vitro.

    Park, Hae-Ryung; Harris, Sean M; Boldenow, Erica; McEachin, Richard C; Sartor, Maureen; Chames, Mark; Loch-Caruso, Rita

    2018-03-01

    Streptococcus agalactiae (group B streptococcus [GBS]) infection in pregnant women is the leading cause of infectious neonatal morbidity and mortality in the United States. Although inflammation during infection has been associated with preterm birth, the contribution of GBS to preterm birth is less certain. Moreover, the early mechanisms by which GBS interacts with the gestational tissue to affect adverse pregnancy outcomes are poorly understood. We hypothesized that short-term GBS inoculation activates pathways related to inflammation and premature birth in human extraplacental membranes. We tested this hypothesis using GBS-inoculated human extraplacental membranes in vitro. In agreement with our hypothesis, a microarray-based transcriptomics analysis of gene expression changes in GBS-inoculated membranes revealed that GBS activated pathways related to inflammation and preterm birth with significant gene expression changes occurring as early as 4 h postinoculation. In addition, pathways related to DNA replication and repair were downregulated with GBS treatment. Conclusions based on our transcriptomics data were further supported by responses of prostaglandin E2 (PGE2), and matrix metalloproteinases 1 (MMP1) and 3 (MMP3), all of which are known to be involved in parturition and premature rupture of membranes. These results support our initial hypothesis and provide new information on molecular targets of GBS infection in human extraplacental membranes.

  13. Oxysterol Restraint of Cholesterol Synthesis Prevents AIM2 Inflammasome Activation.

    Dang, Eric V; McDonald, Jeffrey G; Russell, David W; Cyster, Jason G

    2017-11-16

    Type I interferon restrains interleukin-1β (IL-1β)-driven inflammation in macrophages by upregulating cholesterol-25-hydroxylase (Ch25h) and repressing SREBP transcription factors. However, the molecular links between lipid metabolism and IL-1β production remain obscure. Here, we demonstrate that production of 25-hydroxycholesterol (25-HC) by macrophages is required to prevent inflammasome activation by the DNA sensor protein absent in melanoma 2 (AIM2). We find that in response to bacterial infection or lipopolysaccharide (LPS) stimulation, macrophages upregulate Ch25h to maintain repression of SREBP2 activation and cholesterol synthesis. Increasing macrophage cholesterol content is sufficient to trigger IL-1β release in a crystal-independent but AIM2-dependent manner. Ch25h deficiency results in cholesterol-dependent reduced mitochondrial respiratory capacity and release of mitochondrial DNA into the cytosol. AIM2 deficiency rescues the increased inflammasome activity observed in Ch25h -/- . Therefore, activated macrophages utilize 25-HC in an anti-inflammatory circuit that maintains mitochondrial integrity and prevents spurious AIM2 inflammasome activation. Copyright © 2017 Elsevier Inc. All rights reserved.

  14. Modulation of hyaluronan synthase activity in cellular membrane fractions

    Vigetti, Davide; Genasetti, A; Karousou, Evgenia; Viola, Manuela; Clerici, M; Bartolini, B; Moretto, Paola; DE LUCA, Giancarlo; Hascall, Vc; Passi, Alberto

    2009-01-01

    Hyaluronan (HA), the only non-sulfated glycosaminoglycan, is involved in morphogenesis, wound healing, inflammation, angiogenesis, and cancer. In mammals, HA is synthesized by three homologous HA synthases, HAS1, HAS2, and HAS3, that polymerize the HA chain using UDP-glucuronic acid and UDP-N-acetylglucosamine as precursors. Since the amount of HA is critical in several pathophysiological conditions, we developed a non-radioactive assay for measuring the activity of HA synthases (HASs) in euk...

  15. Beneficial effects of gamma linolenic acid supplementation on nerve conduction velocity, Na+, K+ ATPase activity, and membrane fatty acid composition in sciatic nerve of diabetic rats.

    Coste, T; Pierlovisi, M; Leonardi, J; Dufayet, D; Gerbi, A; Lafont, H; Vague, P; Raccah, D

    1999-07-01

    Metabolic and vascular abnormalities are implicated in the pathogenesis of diabetic neuropathy. Two principal metabolic defects are altered lipid metabolism resulting from the impairment of delta-6-desaturase, which converts linoleic acid (LA) into gamma linolenic acid (GLA), and reduced nerve Na+, K+ ATPase activity. This reduction may be caused by a lack of incorporation of (n-6) fatty acids in membrane phospholipids. Because this ubiquitous enzyme maintains the membrane electrical potential and allows repolarization, disturbances in its activity can alter the process of nerve conduction velocity (NCV). We studied the effects of supplementation with GLA (260 mg per day) on NCV, fatty acid phospholipid composition, and Na+, K+ ATPase activity in streptozotocin-diabetic rats. Six groups of 10 rats were studied. Two groups served as controls supplemented with GLA or sunflower oil (GLA free). Two groups with different durations of diabetes were studied: 6 weeks with no supplementation and 12 weeks supplemented with sunflower oil. To test the ability of GLA to prevent or reverse the effects of diabetes, two groups of diabetic rats were supplemented with GLA, one group for 12 weeks and one group for 6 weeks, starting 6 weeks after diabetes induction. Diabetes resulted in a 25% decrease in NCV (P < 0.0001), a 45% decrease in Na+, K+ ATPase activity (P < 0.0001), and an abnormal phospholipid fatty acid composition. GLA restored NCV both in the prevention and reversal studies and partially restored Na+, K+ ATPase activity in the preventive treatment group (P < 0.0001). These effects were accompanied by a modification of phospholipid fatty acid composition in nerve membranes. Overall, the results suggest that membrane fatty acid composition plays a direct role in NCV and confirm the beneficial effect of GLA supplementation in diabetic neuropathy.

  16. Physical activity in the prevention and rehabilitation of cardiovascular risk

    Jovović Veselin

    2015-01-01

    Full Text Available Cardiovascular diseases (CVD are more widespread today, whereby they take dimensions of global epidemic. They are the leading cause of diseases in the world, of inability to work, of absenteeism and premature mortality up to 65 years of age. Modern lifestyle in which there is not enough physical activity is recognized as one of the major risk factors for health and emergence of CVD. Physical inactivity is responsible for poor health quality, unnecessary illnesses and premature death. The aim of this work is to point out the basic risk factors and importance and the role of physical exercise in the prevention and rehabilitation of CVD. In the analysis of the data, the methods of speculation and introspection are used. Numerous studies have shown that properly practiced physical activity is a powerful and beneficial effect in the prevention, treatment and rehabilitation of cardiovascular diseases (Scrutino et al. 2005; Secco et al. 2000; Jovović, 2008; Šuščević et al. 2011. Physical activity belongs to the concept of numerous factors, which along with the reduction of risk factors, lifestyle changes and medical therapy leads to the reduction of risk for cardiovascular diseases. To achieve the desired effect, a combination of aerobic, interval and isotonic muscle activity of moderate intensity at least four times a week for 45 minutes is recommended. During the secondary prevention and rehabilitation, physical activity adapts to health status, level of individual risk and the estimated functional abilities of patients. Transformational processes can only be achieved through regular exercise. The risk of emergence of complications during physical exercise is negligible, especially if the walking is practiced as a form of physical exercise.

  17. Plasma membrane recruitment of dephosphorylated beta-catenin upon activation of the Wnt pathway

    Hendriksen, Jolita; Jansen, Marnix; Brown, Carolyn M.; van der Velde, Hella; van Ham, Marco; Galjart, Niels; Offerhaus, G. Johan; Fagotto, Francois; Fornerod, Maarten

    2008-01-01

    The standard model of Wnt signaling specifies that after receipt of a Wnt ligand at the membranous receptor complex, downstream mediators inhibit a cytoplasmic destruction complex, allowing beta-catenin to accumulate in the cytosol and nucleus and co-activate Wnt target genes. Unexpectedly, shortly

  18. Membrane localization is critical for activation of the PICK1 BAR domain

    Madsen, Kenneth L; Eriksen, Jacob; Milan-Lobo, Laura

    2008-01-01

    The PSD-95/Discs-large/ZO-1 homology (PDZ) domain protein, protein interacting with C kinase 1 (PICK1) contains a C-terminal Bin/amphiphysin/Rvs (BAR) domain mediating recognition of curved membranes; however, the molecular mechanisms controlling the activity of this domain are poorly understood....

  19. Membrane interactions and biological activity of antimicrobial peptides from Australian scorpion.

    Luna-Ramírez, Karen; Sani, Marc-Antoine; Silva-Sanchez, Jesus; Jiménez-Vargas, Juana María; Reyna-Flores, Fernando; Winkel, Kenneth D; Wright, Christine E; Possani, Lourival D; Separovic, Frances

    2014-09-01

    UyCT peptides are antimicrobial peptides isolated from the venom of the Australian scorpion. The activity of the UyCT peptides against Gram positive and Gram negative bacteria and red blood cells was determined. The membrane interactions of these peptides were evaluated by dye release (DR) of the fluorophore calcein from liposomes and isothermal titration calorimetry (ITC); and their secondary structure was determined by circular dichroism (CD). Three different lipid systems were used to mimic red blood cells, Escherichia coli and Staphylococcus aureus membranes. UyCT peptides exhibited broad spectrum antimicrobial activity with low MIC for S. aureus and multi-drug resistant Gram negative strains. Peptide combinations showed some synergy enhancing their potency but not hemolytic activity. The UyCT peptides adopted a helical structure in lipid environments and DR results confirmed that the mechanism of action is by disrupting the membrane. ITC data indicated that UyCT peptides preferred prokaryotic rather than eukaryotic membranes. The overall results suggest that UyCT peptides could be pharmaceutical leads for the treatment of Gram negative multiresistant bacterial infections, especially against Acinetobacter baumanni, and candidates for peptidomimetics to enhance their potency and minimize hemolysis. This article is part of a Special Issue entitled: Interfacially Active Peptides and Proteins. Guest Editors: William C. Wimley and Kalina Hristova. © 2013.

  20. Mapping membrane activity in undiscovered peptide sequence space using machine learning.

    Lee, Ernest Y; Fulan, Benjamin M; Wong, Gerard C L; Ferguson, Andrew L

    2016-11-29

    There are some ∼1,100 known antimicrobial peptides (AMPs), which permeabilize microbial membranes but have diverse sequences. Here, we develop a support vector machine (SVM)-based classifier to investigate ⍺-helical AMPs and the interrelated nature of their functional commonality and sequence homology. SVM is used to search the undiscovered peptide sequence space and identify Pareto-optimal candidates that simultaneously maximize the distance σ from the SVM hyperplane (thus maximize its "antimicrobialness") and its ⍺-helicity, but minimize mutational distance to known AMPs. By calibrating SVM machine learning results with killing assays and small-angle X-ray scattering (SAXS), we find that the SVM metric σ correlates not with a peptide's minimum inhibitory concentration (MIC), but rather its ability to generate negative Gaussian membrane curvature. This surprising result provides a topological basis for membrane activity common to AMPs. Moreover, we highlight an important distinction between the maximal recognizability of a sequence to a trained AMP classifier (its ability to generate membrane curvature) and its maximal antimicrobial efficacy. As mutational distances are increased from known AMPs, we find AMP-like sequences that are increasingly difficult for nature to discover via simple mutation. Using the sequence map as a discovery tool, we find a unexpectedly diverse taxonomy of sequences that are just as membrane-active as known AMPs, but with a broad range of primary functions distinct from AMP functions, including endogenous neuropeptides, viral fusion proteins, topogenic peptides, and amyloids. The SVM classifier is useful as a general detector of membrane activity in peptide sequences.

  1. Outer Membrane Vesicle Vaccines from Biosafe Surrogates Prevent Acute Lethal Glanders in Mice

    Michael H. Norris

    2018-01-01

    Full Text Available Burkholderia mallei is a host-adapted Gram-negative mammalian pathogen that causes the severe disease glanders. Glanders can manifest as a rapid acute progression or a chronic debilitating syndrome primarily affecting solipeds and humans in close association with infected animals. In USA, B. mallei is classified as one of the most important bacterial biothreat agents. Presently, there is no licensed glanders vaccine available for humans or animals. In this work, outer membrane vesicles (OMVs were isolated from three attenuated biosafe bacterial strains, Burkholderia pseudomallei Bp82, B. thailandensis E555, and B. thailandensis TxDOH and used to vaccinate mice. B. thailandensis OMVs induced significantly higher antibody responses that were investigated. B. mallei specific serum antibody responses were of higher magnitude in mice vaccinated with B. thailandensis OMVs compared to levels in mice vaccinated with B. pseudomallei OMVs. OMVs derived from biosafe strains protected mice from acute lethal glanders with vesicles from the two B. thailandensis strains affording significant protection (>90% up to 35 days post-infection with some up to 60 days. Organ loads from 35-day survivors indicated bacteria colonization of the lungs, liver, and spleen while those from 60 days had high CFUs in the spleens. The highest antibody producing vaccine (B. thailandensis E555 OMVs also protected C57BL/6 mice from acute inhalational glanders with evidence of full protection.

  2. Apricot melanoidins prevent oxidative endothelial cell death by counteracting mitochondrial oxidation and membrane depolarization.

    Annalisa Cossu

    Full Text Available The cardiovascular benefits associated with diets rich in fruit and vegetables are thought to be due to phytochemicals contained in fresh plant material. However, whether processed plant foods provide the same benefits as unprocessed ones is an open question. Melanoidins from heat-processed apricots were isolated and their presence confirmed by colorimetric analysis and browning index. Oxidative injury of endothelial cells (ECs is the key step for the onset and progression of cardiovascular diseases (CVD, therefore the potential protective effect of apricot melanoidins on hydrogen peroxide-induced oxidative mitochondrial damage and cell death was explored in human ECs. The redox state of cytoplasmic and mitochondrial compartments was detected by using the redox-sensitive, fluorescent protein (roGFP, while the mitochondrial membrane potential (MMP was assessed with the fluorescent dye, JC-1. ECs exposure to hydrogen peroxide, dose-dependently induced mitochondrial and cytoplasmic oxidation. Additionally detected hydrogen peroxide-induced phenomena were MMP dissipation and ECs death. Pretreatment of ECs with apricot melanoidins, significantly counteracted and ultimately abolished hydrogen peroxide-induced intracellular oxidation, mitochondrial depolarization and cell death. In this regard, our current results clearly indicate that melanoidins derived from heat-processed apricots, protect human ECs against oxidative stress.

  3. Ultra Structural Characterisation of Tetherin - a Protein Capable of Preventing Viral Release from the Plasma Membrane

    Ravindra K. Gupta

    2010-04-01

    Full Text Available Tetherin is an antiviral restriction factor made by mammalian cells to protect them from viral infection. It prevents newly formed virus particles from leaving infected cells. Its antiviral mechanism appears to be remarkably uncomplicated. In 2 studies published in PLoS Pathogens electron microscopy is used to support the hypothesis that the tethers that link HIV-1 virions to tetherin expressing cells contain tetherin and are likely to contain tetherin alone. They also show that the HIV-1 encoded tetherin antagonist that is known to cause tetherin degradation, Vpu, serves to reduce the amount of tetherin in the particles thereby allowing their release.

  4. Active liquid treatment by a combination of precipitation and membrane processes

    Gutman, R.G.; Cumming, I.W.; Williams, G.H.

    1986-08-01

    New ultrafiltration processes developed for the treatment of low and medium active radioactive wastes, were applied successfully to a variety of simulated and real wastes, including magnesium alloy clad spent storage fuel pond waters, reprocessing plant solvent wash liquors, plutonium production effluents and mixed site effluents. After initial laboratory scale feasibility experiments the process was scaled up successfully, using a variety of different ultrafiltration modules. The information accumulated on membrane performance, membrane fouling and flux restoration techniques, and ancillary equipment performance was used to design a much larger demonstration pilot plant. This plant has been constructed and is now processing continuously each day over 1m 3 of a real radioactive effluent. (author)

  5. Zinc can increase the activity of protein kinase C and contributes to its binding to plasma membranes in T lymphocytes.

    Csermely, P; Szamel, M; Resch, K; Somogyi, J

    1988-05-15

    In the primary structure of protein kinase C, the presence of a putative metal-binding site has been suggested (Parker, P.J., Coussens, L., Totty, N., Rhee, L., Young, S., Chen, E., Stabel, S., Waterfield, M.D., and Ullrich, A. (1986) Science 233, 853-859). In the present report, we demonstrate that the most abundant intracellular heavy metal, zinc, can increase the activity of cytosolic protein kinase C. Zinc reversibly binds the enzyme to plasma membranes, and it may contribute to the calcium-induced binding as well. The intracellular heavy metal chelator N,N,N',N'-tetrakis(2-pyridylmethyl) ethylenediamine prevents the phorbol ester- and antigen-induced translocation of protein kinase C. This effect can be totally reversed by the concomitant addition of Zn2+, while Fe2+ and Mn2+ are only partially counteractive. Our results suggest that zinc can activate protein kinase C and contributes to its binding to plasma membranes in T lymphocytes induced by Ca2+, phorbol ester, or antigen.

  6. Monitoring Ion Activities In and Around Cells Using Ion-Selective Liquid-Membrane Microelectrodes

    Mark D. Parker

    2013-01-01

    Full Text Available Determining the effective concentration (i.e., activity of ions in and around living cells is important to our understanding of the contribution of those ions to cellular function. Moreover, monitoring changes in ion activities in and around cells is informative about the actions of the transporters and/or channels operating in the cell membrane. The activity of an ion can be measured using a glass microelectrode that includes in its tip a liquid-membrane doped with an ion-selective ionophore. Because these electrodes can be fabricated with tip diameters that are less than 1 μm, they can be used to impale single cells in order to monitor the activities of intracellular ions. This review summarizes the history, theory, and practice of ion-selective microelectrode use and brings together a number of classic and recent examples of their usefulness in the realm of physiological study.

  7. Menadione (vitamin K enhances the antibiotic activity of drugs by cell membrane permeabilization mechanism

    Jacqueline C. Andrade

    2017-01-01

    Full Text Available Menadione, vitamin K3, belongs to the class of lipid-soluble vitamins and lipophilic substances as menadione cause disturbances in the bacterial membrane, resulting in damage to the fundamental elements for the integrity of the membrane, thus allowing increased permeability. Accordingly, the aim of this study was to evaluate in vitro the antibiotic-modifying activity of menadione in multiresistant strains of Staphylococcus aureus, Pseudomonas aeruginosa and Escherichia coli, with a gradual increase in its subinhibitory concentration. In addition, menadione was compared with cholesterol and ergosterol for similarity in mechanism of drug modulatory action. Antibiotic-modifying activity and antibacterial effect were determined by the broth microdilution assay. Menadione, cholesterol and ergosterol showed modulatory activity at clinically relevant concentrations, characterizing them as modifiers of bacterial drug resistance, since they lowered the MIC of the antibiotics tested. This is the first report of the antibacterial activity of menadione and its potentiation of aminoglycosides against multiresistant bacteria.

  8. Safety and efficacy of distal perfusion catheterization to prevent limb ischemia after common femoral artery cannulation for extracorporeal membrane oxygenation

    Jeon, Chang Ho; Seong, Nak Jong; Yoon, Chang Jin [Dept. of Radiology, Seoul National University Bundang Hospital, Seongnam (Korea, Republic of)

    2016-06-15

    The extracorporeal membrane oxygenation (ECMO) cannula has the potential for obstructing flow to the lower limb, thus causing severe ischemia and possible limb loss. We evaluated the safety and clinical efficacy of percutaneous distal perfusion catheterization in preventing limb ischemia. Between March 2013 and February 2015, 28 patients with distal perfusion catheterization after ECMO were included in this retrospective study. The technical success was evaluated by Doppler ultrasound at the popliteal level after saline injection via distal perfusion catheter. Clinical success was assessed when at least one of the following conditions was met: restoration of continuous peripheral limb oximetry value or presence of distal arterial pulse on Doppler ultrasound evaluation or resolution of early ischemic sign after connecting the catheter with ECMO. Twenty-six patients with early ischemia were successfully cannulated with a distal perfusion catheter (92.8%). Clinical success was achieved in 12/28 (42.8%) patients; 8/10 (80.0%) patients with survival duration exceeding 7 days and 4/18 (22.2%) patients with survival duration less than 7 days, respectively. A percutaneous distal perfusion catheter placement was a feasible tool with safety and efficacy in preventing lower limb ischemia for patients with prolonged common femoral arterial cannulation for ECMO.

  9. Safety and efficacy of distal perfusion catheterization to prevent limb ischemia after common femoral artery cannulation for extracorporeal membrane oxygenation

    Jeon, Chang Ho; Seong, Nak Jong; Yoon, Chang Jin

    2016-01-01

    The extracorporeal membrane oxygenation (ECMO) cannula has the potential for obstructing flow to the lower limb, thus causing severe ischemia and possible limb loss. We evaluated the safety and clinical efficacy of percutaneous distal perfusion catheterization in preventing limb ischemia. Between March 2013 and February 2015, 28 patients with distal perfusion catheterization after ECMO were included in this retrospective study. The technical success was evaluated by Doppler ultrasound at the popliteal level after saline injection via distal perfusion catheter. Clinical success was assessed when at least one of the following conditions was met: restoration of continuous peripheral limb oximetry value or presence of distal arterial pulse on Doppler ultrasound evaluation or resolution of early ischemic sign after connecting the catheter with ECMO. Twenty-six patients with early ischemia were successfully cannulated with a distal perfusion catheter (92.8%). Clinical success was achieved in 12/28 (42.8%) patients; 8/10 (80.0%) patients with survival duration exceeding 7 days and 4/18 (22.2%) patients with survival duration less than 7 days, respectively. A percutaneous distal perfusion catheter placement was a feasible tool with safety and efficacy in preventing lower limb ischemia for patients with prolonged common femoral arterial cannulation for ECMO

  10. Antibacterial Activity and Membrane-Disruptive Mechanism of 3-p-trans-Coumaroyl-2-hydroxyquinic Acid, a Novel Phenolic Compound from Pine Needles of Cedrus deodara, against Staphylococcus aureus

    Yanping Wu

    2016-08-01

    Full Text Available Recently, we reported that a novel phenolic compound isolated from Cedrus deodara, 3-p-trans-coumaroyl-2-hydroxyquinic acid (CHQA, exhibits a potent antioxidant activity. The present study aimed to evaluate the antibacterial activity of CHQA against eleven food-borne pathogens and to elucidate its mechanism of action against Staphylococcus aureus. The results from minimum inhibitory concentration (MIC determinations showed that CHQA exhibited moderate inhibitory effects on all of the tested pathogens with MIC values ranging from 2.5–10 mg/mL. Membrane potential measurements and flow cytometric analysis demonstrated that CHQA damaged the cytoplasmic membrane of S. aureus, causing a significant membrane hyperpolarization with a loss of membrane integrity. Moreover, CHQA induced an increase in membrane fluidity and conformational changes in membrane protein of S. aureus, suggesting that CHQA probably acts on the cell membrane by interactions with membrane lipid and protein. Transmission electron microscopic observations further confirmed that CHQA disrupted the cell membrane of S. aureus and caused severe morphological changes, which even led to leakage of intracellular constituents. These findings indicated that CHQA could have the potential to serve as a natural antibacterial agent to control and prevent the growth of pathogens in food and in food-processing environments.

  11. Treatment of coal gasification wastewater by membrane bioreactor hybrid powdered activated carbon (MBR–PAC) system.

    Jia, Shengyong; Han, Hongjun; Hou, Baolin; Zhuang, Haifeng; Fang, Fang; Zhao, Qian

    2014-12-01

    A laboratory-scale membrane bioreactor hybrid powdered activated carbon (MBR–PAC) system was developed to treat coal gasification wastewater to enhance the COD, total phenols (TPh), NH4+ removals and migrate the membrane fouling. Since the MBR–PAC system operated with PAC dosage of 4 g L−1, the maximum removal efficiencies of COD, TPh and NH4+ reached 93%, 99% and 63%, respectively with the corresponding influent concentrations of 2.27 g L−1, 497 mg L−1 and 164 mg N L−1; the PAC extraction efficiencies of COD, TPh and NH4+ were 6%, 3% and 13%, respectively; the transmembrane pressure decreased 34% with PAC after 50 d operation. The results demonstrate that PAC played a key role in the enhancement of biodegradability and mitigation of membrane fouling.

  12. Block copolymer hollow fiber membranes with catalytic activity and pH-response

    Hilke, Roland

    2013-08-14

    We fabricated block copolymer hollow fiber membranes with self-assembled, shell-side, uniform pore structures. The fibers in these membranes combined pores able to respond to pH and acting as chemical gates that opened above pH 4, and catalytic activity, achieved by the incorporation of gold nanoparticles. We used a dry/wet spinning process to produce the asymmetric hollow fibers and determined the conditions under which the hollow fibers were optimized to create the desired pore morphology and the necessary mechanical stability. To induce ordered micelle assembly in the doped solution, we identified an ideal solvent mixture as confirmed by small-angle X-ray scattering. We then reduced p-nitrophenol with a gold-loaded fiber to confirm the catalytic performance of the membranes. © 2013 American Chemical Society.

  13. Improved ZIF-8 membrane: Effect of activation procedure and determination of diffusivities of light hydrocarbons

    Pan, Yichang; Liu, Wei; Zhao, Yingjie; Wang, Chongqing; Lai, Zhiping

    2015-01-01

    Zeolitic imidazolate framework ZIF-8 has shown great potential for effective separation of hydrocarbon mixtures based on its intrinsic ultramicroporous feature. In order to explore the permeation and diffusion properties of hydrocarbons through ZIF-8 membrane, high-quality ZIF-8 membranes with a separation factor of ~90 for propylene/propane are successfully prepared via optimizing the activation processes. Single-component permeation data for hydrocarbons (C1–C4) through the improved ZIF-8 membrane are measured and analyzed by Maxwell-Stefan (MS) model to get the transport diffusivities of these hydrocarbons. The diffusivity values of hydrocarbon compare well with those obtained by other experimental techniques. Binary mixture permeation also can be well predicted through single-component adsorption parameters.

  14. Improved ZIF-8 membrane: Effect of activation procedure and determination of diffusivities of light hydrocarbons

    Pan, Yichang

    2015-06-23

    Zeolitic imidazolate framework ZIF-8 has shown great potential for effective separation of hydrocarbon mixtures based on its intrinsic ultramicroporous feature. In order to explore the permeation and diffusion properties of hydrocarbons through ZIF-8 membrane, high-quality ZIF-8 membranes with a separation factor of ~90 for propylene/propane are successfully prepared via optimizing the activation processes. Single-component permeation data for hydrocarbons (C1–C4) through the improved ZIF-8 membrane are measured and analyzed by Maxwell-Stefan (MS) model to get the transport diffusivities of these hydrocarbons. The diffusivity values of hydrocarbon compare well with those obtained by other experimental techniques. Binary mixture permeation also can be well predicted through single-component adsorption parameters.

  15. Block copolymer hollow fiber membranes with catalytic activity and pH-response

    Hilke, Roland; Neelakanda, Pradeep; Madhavan, Poornima; Vainio, Ulla; Behzad, Ali Reza; Sougrat, Rachid; Nunes, Suzana Pereira; Peinemann, Klaus-Viktor

    2013-01-01

    We fabricated block copolymer hollow fiber membranes with self-assembled, shell-side, uniform pore structures. The fibers in these membranes combined pores able to respond to pH and acting as chemical gates that opened above pH 4, and catalytic activity, achieved by the incorporation of gold nanoparticles. We used a dry/wet spinning process to produce the asymmetric hollow fibers and determined the conditions under which the hollow fibers were optimized to create the desired pore morphology and the necessary mechanical stability. To induce ordered micelle assembly in the doped solution, we identified an ideal solvent mixture as confirmed by small-angle X-ray scattering. We then reduced p-nitrophenol with a gold-loaded fiber to confirm the catalytic performance of the membranes. © 2013 American Chemical Society.

  16. Synaptic input correlations leading to membrane potential decorrelation of spontaneous activity in cortex.

    Graupner, Michael; Reyes, Alex D

    2013-09-18

    Correlations in the spiking activity of neurons have been found in many regions of the cortex under multiple experimental conditions and are postulated to have important consequences for neural population coding. While there is a large body of extracellular data reporting correlations of various strengths, the subthreshold events underlying the origin and magnitude of signal-independent correlations (called noise or spike count correlations) are unknown. Here we investigate, using intracellular recordings, how synaptic input correlations from shared presynaptic neurons translate into membrane potential and spike-output correlations. Using a pharmacologically activated thalamocortical slice preparation, we perform simultaneous recordings from pairs of layer IV neurons in the auditory cortex of mice and measure synaptic potentials/currents, membrane potentials, and spiking outputs. We calculate cross-correlations between excitatory and inhibitory inputs to investigate correlations emerging from the network. We furthermore evaluate membrane potential correlations near resting potential to study how excitation and inhibition combine and affect spike-output correlations. We demonstrate directly that excitation is correlated with inhibition thereby partially canceling each other and resulting in weak membrane potential and spiking correlations between neurons. Our data suggest that cortical networks are set up to partially cancel correlations emerging from the connections between neurons. This active decorrelation is achieved because excitation and inhibition closely track each other. Our results suggest that the numerous shared presynaptic inputs do not automatically lead to increased spiking correlations.

  17. Substrate Specificity, Membrane Topology, and Activity Regulation of Human Alkaline Ceramidase 2 (ACER2)*

    Sun, Wei; Jin, Junfei; Xu, Ruijuan; Hu, Wei; Szulc, Zdzislaw M.; Bielawski, Jacek; Obeid, Lina M.; Mao, Cungui

    2010-01-01

    Human alkaline ceramidase 2 (ACER2) plays an important role in cellular responses by regulating the hydrolysis of ceramides in cells. Here we report its biochemical characterization, membrane topology, and activity regulation. Recombinant ACER2 was expressed in yeast mutant cells (Δypc1Δydc1) that lack endogenous ceramidase activity, and microsomes from ACER2-expressiong yeast cells were used to biochemically characterize ACER2. ACER2 catalyzed the hydrolysis of various ceramides and followed...

  18. Characterization of membrane determinant in old T-cells with suppressor activity

    Hendricks, L.C.; Heidrick, M.L.

    1986-01-01

    T-cell function declines with age. Many T-cell functions are initiated at the cell membrane; therefore, age-related membrane alterations may contribute to loss of function. They have previously reported developing a monoclonal antibody, HH-AGE-T(1), which recognizes a cell with suppressor activity and binds to 15-20% of the T-cells from old BC3F 1 mice, but only to 0-4% of young T-cells. To further characterize the determinant recognized by HH-AGE-T(1), they analyzed immunoprecipitates (IP) of young and old T-cell membranes by 2D-SDS PAGE, followed by Western blotting. Immunodetection of the blots showed that HH-AGE-T(1) bound a heterodimer (66 kD, pI 8.44 and 36 kD, pI 5.82-7.12 subunits) in IP from old mice; but not young mice. Monoclonal anti-Lyt 2 antibody did not bind the determinant. When IP of iodinated T-cells were run on SDS-PAGE gels followed by blotting and autoradiography of the blots, very prominent bands were detected in the old sample and faint bands were detected in the young sample. These results suggest that HH-AGE-T(1) recognizes a membrane protein which is present in small amounts on young T-cells but which increases markedly with age. Further studies are needed to determine the significance of this age-related membrane change

  19. Dansyl-Galactoside, a Fluorescent Probe of Active Transport in Bacterial Membrane Vesicles*

    Reeves, John P.; Shechter, Emanuel; Weil, Rudolf; Kaback, H. R.

    1973-01-01

    A fluorescent galactoside, 2-(N-dansyl)-aminoethyl β-D-thiogalactoside (dansyl-galactoside), competitively inhibits lactose transport by membrane vesicles of Escherichia coli, but is not actively transported. An increase in dansyl-galactoside fluorescence is observed upon addition of D-lactate. The fluorescence increase is not observed in membrane vesicles lacking the β-galactoside transport system, and is blocked or rapidly reversed by addition of β-galactosides, sulfhydryl reagents, inhibitors of D-lactate oxidation, or uncoupling agents. The fluorescence increase exhibits an emission maximum at 500 nm and excitation maxima at 345 nm and at 292 nm. The latter excitation maximum is absent unless D-lactate is added, indicating that the bound dansyl-galactoside molecules are excited by energy transfer from the membrane proteins. Titration of vesicles with dansyl-galactoside in the presence of D-lactate demonstrates that the β-galactoside carrier protein represents about 3.3% of the total membrane protein. The data indicate that D-lactate oxidation leads to binding of the fluorescent galactoside to the β-galactoside carrier protein in such a manner that the dansyl group is transferred to a hydrophobic environment within the membrane. PMID:4583021

  20. Adaptation of H+-pumping and plasma membrane H+ ATPase activity in proteoid roots of white lupin under phosphate deficiency.

    Yan, Feng; Zhu, Yiyong; Müller, Caroline; Zörb, Christian; Schubert, Sven

    2002-05-01

    White lupin (Lupinus albus) is able to adapt to phosphorus deficiency by producing proteoid roots that release a huge amount of organic acids, resulting in mobilization of sparingly soluble soil phosphate in rhizosphere. The mechanisms responsible for the release of organic acids by proteoid root cells, especially the trans-membrane transport processes, have not been elucidated. Because of high cytosolic pH, the release of undissociated organic acids is not probable. In the present study, we focused on H+ export by plasma membrane H+ ATPase in active proteoid roots. In vivo, rhizosphere acidification of active proteoid roots was vanadate sensitive. Plasma membranes were isolated from proteoid roots and lateral roots from P-deficient and -sufficient plants. In vitro, in comparison with two types of lateral roots and proteoid roots of P-sufficient plants, the following increase of the various parameters was induced in active proteoid roots of P-deficient plants: (a) hydrolytic ATPase activity, (b) Vmax and Km, (c) H+ ATPase enzyme concentration of plasma membrane, (d) H+-pumping activity, (e) pH gradient across the membrane of plasmalemma vesicles, and (f) passive H+ permeability of plasma membrane. In addition, lower vanadate sensitivity and more acidic pH optimum were determined for plasma membrane ATPase of active proteoid roots. Our data support the hypothesis that in active proteoid root cells, H+ and organic anions are exported separately, and that modification of plasma membrane H+ ATPase is essential for enhanced rhizosphere acidification by active proteoid roots.

  1. Adsorption of pharmaceuticals onto isolated polyamide active layer of NF/RO membranes.

    Liu, Yan-Ling; Wang, Xiao-Mao; Yang, Hong-Wei; Xie, Yuefeng F

    2018-06-01

    Adsorption of trace organic compounds (TrOCs) onto the membrane materials has a great impact on their rejection by nanofiltration (NF) and reverse osmosis (RO) membranes. This study aimed to investigate the difference in adsorption of various pharmaceuticals (PhACs) onto different NF/RO membranes and to demonstrate the necessity of isolating the polyamide (PA) active layer from the polysulfone (PS) support layer for adsorption characterization and quantification. Both the isolated PA layers and the PA+PS layers of NF90 and ESPA1 membranes were used to conduct static adsorption tests. Results showed that apparent differences existed between the PA layer and the PA+PS layer in the adsorption capacity of PhACs as well as the time necessary to reach the adsorption equilibrium. PhACs with different physicochemical properties could be adsorbed to different extents by the isolated PA layer, which was mainly attributed to electrostatic attraction/repulsion and hydrophobic interactions. The PA layer of ESPA1 exhibited apparently higher adsorption capacities for the positively charged PhACs and similar adsorption capacities for the neutral PhACs although it had significantly less total interfacial area (per unit membrane surface area) for adsorption compared to the PA layer of NF90. The higher affinity of the PA layer of ESPA1 for the PhACs could be due to its higher capacity of forming hydrogen bonds with PhACs resulted from the modified chemistry with more -OH groups. This study provides a novel approach to determining the TrOC adsorption onto the active layer of membranes for the ease of investigating adsorption mechanisms. Copyright © 2018 Elsevier Ltd. All rights reserved.

  2. AMPK Activation Prevents and Reverses Drug-Induced Mitochondrial and Hepatocyte Injury by Promoting Mitochondrial Fusion and Function.

    Sun Woo Sophie Kang

    Full Text Available Mitochondrial damage is the major factor underlying drug-induced liver disease but whether conditions that thwart mitochondrial injury can prevent or reverse drug-induced liver damage is unclear. A key molecule regulating mitochondria quality control is AMP activated kinase (AMPK. When activated, AMPK causes mitochondria to elongate/fuse and proliferate, with mitochondria now producing more ATP and less reactive oxygen species. Autophagy is also triggered, a process capable of removing damaged/defective mitochondria. To explore whether AMPK activation could potentially prevent or reverse the effects of drug-induced mitochondrial and hepatocellular damage, we added an AMPK activator to collagen sandwich cultures of rat and human hepatocytes exposed to the hepatotoxic drugs, acetaminophen or diclofenac. In the absence of AMPK activation, the drugs caused hepatocytes to lose polarized morphology and have significantly decreased ATP levels and viability. At the subcellular level, mitochondria underwent fragmentation and had decreased membrane potential due to decreased expression of the mitochondrial fusion proteins Mfn1, 2 and/or Opa1. Adding AICAR, a specific AMPK activator, at the time of drug exposure prevented and reversed these effects. The mitochondria became highly fused and ATP production increased, and hepatocytes maintained polarized morphology. In exploring the mechanism responsible for this preventive and reversal effect, we found that AMPK activation prevented drug-mediated decreases in Mfn1, 2 and Opa1. AMPK activation also stimulated autophagy/mitophagy, most significantly in acetaminophen-treated cells. These results suggest that activation of AMPK prevents/reverses drug-induced mitochondrial and hepatocellular damage through regulation of mitochondrial fusion and autophagy, making it a potentially valuable approach for treatment of drug-induced liver injury.

  3. AMPK Activation Prevents and Reverses Drug-Induced Mitochondrial and Hepatocyte Injury by Promoting Mitochondrial Fusion and Function

    Taniane, Caitlin; Farrell, Geoffrey; Arias, Irwin M.; Lippincott-Schwartz, Jennifer; Fu, Dong

    2016-01-01

    Mitochondrial damage is the major factor underlying drug-induced liver disease but whether conditions that thwart mitochondrial injury can prevent or reverse drug-induced liver damage is unclear. A key molecule regulating mitochondria quality control is AMP activated kinase (AMPK). When activated, AMPK causes mitochondria to elongate/fuse and proliferate, with mitochondria now producing more ATP and less reactive oxygen species. Autophagy is also triggered, a process capable of removing damaged/defective mitochondria. To explore whether AMPK activation could potentially prevent or reverse the effects of drug-induced mitochondrial and hepatocellular damage, we added an AMPK activator to collagen sandwich cultures of rat and human hepatocytes exposed to the hepatotoxic drugs, acetaminophen or diclofenac. In the absence of AMPK activation, the drugs caused hepatocytes to lose polarized morphology and have significantly decreased ATP levels and viability. At the subcellular level, mitochondria underwent fragmentation and had decreased membrane potential due to decreased expression of the mitochondrial fusion proteins Mfn1, 2 and/or Opa1. Adding AICAR, a specific AMPK activator, at the time of drug exposure prevented and reversed these effects. The mitochondria became highly fused and ATP production increased, and hepatocytes maintained polarized morphology. In exploring the mechanism responsible for this preventive and reversal effect, we found that AMPK activation prevented drug-mediated decreases in Mfn1, 2 and Opa1. AMPK activation also stimulated autophagy/mitophagy, most significantly in acetaminophen-treated cells. These results suggest that activation of AMPK prevents/reverses drug-induced mitochondrial and hepatocellular damage through regulation of mitochondrial fusion and autophagy, making it a potentially valuable approach for treatment of drug-induced liver injury. PMID:27792760

  4. Transferred nuclear Overhauser effect analyses of membrane-bound enkephalin analogues by sup 1 H nuclear magnetic resonance: Correlation between activities and membrane-bound conformations

    Milon, Alain; Miyazawa, Tatsuo; Higashijima, Tsutomu (Univ. of Tokyo (Japan))

    1990-01-09

    Leu-enkephalin, (D-Ala{sup 2})Leu-enkephalin, and (D-Ala{sup 2})Leu-enkephalinamide (agonists) and (L-Ala{sup 2})Leu-enkephalin (inactive analogue) bind to lipid bilayer consisting of phosphatidylcholine and phosphatidylserine. The conformations that these compounds assume, once bound to perdeuterated phospholipid bilayer, have been shown to be unique, as shown by the transferred nuclear Overhauser effect (TRNOE) of {sup 1}H NMR spectroscopy. In addition, their location in the bilayer was analyzed by TRNOE in the presence of spin-labeled phospholipids. These analyses showed a clear relationship between the activity and the peptide-membrane interaction. The three active peptides, when bound to membranes, adopt the same conformation, characterized by a type II{prime} {beta}-turn around Gly{sup 3}-Phe and a {gamma}-turn around Gly{sup 2} (or D-Ala{sup 2}). The inactive analogue, (L-Ala{sup 2})Leu-enkephalin, displayed a completely different TRNOE pattern corresponding to a different conformation in the membrane-bound state. The tyrosine residue of the active compounds is not inserted into the interior of membrane, but it is inserted into the bilayer for the L-Ala{sup 2} analogue. According to these results, (L-Ala{sup 2})Leu-enkephalin may be explained to be inactive because the mode of binding to the membranes is different from that of active compounds.

  5. Diet and physical activity in the prevention of colorectal cancer.

    Mehta, Mamta; Shike, Moshe

    2014-12-01

    Diet has been linked to the prevention of colorectal cancer (CRC) and may explain some of the differences in incidence and mortality among various populations. Evidence suggests that a high intake of red and processed meats is associated with an increased risk of CRC. The protective benefits of fiber are unclear, although in some studies fiber is associated with reduced CRC risk. The role of supplements, such as calcium, vitamin D, and folic acid, remains uncertain, and these nutrients cannot be currently recommended for chemoprevention. Obesity and sedentary lifestyle have been associated with an increased risk for colon cancer. Because of the inherent difficulty in studying the effects of specific nutrients, dietary pattern analysis may be a preferable approach to the investigation of the relationship between diet and risk for human diseases. Lifestyle modifications, such as increasing physical activity and consumption of a diet rich in fiber, fruits, vegetables, fish, and poultry and low in red and processed meats, have been advocated for primary prevention of several chronic diseases, and may in fact be beneficial for cancer prevention, particularly CRC. Copyright © 2014 by the National Comprehensive Cancer Network.

  6. Investigation on degradation mechanism of ion exchange membrane immersed in highly concentrated tritiated water under the Broader Approach Activities

    Iwai, Yasunori, E-mail: iwai.yasunori@jaea.go.jp; Sato, Katsumi; Yamanishi, Toshihiko

    2014-10-15

    Highlights: • Endurance of Nafion ion exchange membrane immersed into 1.38 × 10{sup 12} Bq/kg of highly concentrated tritiated water has been demonstrated. • The formation of free hydrophobic free products by reactions between radicals on the membrane and oxygen caused the decrease in ionic conductivity. • From the {sup 19}F NMR spectrum, no distinctive degradation in the membrane structure by interaction with tritium was measured. - Abstract: The ion exchange membrane is a key material for electrolysis cells of the water detritiation system. Durability of ion exchange membrane has been demonstrated under the Broader Approach Activities. Long-term exposure of Nafion{sup ®} ion exchange membrane in 1.38 × 10{sup 12} Bq/kg of tritiated water was conducted at room temperature for up to 2 years. The ionic conductivity of Nafion{sup ®} membrane after immersed in tritiated water was changed. The change in color of membrane from colorless to yellowish was caused by reactions of radicals on the polymer and oxygen molecules in air. Infrared Fourier transform spectrum of a yellowish membrane revealed a small peak for bending vibration of C-H situated at 1437 cm{sup −1}, demonstrating the formation of hydrophobic functional group in the membrane. The hydrophilic network in Nafion{sup ®} membrane was partially obstructed by the hydrophobic free products. This caused the decrease in ionic conductivity. The peak for bending vibration was clearly eliminated in the spectrum of the membrane after treatment by acid for removal of free products. The high-resolution solid state {sup 19}F NMR spectrum of a membrane after immersed in tritiated water was similar to that of a membrane irradiated with gamma-rays. From the {sup 19}F NMR spectrum, no distinctive degradation in the membrane structure by interaction with tritium was measured.

  7. Investigation on degradation mechanism of ion exchange membrane immersed in highly concentrated tritiated water under the Broader Approach Activities

    Iwai, Yasunori; Sato, Katsumi; Yamanishi, Toshihiko

    2014-01-01

    Highlights: • Endurance of Nafion ion exchange membrane immersed into 1.38 × 10 12 Bq/kg of highly concentrated tritiated water has been demonstrated. • The formation of free hydrophobic free products by reactions between radicals on the membrane and oxygen caused the decrease in ionic conductivity. • From the 19 F NMR spectrum, no distinctive degradation in the membrane structure by interaction with tritium was measured. - Abstract: The ion exchange membrane is a key material for electrolysis cells of the water detritiation system. Durability of ion exchange membrane has been demonstrated under the Broader Approach Activities. Long-term exposure of Nafion ® ion exchange membrane in 1.38 × 10 12 Bq/kg of tritiated water was conducted at room temperature for up to 2 years. The ionic conductivity of Nafion ® membrane after immersed in tritiated water was changed. The change in color of membrane from colorless to yellowish was caused by reactions of radicals on the polymer and oxygen molecules in air. Infrared Fourier transform spectrum of a yellowish membrane revealed a small peak for bending vibration of C-H situated at 1437 cm −1 , demonstrating the formation of hydrophobic functional group in the membrane. The hydrophilic network in Nafion ® membrane was partially obstructed by the hydrophobic free products. This caused the decrease in ionic conductivity. The peak for bending vibration was clearly eliminated in the spectrum of the membrane after treatment by acid for removal of free products. The high-resolution solid state 19 F NMR spectrum of a membrane after immersed in tritiated water was similar to that of a membrane irradiated with gamma-rays. From the 19 F NMR spectrum, no distinctive degradation in the membrane structure by interaction with tritium was measured

  8. Covalent glycoinositolphospholipid (GPI binding to hemoglobin is associated with insulin-activation of erythrocyte membrane protease

    VESNA NIKETIC

    2004-05-01

    Full Text Available Recently, it was demonstrated that prolonged hyperinsulinism associated with hypoglycemia, both in vivo and in vitro, caused covalent glycoinositolphospholipid (GPI binding to the C termini of both hemoglobin b-chains, which resulted in the formation of a novel, hitherto unrecognized, minor hemoglobin fraction (GPI-Hb (Niketic et al., Biochem. Biophys. Res. Commun. 239 (1997 435. In this study it was demonstrated that exposure of erythrocyte membranes to insulin causes the activation of membrane protease as well as that the formation of GPI-Hb parallels its activity. It is suggested that the insulin-activated protease is able to catalyze, albeit slowly, the transpeptidation, i.e., the replacement of the carboxy-terminal amino acid(s residues of the Hb b-chains with GPI as an exogenous nucleophile. To our knowledge the present results show for the first time that insulin stimulates protease activity in erythrocyte membranes, as well as that insulin-activated protease may be involved in post-translational GPI binding to proteins.

  9. Shape-dependent bactericidal activity of copper oxide nanoparticle mediated by DNA and membrane damage

    Laha, Dipranjan; Pramanik, Arindam; Laskar, Aparna; Jana, Madhurya; Pramanik, Panchanan; Karmakar, Parimal

    2014-01-01

    Highlights: • Spherical and sheet shaped copper oxide nanoparticles were synthesized. • Physical characterizations of these nanoparticles were done by TEM, DLS, XRD, FTIR. • They showed shape dependent antibacterial activity on different bacterial strain. • They induced both membrane damage and ROS mediated DNA damage in bacteria. - Abstract: In this work, we synthesized spherical and sheet shaped copper oxide nanoparticles and their physical characterizations were done by the X-ray diffraction, fourier transform infrared spectroscopy, transmission electron microscopy and dynamic light scattering. The antibacterial activity of these nanoparticles was determined on both gram positive and gram negative bacterial. Spherical shaped copper oxide nanoparticles showed more antibacterial property on gram positive bacteria where as sheet shaped copper oxide nanoparticles are more active on gram negative bacteria. We also demonstrated that copper oxide nanoparticles produced reactive oxygen species in both gram negative and gram positive bacteria. Furthermore, they induced membrane damage as determined by atomic force microscopy and scanning electron microscopy. Thus production of and membrane damage are major mechanisms of the bactericidal activity of these copper oxide nanoparticles. Finally it was concluded that antibacterial activity of nanoparticles depend on physicochemical properties of copper oxide nanoparticles and bacterial strain

  10. Antioxidant activities of bambara groundnut (Vigna subterranea) protein hydrolysates and their membrane ultrafiltration fractions.

    Arise, Abimbola K; Alashi, Adeola M; Nwachukwu, Ifeanyi D; Ijabadeniyi, Oluwatosin A; Aluko, Rotimi E; Amonsou, Eric O

    2016-05-18

    In this study, the bambara protein isolate (BPI) was digested with three proteases (alcalase, trypsin and pepsin), to produce bambara protein hydrolysates (BPHs). These hydrolysates were passed through ultrafiltration membranes to obtain peptide fractions of different sizes (fractions were investigated for antioxidant activities. The membrane fractions showed that peptides with sizes 3 kDa. This is in agreement with the result obtained for the ferric reducing power, metal chelating and hydroxyl radical scavenging activities where higher molecular weight peptides exhibited better activity (p fractions. However, for all the hydrolysates, the low molecular weight peptides were more effective diphenyl-1-picrylhydrazyl (DPPH) radical scavengers but not superoxide radicals when compared to the bigger peptides. In comparison with glutathione (GSH), BPHs and their membrane fractions had better (p fractions that did not show any metal chelating activity. However, the 5-10 kDa pepsin hydrolysate peptide fractions had greater (88%) hydroxyl scavenging activity than GSH, alcalase and trypsin hydrolysates (82%). These findings show the potential use of BPHs and their peptide fraction as antioxidants in reducing food spoilage or management of oxidative stress-related metabolic disorders.

  11. Shape-dependent bactericidal activity of copper oxide nanoparticle mediated by DNA and membrane damage

    Laha, Dipranjan; Pramanik, Arindam [Department of Life Science and Biotechnology, Jadavpur University, 188, Raja S C Mallick Road, Kolkata 700032 (India); Laskar, Aparna [CSIR-Indian Institute of Chemical Biology, Kolkata 700032 (India); Jana, Madhurya [Department of Life Science and Biotechnology, Jadavpur University, 188, Raja S C Mallick Road, Kolkata 700032 (India); Pramanik, Panchanan [Department of Chemistry, Indian Institute of Technology, Kharagpur 721302 (India); Karmakar, Parimal, E-mail: pkarmakar_28@yahoo.co.in [Department of Life Science and Biotechnology, Jadavpur University, 188, Raja S C Mallick Road, Kolkata 700032 (India)

    2014-11-15

    Highlights: • Spherical and sheet shaped copper oxide nanoparticles were synthesized. • Physical characterizations of these nanoparticles were done by TEM, DLS, XRD, FTIR. • They showed shape dependent antibacterial activity on different bacterial strain. • They induced both membrane damage and ROS mediated DNA damage in bacteria. - Abstract: In this work, we synthesized spherical and sheet shaped copper oxide nanoparticles and their physical characterizations were done by the X-ray diffraction, fourier transform infrared spectroscopy, transmission electron microscopy and dynamic light scattering. The antibacterial activity of these nanoparticles was determined on both gram positive and gram negative bacterial. Spherical shaped copper oxide nanoparticles showed more antibacterial property on gram positive bacteria where as sheet shaped copper oxide nanoparticles are more active on gram negative bacteria. We also demonstrated that copper oxide nanoparticles produced reactive oxygen species in both gram negative and gram positive bacteria. Furthermore, they induced membrane damage as determined by atomic force microscopy and scanning electron microscopy. Thus production of and membrane damage are major mechanisms of the bactericidal activity of these copper oxide nanoparticles. Finally it was concluded that antibacterial activity of nanoparticles depend on physicochemical properties of copper oxide nanoparticles and bacterial strain.

  12. In vitro and in vivo activation of mitochondrial membrane permeability transition pore using triiodothyronine.

    Endlicher, R; Drahota, Z; Červinková, Z

    2016-06-20

    Using a novel method for evaluating mitochondrial swelling (Drahota et al. 2012a) we studied the effect of calcium (Ca(2+)), phosphate (P(i)), and triiodothyronine (T(3)) on the opening of mitochondrial membrane permeability transition pore and how they interact in the activation of swelling process. We found that 0.1 mM P(i), 50 microM Ca(2+) and 25 microM T(3) when added separately increase the swelling rate to about 10 % of maximal values when all three factors are applied simultaneously. Our findings document that under experimental conditions in which Ca(2+) and P(i) are used as activating factors, the addition of T(3) doubled the rate of swelling. T(3) has also an activating effect on mitochondrial membrane potential. The T(3) activating effect was also found after in vivo application of T(3). Our data thus demonstrate that T(3) has an important role in opening the mitochondrial membrane permeability pore and activates the function of the two key physiological swelling inducers, calcium and phosphate ions.

  13. Chlorovirus-mediated membrane depolarization of Chlorella alters secondary active transport of solutes.

    Agarkova, Irina; Dunigan, David; Gurnon, James; Greiner, Timo; Barres, Julia; Thiel, Gerhard; Van Etten, James L

    2008-12-01

    Paramecium bursaria chlorella virus 1 (PBCV-1) is the prototype of a family of large, double-stranded DNA, plaque-forming viruses that infect certain eukaryotic chlorella-like green algae from the genus Chlorovirus. PBCV-1 infection results in rapid host membrane depolarization and potassium ion release. One interesting feature of certain chloroviruses is that they code for functional potassium ion-selective channel proteins (Kcv) that are considered responsible for the host membrane depolarization and, as a consequence, the efflux of potassium ions. This report examines the relationship between cellular depolarization and solute uptake. Annotation of the virus host Chlorella strain NC64A genome revealed 482 putative transporter-encoding genes; 224 are secondary active transporters. Solute uptake experiments using seven radioactive compounds revealed that virus infection alters the transport of all the solutes. However, the degree of inhibition varied depending on the solute. Experiments with nystatin, a drug known to depolarize cell membranes, produced changes in solute uptake that are similar but not identical to those that occurred during virus infection. Therefore, these studies indicate that chlorovirus infection causes a rapid and sustained depolarization of the host plasma membrane and that this depolarization leads to the inhibition of secondary active transporters that changes solute uptake.

  14. Effect of Powdered Activated Carbon to Reduce Fouling in Membrane Bioreactors: A Sustainable Solution. Case Study

    Giuseppe Mancini

    2013-04-01

    Full Text Available Membrane Bio Reactors (MBRs are mainly used for industrial wastewaters applications where their costs can be more easily afforded. High costs are basically due to energy consumption and membrane cleaning or replacement. Membrane fouling is responsible for reducing treated water production and increasing maintenance as well as operation costs. According to previous researches, the addition of Powdered Activated Carbon (PAC in high dosages could reduce membrane fouling; but such concentrations are economically unsustainable for operative conditions. A MBR pilot plant, fed by mixed liquor of a full-scale activated sludge process from a municipal wastewater treatment plant, was operated dosing low PAC concentrations (0, 2, 5, 10 and 20 mg·L−1, respectively. Experiments were also carried out at two different temperatures corresponding to summer and winter conditions. Results indicated that PAC addition was effective at the low dosages (2 and 5 mg·L−1 by reducing the permeate flux loss (from 16 up to 27%, respectively while higher PAC concentrations turns out in a useless cost increase.

  15. [Adapted physical activity in the prevention and therapy of osteoporosis].

    Bosković, Ksenija; Gava, Branka Protić; Grajić, Mirko; Madić, Dejan; Obradović, Borislav; Todorović, Snezana Tomasević

    2013-01-01

    Osteoporosis, a disease characterized by the progressive loss of bone tissue, is one of the most common complications of aging. According to some calculations, there were 25% of women and 4% of men older than 50 years with osteoporosis in the world in 2010. It is assumed that the number of patients with osteoporosis will increase by 30% in every 10 years in the 21st century. There are many reasons for that: the world's population is growing older, diet is getting poorer in vitamins and minerals and physical activity is decreasing. THE QUALITY AND QUANTITY OF BONE TISSUE: Developing bones are much more responsive to mechanical loading and physical activity than mature bones. This suggests that training in early childhood may be an important factor in the prevention of osteoporosis in later life. It is important to note that the quality of bone achieved by training at younger age cannot be maintained permanently if it is not supported by physical activity later in life. Adapted physical activity represents physical activity individually tailored according to the psychosomatic capabilities of a person and the goal to be achieved. It can be applied at any age in order to maintain strong bones and reduce the risk of fracture. Adapted physical activity is different for men and women, for different age, as well as for the individuals. Aerobic exercises, which lead to an acceleration of breathing, increased heart rate and mild perspiration, as well as resistance exercises and exercises against resistance done by stretching elastic bands, for hands, legs and torso have been proven to increase bone density and improve bone strength. Coordination and balance exercises are important in an individual workout program. An explanation of the action of adapted physical activity is the basis for the theory of control and modulation of bone loss, muscle strength, coordination and balance. Physical activity is very effective in reducing sclerostin, which is known to inhibit bone

  16. Membrane fusion and exocytosis.

    Jahn, R; Südhof, T C

    1999-01-01

    Membrane fusion involves the merger of two phospholipid bilayers in an aqueous environment. In artificial lipid bilayers, fusion proceeds by means of defined transition states, including hourglass-shaped intermediates in which the proximal leaflets of the fusing membranes are merged whereas the distal leaflets are separate (fusion stalk), followed by the reversible opening of small aqueous fusion pores. Fusion of biological membranes requires the action of specific fusion proteins. Best understood are the viral fusion proteins that are responsible for merging the viral with the host cell membrane during infection. These proteins undergo spontaneous and dramatic conformational changes upon activation. In the case of the paradigmatic fusion proteins of the influenza virus and of the human immunodeficiency virus, an amphiphilic fusion peptide is inserted into the target membrane. The protein then reorients itself, thus forcing the fusing membranes together and inducing lipid mixing. Fusion of intracellular membranes in eukaryotic cells involves several protein families including SNAREs, Rab proteins, and Sec1/Munc-18 related proteins (SM-proteins). SNAREs form a novel superfamily of small and mostly membrane-anchored proteins that share a common motif of about 60 amino acids (SNARE motif). SNAREs reversibly assemble into tightly packed helical bundles, the core complexes. Assembly is thought to pull the fusing membranes closely together, thus inducing fusion. SM-proteins comprise a family of soluble proteins that bind to certain types of SNAREs and prevent the formation of core complexes. Rab proteins are GTPases that undergo highly regulated GTP-GDP cycles. In their GTP form, they interact with specific proteins, the effector proteins. Recent evidence suggests that Rab proteins function in the initial membrane contact connecting the fusing membranes but are not involved in the fusion reaction itself.

  17. Location and activity of ubiquinone 10 and ubiquinone analogues in model and biological membranes

    Cornell, B.A.; Keniry, M.A.; Post, A.; Roberston, R.N.; Weir, L.E.; Westerman, P.W.

    1987-01-01

    Deuteriated analogues of ubiquinone 10 (Q 10 ) have been dispersed with plasma membranes of Escherichia coli and with the inner membranes of beetroot mitochondria. Orientational order at various deuteriated sites was measured by solid-state deuterium nuclear magnetic resonance ( 2 H NMR). Similar measurements were made, using the compounds dispersed in dimyristoylphosphatidylcholine (DMPC) and egg yolk lecithin and dispersions prepared from the lipid extracts of beetroot mitochondria. In all cases only a single unresolved 2 H NMR spectrum (typically 1000-Hz full width at half-height) was observed at concentrations down to 0.02 mol % Q 10 per membrane lipid. This result shows that most Q 10 is in a mobile environment which is physically separate from the orientational constraints of the bilayer lipid chains. In contrast, a short-chain analog of Q 10 , in which the 10 isoprene groups have been replaced by a perdeuteriated tridecyl chain, showed 2 H NMR spectra with quadrupolar splittings typical of an ordered lipid that is intercalated into the bilayer. The NADH oxidase activity and O 2 uptake in Escherichia coli and in mitochondria were independent of which analog was incorporated into the membrane. Thus, despite the major difference in their physical association with membranes, or their lipid extracts, the electron transport function of the long- and short-chain ubiquinones is similar, suggesting that the bulk of the long-chain ubiquinone does not have a direct function in electron transporting activity. The physiologically active Q 10 may only be a small fraction of the total ubiquinone, a fraction that is below the level of detection of the present NMR equipment. However, our results do not support any model of Q 10 electron transport action that includes intercalation of the long isoprenoid chain in lipid

  18. Instabilities and diffusion in a hydrodynamic model of a fluid membrane coupled to a thin active fluid layer.

    Sarkar, N; Basu, A

    2012-11-01

    We construct a coarse-grained effective two-dimensional (2d hydrodynamic theory as a theoretical model for a coupled system of a fluid membrane and a thin layer of a polar active fluid in its ordered state that is anchored to the membrane. We show that such a system is prone to generic instabilities through the interplay of nonequilibrium drive, polar order and membrane fluctuation. We use our model equations to calculate diffusion coefficients of an inclusion in the membrane and show that their values depend strongly on the system size, in contrast to their equilibrium values. Our work extends the work of S. Sankararaman and S. Ramaswamy (Phys. Rev. Lett., 102, 118107 (2009)) to a coupled system of a fluid membrane and an ordered active fluid layer. Our model is broadly inspired by and should be useful as a starting point for theoretical descriptions of the coupled dynamics of a cell membrane and a cortical actin layer anchored to it.

  19. Generation and characterization of tabalumab, a human monoclonal antibody that neutralizes both soluble and membrane-bound B-cell activating factor

    Manetta J

    2014-08-01

    Full Text Available Joseph Manetta, Holly Bina, Paul Ryan, Niles Fox, Derrick R Witcher, Kristine Kikly Biotechnology Discovery Research, Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, IN, USA Abstract: B-cell activating factor (BAFF is a B-cell survival factor with a key role in B-cell homeostasis and tolerance. Dysregulated BAFF expression may contribute to autoimmune diseases or B-cell malignancies via effects on abnormal B-lymphocyte activation, proliferation, survival, and immunoglobulin secretion. Monoclonal antibodies were generated against human BAFF, characterized for species specificity and affinity, and screened for the ability to neutralize both membrane-bound and soluble BAFF. In addition, studies were undertaken to determine the relative potency of membrane-bound and soluble BAFF. Tabalumab has a high affinity for human, cynomolgus monkey, and rabbit BAFF. No binding to mouse BAFF was detected. Tabalumab was able to neutralize soluble human, cynomolgus monkey, or rabbit BAFF with equal potency. Our data demonstrate that membrane-bound BAFF can be a more potent stimulus for B-cells than soluble BAFF, and tabalumab also neutralized membrane-bound BAFF. Tabalumab prevented BAFF from binding to BAFF receptors and demonstrated pharmacodynamic effects in human BAFF transgenic mice. Tabalumab is a high-affinity human antibody with neutralizing activity against membrane-bound and soluble BAFF. Given our findings that membrane-bound BAFF can have greater in vitro potency than soluble BAFF, neutralization of both forms of BAFF is likely to be important for optimal therapeutic effect. Keywords: autoimmunity, B-cell malignancies, B-cell survival factor, BAFF

  20. Fouling prevention of peptides from a tryptic whey hydrolysate during electromembrane processes by use of monovalent ion permselective membranes

    Persico, Mathieu; Bazinet, Laurent

    2017-01-01

    Peptide adsorption occurring on conventional anion- and cation-exchange membranes is one of the main technological locks in electrodialysis (ED) for hydrolysate demineralization. Hence, the peptide fouling of monovalent anion (MAP) and monovalent cation (MCP) permselective membranes was studied and compared to conventional membranes (AMX-SB and CMX-SB). It appeared that the main peptide sequences responsible for fouling were TPEVDDEALEKFDK, VAGTWY and VLVLDTDYK for both anionic membranes; and...

  1. Abnormal reticuloendothelial function in patients with active vasculitis and idiopathic membranous glomerulopathy

    van der Woude, F J; Piers, D A; van der Giessen, M; Hoedemaeker, P J; Hauw The, T; van der Hem, G K

    1983-02-01

    Reticuloendothelial function was assessed in 11 patients with systemic lupus erythematosus, 8 patients with Wegener's granulomatosus, and 20 patients with idiopathic membranous sup(99m)Tc-labeled heat-damaged red blood cells. With this method organ uptake could be measured by quantitative scintigraphy. There was no relation between the Tsub(1/2) of the blood disappearance curve and the Tsub(1/2) of the splenic uptake curve. The Tsub(1/2) of the blood disappearance curve was normal in all the patient groups. However, there was a significant shift from spleen to liver uptake in patients with active systemic lupus erythematosus, active Wegener's granulomatosus, and membranous glomerulopathy in comparison with a control group. There was no relation with age, level of circulating immune complexes, complement level, kidney function, or immunosuppressive treatment. We conclude that an increase of the liver component of reticuloendothelial function may compensate abnormalities in splenic function. This stresses the importance of quantitative scanning to detect such abnormalities. The study provides evidence for disease related hyposplenism in patients with active systemical lupus erythematosus, active Wegener's granulamatosus, and membranous glomerulopathy.

  2. Abnormal reticuloendothelial function in patients with active vasculitis and idiopathic membranous glomerulopathy

    Woude, F.J. van der; Piers, D.A.; Giessen, M. van der; Hoedemaeker, P.J.; Hauw The, T.; Hem, G.K. van der; Rijksuniversiteit Groningen; Rijksuniversiteit Groningen

    1983-01-01

    Reticuloendothelial function was assessed in 11 patients with systemic lupus erythematosus, 8 patients with Wegener's granulomatosus, and 20 patients with idiopathic membranous sup(99m)Tc-labeled heat-damaged red blood cells. With this method organ uptake could be measured by quantitative scintigraphy. There was no relation between the Tsub(1/2) of the blood disappearance curve and the Tsub(1/2) of the splenic uptake curve. The Tsub(1/2) of the blood disappearance curve was normal in all the patient groups. However, there was a significant shift from spleen to liver uptake in patients with active systemic lupus erythematosus, active Wegener's granulomatosus, and membranous glomerulopathy in comparison with a control group. There was no relation with age, level of circulating immune complexes, complement level, kidney function, or immunosuppressive treatment. We conclude that an increase of the liver component of reticuloen-dothelial function may compensate abnormalities in splenic function. This stresses the importance of quantitative scanning to detect such abnormalities. The study provides evidence for disaase related hyposplenism in patients with active systemical lupus erythematosus, active Wegener's granulamatosus, and membranous glomerulopathy. (orig.)

  3. Development of a novel multi-functional active membrane capping barrier for the remediation of nitrobenzene-contaminated sediment.

    Wang, Qing; Li, Yi; Wang, Chao; Wu, Yue; Wang, Peifang

    2014-07-15

    A novel bio-reactive capping barrier composed of polysulfone/granular activated carbon (PS/GAC) hybrid membranes immobilized with microorganism was developed for the remediation of nitrobenzene in sediments. The SEM observation demonstrated that all the membranes had a dense top layer and a porous sublayer, this structure can block the transfer of nitrobenzene from sediment to the water and enhance nitrobenzene degradation. Adsorption behaviors of nitrobenzene on membranes showed that the membrane impregnated with GAC had better performance than the pure PS membrane. The values of Kads increased from 4.64 (without GAC) to 6.19 (1:2 GAC). 20mg/L nitrobenzene can be completely degraded by Pseudomonas putida immobilized on membranes. The biodegradation rate of activated carbon-filled membrane system was little higher than that of pure PS membrane system. For remediation experiments, only about 21.7, 28.3 and 43.9% of nitrobenzene in the sediment was removed by the end of the experiments for PS/GAC membrane, sand-alone and sand amended with activated carbon capping systems, respectively. While for PS/GAC+microorganisms capping system, more than 70% of nitrobenzene loss was observed. This demonstrated that nitrobenzene can be effectively removed from contaminated sediments by microbial degradation in the bio-reactive capping system. Copyright © 2014 Elsevier B.V. All rights reserved.

  4. Membrane omega-3 Fatty Acid deficiency as a preventable risk factor for comorbid coronary heart disease in major depressive disorder.

    McNamara, Robert K

    2009-01-01

    Major depression disorder (MDD) significantly increases the risk for coronary heart disease (CHD) which is a leading cause of mortality in patients with MDD. Moreover, depression is frequently observed in a subset of patients following acute coronary syndrome (ACS) and increases risk for mortality. Here evidence implicating omega-3 (n-3) fatty acid deficiency in the pathoaetiology of CHD and MDD is reviewed, and the hypothesis that n-3 fatty acid deficiency is a preventable risk factor for CHD comorbidity in MDD patients is evaluated. This hypothesis is supported by cross-national and cross-sectional epidemiological surveys finding an inverse correlation between n-3 fatty acid status and prevalence rates of both CHD and MDD, prospective studies finding that lower dietary or membrane EPA+DHA levels increase risk for both MDD and CHD, case-control studies finding that the n-3 fatty acid status of MDD patients places them at high risk for emergent CHD morbidity and mortality, meta-analyses of controlled n-3 fatty acid intervention studies finding significant advantage over placebo for reducing depression symptom severity in MDD patients, and for secondary prevention of cardiac events in CHD patients, findings that n-3 fatty acid status is inversely correlated with other documented CHD risk factors, and patients diagnosed with MDD after ACS exhibit significantly lower n-3 fatty acid status compared with nondepressed ACS patients. This body of evidence provides strong support for future studies to evaluate the effects of increasing dietary n-3 fatty acid status on CHD comorbidity and mortality in MDD patients.

  5. Predicting and preventing the future: actively managing multiple sclerosis.

    Hutchinson, Michael

    2012-02-01

    Relapsing-remitting multiple sclerosis (MS) has a highly variable clinical course but a number of demographic, clinical and MRI features can guide the clinician in the assessment of disease activity and likely disability outcome. It is also clear that the inflammatory activity in the first five years of relapsing-remitting MS results in the neurodegenerative changes seen in secondary progressive MS 10-15 years later. While conventional first-line disease modifying therapy has an effect on relapses, about one third of patients have a suboptimal response to treatment. With the advent of highly active second-line therapies with their evident marked suppression of inflammation, the clinician now has the tools to manage the course of relapsing-remitting MS more effectively. The development of treatment optimisation recommendations based on the clinical response to first-line therapies can guide the neurologist in more active management of the early course of relapsing-remitting MS, with the aim of preventing both acute inflammatory axonal injury and the neurodegenerative process which leads to secondary progressive MS.

  6. Virulent poxviruses inhibit DNA sensing by preventing STING activation.

    Georgana, Iliana; Sumner, Rebecca P; Towers, Greg J; Maluquer de Motes, Carlos

    2018-02-28

    efficient oncolytics in virotherapy. The successful therapeutic use of VACV depends on a detailed understanding of its ability to modulate host innate immune responses. DNA sensing is a critical cellular mechanism for pathogen detection and activation of innate immunity that is centrally coordinated by the ER-resident protein STING. Here STING is shown to mediate immune activation in response to MVA, but not to virulent VACV strains or other virulent poxviruses, which prevent STING activation and DNA sensing during infection and after DNA transfection. These results provide new insights into poxvirus immune evasion and have implications in the rational design of VACV-based therapeutics. Copyright © 2018 Georgana et al.

  7. Active Gating, Molecular Pumping, and Turnover Determination in Biomimetic Lipidic Cubic Mesophases with Reconstituted Membrane Proteins.

    Speziale, Chiara; Zabara, Alexandru Florian; Drummond, Calum John; Mezzenga, Raffaele

    2017-11-28

    Understanding the mechanisms controlling molecular transport in bioinspired materials is a central topic in many branches of nanotechnology. In this work, we show that biomolecules of fundamental importance in biological processes, such as glucose, can be transported in an active, controlled, and selective manner across macroscopic lipidic cubic mesophases, by correctly reconstituting within them their corresponding membrane protein transporters, such as Staphylococcus epidermidis (GlcP Se ). Importantly, by duly exploiting the symporter properties of GlcP Se of coupled glucose/H + transport, the diffusion of glucose can further be tuned by independent physiological stimuli, such as parallel or antiparallel pH gradients, offering an important model to study molecular exchange processes in cellular machinery. We finally show that by measuring the transport properties of the lipidic mesophases with and without the GlcP Se membrane protein reconstituted within, it becomes possible to determine its intrinsic conductance. We generalize these findings to other membrane proteins from the antiporters family, such as the bacterial ClC exchanger from Escherichia coli (EcClC), providing a robust method for evaluating the turnover rate of the membrane proteins in general.

  8. Light-activated control of protein channel assembly mediated by membrane mechanics

    Miller, David M.; Findlay, Heather E.; Ces, Oscar; Templer, Richard H.; Booth, Paula J.

    2016-12-01

    Photochemical processes provide versatile triggers of chemical reactions. Here, we use a photoactivated lipid switch to modulate the folding and assembly of a protein channel within a model biological membrane. In contrast to the information rich field of water-soluble protein folding, there is only a limited understanding of the assembly of proteins that are integral to biological membranes. It is however possible to exploit the foreboding hydrophobic lipid environment and control membrane protein folding via lipid bilayer mechanics. Mechanical properties such as lipid chain lateral pressure influence the insertion and folding of proteins in membranes, with different stages of folding having contrasting sensitivities to the bilayer properties. Studies to date have relied on altering bilayer properties through lipid compositional changes made at equilibrium, and thus can only be made before or after folding. We show that light-activation of photoisomerisable di-(5-[[4-(4-butylphenyl)azo]phenoxy]pentyl)phosphate (4-Azo-5P) lipids influences the folding and assembly of the pentameric bacterial mechanosensitive channel MscL. The use of a photochemical reaction enables the bilayer properties to be altered during folding, which is unprecedented. This mechanical manipulation during folding, allows for optimisation of different stages of the component insertion, folding and assembly steps within the same lipid system. The photochemical approach offers the potential to control channel assembly when generating synthetic devices that exploit the mechanosensitive protein as a nanovalve.

  9. Effect of ozone on the performance of a hybrid ceramic membrane-biological activated carbon process.

    Guo, Jianning; Hu, Jiangyong; Tao, Yi; Zhu, Jia; Zhang, Xihui

    2014-04-01

    Two hybrid processes including ozonation-ceramic membrane-biological activated carbon (BAC) (Process A) and ceramic membrane-BAC (Process B) were compared to treat polluted raw water. The performance of hybrid processes was evaluated with the removal efficiencies of turbidity, ammonia and organic matter. The results indicated that more than 99% of particle count was removed by both hybrid processes and ozonation had no significant effect on its removal. BAC filtration greatly improved the removal of ammonia. Increasing the dissolved oxygen to 30.0 mg/L could lead to a removal of ammonia with concentrations as high as 7.80 mg/L and 8.69 mg/L for Processes A and B, respectively. The average removal efficiencies of total organic carbon and ultraviolet absorbance at 254 nm (UV254, a parameter indicating organic matter with aromatic structure) were 49% and 52% for Process A, 51% and 48% for Process B, respectively. Some organic matter was oxidized by ozone and this resulted in reduced membrane fouling and increased membrane flux by 25%-30%. However, pre-ozonation altered the components of the raw water and affected the microorganisms in the BAC, which may impact the removals of organic matter and nitrite negatively. Copyright © 2014 The Research Centre for Eco-Environmental Sciences, Chinese Academy of Sciences. Published by Elsevier B.V. All rights reserved.

  10. Delta-Opioid receptors exhibit high efficiency when activating trimeric G proteins in membrane domains

    Bouřová, Lenka; Koštrnová, Alexandra; Hejnová, Lucie; Moravcová, Zuzana; Moon, H. E.; Novotný, Jiří; Milligan, G.; Svoboda, Petr

    2003-01-01

    Roč. 85, č. 1 (2003), s. 34-49 ISSN 0022-3042 R&D Projects: GA MŠk LN00A026; GA ČR GA309/01/0255 Grant - others:Welcome Trust(GB) xx Institutional research plan: CEZ:AV0Z5011922 Keywords : membrane domains * GTPase activity * G protein coupling Subject RIV: CE - Biochemistry Impact factor: 4.825, year: 2003

  11. Auxin-activated NADH oxidase activity of soybean plasma membranes is distinct from the constitutive plasma membrane NADH oxidase and exhibits prion-like properties

    Morre, D. James; Morre, Dorothy M.; Ternes, Philipp

    2003-01-01

    The hormone-stimulated and growth-related cell surface hydroquinone (NADH) oxidase activity of etiolated hypocotyls of soybeans oscillates with a period of about 24 min or 60 times per 24-h day. Plasma membranes of soybean hypocotyls contain two such NADH oxidase activities that have been resolved by purification on concanavalin A columns. One in the apparent molecular weight range of 14-17 kDa is stimulated by the auxin herbicide 2,4-dichlorophenoxyacetic acid (2,4-D). The other is larger and unaffected by 2,4-D. The 2,4-D-stimulated activity absolutely requires 2,4-D for activity and exhibits a period length of about 24 min. Also exhibiting 24-min oscillations is the rate of cell enlargement induced by the addition of 2,4-D or the natural auxin indole-3-acetic acid (IAA). Immediately following 2,4-D or IAA addition, a very complex pattern of oscillations is frequently observed. However, after several hours a dominant 24-min period emerges at the expense of the constitutive activity. A recruitment process analogous to that exhibited by prions is postulated to explain this behavior.

  12. Inhibition of Matrix Metalloproteinase Activity Prevents Increases in Myocardial Tumor Necrosis Factor-α

    Murray, David B.; Levick, Scott P; Brower, Gregory L.; Janicki, Joseph S.

    2010-01-01

    Aim TNF-α is known to cause adverse myocardial remodeling. While we have previously shown a role for cardiac mast cells in mediating myocardial TNF-α, matrix metalloproteinases (MMP) activation of TNF-α may also be contributory. We sought to determine the relative roles of MMPs and cardiac mast cells in the activation of TNF-α in the hearts of rats subjected to chronic volume overload. Methods Interventions with the broad spectrum MMP inhibitor, GM6001, or the mast cell stabilizer, nedocromil, were performed in the rat aortocaval fistula (ACF) model of volume overload. Results Myocardial TNF-α levels were significantly increased in the ACF. This increase was prevented by MMP inhibition with GM6001 (p ≤ 0.001 vs. ACF). Conversely, myocardial TNF-α levels were increased in the ACF + nedocromil treated fistula groups (p ≤ 0.001 vs. sham). The degradation of interstitial collagen volume fraction seen in the untreated ACF group was prevented in both the GM6001 and nedocromil treated hearts. Significant increases in LV myocardial ET-1 levels also occurred in the ACF group at 3 days post-fistula. Whereas administration of GM6001 significantly attenuated this increase, mast cell stabilization with nedocromil markedly exacerbated the increase, producing ET-1 levels 6.5 fold and 2 fold greater than that in the sham-operated control and ACF group, respectively. Conclusion The efficacy of the MMP inhibitor, GM6001, to prevent increased levels of myocardial TNF-α is indicative of MMP-mediated cleavage of latent extracellular membrane bound TNF-α protein as the primary source of bioactive TNF-α in the myocardium of the volume-overload heart. PMID:20403361

  13. Silver nanoparticles incorporated into nanostructured biopolymer membranes produced by electrospinning: a study of antimicrobial activity

    Karen Segala

    2015-12-01

    Full Text Available abstract This study examines the antimicrobial activity of silver nanoparticles incorporated into nanostructured membranes made of cellulose acetate (CA and blends of chitosan/poly-(ethylene oxide, CTS/PEO and prepared by electrospinning. The formation of chemically synthesized Ag nanoparticles (AgNPs was monitored by UV-visible spectroscopy (UV-Vis and characterized by transmission electron microscopy (TEM. The size distribution of the AgNPs was measured by dynamic light scattering (DLS, with an average size of approximately 20 nm. The presence of AgNPs on the surface of electrospun nanofibers was observed by field emission electron microscopy (FEG and confirmed by TEM. The antimicrobial activity of AgNPs incorporated into nanostructured membranes made of CA and CTS/PEO electrospun nanofibers was evaluated in the presence of both Gram-positive bacteria, such as Staphylococcus aureus ATCC 29213 and Propionibacterium acnes ATCC 6919, and Gram-negative bacteria, such as Escherichia coli ATCC 25992 and Pseudomonas aeruginosa ATCC 17933. Microbiological results showed that the presence of AgNPs in CA and CTS/PEO nanostructured membranes has significant antimicrobial activity for the Gram-positive bacteria Escherichia coli and Propionibacterium acnes.

  14. Membrane-active mechanism of LFchimera against Burkholderia pseudomallei and Burkholderia thailandensis.

    Kanthawong, Sakawrat; Puknun, Aekkalak; Bolscher, Jan G M; Nazmi, Kamran; van Marle, Jan; de Soet, Johannes J; Veerman, Enno C I; Wongratanacheewin, Surasakdi; Taweechaisupapong, Suwimol

    2014-10-01

    LFchimera, a construct combining two antimicrobial domains of bovine lactoferrin, lactoferrampin265-284 and lactoferricin17-30, possesses strong bactericidal activity. As yet, no experimental evidence was presented to evaluate the mechanisms of LFchimera against Burkholderia isolates. In this study we analyzed the killing activity of LFchimera on the category B pathogen Burkholderia pseudomallei in comparison to the lesser virulent Burkholderia thailandensis often used as a model for the highly virulent B. pseudomallei. Killing kinetics showed that B. thailandensis E264 was more susceptible for LFchimera than B. pseudomallei 1026b. Interestingly the bactericidal activity of LFchimera appeared highly pH dependent; B. thailandensis killing was completely abolished at and below pH 6.4. FITC-labeled LFchimera caused a rapid accumulation within 15 min in the cytoplasm of both bacterial species. Moreover, freeze-fracture electron microscopy demonstrated extreme effects on the membrane morphology of both bacterial species within 1 h of incubation, accompanied by altered membrane permeability monitored as leakage of nucleotides. These data indicate that the mechanism of action of LFchimera is similar for both species and encompasses disruption of the plasma membrane and subsequently leakage of intracellular nucleotides leading to cell dead.

  15. Studies on the membrane-associated proteolytic activities of Escherichia coli

    Palmer, S.M.

    1986-01-01

    The membrane fraction contains three proteolytic activities which can be resolved from whole membrane detergent extracts by DEAE-cellulose chromatography. The first two eluting activities have been previously reported as protease V and protease IV. These two enzymes were further purified by gel permeation HPLC. Protease V has a M. W. of 31,000 in SDS-PAGE gels. Protease IV has a M. W. of 62,000 and exists in two distinct isoforms of pl ≅ 6.7 and 6.9. The third enzyme eluting from the DEAE-cellulose column was further purified by affinity chromatography on Benzamidine-Sepharose 6B. This enzyme, referred to herein as protease VI, is a 43 kdal protein which has not been previously characterized. Protease VI was sensitive to inhibition by the serine protease inhibitors phenylmethylsulfony fluoride (PMSF), diisopropylfluorophosphate (DFP) and p-amino-benzamidine (PAB). Additionally, this enzyme exhibited maximal activity at pH ≅ 8.0. Incubation of whole membrane preparations with [ 3 H] DFP resulted in 11 specific proteins acquiring the radioactive label, included in this group of proteins were proteases IV, V, and VI. Several of the DFP-reactive proteins were also shown to bind [ 125 I] ampicillin

  16. New separation technique. Catalytically functionated separation membrane

    Urgami, Tadashi [Kansai Univ., Osaka (Japan)

    1989-02-01

    This report introduces research examples, showing the fundamental principle of the membrane by separating the catalytically functionated separation membrane into enzyme fixing separation membrane, polymerized metal complex separation membrane and polymer catalyst separation membrane. This membrane can achieve both functions of separation and catalytic reaction simultaneously and has sufficient possibility to combine powerful functions. Enzyme fixing separation membrane is prepared by carrier combination method, bridging method or covering method and the enzyme fixing method with polymerized complex in which enzyme is controlled to prevent the activity lowering as much as possible and enzyme is fixed from an aqueous solution into polymer membrane. This membrane is applied to the continuous manufacturing of invert sugar from cane sugar and adsorption and removing of harmful substances from blood by utilizing both micro-capsuled urease and active carbon. Alginic acid-copper (II) complex membrane is used for the polymerized metal complex membrane and polystyrene sulfonate membrane is used for the polymer catalyst separation membrane. 28 refs., 4 figs., 1 tabs.

  17. Nicotinic Acid-Mediated Activation of Both Membrane and Nuclear Receptors towards Therapeutic Glucocorticoid Mimetics for Treating Multiple Sclerosis

    W. Todd Penberthy

    2009-01-01

    Full Text Available Acute attacks of multiple sclerosis (MS are most commonly treated with glucocorticoids, which can provide life-saving albeit only temporary symptomatic relief. The mechanism of action (MOA is now known to involve induction of indoleamine 2,3-dioxygenase (IDO and interleukin-10 (IL-10, where IL-10 requires subsequent heme oxygenase-1 (HMOX-1 induction. Ectopic expression studies reveal that even small changes in expression of IDO, HMOX-1, or mitochondrial superoxide dismutase (SOD2 can prevent demyelination in experimental autoimmune encephalomyelitis (EAE animal models of MS. An alternative to glucocorticoids is needed for a long-term treatment of MS. A distinctly short list of endogenous activators of both membrane G-protein-coupled receptors and nuclear peroxisome proliferating antigen receptors (PPARs demonstrably ameliorate EAE pathogenesis by MOAs resembling that of glucocorticoids. These dual activators and potential MS therapeutics include endocannabinoids and the prostaglandin 15-deoxy-Δ12,14-PGJ2. Nicotinamide profoundly ameliorates and prevents autoimmune-mediated demyelination in EAE via maintaining levels of nicotinamide adenine dinucleotide (NAD, without activating PPAR nor any G-protein-coupled receptor. By comparison, nicotinic acid provides even greater levels of NAD than nicotinamide in many tissues, while additionally activating the PPAR-dependent pathway already shown to provide relief in animal models of MS after activation of GPR109a/HM74a. Thus nicotinic acid is uniquely suited for providing therapeutic relief in MS. However nicotinic acid is unexamined in MS research. Nicotinic acid penetrates the blood brain barrier, cures pellagric dementia, has been used for over 50 years clinically without toxicity, and raises HDL concentrations to a greater degree than any pharmaceutical, thus providing unparalleled benefits against lipodystrophy. Summary analysis reveals that the expected therapeutic benefits of high-dose nicotinic

  18. Antibacterial activity of alkyl gallates is a combination of direct targeting of FtsZ and permeabilization of bacterial membranes

    Krol, Ewa; de Sousa Borges, Anabela; da Silva, Isabel; Polaquini, Carlos; Regasini, Luis; Ferreira, Henrique; Scheffers, Dirk

    2015-01-01

    Alkyl gallates are compounds with reported antibacterial activity. One of the modes of action is binding of the alkyl gallates to the bacterial membrane and interference with membrane integrity. However, alkyl gallates also cause cell elongation and disruption of cell division in the important plant

  19. Activation of stretch-activated channels and maxi-K+ channels by membrane stress of human lamina cribrosa cells.

    Irnaten, Mustapha

    2009-01-01

    The lamina cribrosa (LC) region of the optic nerve head is considered the primary site of damage in glaucomatous optic neuropathy. Resident LC cells have a profibrotic potential when exposed to cyclical stretch. However, the mechanosensitive mechanisms of these cells remain unknown. Here the authors investigated the effects of membrane stretch on cell volume change and ion channel activity and examined the associated changes in intracellular calcium ([Ca(2+)](i)).

  20. Activity of adenylate cyclase in plasma membranes of pulmonary tissue remote times following nonlethal gamma-irradiation of rats

    Slozhenkina, L.V.; Ruda, V.P.; Ushakova, T.E.; Kuzin, A.M.

    1990-01-01

    Basal and stimulated activity of adenylate cyclase (cyclizing ATP-pyrophosphate lyase, E.C. 4.6.1.1., AC) in plasma membranes of pumonary tissye was being studied during a year after fractionated irradiation of rats (2 Gyx3). Basal and hormone-stimulated activity of AC was shown to vary significantly from normal 6 and 12 months after irradiation. The exposed membranes responded differently to AC activation by isoproterenol and F -

  1. Protective Effects of Ferulic Acid on High Glucose-Induced Protein Glycation, Lipid Peroxidation, and Membrane Ion Pump Activity in Human Erythrocytes.

    Weerachat Sompong

    Full Text Available Ferulic acid (FA is the ubiquitous phytochemical phenolic derivative of cinnamic acid. Experimental studies in diabetic models demonstrate that FA possesses multiple mechanisms of action associated with anti-hyperglycemic activity. The mechanism by which FA prevents diabetes-associated vascular damages remains unknown. The aim of study was to investigate the protective effects of FA on protein glycation, lipid peroxidation, membrane ion pump activity, and phosphatidylserine exposure in high glucose-exposed human erythrocytes. Our results demonstrated that FA (10-100 μM significantly reduced the levels of glycated hemoglobin (HbA1c whereas 0.1-100 μM concentrations inhibited lipid peroxidation in erythrocytes exposed to 45 mM glucose. This was associated with increased glucose consumption. High glucose treatment also caused a significant reduction in Na+/K+-ATPase activity in the erythrocyte plasma membrane which could be reversed by FA. Furthermore, we found that FA (0.1-100 μM prevented high glucose-induced phosphatidylserine exposure. These findings provide insights into a novel mechanism of FA for the prevention of vascular dysfunction associated with diabetes.

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

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

    2017-08-15

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

  3. Startup of the Anammox Process in a Membrane Bioreactor (AnMBR) from Conventional Activated Sludge.

    Gutwiński, P; Cema, G; Ziembińska-Buczyńska, A; Surmacz-Górska, J; Osadnik, M

    2016-12-01

      In this study, a laboratory-scale anammox process in a membrane bioreactor (AnMBR) was used to startup the anaerobic ammonium oxidation (anammox) process from conventional activated sludge. Stable operation was achieved after 125 days. From that time, nitrogen load was gradually increased. After six months, the average nitrogen removal efficiency exceeded 80%. The highest obtained special anammox activity (SAA) achieved was 0.17 g (-N + -N) (g VSS × d)-1. Fluorescent in situ hybridization also proved the presence of the anammox bacteria, typically a genus of Brocadia anammoxidans and Kuenenia stuttgartiensis.

  4. Development of adsorptive membranes by confinement of activated biochar into electrospun nanofibers

    Mehrdad Taheran

    2016-11-01

    Full Text Available Adsorptive membranes have many applications in removal of contaminants, such as heavy metals and organic contaminants from water. Recently, increasing concentrations of pharmaceutically active compounds, especially antibiotics, such as chlortetracycline in water and wastewater sources has raised concerns about their potentially adverse impacts on environment and human health. In this study, a series of polyacrylonitrile (PAN/activated biochar nanofibrous membranes (NFMs with different loadings of biochar (0–2%, w/w were fabricated using electrospinning. The morphology and structure of fabricated membranes was investigated by scanning electron microscopy, Fourier transform infrared and thermogravimetric analysis. The results showed that at 1.5% of biochar loading, the surface area reached the maximum value of 12.4 m2/g and beyond this loading value, agglomeration of particles inhibited fine interaction with nanofibrous matrix. Also, the adsorption tests using chlortetracycline showed that, under environmentally relevant concentrations, the fabricated adsorptive NFMs had a potential for removal of these types of emerging contaminants from water and wastewaters.

  5. Neuronal sphingolipidoses: Membrane lipids and sphingolipid activator proteins regulate lysosomal sphingolipid catabolism.

    Sandhoff, Konrad

    2016-11-01

    Glycosphingolipids and sphingolipids of cellular plasma membranes (PMs) reach luminal intra-lysosomal vesicles (LVs) for degradation mainly by pathways of endocytosis. After a sorting and maturation process (e.g. degradation of sphingomyelin (SM) and secretion of cholesterol), sphingolipids of the LVs are digested by soluble enzymes with the help of activator (lipid binding and transfer) proteins. Inherited defects of lipid-cleaving enzymes and lipid binding and transfer proteins cause manifold and fatal, often neurodegenerative diseases. The review summarizes recent findings on the regulation of sphingolipid catabolism and cholesterol secretion from the endosomal compartment by lipid modifiers, an essential stimulation by anionic membrane lipids and an inhibition of crucial steps by cholesterol and SM. Reconstitution experiments in the presence of all proteins needed, hydrolase and activator proteins, reveal an up to 10-fold increase of ganglioside catabolism just by the incorporation of anionic lipids into the ganglioside carrying membranes, whereas an additional incorporation of cholesterol inhibits GM2 catabolism substantially. It is suggested that lipid and other low molecular modifiers affect the genotype-phenotype relationship observed in patients with lysosomal diseases. Copyright © 2016 Elsevier B.V. and Société Française de Biochimie et Biologie Moléculaire (SFBBM). All rights reserved.

  6. PF-1355, a mechanism-based myeloperoxidase inhibitor, prevents immune complex vasculitis and anti-glomerular basement membrane glomerulonephritis.

    Zheng, Wei; Warner, Roscoe; Ruggeri, Roger; Su, Chunyan; Cortes, Christian; Skoura, Athanasia; Ward, Jessica; Ahn, Kay; Kalgutkar, Amit; Sun, Dexue; Maurer, Tristan S; Bonin, Paul D; Okerberg, Carlin; Bobrowski, Walter; Kawabe, Thomas; Zhang, Yanwei; Coskran, Timothy; Bell, Sammy; Kapoor, Bhupesh; Johnson, Kent; Buckbinder, Leonard

    2015-05-01

    Small vessel vasculitis is a life-threatening condition and patients typically present with renal and pulmonary injury. Disease pathogenesis is associated with neutrophil accumulation, activation, and oxidative damage, the latter being driven in large part by myeloperoxidase (MPO), which generates hypochlorous acid among other oxidants. MPO has been associated with vasculitis, disseminated vascular inflammation typically involving pulmonary and renal microvasculature and often resulting in critical consequences. MPO contributes to vascular injury by 1) catabolizing nitric oxide, impairing vasomotor function; 2) causing oxidative damage to lipoproteins and endothelial cells, leading to atherosclerosis; and 3) stimulating formation of neutrophil extracellular traps, resulting in vessel occlusion and thrombosis. Here we report a selective 2-thiouracil mechanism-based MPO inhibitor (PF-1355 [2-(6-(2,5-dimethoxyphenyl)-4-oxo-2-thioxo-3,4-dihydropyrimidin-1(2H)-yl)acetamide) and demonstrate that MPO is a critical mediator of vasculitis in mouse disease models. A pharmacokinetic/pharmacodynamic response model of PF-1355 exposure in relation with MPO activity was derived from mouse peritonitis. The contribution of MPO activity to vasculitis was then examined in an immune complex model of pulmonary disease. Oral administration of PF-1355 reduced plasma MPO activity, vascular edema, neutrophil recruitment, and elevated circulating cytokines. In a model of anti-glomerular basement membrane disease, formerly known as Goodpasture disease, albuminuria and chronic renal dysfunction were completely suppressed by PF-1355 treatment. This study shows that MPO activity is critical in driving immune complex vasculitis and provides confidence in testing the hypothesis that MPO inhibition will provide benefit in treating human vasculitic diseases. Copyright © 2015 by The American Society for Pharmacology and Experimental Therapeutics.

  7. Antioxidant activity of membrane-fractionated coffee extracts in dependence of the storage conditions

    Mitev, D; Peshev, D; Peev, G; Peeva, L

    2016-01-01

    Present paper aims at one of the important aspects of the application of products with antioxidant activity: namely the preservation and change of their properties during the storage in different conditions, as well as their reliable characterisation. The tests of antioxidant properties were conducted with membrane-separated coffee extracts, isolated using a “Microdyn Nadir NP030P” type of commercial nanofiltration membrane (30% retention of NaCl; MWCO∼400). Prepared coffee permeates and retentates were stored 0÷10 days in cool/warm conditions, with/without air access and at different illumination conditions. The kinetics of content changes was evaluated according to Folin-Ciocalteu method of total phenolic/reducing content determination. (paper)

  8. Ceramic Membrane combined with Powdered Activated Carbon (PAC) or Coagulation for Treatment of Impaired Quality Waters

    Hamad, Juma Z.

    2013-08-29

    Ceramic membranes (CM) are robust membranes attributed with high production, long life span and stability against critical conditions. While capital costs are high, these are partially offset by lower operation and maintenance costs compared to polymeric membranes. Like any other low-pressure membrane (LPM), CM faces problems of fouling, low removal of organic matter and poor removal of trace organic compounds (TOrCs). Current pretreatment approaches that are mainly based on coagulation and adsorption can remove some organic matter but with a low removal of the biopolymers component which is responsible for fouling. Powdered activated carbon (PAC) accompanied with a LPM maintains good removal of TOrCs. However, enhanced removal of TOrCs to higher level is required. Submicron powdered activated carbon (SPAC), obtained after crushing commercial activated carbon into very fine particle, and novel activated carbon (KCU 6) which is characterized with larger pores and high surface area were employed. A pre-coating approach, which provides intimated contact between PAC and contaminants, was adopted for wastewater and (high DOC) surface water treatment. For seawater, in-line coagulation with iron III chloride was adopted. Both SPAC and KCU 6 showed good removal of biopolymers at a dose of 30 mg/L with > 85 % and 90 %, respectively. A dose of 40 mg/L of SPAC and 30 mg/L KCU 6 pre-coats were successful used in controlling membrane fouling. SPAC is suggested to remove biopolymers by physical means and adsorption while KCU 6 removed biopolymers through adsorption. Both KCU 6 and SPAC attained high removal of TOrCs whereas KCU 6 showed outstanding performance. Out of 29 TOrCs investigated, KCU 6 showed > 87 % TOrCs rejection for 28 compounds. In seawater pretreatment, transparent exopolymer particles (TEP) were found to be an important foulant. TEP promoted both reversible and irreversible fouling. TEP are highly electronegative while alumina CM is positively charged which

  9. A protein chip membrane-capture assay for botulinum neurotoxin activity

    Marconi, Severine; Ferracci, Geraldine; Berthomieu, Maelys; Kozaki, Shunji; Miquelis, Raymond; Boucraut, Jose; Seagar, Michael

    2008-01-01

    Botulinum neurotoxins A and B (BoNT/A and B) are neuromuscular blocking agents which inhibit neurotransmission by cleaving the intra-cellular presynaptic SNARE proteins SNAP-25 and VAMP2, localized respectively in plasma membrane and synaptic vesicles. These neurotoxins are both dangerous pathogens and powerful therapeutic agents with numerous clinical and cosmetic applications. Consequently there is a need for in vitro assays of their biological activity to screen for potential inhibitors and to replace the widely used in vivo mouse assay. Surface plasmon resonance (SPR) was used to measure membrane vesicle capture by antibodies against SNAP-25 and VAMP2. Substrate cleavage by BoNTs modified capture providing a method to assay toxin activity. Firstly using synaptic vesicles as a substrate, a comparison of the EC 50 s for BoNT/B obtained by SPR, ELISA or flow cytometry indicated similar sensitivity although SPR assays were more rapid. Sonication of brain or neuronal cultures generated plasma membrane fragments with accessible intra-cellular epitopes adapted to measurement of BoNT/A activity. SPR responses were proportional to antigen concentration permitting detection of as little as 4 pM SNAP-25 in crude lysates. BoNT/A activity was assayed using monoclonal antibodies that specifically recognize a SNAP-25 epitope generated by the proteolytic action of the toxin. Incubation of intact primary cultured neurons with BoNT/A yielded an EC 50 of 0.5 pM. The SPR biosensor method was sensitive enough to monitor BoNT/A and B activity in cells cultured in a 96-well format providing an alternative to experimental animals for toxicological assays

  10. Antibacterial activity of polyphenolic fraction of Kombucha against Vibrio cholerae: targeting cell membrane.

    Bhattacharya, D; Ghosh, D; Bhattacharya, S; Sarkar, S; Karmakar, P; Koley, H; Gachhui, R

    2018-02-01

    The present study was undertaken to determine the mechanism of antibacterial activity of a polyphenolic fraction, composed of mainly catechin and isorhamnetin, previously isolated from Kombucha, a 14-day fermented beverage of sugared black tea, against the enteropathogen Vibrio cholerae N16961. Bacterial growth was found to be seriously impaired by the polyphenolic fraction in a dose-dependent manner. Scanning Electron Microscopy demonstrated morphological alterations in bacterial cells when exposed to the polyphenolic fraction in a concentration-dependent manner. Permeabilization assays confirmed that the fraction disrupted bacterial membrane integrity in both time- and dose-dependent manners, which were proportional to the production of intracellular reactive oxygen species (ROS). Furthermore, each of the polyphenols catechin and isorhamnetin showed the ability to permeate bacterial cell membranes by generating oxidative stress, thereby suggesting their role in the antibacterial potential of Kombucha. Thus, the basic mechanism of antibacterial activity of the Kombucha polyphenolic fraction against V. cholerae involved bacterial membrane permeabilization and morphological changes, which might be due to the generation of intracellular ROS. To the best of our knowledge, this is the first report on the investigation of antibacterial mechanism of Kombucha, which is mostly attributed to its polyphenolic content. The emergence of multidrug-resistant Vibrio cholerae strains has hindered an efficient anti-Vibrio therapy. This study has demonstrated the membrane damage-mediated antibacterial mechanism of Kombucha, a popular fermented beverage of sugared tea, which is mostly attributed to its polyphenolic content. This study also implies the exploitation of Kombucha as a potential new source of bioactive polyphenols against V. cholerae. © 2017 The Society for Applied Microbiology.

  11. Integration of biotechnology in remediation and pollution prevention activities

    Strong-Gunderson, J.M.

    1996-01-01

    The North American Free Trade Agreement/North American Agreement on Environmental Cooperation provides a mechanism for an international collaboration between the US, Canada, and Mexico to jointly develop, modify, or refine technologies that remediate or protect the environment. These countries have a vested interest in this type of collaboration because contaminants do not respect the boundaries of a manufacturing site, region, city, state, or country. The Environmental Sciences Division (ESD) at Oak Ridge National Laboratory (ORNL) consists of a diverse group of individuals who address a variety of environmental issues. ESD is involved in basic and applied research on the fate, transport, and remediation of contaminants; environmental assessment; environmental engineering; and demonstrations of advanced remediation technologies. The remediation and protection of the environment includes water, air, and soils for organic, inorganic, and radioactive contaminants. In addition to remediating contaminated sites, research also focuses on life-cycle analyses of industrial processes and the production of green technologies. The author focuses this discussion on subsurface remediation and pollution prevention; however, the research activities encompass water, soil and air and many of the technologies are applicable to all environments. The discussion focuses on the integration of biotechnology with remediation activities and subsequently linking these biological processes to other remediation technologies

  12. Ghrelin treatment prevents development of activity based anorexia in mice.

    Legrand, Romain; Lucas, Nicolas; Breton, Jonathan; Azhar, Saïda; do Rego, Jean-Claude; Déchelotte, Pierre; Coëffier, Moïse; Fetissov, Sergueï O

    2016-06-01

    Stimulation of feeding is necessary for treatment of pathological conditions of chronic malnutrition due to anorexia. Ghrelin, a hunger hormone, is one of the candidate for pharmacological treatments of anorexia, but because of its instability in plasma has limited efficacy. We previously showed that plasmatic IgG protect ghrelin from degradation and that IgG from obese subjects and mice may increase ghrelin׳s orexigenic effect. In this study we tested if ghrelin alone or combined with IgG may improve feeding in chronically food-restricted mice with or without physical activity-based anorexia (ABA) induced by free access to a running wheel. Mice received a single daily intraperitoneal injection of ghrelin (1nM) together or not with total IgG (1nM) from obese ob/ob or lean mice before access to food during 8 days of 3h/day feeding time. We found that both ghrelin and ghrelin combined with IgG from obese, but not lean mice, prevented ABA, however, they were not able to diminish body weight loss. Physical activity was lower during the feeding period and was increased shortly after feeding in mice receiving ghrelin together with IgG from obese mice. In food-restricted mice without ABA, ghrelin treatments did not have significant effects on food intake. Thus, this study supports pharmacological use of ghrelin or ghrelin combined with IgG from obese animals for treatment of anorexia accompanied by elevated physical activity. The utility of combining ghrelin with protective IgG should be further determined in animal models of anorexia with unrestricted access to food. Copyright © 2016 Elsevier B.V. and ECNP. All rights reserved.

  13. Carbonic anhydrase activity of integral-functional complexes of thylakoid membranes of spinach chloroplasts

    A. V. Semenihin

    2015-06-01

    Full Text Available Isolated thylakoid membranes were disrupted by treatment with nonionic detergents digitonin or dodecyl maltoside. Solubilized polypeptide complexes were separated by native gel charge shift electrophoresis. The position of ATP-synthase complex and its isolated catalytic part (CF1 within gel was determined using the color reaction for ATPase activity. Due to the presence of cytochromes, the red band in unstained gels corresponded to the cytochrome b6f complex. Localization of the cytochrome b6f complex, ATP synthase and coupling CF1 in the native gel was confirmed by their subunit composition determined after SDS-electrophoretic analysis. Carbonic anhydrase (CA activity in polypeptide zones of PS II, cytochrome b6f complex, and ATP-synthase CF1 was identified in native gels using indicator bromothymol blue. CA activity of isolated CF1 in solution was determined by infrared gas analysis as the rate of bicarbonate dehydration. The water-soluble acetazolamide, an inhibitor of CA, unlike lipophilic ethoxyzolamide inhibited CA activity of CF1. Thus, it was shown for the first time that ATP-synthase has a component which is capable of catalyzing the interconversion of forms of carbonic acid associated with proton exchange. The data obtained suggest the presence of multiple forms of carbonic anhydrase in the thylakoid membranes of spinach chloroplasts and confirm their involvement in the proton transfer to the ATP synthase.

  14. Milrinone and thyroid hormone stimulate myocardial membrane Ca2+-ATPase activity and share structural homologies.

    Mylotte, K M; Cody, V; Davis, P J; Davis, F B; Blas, S D; Schoenl, M

    1985-01-01

    We have recently shown that thyroid hormone in physiological concentrations stimulates sarcolemma-enriched rabbit-myocardial-membrane Ca2+-ATPase in vitro. In this study, milrinone [2-methyl-5-cyano-(3,4'-bipyridin)-6(1H)-one], a cardiac inotropic agent, was thyromimetic in the same system. At clinically achievable concentrations (50-500 nM), milrinone significantly stimulated membrane Ca2+-ATPase in vitro. This action was antagonized by W-7 [N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide], an agent that also blocks thyroid hormone action on the Ca2+-ATPase, at concentrations as low as 5 microM. Progressive additions of milrinone to membranes incubated with a fixed concentration of thyroxine (0.10 nM) or triiodothyronine resulted in a progressive obliteration of the thyroid hormone effect on Ca2+-ATPase. Amrinone [5-amino-(3,4'-bipyridin)-6(1H)-one], the parent bipyridine of milrinone, had no effect on myocardial Ca2+-ATPase activity. X-ray crystallographic analysis of milrinone and amrinone revealed structural homologies between the phenolic ring of thyroxine and the substituted ring of milrinone, whereas amrinone did not share these homologies. The mechanism(s) of the inotropic actions of thyroxine and of milrinone is not clearly understood, but these observations implicate Ca2+-ATPase, a calcium pump-associated enzyme, as one mediator of the effects on the heart of these two compounds. PMID:2933747

  15. Polyurethane Ionophore-Based Thin Layer Membranes for Voltammetric Ion Activity Sensing.

    Cuartero, Maria; Crespo, Gaston A; Bakker, Eric

    2016-06-07

    We report on a plasticized polyurethane ionophore-based thin film material (of hundreds of nanometer thickness) for simultaneous voltammetric multianalyte ion activity detection triggered by the oxidation/reduction of an underlying poly(3-octylthiophene) film. This material provides excellent mechanical, physical, and chemical robustness compared to other polymers. Polyurethane films did not exhibit leaching of lipophilic additives after rinsing with a direct water jet and exhibited resistance to detachment from the underlying electrode surface, resulting in a voltammetric current response with less than acrylate) ionophore-based membranes of the same thickness and composition exhibited a significant deterioration of the signal after identical treatment. While previously reported works emphasized fundamental advancement of multi-ion detection with multi-ionophore-based thin films, polyurethane thin membranes allow one to achieve real world measurements without sacrificing analytical performance. Indeed, polyurethane membranes are demonstrated to be useful for the simultaneous determination of potassium and lithium in undiluted human serum and blood with attractive precision.

  16. Membrane Localization is Critical for Activation of the PICK1 BAR Domain

    Madsen, Kenneth L.; Eriksen, Jacob; Milan-Lobo, Laura; Han, Daniel S.; Niv, Masha Y.; Ammendrup-Johnsen, Ina; Henriksen, Ulla; Bhatia, Vikram K.; Stamou, Dimitrios; Sitte, Harald H.; McMahon, Harvey T.; Weinstein, Harel; Gether, Ulrik

    2013-01-01

    The PSD-95/Discs-large/ZO-1 homology (PDZ) domain protein, protein interacting with C kinase 1 (PICK1) contains a C-terminal Bin/amphiphysin/Rvs (BAR) domain mediating recognition of curved membranes; however, the molecular mechanisms controlling the activity of this domain are poorly understood. In agreement with negative regulation of the BAR domain by the N-terminal PDZ domain, PICK1 distributed evenly in the cytoplasm, whereas truncation of the PDZ domain caused BAR domain-dependent redistribution to clusters colocalizing with markers of recycling endosomal compartments. A similar clustering was observed both upon truncation of a short putative α-helical segment in the linker between the PDZ and the BAR domains and upon coexpression of PICK1 with a transmembrane PDZ ligand, including the alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor GluR2 subunit, the GluR2 C-terminus transferred to the single transmembrane protein Tac or the dopamine transporter C-terminus transferred to Tac. In contrast, transfer of the GluR2 C-terminus to cyan fluorescent protein, a cytosolic protein, did not elicit BAR domain-dependent clustering. Instead, localizing PICK1 to the membrane by introducing an N-terminal myristoylation site produced BAR domain-dependent, but ligand-independent, PICK1 clustering. The data support that in the absence of PDZ ligand, the PICK1 BAR domain is inhibited through a PDZ domain-dependent and linker-dependent mechanism. Moreover, they suggest that unmasking of the BAR domain’s membrane-binding capacity is not a consequence of ligand binding to the PDZ domain per se but results from, and coincides with, recruitment of PICK1 to a membrane compartment. PMID:18466293

  17. A comparison of BNR activated sludge systems with membrane and settling tank solid-liquid separation.

    Ramphao, M C; Wentzel, M C; Ekama, G A; Alexander, W V

    2006-01-01

    Installing membranes for solid-liquid separation into biological nutrient removal (BNR) activated sludge (AS) systems makes a profound difference not only to the design of the membrane bio-reactor (MBR) BNR system itself, but also to the design approach for the whole wastewater treatment plant (WWTP). In multi-zone BNR systems with membranes in the aerobic reactor and fixed volumes for the anaerobic, anoxic and aerobic zones (i.e. fixed volume fractions), the mass fractions can be controlled (within a range) with the inter-reactor recycle ratios. This zone mass fraction flexibility is a significant advantage of MBR BNR systems over BNR systems with secondary settling tanks (SSTs), because it allows changing the mass fractions to optimise biological N and P removal in conformity with influent wastewater characteristics and the effluent N and P concentrations required. For PWWF/ADWF ratios (fq) in the upper range (fq approximately 2.0), aerobic mass fractions in the lower range (f(maer) settling and long sludge age). However, the volume reduction compared with equivalent BNR systems with SSTs will not be large (40-60%), but the cost of the membranes can be offset against sludge thickening and stabilisation costs. Moving from a flow unbalanced raw wastewater system to a flow balanced (fq = 1) low (usually settled) wastewater strength system can double the ADWF capacity of the biological reactor, but the design approach of the WWTP changes away from extended aeration to include primary sludge stabilisation. The cost of primary sludge treatment then has to be offset against the savings of the increased WWTP capacity.

  18. Modulation by endothelin-1 of spontaneous activity and membrane currents of atrioventricular node myocytes from the rabbit heart.

    Stéphanie C Choisy

    Full Text Available The atrioventricular node (AVN is a key component of the cardiac pacemaker-conduction system. Although it is known that receptors for the peptide hormone endothelin-1 (ET-1 are expressed in the AVN, there is very little information available on the modulatory effects of ET-1 on AVN electrophysiology. This study characterises for the first time acute modulatory effects of ET-1 on AVN cellular electrophysiology.Electrophysiological experiments were conducted in which recordings were made from rabbit isolated AVN cells at 35-37°C using the whole-cell patch clamp recording technique.Application of ET-1 (10 nM to spontaneously active AVN cells led rapidly (within ~13 s to membrane potential hyperpolarisation and cessation of spontaneous action potentials (APs. This effect was prevented by pre-application of the ET(A receptor inhibitor BQ-123 (1 µM and was not mimicked by the ET(B receptor agonist IRL-1620 (300 nM. In whole-cell voltage-clamp experiments, ET-1 partially inhibited L-type calcium current (I(Ca,L and rapid delayed rectifier K(+ current (I(Kr, whilst it transiently activated the hyperpolarisation-activated current (I(f at voltages negative to the pacemaking range, and activated an inwardly rectifying current that was inhibited by both tertiapin-Q (300 nM and Ba(2+ ions (2 mM; each of these effects was sensitive to ET(A receptor inhibition. In cells exposed to tertiapin-Q, ET-1 application did not produce membrane potential hyperpolarisation or immediate cessation of spontaneous activity; instead, there was a progressive decline in AP amplitude and depolarisation of maximum diastolic potential.Acutely applied ET-1 exerts a direct modulatory effect on AVN cell electrophysiology. The dominant effect of ET-1 in this study was activation of a tertiapin-Q sensitive inwardly rectifying K(+ current via ET(A receptors, which led rapidly to cell quiescence.

  19. 75 FR 17956 - Office of Juvenile Justice and Delinquency Prevention; Agency Information Collection Activities...

    2010-04-08

    ... and Delinquency Prevention; Agency Information Collection Activities: Proposed Collection; Comments... of Justice Programs, Office of Juvenile Justice and Delinquency Prevention, will be submitting the... information, please contact Janet Chiancone, (202) 353-9258, Office of Juvenile Justice and Delinquency...

  20. Nrf2 activation prevents cadmium-induced acute liver injury

    Wu, Kai C.; Liu, Jie J.; Klaassen, Curtis D.

    2012-01-01

    Oxidative stress plays an important role in cadmium-induced liver injury. Nuclear factor erythroid 2-related factor 2 (Nrf2) is a transcription factor that up-regulates cytoprotective genes in response to oxidative stress. To investigate the role of Nrf2 in cadmium-induced hepatotoxicity, Nrf2-null mice, wild-type mice, kelch-like ECH-associated protein 1-knockdown (Keap1-KD) mice with enhanced Nrf2, and Keap1-hepatocyte knockout (Keap1-HKO) mice with maximum Nrf2 activation were treated with cadmium chloride (3.5 mg Cd/kg, i.p.). Blood and liver samples were collected 8 h thereafter. Cadmium increased serum alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) activities, and caused extensive hepatic hemorrhage and necrosis in the Nrf2-null mice. In contrast, Nrf2-enhanced mice had lower serum ALT and LDH activities and less morphological alternations in the livers than wild-type mice. H 2 DCFDA (2′,7′-dichlorodihydrofluoresein diacetate) staining of primary hepatocytes isolated from the four genotypes of mice indicated that oxidative stress was higher in Nrf2-null cells, and lower in Nrf2-enhanced cells than in wild-type cells. To further investigate the mechanism of the protective effect of Nrf2, mRNA of metallothionein (MT) and other cytoprotective genes were determined. Cadmium markedly induced MT-1 and MT-2 in livers of all four genotypes of mice. In contrast, genes involved in glutathione synthesis and reducing reactive oxygen species, including glutamate-cysteine ligase (Gclc), glutathione peroxidase-2 (Gpx2), and sulfiredoxin-1 (Srxn-1) were only induced in Nrf2-enhanced mice, but not in Nrf2-null mice. In conclusion, the present study shows that Nrf2 activation prevents cadmium-induced oxidative stress and liver injury through induction of genes involved in antioxidant defense rather than genes that scavenge Cd. -- Highlights: ► Cadmium caused extensive hepatic hemorrhage and necrosis in Nrf2-null mice. ► Keap1-KD and Keap1-HKO mice were

  1. Nrf2 activation prevents cadmium-induced acute liver injury

    Wu, Kai C. [Department of Pharmacology, Toxicology, and Therapeutics, University of Kansas Medical Center, Kansas City, KS (United States); Liu, Jie J. [Department of Internal Medicine, University of Kansas Medical Center, Kansas City, KS (United States); Klaassen, Curtis D., E-mail: cklaasse@kumc.edu [Department of Internal Medicine, University of Kansas Medical Center, Kansas City, KS (United States)

    2012-08-15

    Oxidative stress plays an important role in cadmium-induced liver injury. Nuclear factor erythroid 2-related factor 2 (Nrf2) is a transcription factor that up-regulates cytoprotective genes in response to oxidative stress. To investigate the role of Nrf2 in cadmium-induced hepatotoxicity, Nrf2-null mice, wild-type mice, kelch-like ECH-associated protein 1-knockdown (Keap1-KD) mice with enhanced Nrf2, and Keap1-hepatocyte knockout (Keap1-HKO) mice with maximum Nrf2 activation were treated with cadmium chloride (3.5 mg Cd/kg, i.p.). Blood and liver samples were collected 8 h thereafter. Cadmium increased serum alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) activities, and caused extensive hepatic hemorrhage and necrosis in the Nrf2-null mice. In contrast, Nrf2-enhanced mice had lower serum ALT and LDH activities and less morphological alternations in the livers than wild-type mice. H{sub 2}DCFDA (2′,7′-dichlorodihydrofluoresein diacetate) staining of primary hepatocytes isolated from the four genotypes of mice indicated that oxidative stress was higher in Nrf2-null cells, and lower in Nrf2-enhanced cells than in wild-type cells. To further investigate the mechanism of the protective effect of Nrf2, mRNA of metallothionein (MT) and other cytoprotective genes were determined. Cadmium markedly induced MT-1 and MT-2 in livers of all four genotypes of mice. In contrast, genes involved in glutathione synthesis and reducing reactive oxygen species, including glutamate-cysteine ligase (Gclc), glutathione peroxidase-2 (Gpx2), and sulfiredoxin-1 (Srxn-1) were only induced in Nrf2-enhanced mice, but not in Nrf2-null mice. In conclusion, the present study shows that Nrf2 activation prevents cadmium-induced oxidative stress and liver injury through induction of genes involved in antioxidant defense rather than genes that scavenge Cd. -- Highlights: ► Cadmium caused extensive hepatic hemorrhage and necrosis in Nrf2-null mice. ► Keap1-KD and Keap1-HKO mice

  2. Punica granatum (Pomegranate activity in health promotion and cancer prevention

    Shahindokht Bassiri-Jahromi

    2018-01-01

    Full Text Available Cancer has become one of the most fatal diseases in most countries. In spite of the medical care developing, cancer still remains a significant problem. The majority of the cancers are resistant to treatment. Thus, the research for novel, more efficient and less side effect treatment methods continues. Pomegranate contains strong antioxidant activity, with potential health interests. Research concern in pomegranate is increasing because of their anticancer potential due to possess rich in polyphenols. We highlight the pomegranate potential health benefits and mechanism of cancer progression inhibition. Pomegranate has indicated antiproliferative, anti-metastatic and anti-invasive effects on different cancer cell line in vitro, in vivo and clinical trial. The aim of this review is to evaluate functional properties and the medical benifits of pomegranate against various cancer diseases. In addition, pomegranate properties in in vitro and in vivo experimental human and animal clinical trials and its future use are explored. The available data suggest that Punica granatum (pomegranate might be used in the control and potential therapeutic for some disease conditions and benefits human health status. This review summarizes in vitro, in vivo and clinical trial studies highlighting the pomegranate role in prevent and treatment of breast, prostate, lung, colon, skin and hepatocellular cell cancers.

  3. Punica granatum (Pomegranate) activity in health promotion and cancer prevention

    2018-01-01

    Cancer has become one of the most fatal diseases in most countries. In spite of the medical care developing, cancer still remains a significant problem. The majority of the cancers are resistant to treatment. Thus, the research for novel, more efficient and less side effect treatment methods continues. Pomegranate contains strong antioxidant activity, with potential health interests. Research concern in pomegranate is increasing because of their anticancer potential due to possess rich in polyphenols. We highlight the pomegranate potential health benefits and mechanism of cancer progression inhibition. Pomegranate has indicated antiproliferative, anti-metastatic and anti-invasive effects on different cancer cell line in vitro, in vivo and clinical trial. The aim of this review is to evaluate functional properties and the medical benifits of pomegranate against various cancer diseases. In addition, pomegranate properties in in vitro and in vivo experimental human and animal clinical trials and its future use are explored. The available data suggest that Punica granatum (pomegranate) might be used in the control and potential therapeutic for some disease conditions and benefits human health status. This review summarizes in vitro, in vivo and clinical trial studies highlighting the pomegranate role in prevent and treatment of breast, prostate, lung, colon, skin and hepatocellular cell cancers. PMID:29441150

  4. Punica granatum (Pomegranate) activity in health promotion and cancer prevention.

    Bassiri-Jahromi, Shahindokht

    2018-01-30

    Cancer has become one of the most fatal diseases in most countries. In spite of the medical care developing, cancer still remains a significant problem. The majority of the cancers are resistant to treatment. Thus, the research for novel, more efficient and less side effect treatment methods continues. Pomegranate contains strong antioxidant activity, with potential health interests. Research concern in pomegranate is increasing because of their anticancer potential due to possess rich in polyphenols. We highlight the pomegranate potential health benefits and mechanism of cancer progression inhibition. Pomegranate has indicated antiproliferative, anti-metastatic and anti-invasive effects on different cancer cell line in vitro , in vivo and clinical trial. The aim of this review is to evaluate functional properties and the medical benifits of pomegranate against various cancer diseases. In addition, pomegranate properties in in vitro and in vivo experimental human and animal clinical trials and its future use are explored. The available data suggest that Punica granatum (pomegranate) might be used in the control and potential therapeutic for some disease conditions and benefits human health status. This review summarizes in vitro , in vivo and clinical trial studies highlighting the pomegranate role in prevent and treatment of breast, prostate, lung, colon, skin and hepatocellular cell cancers.

  5. Effects of coconut granular activated carbon pretreatment on membrane filtration in a gravitational driven process to improve drinking water quality.

    da Silva, Flávia Vieira; Yamaguchi, Natália Ueda; Lovato, Gilselaine Afonso; da Silva, Fernando Alves; Reis, Miria Hespanhol Miranda; de Amorim, Maria Teresa Pessoa Sousa; Tavares, Célia Regina Granhen; Bergamasco, Rosângela

    2012-01-01

    This study evaluates the performance of a polymeric microfiltration membrane, as well as its combination with a coconut granular activated carbon (GAC) pretreatment, in a gravitational filtration module, to improve the quality of water destined to human consumption. The proposed membrane and adsorbent were thoroughly characterized using instrumental techniques, such as contact angle, Brunauer-Emmett-Teller) and Fourier transform infrared spectroscopy analyses. The applied processes (membrane and GAC + membrane) were evaluated regarding permeate flux, fouling percentage, pH and removal of Escherichia coli, colour, turbidity and free chlorine. The obtained results for filtrations with and without GAC pretreatment were similar in terms of water quality. GAC pretreatment ensured higher chlorine removals, as well as higher initial permeate fluxes. This system, applying GAC as a pretreatment and a gravitational driven membrane filtration, could be considered as an alternative point-of-use treatment for water destined for human consumption.

  6. Antioxidant activity and haemolysis prevention efficiency of polyaniline nanofibers

    Banerjee, Somik; Kumar, A [Materials Research Laboratory, Department of Physics, Tezpur University, Tezpur 784028, Assam (India); Saikia, Jyoti P; Konwar, B K, E-mail: ask@tezu.ernet.in [Department of Molecular Biology and Biotechnology, Tezpur University, Tezpur 784028, Assam (India)

    2010-01-29

    Polyaniline (PAni) nanofibers have been synthesized by interfacial polymerization using hydrochloric acid (HCl) and camphor sulfonic acid (CSA) as dopants. The powder x-ray diffraction pattern of bulk polyaniline reveals ES I structure and has been indexed in a pseudo-orthorhombic lattice. The broadening of (110) reflection in the nanofiber samples has been analysed in terms of domain length and strain using a convolution method employing a Voigt function. The increase in d spacing for the (110) reflection in HCl-doped PAni nanofibers have been assigned to the change in structural conformation due to the increase in the tilt angle of the polymer chain, which is also evident from microRaman spectra. UV-vis spectra of the PAni nanofibers exhibit a remarkable blueshift in the absorption bands attributed to {pi}-{pi}{sup *} and {pi}-polaron band transitions indicating a reduction in particle size, which is also observed in TEM micrographs. The antioxidant activity of the polyaniline nanofiber samples has been investigated using 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging assay by employing UV-visible spectroscopy. It has also been observed that polyaniline nanofibers are able to protect the haemolysis of red blood cells (RBCs) from cytotoxic agents, namely H{sub 2}O{sub 2}. The observed enhancement in the antioxidant and haemolysis prevention activity of the PAni nanofibers as compared to bulk has been attributed to the reduction in particle size and changes in structural conformation, as evident from TEM, XRD and microRaman spectroscopy.

  7. Membrane Localization is Critical for Activation of the PICK1 BAR Domain

    Madsen, Kenneth L.; Eriksen, Jacob; Milan-Lobo, Laura; Han, Daniel S.; Niv, Masha Y.; Ammendrup-Johnsen, Ina; Henriksen, Ulla; Bhatia, Vikram K.; Stamou, Dimitrios; Sitte, Harald H.; McMahon, Harvey T.; Weinstein, Harel; Gether, Ulrik

    2008-01-01

    The PSD-95/Discs-large/ZO-1 homology (PDZ) domain protein, protein interacting with C kinase 1 (PICK1) contains a C-terminal Bin/amphiphysin/Rvs (BAR) domain mediating recognition of curved membranes; however, the molecular mechanisms controlling the activity of this domain are poorly understood. In agreement with negative regulation of the BAR domain by the N-terminal PDZ domain, PICK1 distributed evenly in the cytoplasm, whereas truncation of the PDZ domain caused BAR domain-dependent redis...

  8. Novel synthesis of highly durable and active Pt catalyst encapsulated in nitrogen containing carbon for polymer electrolyte membrane fuel cell

    Lee, Hyunjoon; Sung, Yung-Eun; Choi, Insoo; Lim, Taeho; Kwon, Oh Joong

    2017-09-01

    Novel synthesis of a Pt catalyst encapsulated in a N-containing carbon layer for use in a polymer electrolyte membrane fuel cell is described in this study. A Pt-aniline complex, formed by mixing Pt precursor and aniline monomer, was used as the source of Pt, C, and N. Heat treatment of the Pt-aniline complex with carbon black yielded 5 nm Pt nanoparticles encapsulated by a N-containing carbon layer originating from aniline carbonization. The synthesized Pt catalyst exhibited higher mass specific activity to oxygen reduction reaction than that shown by conventional Pt/C catalyst because pyridinic N with graphitic carbon in the carbon layer provided active sites for oxygen reduction reaction in addition to those provided by Pt. In single cell testing, initial performance of the synthesized catalyst was limited because the thick catalyst layer increased resistance related to mass transfer. However, it was observed that the carbon layer successfully prevented Pt nanoparticles from growing via agglomeration and Ostwald ripening under fuel cell operation, thereby improving durability. Furthermore, a mass specific performance of the synthesized catalyst higher than that of a conventional Pt/C catalyst was achieved by modifying the synthesized catalyst's layer thickness.

  9. Modelling porous active layer electrodes of proton exchange membrane fuel cells; Modelisation des couches actives d'electrodes volumiques de piles a combustible a membrane echangeuse de protons

    Bultel, Yann

    1997-07-01

    This work focusses on the modeling of mass, charge and heat transfer in the active layers of the volume electrodes of proton exchange membrane fuel cells (PEMFC). A first part describes the structure of fuel cells and the physico-chemical processes taking place at the electrodes. An analysis of the classical models encountered in the literature shows that they all assume that the electro-catalysts is uniformly distributed in a plane or in volume. In a second part, the modeling of mass and charge transport phenomena has been carried out with a numerical calculation software which uses the finite-elements method and which allows to take into consideration the discrete distribution of the catalyst in nano-particulates. The simulations show the limitations of the catalyst use because of the diffusion and ionic ohmic drop both at the electrolyte and particulates scale. In order to improve the modeling of PEMFC fuel cells, the classical models have been modified to consider these local contributions. They require only simple numerical methods, like the finite-differences one. When applied to the oxygen reduction at the cathode or to the hydrogen oxidation at the anode, these models allow to determine the kinetics parameters (exchange current densities and slopes of the Tafel lines) after correction of the active layer diffusion. A modeling of the heat transfers at the active layers scale is proposed. The model takes into account the convective heat transfers between the solid phases and the gas, the electro-osmosis water transfer, and the generation of heat by joule effect and by the electrochemical reactions. Finally, the last chapter presents a study of the reaction mechanisms in the case of porous electrodes using the impedances method. Numerical and analytical models have been developed to calculate the electrode impedances and are applied to the study of oxygen reduction and hydrogen oxidation. (J.S.)

  10. Biological Activity Alterations of Human Amniotic Membrane Pre and Post Irradiation Tissue Banking.

    Nemr, Waleed; Bashandy, A S; Araby, Eman; Khamiss, O

    Innate immunity of Human Amniotic Membrane (HAM) and its highly active secretome that rich with various types of growth factors and anti-inflammatory substances proposed it as a promising material for many medical studies and applications. This study evaluate the biological activity of cultivated HAM pre and post tissue banking process in which freeze-dried HAM was sterilized by 25 KGray (kGy) dose of γ radiation. The HAM's antimicrobial activity, viability, growth of isolated human amniotic epithelial cells (HAECs), hematopoietic stimulation of co-cultivated murine bone marrow cells (mammalian model), scaffold efficiency for fish brain building up (non-mammalian model) and self re-epithelialization after trypsin denuding treatment were examined as supposed biological activity features. Native HAM revealed viability indications and was active to kill all tested microorganisms; 6 bacterial species (3 Gram-positive and 3 Gram-negative) and Candida albicans as a pathogenic fungus. Also, HAM activity promoted colony formation of murine hematopoietic cells, Tilapia nilotica brain fragment building-up and self re-epithelialization after trypsin treatment. In contrary, radiation-based tissue banking of HAM caused HAM cellular death and consequently lacked almost all of examined biological activity features. Viable HAM was featured with biological activity than fixed HAM prepared by irradiation tissue banking.

  11. Effects of deuterated water upon specific activity of some marker enzymes for cytosol and plasmatic membrane

    Buzgariu, Wanda; Coroiu, Viorica; Moldovan, Lucia; Titescu, G.; Stefanescu, I.

    2004-01-01

    Recently, numerous studies were devoted to the effects of an increased environmental deuterium concentration on physiological characteristics of various biological systems, from monocellular organisms up to mammals. Within these preoccupations the experiments on enzyme activity and parameters are of special interest since they throw light upon the mechanisms in metabolic biochemical reactions (glycolysis, photosynthesis, transport across membranes, etc). The present work concerns the effects of heavy water upon the activity of some enzymes (dehydrogenase-LDH lactate and 5' nucleotidase) implied in different metabolic pathways, serving as functional indicators for some cellular compartments such as the cytosols and cellular membranes. Enzyme activity was determined by growing for 6 days the cells (Hep 2, CHO, fibroblasts) in deuterated culture media at different concentration levels (20%, 40%, 65% si 90%), as well as in a reaction medium deuterated at 99.96%. In case of the first experimental run the LDH activity was monitored for the three cellular lines (Hep 2, CHO, fibroblasts) for different time intervals (1 d, 3 d and 6 d). After the first 24 h of cells' exposure the activity values were similar regardless of the heavy water concentration in the medium. Exposing the cells for longer time (6 days) led to modifications of LDH activity. In contrast to the case of media with relatively moderate D 2 O content, cell growing in conditions of intense deuteration 65% and 90 % D 2 O) led to an increase of cytosolic enzyme activity of about 50%. In case of 5' nucleotidase after 6 days of cell cultivation in deuteration conditions the activity decreased to 50% and 70% from the value corresponding to normal conditions for cell growth. This diminution of the activity was characteristic for the media with 65% and 90% D 2 O. In the second experimental run the activities of dehydrogenase lactate and 5' nucleotidase from the cellular homogenate obtained from cells grown in

  12. The Balance of Fluid and Osmotic Pressures across Active Biological Membranes with Application to the Corneal Endothelium.

    Xi Cheng

    Full Text Available The movement of fluid and solutes across biological membranes facilitates the transport of nutrients for living organisms and maintains the fluid and osmotic pressures in biological systems. Understanding the pressure balances across membranes is crucial for studying fluid and electrolyte homeostasis in living systems, and is an area of active research. In this study, a set of enhanced Kedem-Katchalsky (KK equations is proposed to describe fluxes of water and solutes across biological membranes, and is applied to analyze the relationship between fluid and osmotic pressures, accounting for active transport mechanisms that propel substances against their concentration gradients and for fixed charges that alter ionic distributions in separated environments. The equilibrium analysis demonstrates that the proposed theory recovers the Donnan osmotic pressure and can predict the correct fluid pressure difference across membranes, a result which cannot be achieved by existing KK theories due to the neglect of fixed charges. The steady-state analysis on active membranes suggests a new pressure mechanism which balances the fluid pressure together with the osmotic pressure. The source of this pressure arises from active ionic fluxes and from interactions between solvent and solutes in membrane transport. We apply the proposed theory to study the transendothelial fluid pressure in the in vivo cornea, which is a crucial factor maintaining the hydration and transparency of the tissue. The results show the importance of the proposed pressure mechanism in mediating stromal fluid pressure and provide a new interpretation of the pressure modulation mechanism in the in vivo cornea.

  13. The Sur7 Protein Regulates Plasma Membrane Organization and Prevents Intracellular Cell Wall Growth in Candida albicans

    Alvarez, Francisco J.; Douglas, Lois M.; Rosebrock, Adam; Konopka, James B.

    2008-01-01

    The Candida albicans plasma membrane plays important roles in cell growth and as a target for antifungal drugs. Analysis of Ca-Sur7 showed that this four transmembrane domain protein localized to stable punctate patches, similar to the plasma membrane subdomains known as eisosomes or MCC that were discovered in S. cerevisiae. The localization of Ca-Sur7 depended on sphingolipid synthesis. In contrast to S. cerevisiae, a C. albicans sur7Δ mutant displayed defects in endocytosis and morphogenes...

  14. Multi-functional electrospun antibacterial core-shell nanofibrous membranes for prolonged prevention of post-surgical tendon adhesion and inflammation.

    Shalumon, K T; Sheu, Chialin; Chen, Chih-Hao; Chen, Shih-Heng; Jose, Gils; Kuo, Chang-Yi; Chen, Jyh-Ping

    2018-05-01

    The possibility of endowing an electrospun anti-adhesive barrier membrane with multi-functionality, such as lubrication, prevention of fibroblast attachment and anti-infection and anti-inflammation properties, is highly desirable for the management of post-surgical tendon adhesion. To this end, we fabricated core-shell nanofibrous membranes (CSNMs) with embedded silver nanoparticles (Ag NPs) in the poly(ethylene glycol) (PEG)/poly(caprolactone) (PCL) shell and hyaluronic acid (HA)/ibuprofen in the core. HA imparted a lubrication effect for smooth tendon gliding and reduced fibroblast attachment, while Ag NPs and ibuprofen functioned as anti-infection and anti-inflammation agents, respectively. CSNMs with a PEG/PCL/Ag shell (PPA) and HA core containing 0% (H/PPA), 10% (HI10/PPA), 30% (HI30/PPA) and 50% (HI50/PPA) ibuprofen were fabricated through co-axial electrospinning and assessed through microscopic, spectroscopic, thermal, mechanical and drug release analyses. Considering nutrient passage through the barrier, the microporous CSNMs exerted the same barrier effect but drastically increased the mass transfer coefficients of bovine serum albumin compared with the commercial anti-adhesive membrane SurgiWrap®. Cell attachment/focal adhesion formation of fibroblasts revealed effective reduction of initial cell attachment on the CSNM surface with minimum cytotoxicity (except HI50/PPA). The anti-bacterial effect against both Gram-negative and Gram-positive bacteria was verified to be due to the Ag NPs in the membranes. In vivo studies using H/PPA and HI30/PPA CSNMs and SurgiWrap® in a rabbit flexor tendon rupture model demonstrated the improved efficacy of HI30/PPA CSNMs in reducing inflammation and tendon adhesion formation based on gross observation, histological analysis and functional assays. We conclude that HI30/PPA CSNMs can act as a multifunctional barrier membrane to prevent peritendinous adhesion after tendon surgery. A multi-functional anti-adhesion barrier

  15. Community-based osteoporosis prevention: Physical activity in relation to bone density, fall prevention, and the effect of training programmes : The Vadstena Osteoporosis Prevention Project

    Grahn Kronhed, Ann-Charlotte

    2003-01-01

    This thesis is based on studies of the ten-year community-based intervention programme entitled, the Vadstena Osteoporosis Prevention Project (VOPP). The specific aims of the research were to describe the effects of physical activity and training programmes on bone mass and balance performance in adults, to determine whether a fall risk prevention programme could motivate personal actions among the elderly, to ascertain whether the intervention programme could reduce the incidence of forearm ...

  16. [Fundamental role of the workers' representative in preventive safety activity].

    Ossicini, A; Bindi, L; Casale, M C

    2003-01-01

    With the arrival of Legislative Decree 626/94 which brought into Italian law the EU directives on workers' health and safety at the workplace, our country has also introduced rules that make a break with the past in this area, with the creation of new professional roles. The workers' safety representative takes on a fundamentally important role in the management of prevention, safety and health for workers in their place of employment in accordance with article 19. In fact, before the introduction of this Legislative Decree, the "protection" of workers' health was essentially based on rules and regulations the application of which was left to the exclusive and direct responsibility of the relationship between the employer and doctor, leaving out any participation by the worker. Whereas in the past workers could only be considered the final receivers of instructions about the security measures to apply, with Law 626 the workers themselves became active participants in the assessment of risks at work and consequently in the implementing of all the safety and hygiene measures contributing to the reduction of risk levels. The new regulations now in force assign important tasks to the workers' safety representative; all tasks and responsibilities associated with that role are examined and discussed, especially those relating to rights to information and training, consultation and participation in the process of designing and promoting safety measures. The job of workers' representative today takes on a fundamentally important meaning and role in a self-regulating system of work safety, where he or she has a proper area responsibility, so becoming a reference point for the workers generally.

  17. Resveratrol induces membrane and DNA disruption via pro-oxidant activity against Salmonella typhimurium.

    Lee, Wonjong; Lee, Dong Gun

    2017-07-22

    Resveratrol is a flavonoid found in various plants including grapes, which has been reported to be active against various pathogenic bacteria. However, antibacterial effects and mechanisms via pro-oxidant property of resveratrol remain unknown and speculative. This research investigated antibacterial mechanism of resveratrol against a food-borne human pathogen Salmonella typhimurium, and confirmed the cell death associated oxidative damage. Resveratrol increased outer membrane permeability and membrane depolarization. It also was observed DNA injury responses such as DNA fragmentation, increasing DNA contents and cell division inhibition. Intracellular ROS accumulation, GSH depletion and significant increased malondialdehyde levels were confirmed, which indicated pro-oxidant activity of resveratrol and oxidative stress. Furthermore, the observed lethal damages were reduced by antioxidant N-acetylcysteine treatment supported the view that resveratrol-induced oxidative stress stimulated S. typhimurium cell death. In conclusion, this study expands understanding on role of pro-oxidant property and insight into previously unrecognized oxygen-dependent anti-Salmonella mechanism on resveratrol. Copyright © 2017 Elsevier Inc. All rights reserved.

  18. Activity targets for nanostructured platinum-group-metal-free catalysts in hydroxide exchange membrane fuel cells

    Setzler, Brian P.; Zhuang, Zhongbin; Wittkopf, Jarrid A.; Yan, Yushan

    2016-12-01

    Fuel cells are the zero-emission automotive power source that best preserves the advantages of gasoline automobiles: low upfront cost, long driving range and fast refuelling. To make fuel-cell cars a reality, the US Department of Energy has set a fuel cell system cost target of US$30 kW-1 in the long-term, which equates to US$2,400 per vehicle, excluding several major powertrain components (in comparison, a basic, but complete, internal combustion engine system costs approximately US$3,000). To date, most research for automotive applications has focused on proton exchange membrane fuel cells (PEMFCs), because these systems have demonstrated the highest power density. Recently, however, an alternative technology, hydroxide exchange membrane fuel cells (HEMFCs), has gained significant attention, because of the possibility to use stable platinum-group-metal-free catalysts, with inherent, long-term cost advantages. In this Perspective, we discuss the cost profile of PEMFCs and the advantages offered by HEMFCs. In particular, we discuss catalyst development needs for HEMFCs and set catalyst activity targets to achieve performance parity with state-of-the-art automotive PEMFCs. Meeting these targets requires careful optimization of nanostructures to pack high surface areas into a small volume, while maintaining high area-specific activity and favourable pore-transport properties.

  19. Latent nitrate reductase activity is associated with the plasma membrane of corn roots

    Ward, M. R.; Grimes, H. D.; Huffaker, R. C.

    1989-01-01

    Latent nitrate reductase activity (NRA) was detected in corn (Zea mays L., Golden Jubilee) root microsome fractions. Microsome-associated NRA was stimulated up to 20-fold by Triton X-100 (octylphenoxy polyethoxyethanol) whereas soluble NRA was only increased up to 1.2-fold. Microsome-associated NRA represented up to 19% of the total root NRA. Analysis of microsomal fractions by aqueous two-phase partitioning showed that the membrane-associated NRA was localized in the second upper phase (U2). Analysis with marker enzymes indicated that the U2 fraction was plasma membrane (PM). The PM-associated NRA was not removed by washing vesicles with up to 1.0 M NACl but was solubilized from the PM with 0.05% Triton X-100. In contrast, vanadate-sensitive ATPase activity was not solubilized from the PM by treatment with 0.1% Triton X-100. The results show that a protein capable of reducing nitrate is embedded in the hydrophobic region of the PM of corn roots.

  20. The Membrane Topology of ALMT1, an Aluminum-Activated Malate Transport Protein in Wheat (Triticum aestivum)

    Motoda, Hirotoshi; Sasaki, Takayuki; Kano, Yoshio; Ryan, Peter R; Delhaize, Emmanuel; Matsumoto, Hideaki; Yamamoto, Yoko

    2007-01-01

    The wheat ALMT1 gene encodes an aluminum (Al)-activated malate transport protein which confers Al-resistance. We investigated the membrane topology of this plasma-membrane localized protein with immunocytochemical techniques. Several green fluorescent protein (GFP)-fused and histidine (His)-tagged chimeras of ALMT1 were prepared based on a computer-predicted secondary structure and transiently expressed in cultured mammalian cells. Antibodies raised to polypeptide epitopes of ALMT1 were used ...

  1. Unique battery with an active membrane separator having uniform physico-chemically functionalized ion channels and a method making the same

    Gerald, II, Rex E.; Ruscic, Katarina J [Chicago, IL; Sears, Devin N [Spruce Grove, CA; Smith, Luis J [Natick, MA; Klingler, Robert J [Glenview, IL; Rathke, Jerome W [Homer Glen, IL

    2012-02-21

    The invention relates to a unique battery having an active, porous membrane and method of making the same. More specifically the invention relates to a sealed battery system having a porous, metal oxide membrane with uniform, physicochemically functionalized ion channels capable of adjustable ionic interaction. The physicochemically-active porous membrane purports dual functions: an electronic insulator (separator) and a unidirectional ion-transporter (electrolyte). The electrochemical cell membrane is activated for the transport of ions by contiguous ion coordination sites on the interior two-dimensional surfaces of the trans-membrane unidirectional pores. The membrane material is designed to have physicochemical interaction with ions. Control of the extent of the interactions between the ions and the interior pore walls of the membrane and other materials, chemicals, or structures contained within the pores provides adjustability of the ionic conductivity of the membrane.

  2. Outer Membrane Protein 25 of Brucella Activates Mitogen-Activated Protein Kinase Signal Pathway in Human Trophoblast Cells

    Jing Zhang

    2017-12-01

    Full Text Available Outer membrane protein 25 (OMP25, a virulence factor from Brucella, plays an important role in maintaining the structural stability of Brucella. Mitogen-activated protein kinase (MAPK signal pathway widely exists in eukaryotic cells. In this study, human trophoblast cell line HPT-8 and BALB/c mice were infected with Brucella abortus 2308 strain (S2308 and 2308ΔOmp25 mutant strain. The expression of cytokines and activation of MAPK signal pathway were detected. We found that the expressions of tumor necrosis factor-α, interleukin-1, and interleukin-10 (IL-10 were increased in HPT-8 cells infected with S2308 and 2308ΔOmp25 mutant. S2308 also activated p38 phosphorylation protein, extracellular-regulated protein kinases (ERK, and Jun-N-terminal kinase (JNK from MAPK signal pathway. 2308ΔOmp25 could not activate p38, ERK, and JNK branches. Immunohistochemistry experiments showed that S2308 was able to activate phosphorylation of p38 and ERK in BABL/c mice. However, 2308ΔOmp25 could weakly activate phosphorylation of p38 and ERK. These results suggest that Omp25 played an important role in the process of Brucella activation of the MAPK signal pathway.

  3. Exploring in vivo cholesterol-mediated interactions between activated EGF receptors in plasma membrane with single-molecule optical tracking

    Lin, Chien Y.; Huang, Jung Y.; Lo, Leu-Wei

    2016-01-01

    The first step in many cellular signaling processes occurs at various types of receptors in the plasma membrane. Membrane cholesterol can alter these signaling pathways of living cells. However, the process in which the interaction of activated receptors is modulated by cholesterol remains unclear. In this study, we measured single-molecule optical trajectories of epidermal growth factor receptors moving in the plasma membranes of two cancerous cell lines and one normal endothelial cell line. A stochastic model was developed and applied to identify critical information from single-molecule trajectories. We discovered that unliganded epidermal growth factor receptors may reside nearby cholesterol-riched regions of the plasma membrane and can move into these lipid domains when subjected to ligand binding. The amount of membrane cholesterol considerably affects the stability of correlated motion of activated epidermal growth factor receptors. Our results provide single-molecule evidence of membrane cholesterol in regulating signaling receptors. Because the three cell lines used for this study are quite diverse, our results may be useful to shed light on the mechanism of cholesterol-mediated interaction between activated receptors in live cells

  4. Mechanical properties and osteogenic activity of poly(l-lactide) fibrous membrane synergistically enhanced by chitosan nanofibers and polydopamine layer.

    Liu, Hua; Li, Wenling; Wen, Wei; Luo, Binghong; Liu, Mingxian; Ding, Shan; Zhou, Changren

    2017-12-01

    To synergistically improve the mechanical properties and osteogenic activity of electrospinning poly(l-lactide) (PLLA) membrane, chitosan (CS) nanofibers were firstly introduced to prepare sub-micro and nanofibers interpenetrated PLLA/CS membrane, which was further surface modified with a polydopamine (PDA) layer to obtain PLLA/CS-PDA. Surface morphology, porosity, surface area and hydrophilicity of the obtained fibrous membranes were studied in detail. As compared to pure PLLA, the significant increase in the mechanical properties of the PLLA/CS, and especially of the PLLA/CS-PDA, was confirmed by tensile testing both in dry and wet states. Cells culture results indicated that both the PLLA/CS and PLLA/CS-PDA membranes, especially the latter, were more beneficial to adhesion, spreading and proliferation, as well as up-regulating alkaline phosphate activity and calcium deposition of MC3T3-E1 cells than PLLA membrane. Results suggested there was a synergistic effect of the CS nanofibers and PDA layer on the mechanical properties and osteogenic activity of PLLA membrane. Copyright © 2017 Elsevier B.V. All rights reserved.

  5. Active fragments from pro- and antiapoptotic BCL-2 proteins have distinct membrane behavior reflecting their functional divergence.

    Yannis Guillemin

    Full Text Available BACKGROUND: The BCL-2 family of proteins includes pro- and antiapoptotic members acting by controlling the permeabilization of mitochondria. Although the association of these proteins with the outer mitochondrial membrane is crucial for their function, little is known about the characteristics of this interaction. METHODOLOGY/PRINCIPAL FINDINGS: Here, we followed a reductionist approach to clarify to what extent membrane-active regions of homologous BCL-2 family proteins contribute to their functional divergence. Using isolated mitochondria as well as model lipid Langmuir monolayers coupled with Brewster Angle Microscopy, we explored systematically and comparatively the membrane activity and membrane-peptide interactions of fragments derived from the central helical hairpin of BAX, BCL-xL and BID. The results show a connection between the differing abilities of the assayed peptide fragments to contact, insert, destabilize and porate membranes and the activity of their cognate proteins in programmed cell death. CONCLUSION/SIGNIFICANCE: BCL-2 family-derived pore-forming helices thus represent structurally analogous, but functionally dissimilar membrane domains.

  6. 76 FR 9637 - Proposed Information Collection (Veteran Suicide Prevention Online Quantitative Surveys) Activity...

    2011-02-18

    ... Collection (Veteran Suicide Prevention Online Quantitative Surveys) Activity: Comment Request AGENCY... prevention of suicide among Veterans and their families. DATES: Written comments and recommendations on the.... Abstract: VA's top priority is the prevention of Veterans suicide. It is imperative to reach these at-risk...

  7. 76 FR 27384 - Agency Information Collection Activity (Veteran Suicide Prevention Online Quantitative Surveys...

    2011-05-11

    ... Collection Activity (Veteran Suicide Prevention Online Quantitative Surveys) Under OMB Review AGENCY.... Abstract: VA's top priority is the prevention of Veterans suicide. It is imperative to reach these at-risk... families' awareness of VA's suicide prevention and mental health support services. In addition, the surveys...

  8. 77 FR 52341 - Information Collection Activity: Subpart C, Pollution Prevention and Control; Submitted for...

    2012-08-29

    ...-0001; OMB Number 1014-NEW] Information Collection Activity: Subpart C, Pollution Prevention and Control... the regulations under Subpart C, Pollution Prevention and Control. This notice also provides the... information. SUPPLEMENTARY INFORMATION: Title: 30 CFR 250, Subpart C, Pollution Prevention and Control. OMB...

  9. Contribution of NADPH Oxidase to Membrane CD38 Internalization and Activation in Coronary Arterial Myocytes

    Xu, Ming; Li, Xiao-Xue; Ritter, Joseph K.; Abais, Justine M.; Zhang, Yang; Li, Pin-Lan

    2013-01-01

    The CD38-ADP-ribosylcyclase-mediated Ca2+ signaling pathway importantly contributes to the vasomotor response in different arteries. Although there is evidence indicating that the activation of CD38-ADP-ribosylcyclase is associated with CD38 internalization, the molecular mechanism mediating CD38 internalization and consequent activation in response to a variety of physiological and pathological stimuli remains poorly understood. Recent studies have shown that CD38 may sense redox signals and is thereby activated to produce cellular response and that the NADPH oxidase isoform, NOX1, is a major resource to produce superoxide (O2 ·−) in coronary arterial myocytes (CAMs) in response to muscarinic receptor agonist, which uses CD38-ADP-ribosylcyclase signaling pathway to exert its action in these CAMs. These findings led us hypothesize that NOX1-derived O2 ·− serves in an autocrine fashion to enhance CD38 internalization, leading to redox activation of CD38-ADP-ribosylcyclase activity in mouse CAMs. To test this hypothesis, confocal microscopy, flow cytometry and a membrane protein biotinylation assay were used in the present study. We first demonstrated that CD38 internalization induced by endothelin-1 (ET-1) was inhibited by silencing of NOX1 gene, but not NOX4 gene. Correspondingly, NOX1 gene silencing abolished ET-1-induced O2 ·− production and increased CD38-ADP-ribosylcyclase activity in CAMs, while activation of NOX1 by overexpression of Rac1 or Vav2 or administration of exogenous O2 ·− significantly increased CD38 internalization in CAMs. Lastly, ET-1 was found to markedly increase membrane raft clustering as shown by increased colocalization of cholera toxin-B with CD38 and NOX1. Taken together, these results provide direct evidence that Rac1-NOX1-dependent O2 ·− production mediates CD38 internalization in CAMs, which may represent an important mechanism linking receptor activation with CD38 activity in these cells. PMID:23940720

  10. Effect of vanadate on glucose transporter (GLUT4) intrinsic activity in skeletal muscle plasma membrane giant vesicles

    Kristiansen, S; Youn, J; Richter, Erik

    1996-01-01

    of vanadate (NaVO3) on glucose transporter (GLUT4) intrinsic activity (V(max) = intrinsic activity x [GLUT4 protein]) was studied in muscle plasma membrane giant vesicles. Giant vesicles (average diameter 7.6 microns) were produced by collagenase treatment of rat skeletal muscle. The vesicles were incubated......) 55% and 60%, respectively, compared with control. The plasma membrane GLUT4 protein content was not changed in response to vanadate. It is concluded that vanadate decreased glucose transport per GLUT4 (intrinsic activity). This finding suggests that regulation of glucose transport in skeletal muscle...

  11. Analysis of long-chain fatty acid binding activity in vesicles of the outer membrane generated from Escherchia coli

    Black, P.N.

    1987-01-01

    Escherichia coli transports long-chain fatty acids across the dual membrane by a high affinity, saturable, energy-dependent process. The fadL gene codes for an outer membrane protein which appears to act specifically as a long-chain fatty acid binding protein when fatty acid utilization is blocked by mutation. In an effort to understand the function of the fadL gene product, FLP, membranes have been isolated from fadL + and fadL - strains following osmotic lysis. Following isolation, total membranes were separated into inner and outer membrane fractions and assayed for long-chain fatty acid binding activity. Outer membrane vesicles were incubated 2-5 min at 37 0 C with 3 H oleate (C/sub 18:1/), cooled to 0 0 C, and centrifuged through a Lipidex 100 column for 3 min to remove the unbound fatty acid. The level of fatty acid binding was quantitated by scintillation counting of the eluate. Outer membrane vesicles generated from a fadL + strain bind 325 pmol fatty acid/mg protein whereas vesicles generated for a mutant strain bind 175 pmol fatty acid/mg protein. These data suggest that FLP acts at least as a long-chain fatty acid binding protein on the surface of the cell

  12. Characterization of antibacterial polyethersulfone membranes using the respiration activity monitoring system (RAMOS)

    Kochan, J.; Scheidle, M.; Erkel, J. van; Bikel, M.; Büchs, J.; Wong, J.E.; Melin, T.; Wessling, M.

    2012-01-01

    Membranes with antibacterial properties were developed using surface modification of polyethersulfone ultrafiltration membranes. Three different modification strategies using polyelectrolyte layer-by-layer (LbL) technique are described. The first strategy relying on the intrinsic antibacterial

  13. Biotechnological approach for systemic delivery of membrane Receptor Activator of NF-κB Ligand (RANKL) active domain into the circulation

    Cappariello, Alfredo; Paone, Riccardo; Maurizi, Antonio; Capulli, Mattia; Rucci, Nadia; Muraca, Maurizio; Teti, Anna

    2015-01-01

    Deficiency of Receptor Activator of NF-κB Ligand (RANKL) prevents osteoclast formation causing osteopetrosis. RANKL is a membrane-bound protein cleaved into active soluble (s)RANKL by metalloproteinase 14 (MMP14). We created a bio-device that harbors primary osteoblasts, cultured on 3D hydroxyapatite scaffolds carrying immobilized MMP14 catalytic domain. Scaffolds were sealed in diffusion chambers and implanted in RANKL-deficient mice. Mice received 1 or 2 diffusion chambers, once or twice and were sacrificed after 1 or 2 months from implants. A progressive increase of body weight was observed in the implanted groups. Histological sections of tibias of non-implanted mice were negative for the osteoclast marker Tartrate-Resistant Acid Phosphatase (TRAcP), consistent with the lack of osteoclasts. In contrast, tibias excised from implanted mice showed TRAcP-positive cells in the bone marrow and on the bone surface, these latter morphologically similar to mature osteoclasts. In mice implanted with 4 diffusion chambers total, we noted the highest number and size of TRAcP-positive cells, with quantifiable eroded bone surface and significant reduction of trabecular bone volume. These data demonstrate that our bio-device delivers effective sRANKL, inducing osteoclastogenesis in RANKL-deficient mice, supporting the feasibility of an innovative experimental strategy to treat systemic cytokine deficiencies. PMID:25678116

  14. Continued Advancement of Supported Liquid Membranes for Carbon Dioxide Control in Extravehicular Activity Applications

    Wickham, David T.; Gleason, Kevin J.; Engel, Jeffrey R.; Cowley, Scott W.; Chullen, Cinda

    2015-01-01

    The Development of a new, robust, portable life support system (PLSS) is currently a high NASA priority in order to support longer and safer extravehicular activity (EVA) missions that will be necessary as space travel extends to near-Earth asteroids and eventually Mars. One of the critical PLSS functions is maintaining the carbon dioxide (CO2) concentration in the suit at acceptable levels. The Metal Oxide (MetOx) canister has a finite CO2 adsorption capacity and therefore in order to extend mission times, the unit would have to be larger and heavier, which is undesirable; therefore new CO2 control technologies must be developed. While recent work has centered on the use of alternating sorbent beds that can be regenerated during the EVA, this strategy increases the system complexity and power consumption. A simpler approach is to use a membrane that vents CO2 to space but retains oxygen(O2). A membrane has many advantages over current technology: it is a continuous system with no theoretical capacity limit, it requires no consumables, and it requires no hardware for switching beds between absorption and regeneration. Conventional gas separation membranes do not have adequate selectivity for use in the PLSS, but the required performance could be obtained with a supported liquid membrane (SLM), which consists of a microporous film filled with a liquid that selectively reacts with CO2 over oxygen (O2). In a recently completed Phase II Small Business Innovative Research project, Reaction Systems developed a new reactive liquid that has effectively zero vapor pressure, making it an ideal candidate for use in an SLM. Results obtained with the SLM in a flat sheet configuration with representative pressures of CO2, O2, and water (H2O) have shown that the CO2 permeation rate and CO2/O2 selectivity requirements have been met. In addition, the SLM vents moisture to space very effectively. The SLM has also been prepared and tested in a hollow fiber form, which will be

  15. Systematic approach to development of pressure sensors using dielectric electro-active polymer membranes

    York, A; Seelecke, S; Dunn, J

    2013-01-01

    Dielectric electro-active polymers (DEAPs) have become attractive materials for various actuation and sensing applications due to their high energy and power density, high efficiency, light weight, and fast response speed. However, commercial development has been hindered due to a variety of constraints such as reliability, non-linear behavior, cost of driving electronics, and form factor requirements. This paper presents the systematic development from laboratory concept to commercial readiness of a novel pressure sensing system using a DEAP membrane. The pressure sensing system was designed for in-line pressure measurements for low pressure applications such as health systems monitoring. A first generation sensor was designed, built and tested with a focus on the qualitative capabilities of EAP membranes as sensors. Experimental measurements were conducted that demonstrated the capability of the sensor to output a voltage signal proportional to a changing pressure. Several undesirable characteristics were observed during these initial tests such as strong hysteresis, non-linearity, very limited pressure range, and low fatigue life. A second generation prototype was then designed to remove or compensate for these undesirable characteristics. This prototype was then built and tested. The new design showed an almost complete removal of hysteretic non-linear effects and was capable of operating at 10 × the pressure range of the initial generation. This new design is the framework for a novel DEAP based pressure sensor ready for commercial applications. (paper)

  16. Novel Biosensor of Membrane Protein Proximity Based on Fluorogen Activated Proteins.

    Vasilev, Kalin V; Gallo, Eugenio; Shank, Nathaniel; Jarvik, Jonathan W

    2016-01-01

    We describe a novel biosensor system for reporting proximity between cell surface proteins in live cultured cells. The biosensor takes advantage of recently developed fluorogen-activating proteins (FAPs) that display fluorescence only when bound to otherwise-nonfluorescent fluorogen molecules. To demonstrate feasibility for the approach, two recombinant rapamycin-binding proteins were expressed as single-pass plasma membrane proteins in HeLa cells; one of the proteins (scAvd- FRB) carried an extracellular avidin tag; the other (HL1-TO1-FKBP) carried an extracellular FAP. Cells were incubated with a membrane-impermeable bivalent ligand (biotin-PEG2000-DIR) consisting of biotin joined to a dimethyl-indole red (DIR) fluorogen by a polyethylene glycol linker, thus tethering the fluorogen to the scAvd-FRB fusion protein. Addition of rapamycin, which promotes FKBP-FRB dimerization and thereby brings the FAP in close proximity to the tethered fluorogen, led to a significant increase in DIR fluorescence. We call the new proximity assay TEFLA, for tethered fluorogen assay.

  17. Active generation and propagation of Ca2+ signals within tunneling membrane nanotubes.

    Smith, Ian F; Shuai, Jianwei; Parker, Ian

    2011-04-20

    A new mechanism of cell-cell communication was recently proposed after the discovery of tunneling nanotubes (TNTs) between cells. TNTs are membrane protrusions with lengths of tens of microns and diameters of a few hundred nanometers that permit the exchange of membrane and cytoplasmic constituents between neighboring cells. TNTs have been reported to mediate intercellular Ca(2+) signaling; however, our simulations indicate that passive diffusion of Ca(2+) ions alone would be inadequate for efficient transmission between cells. Instead, we observed spontaneous and inositol trisphosphate (IP(3))-evoked Ca(2+) signals within TNTs between cultured mammalian cells, which sometimes remained localized and in other instances propagated as saltatory waves to evoke Ca(2+) signals in a connected cell. Consistent with this, immunostaining showed the presence of both endoplasmic reticulum and IP(3) receptors along the TNT. We propose that IP(3) receptors may actively propagate intercellular Ca(2+) signals along TNTs via Ca(2+)-induced Ca(2+) release, acting as amplification sites to overcome the limitations of passive diffusion in a chemical analog of electrical transmission of action potentials. Copyright © 2011 Biophysical Society. Published by Elsevier Inc. All rights reserved.

  18. Membrane composition and physiological activity of plastids from an oenothera plastome mutator-induced chloroplast mutant.

    Johnson, E M; Sears, B B

    1990-01-01

    Plastids were isolated from a plastome mutator-induced mutant (pm7) of Oenothera hookeri and were analyzed for various physiological and biochemical attributes. No photosynthetic electron transport activity was detected in the mutant plastids. This is consistent with previous ultrastructural analysis showing the absence of thylakoid membranes in the pm7 plastids and with the observation of aberrant processing and accumulation of chloroplast proteins in the mutant. In comparison to wild type, the mutant tissue lacks chlorophyll, and has significant differences in levels of four fatty acids. The analyses did not reveal any differences in carotenoid levels nor in the synthesis of several chloroplast lipids. The consequences of the altered composition of the chloroplast membrane are discussed in terms of their relation to the aberrant protein processing of the pm7 plastids. The pigment, fatty acid, and lipid measurements were also performed on two distinct nuclear genotypes (A/A and A/C) which differ in their compatibility with the plastid genome (type I) contained in these lines. In these cases, only chlorophyll concentrations differed significantly.

  19. Lipotoxicity Mediated Cell Dysfunction and Death Involves Lysosomal Membrane Permeabilization and Cathepsin L Activity

    Almaguel, Frankis G.; Liu, Jo-Wen; Pacheco, Fabio J.; De Leon, Daisy; Casiano, Carlos A.; De Leon, Marino

    2010-01-01

    Lipotoxicity, which is triggered when cells are exposed to elevated levels of free fatty acids, involves cell dysfunction and apoptosis and is emerging as an underlying factor contributing to various pathological conditions including disorders of the central nervous system and diabetes. We have shown that palmitic acid (PA)-induced lipotoxicity (PA-LTx) in nerve growth factor-differentiated PC12 (NGFDPC12) cells is linked to an augmented state of cellular oxidative stress (ASCOS) and apoptosis, and that these events are inhibited by docosahexanoic acid (DHA). The mechanisms of PA-LTx in nerve cells are not well understood, but our previous findings indicate that it involves ROS generation, mitochondrial membrane permeabilization (MMP), and caspase activation. The present study used nerve growth factor differentiated PC12 cells (NGFDPC12 cells) and found that lysosomal membrane permeabilization (LMP) is an early event during PA-induced lipotoxicity that precedes MMP and apoptosis. Cathepsin L, but not cathepsin B, is an important contributor in this process since its pharmacological inhibition significantly attenuated LMP, MMP, and apoptosis. In addition, co-treatment of NGFDPC12 cells undergoing lipotoxicity with DHA significantly reduced LMP, suggesting that DHA acts by antagonizing upstream signals leading to lysosomal dysfunction. These results suggest that LMP is a key early mediator of lipotoxicity, and underscore the value of interventions targeting upstream signals leading to LMP for the treatment of pathological conditions associated with lipotoxicity. PMID:20043885

  20. Antioxidant activity of erythrocyte membranes of rats exposed to X-radiation and injected with α-tocopheral acetate

    Tsvetkova, T.V.; Tsokur, Eh.V.

    1988-01-01

    Injection of α-tocopherol acetate to albino mongrel rats potentiates antioxidant activity (AOA) that involves water-soluble factors of the antioxidant system in erythrocyte membranes. The activation of AOA by a α-tocopherol takes place immediately after irradiation and drug injection and persists during the first 24 h following irradiation

  1. New Insight Into the Roles of Membrane Microdomains in Physiological Activities of Fungal Cells.

    Malínský, Jan; Opekarová, Miroslava

    2016-01-01

    Roč. 325, mar. (2016), s. 119-180 ISSN 1937-6448 R&D Projects: GA ČR(CZ) GA15-10641S Institutional support: RVO:68378041 Keywords : membrane microdomain * membrane structure * fungi * membrane contact sites Subject RIV: EA - Cell Biology Impact factor: 3.752, year: 2015

  2. Arabidopsis protein kinase PKS5 inhibits the plasma membrane H+ -ATPase by preventing interaction with 14-3-3 protein

    Fuglsang, Anja Thoe; Guo, Yan; Cuin, Tracey A.

    2007-01-01

    Regulation of the trans-plasma membrane pH gradient is an important part of plant responses to several hormonal and environmental cues, including auxin, blue light, and fungal elicitors. However, little is known about the signaling components that mediate this regulation. Here, we report...... that an Arabidopsis thaliana Ser/Thr protein kinase, PKS5, is a negative regulator of the plasma membrane proton pump (PM Hþ-ATPase). Loss-of-function pks5 mutant plants are more tolerant of high external pH due to extrusion of protons to the extracellular space. PKS5 phosphorylates the PM Hþ-ATPase AHA2 at a novel...

  3. Prevention

    Halken, S; Høst, A

    2001-01-01

    , breastfeeding should be encouraged for 4-6 months. In high-risk infants a documented extensively hydrolysed formula is recommended if exclusive breastfeeding is not possible for the first 4 months of life. There is no evidence for preventive dietary intervention neither during pregnancy nor lactation...... populations. These theories remain to be documented in proper, controlled and prospective studies. Breastfeeding and the late introduction of solid foods (>4 months) is associated with a reduced risk of food allergy, atopic dermatitis, and recurrent wheezing and asthma in early childhood. In all infants....... Preventive dietary restrictions after the age of 4-6 months are not scientifically documented....

  4. Fibronectin-synthesizing activity of free and membrane-bound polyribosomes from human embryonic fibroblasts and chick embryos

    Belkin, V.M.; Volodarskaya, S.M.

    1986-01-01

    The fibronectin-synthesizing activity of membrane-bound and free polyribosomes in a cell-free system was studied using immunochemical methods. It was found that fibronectin biosynthesis on membrane-bound polyribosomes from human embryonic fibroblasts accounts for 4.9% and those from 10-day-old chick embryos for 1.1% of the total amount of newly synthesized proteins, whereas on free polyribosomes it is 1.0 and 0.3%, respectively. Fibronectin monomers with a molecular weight of 220,000 were found only in the material of the cell-free system containing heavy fractions of membrane-bound polyribosomes newly synthesized in the presence of spermidine. Thus, it was shown that fibronectin is synthesized primarily on membrane-bound polyribosomes

  5. The effect of activated carbon addition on membrane bioreactor processes for wastewater treatment and reclamation - A critical review.

    Skouteris, George; Saroj, Devendra; Melidis, Paraschos; Hai, Faisal I; Ouki, Sabèha

    2015-06-01

    This review concentrates on the effect of activated carbon (AC) addition to membrane bioreactors (MBRs) treating wastewaters. Use of AC-assisted MBRs combines adsorption, biodegradation and membrane filtration. This can lead to advanced removal of recalcitrant pollutants and mitigation of membrane fouling. The relative contribution of adsorption and biodegradation to overall removal achieved by an AC-assisted MBR process can vary, and "biological AC" may not fully develop due to competition of target pollutants with bulk organics in wastewater. Thus periodic replenishment of spent AC is necessary. Sludge retention time (SRT) governs the frequency of spent AC withdrawal and addition of fresh AC, and is an important parameter that significantly influences the performance of AC-assisted MBRs. Of utmost importance is AC dosage because AC overdose may aggravate membrane fouling, increase sludge viscosity, impair mass transfer and reduce sludge dewaterability. Copyright © 2015 Elsevier Ltd. All rights reserved.

  6. Improving the performance of an aerobic membrane bioreactor (MBR) treating pharmaceutical wastewater with powdered activated carbon (PAC) addition.

    Kaya, Yasemin; Bacaksiz, A Murat; Golebatmaz, Ugur; Vergili, Ilda; Gönder, Z Beril; Yilmaz, Gulsum

    2016-04-01

    In this study, the effects of organic loading rate (OLR) and the addition of powdered activated carbon (PAC) on the performance and membrane fouling of MBR were conducted to treat real pharmaceutical process wastewater. Over 145 days of operation, the MBR system was operated at OLRs ranging from 1 to 2 kg COD m(-3) day(-1) without sludge wasting. The addition of PAC provided an improvement in the flux, despite an increase in the OLR:PAC ratio. The results demonstrated that the hybrid PAC-MBR system maintained a reduced amount of membrane fouling and steadily increased the removal performance of etodolac. PAC addition reduced the deposition of extracellular polymeric substance and organic matter on the membrane surface and resulted an increase in COD removal even at higher OLRs with low PAC addition. Membrane fouling mechanisms were investigated using combined adsorption fouling models. Modified fouling index values and normalized mass transfer coefficient values indicated that predominant fouling mechanism was cake adsorption.

  7. ROCK1-directed basement membrane positioning coordinates epithelial tissue polarity.

    Daley, William P; Gervais, Elise M; Centanni, Samuel W; Gulfo, Kathryn M; Nelson, Deirdre A; Larsen, Melinda

    2012-01-01

    The basement membrane is crucial for epithelial tissue organization and function. However, the mechanisms by which basement membrane is restricted to the basal periphery of epithelial tissues and the basement membrane-mediated signals that regulate coordinated tissue organization are not well defined. Here, we report that Rho kinase (ROCK) controls coordinated tissue organization by restricting basement membrane to the epithelial basal periphery in developing mouse submandibular salivary glands, and that ROCK inhibition results in accumulation of ectopic basement membrane throughout the epithelial compartment. ROCK-regulated restriction of PAR-1b (MARK2) localization in the outer basal epithelial cell layer is required for basement membrane positioning at the tissue periphery. PAR-1b is specifically required for basement membrane deposition, as inhibition of PAR-1b kinase activity prevents basement membrane deposition and disrupts overall tissue organization, and suppression of PAR-1b together with ROCK inhibition prevents interior accumulations of basement membrane. Conversely, ectopic overexpression of wild-type PAR-1b results in ectopic interior basement membrane deposition. Significantly, culture of salivary epithelial cells on exogenous basement membrane rescues epithelial organization in the presence of ROCK1 or PAR-1b inhibition, and this basement membrane-mediated rescue requires functional integrin β1 to maintain epithelial cell-cell adhesions. Taken together, these studies indicate that ROCK1/PAR-1b-dependent regulation of basement membrane placement is required for the coordination of tissue polarity and the elaboration of tissue structure in the developing submandibular salivary gland.

  8. An ABC-transporter and an outer membrane lipoprotein participate in posttranslational activation of type VI secretion in Pseudomonas aeruginosa

    Casabona, Maria G.; Silverman, Julie M.; Sall, Khady M.; Boyer, Frédéric; Couté, Yohann; Poirel, Jessica; Grunwald, Didier; Mougous, Joseph D.; Elsen, Sylvie; Attree, Ina

    2012-01-01

    Pseudomonas aeruginosa is capable of injecting protein toxins into other bacterial cells through one of its three type VI secretion systems (T6SS). The activity of this T6SS is tightly regulated on the posttranslational level by phosphorylation-dependent and -independent pathways. The phosphorylation-dependent pathway consists of a Thr kinase/phosphatase pair (PpkA/PppA) that acts on a forkhead domain-containing protein Fha1, and a periplasmic protein, TagR, that positively regulates PpkA. In the present work, we biochemically and functionally characterize three additional proteins of the phosphorylation-dependent regulatory cascade that controls T6S activation: TagT, TagS and TagQ. We show that similar to TagR, these proteins act upstream of the PpkA/PppA checkpoint and influence phosphorylation of Fha1 and export of Hcp1 and Tse1. Localization studies demonstrate that TagQ is an outer membrane lipoprotein and TagR is associated with the outer membrane. Consistent with their homology to lipoprotein outer membrane localization (Lol) components, TagT and TagS form a stable inner membrane complex with ATPase activity. However, we find that outer membrane association of T6SS lipoproteins TagQ and TssJ1, and TagR, is unaltered in a ΔtagTS background. Notably, we found that TagQ is indispensible for anchoring of TagR to the outer membrane fraction. As T6S-dependent fitness of P. aeruginosa requires TagT, S, R and Q, we conclude that these proteins likely participate in a trans-membrane signaling pathway that promotes H1-T6SS activity under optimal environmental conditions. PMID:22765374

  9. An ABC transporter and an outer membrane lipoprotein participate in posttranslational activation of type VI secretion in Pseudomonas aeruginosa.

    Casabona, Maria G; Silverman, Julie M; Sall, Khady M; Boyer, Frédéric; Couté, Yohann; Poirel, Jessica; Grunwald, Didier; Mougous, Joseph D; Elsen, Sylvie; Attree, Ina

    2013-02-01

    Pseudomonas aeruginosa is capable of injecting protein toxins into other bacterial cells through one of its three type VI secretion systems (T6SSs). The activity of this T6SS is tightly regulated on the posttranslational level by phosphorylation-dependent and -independent pathways. The phosphorylation-dependent pathway consists of a Threonine kinase/phosphatase pair (PpkA/PppA) that acts on a forkhead domain-containing protein, Fha1, and a periplasmic protein, TagR, that positively regulates PpkA. In the present work, we biochemically and functionally characterize three additional proteins of the phosphorylation-dependent regulatory cascade that controls T6S activation: TagT, TagS and TagQ. We show that similar to TagR, these proteins act upstream of the PpkA/PppA checkpoint and influence phosphorylation of Fha1 and, apparatus assembly and effector export. Localization studies demonstrate that TagQ is an outer membrane lipoprotein and TagR is associated with the outer membrane. Consistent with their homology to lipoprotein outer membrane localization (Lol) components, TagT and TagS form a stable inner membrane complex with ATPase activity. However, we find that outer membrane association of T6SS lipoproteins TagQ and TssJ1, and TagR, is unaltered in a ΔtagTS background. Notably, we found that TagQ is indispensible for anchoring of TagR to the outer membrane fraction. As T6S-dependent fitness of P. aeruginosa requires TagT, S, R and Q, we conclude that these proteins likely participate in a trans-membrane signalling pathway that promotes H1-T6SS activity under optimal environmental conditions. © 2012 Society for Applied Microbiology and Blackwell Publishing Ltd.

  10. Comparative study of the active cadmium efflux systems operating at the plasma membrane and tonoplast of cucumber root cells.

    Migocka, Magdalena; Papierniak, Anna; Kosatka, Ewelina; Klobus, Grazyna

    2011-10-01

    The strategies developed by plants to avoid the toxicity of cadmium (Cd) and other heavy metals involve active sequestration of metals into the apoplast and vacuoles. The protein systems excluding heavy metals from the cell cytosol localize to the plasma membrane and tonoplast and are energized either by ATP or by the electrochemical gradient generated by H(+)-ATPase or by V-ATPase and pyrophosphatase (PPase), respectively. In this work, a comparative study on the contribution of both the plasma membrane and tonoplast in the active detoxification of plant cells after treatment with Cd was performed. The studies using plants treated and untreated with Cd reveal that both, H(+)-coupled and MgATP-driven efflux of Cd across plasma membranes and tonoplast is markedly stimulated in the presence of Cd in the environment. Previous studies on plasma-membrane localized H(+)-coupled Cd efflux together with the present data demonstrating tonoplast H(+)/Cd(2+) antiport activity suggest that H(+)-coupled secondary transport of Cd displays a lower affinity for Cd when compared with Cd primary pumps driven by MgATP. In addition, it is shown that MgATP-energized Cd efflux across both membranes is significantly enhanced by cysteine, dithiothreitol, and glutathione. These results suggest that Cd is excluded from the cytosol through an energy-dependent system as a free ion as well as a complexed form. Although both membranes contribute in the active exclusion of ionized and complexed Cd from the cytosol, the overall calculation of Cd accumulation in the everted plasma membranes and vacuolar vesicles suggests that the tonoplast and vacuole have a major function in Cd efflux from the cytosol in the roots of cucumber subjected to Cd stress.

  11. Activating College Men to Prevent Sexual Violence: A Qualitative Investigation

    Christensen, M. Candace

    2015-01-01

    This study explores the experiences of male college students who participated in a theatre-based, peer-education, sexual assault prevention presentation. The program was established through the use of Pedagogy of the Oppressed and Theatre of the Oppressed, as well as multicultural feminist theory and approaches. These models emphasize subverting…

  12. Active Nutritional Science Grants | Division of Cancer Prevention

    The Division of Cancer Prevention (DCP) conducts and supports research to determine a person's risk of cancer and to find ways to reduce the risk. This knowledge is critical to making progress against cancer because risk varies over the lifespan as genetic and epigenetic changes can transform healthy tissue into invasive cancer.

  13. Preventing Crystal Agglomeration of Pharmaceutical Crystals Using Temperature Cycling and a Novel Membrane Crystallization Procedure for Seed Crystal Generation

    Elena Simone

    2018-01-01

    Full Text Available In this work, a novel membrane crystallization system was used to crystallize micro-sized seeds of piroxicam monohydrate by reverse antisolvent addition. Membrane crystallization seeds were compared with seeds produced by conventional antisolvent addition and polymorphic transformation of a fine powdered sample of piroxicam form I in water. The membrane crystallization process allowed for a consistent production of pure monohydrate crystals with narrow size distribution and without significant agglomeration. The seeds were grown in 350 g of 20:80 w/w acetone-water mixture. Different seeding loads were tested and temperature cycling was applied in order to avoid agglomeration of the growing crystals during the process. Focused beam reflectance measurement (FBRM; and particle vision and measurement (PVM were used to monitor crystal growth; nucleation and agglomeration during the seeded experiments. Furthermore; Raman spectroscopy was used to monitor solute concentration and estimate the overall yield of the process. Membrane crystallization was proved to be the most convenient and consistent method to produce seeds of highly agglomerating compounds; which can be grown via cooling crystallization and temperature cycling.

  14. Influence of fermentation liquid from waste activated sludge on anoxic/oxic- membrane bioreactor performance: Nitrogen removal, membrane fouling and microbial community.

    Han, Xiaomeng; Zhou, Zhen; Mei, Xiaojie; Ma, Yan; Xie, Zhenfang

    2018-02-01

    In order to investigate effects of waste activated sludge (WAS) fermentation liquid on anoxic/oxic- membrane bioreactor (A/O-MBR), two A/O-MBRs with and without WAS fermentation liquid addition were operated in parallel. Results show that addition of WAS fermentation liquid clearly improved denitrification efficiency without deterioration of nitrification, while severe membrane fouling occurred. WAS fermentation liquid resulted in an elevated production of proteins and humic acids in bound extracellular polymeric substance (EPS) and release of organic matter with high MW fractions in soluble microbial product (SMP) and loosely bound EPS (LB-EPS). Measurement of deposition rate and fluid structure confirmed increased fouling potential of SMP and LB-EPS. γ-Proteobacteria and Ferruginibacter, which can secrete and export EPS, were also found to be abundant in the MBR with WAS fermentation liquid. It is implied that when WAS fermentation liquid was applied, some operational steps to control membrane fouling should be employed. Copyright © 2017 Elsevier Ltd. All rights reserved.

  15. Antimicrobial properties and membrane-active mechanism of a potential α-helical antimicrobial derived from cathelicidin PMAP-36.

    Yinfeng Lv

    Full Text Available Antimicrobial peptides (AMPs, which present in the non-specific immune system of organism, are amongst the most promising candidates for the development of novel antimicrobials. The modification of naturally occurring AMPs based on their residue composition and distribution is a simple and effective strategy for optimization of known AMPs. In this study, a series of truncated and residue-substituted derivatives of antimicrobial peptide PMAP-36 were designed and synthesized. The 24-residue truncated peptide, GI24, displayed antimicrobial activity comparable to the mother peptide PMAP-36 with MICs ranging from 1 to 4 µM, which is lower than the MICs of bee venom melittin. Although GI24 displayed high antimicrobial activity, its hemolytic activity was much lower than melittin, suggesting that GI24 have optimal cell selectivity. In addition, the crucial site of GI24 was identified through single site-mutation. An amino acid with high hydrophobicity at position 23 played an important role in guaranteeing the high antimicrobial activity of GI24. Then, lipid vesicles and whole bacteria were employed to investigate the membrane-active mechanisms. Membrane-simulating experiments showed that GI24 interacted strongly with negatively charged phospholipids and weakly with zwitterionic phospholipids, which corresponded well with the data of its biological activities. Membrane permeabilization and flow cytometry provide the evidence that GI24 killed microbial cells by permeabilizing the cell membrane and damaging membrane integrity. GI24 resulted in greater cell morphological changes and visible pores on cell membrane as determined using scanning electron microscopy (SEM and transmission electron microscope (TEM. Taken together, the peptide GI24 may provide a promising antimicrobial agent for therapeutic applications against the frequently-encountered bacteria.

  16. Effluent quality of a conventional activated sludge and a membrane bioreactor system treating hospital wastewater

    Pauwels, B.; Ngwa, F.; Deconinck, S.; Verstraete, W.

    2005-01-01

    Two lab scale wastewater treatment plants treating hospital wastewater in parallel were compared in terms of performance characteristics. One plant consisted of a conventional activated sludge system (CAS) and comprised In anoxic and aerobic compartment followed by a settling tank with recycle loop. The second pilot plant was a -late membrane bioreactor (MBR). The wastewater as obtained from the hospital had a variable COD (Chemical Oxygen Demand) ranging from 250 to 2300 mg/L. Both systems were operated at a similar hydraulic residence time of 12 hours. The reference conventional activated sludge system did not meet the regulatory standard for effluent COD of 125 mg /L most of the time. Its COD removal efficiency was 88%. The plate MBR delivered an effluent with a COD value of 50 mg/L or less, and attained an efficiency of 93%. The effluent contained no suspended particles. In addition, the MBR resulted in consistent operational parameters with a flux remaining around 8 -10 L/m/sup 2/.h and a trans membrane pressure <0.1 bar without the need for backwash or chemical cleaning. The CAS and the MBR system performed equally good in terms of TAN removal and EE2 removal. The CAS system typically decreased bacterial groups for about 1 log unit, whereas the MBR decreased these groups for about 3 log units. Enterococci were decreased below the detection limit in the MBR and indicator organisms such as fecal coliforms were decreased for 1.4 log units in the CAS system compared to a 3.6 log removal in the MBR. (author)

  17. Interplay between membrane elasticity and active cytoskeleton forces regulates the aggregation dynamics of the immunological synapse

    Dharan, Nadiv; Farago, Oded

    Adhesion between a T cell and an antigen presenting cell is achieved by TCR-pMHC and LFA1-ICAM1 protein complexes. These segregate to form a special pattern, known as the immunological synapse (IS), consisting of a central quasi-circular domain of TCR-pMHC bonds surrounded by a peripheral domain of LFA1-ICAM1 complexes. Insights gained from imaging studies had led to the conclusion that the formation of the central adhesion domain in the IS is driven by active (ATP-driven) mechanisms. Recent studies, however, suggested that passive (thermodynamic) mechanisms may also play an important role in this process. Here, we present a simple physical model, taking into account the membrane-mediated thermodynamic attraction between the TCR-pMHC bonds and the effective forces that they experience due to ATP-driven actin retrograde flow and transport by dynein motor proteins. Monte Carlo simulations of the model exhibit a good spatio-temporal agreement with the experimentally observed pattern evolution of the TCR-pMHC microclusters. The agreement is lost when one of the aggregation mechanisms is "muted", which helps to identify the respective roles in the process. We conclude that actin retrograde flow drives the centripetal motion of TCR-pMHC bonds, while the membrane-mediated interactions facilitate microcluster formation and growth. In the absence of dynein motors, the system evolves into a ring-shaped pattern, which highlights the role of dynein motors in the formation of the final concentric pattern. The interplay between the passive and active mechanisms regulates the rate of the accumulation process, which in the absence of one them proceeds either too quickly or slowly.

  18. Comparative catalytic activity of PET track-etched membranes with embedded silver and gold nanotubes

    Mashentseva, Anastassiya; Borgekov, Daryn; Kislitsin, Sergey; Zdorovets, Maxim; Migunova, Anastassiya

    2015-12-01

    Irradiated by heavy ions nanoporous polyethylene terephthalate track-etched membranes (PET TeMs) after +15Kr84 ions bombardment (1.75 MeV/nucl with the ion fluency of 1 × 109 cm-2) and sequential etching was applied in this research as a template for development of composites with catalytically enriched properties. A highly ordered silver and gold nanotubes arrays were embedded in 100 nm pores of PET TeMs via electroless deposition technique at 4 °C during 1 h. All "as-prepared" composites were examined for catalytic activity using reduction of 4-nitrophenol (4-NP) to 4-aminophenol (4-AP) by sodium borohydride as a common reaction to test metallic nanostructures catalysts. The effect of temperature on the catalytic activity was investigated in range of 292-313 K and activation energy were calculated. Kapp of Ag/PET composites linearly increase with an increase of the temperature thus normal Arrhenius behavior have been seen and the activation energy was calculated to be 42.13 kJ/mol. Au/PET composites exhibit not only more powerful catalytic activity but also non-linear dependence of rate constant from temperature. Kapp increased with increasing temperature throughout the 292-308 K temperature range; the reaction had an activation energy 65.32 kJ/mol. In range 311-313 K rate constant dramatically decreased and the apparent activation energy at this temperature rang was -91.44 kJ/mol due some structural changes, i.e. agglomeration of Au nanoparticles on the surface of composite.

  19. Pressure Modulation of the Enzymatic Activity of Phospholipase A2, A Putative Membrane-Associated Pressure Sensor.

    Suladze, Saba; Cinar, Suleyman; Sperlich, Benjamin; Winter, Roland

    2015-10-07

    Phospholipases A2 (PLA2) catalyze the hydrolysis reaction of sn-2 fatty acids of membrane phospholipids and are also involved in receptor signaling and transcriptional pathways. Here, we used pressure modulation of the PLA2 activity and of the membrane's physical-chemical properties to reveal new mechanistic information about the membrane association and subsequent enzymatic reaction of PLA2. Although the effect of high hydrostatic pressure (HHP) on aqueous soluble and integral membrane proteins has been investigated to some extent, its effect on enzymatic reactions operating at the water/lipid interface has not been explored, yet. This study focuses on the effect of HHP on the structure, membrane binding and enzymatic activity of membrane-associated bee venom PLA2, covering a pressure range up to 2 kbar. To this end, high-pressure Fourier-transform infrared and high-pressure stopped-flow fluorescence spectroscopies were applied. The results show that PLA2 binding to model biomembranes is not significantly affected by pressure and occurs in at least two kinetically distinct steps. Followed by fast initial membrane association, structural reorganization of α-helical segments of PLA2 takes place at the lipid water interface. FRET-based activity measurements reveal that pressure has a marked inhibitory effect on the lipid hydrolysis rate, which decreases by 75% upon compression up to 2 kbar. Lipid hydrolysis under extreme environmental conditions, such as those encountered in the deep sea where pressures up to the kbar-level are encountered, is hence markedly affected by HHP, rendering PLA2, next to being a primary osmosensor, a good candidate for a sensitive pressure sensor in vivo.

  20. TFEB activation promotes the recruitment of lysosomal glycohydrolases β-hexosaminidase and β-galactosidase to the plasma membrane

    Magini, Alessandro [Department of Experimental Medicine and Biochemical Sciences, University of Perugia, Perugia (Italy); Department of Medical and Biological Sciences (DSMB), University of Udine, Udine (Italy); Polchi, Alice; Urbanelli, Lorena [Department of Experimental Medicine and Biochemical Sciences, University of Perugia, Perugia (Italy); Cesselli, Daniela; Beltrami, Antonio [Department of Medical and Biological Sciences (DSMB), University of Udine, Udine (Italy); Tancini, Brunella [Department of Experimental Medicine and Biochemical Sciences, University of Perugia, Perugia (Italy); Emiliani, Carla, E-mail: carla.emiliani@unipg.it [Department of Experimental Medicine and Biochemical Sciences, University of Perugia, Perugia (Italy)

    2013-10-18

    Highlights: •TFEB activation promotes the increase of Hex and Gal activities. •The increase of Hex and Gal activities is related to transcriptional regulation. •TFEB promotes the recruitment of mature Hex and Gal on cell surface. -- Abstract: Lysosomes are membrane-enclosed organelles containing acid hydrolases. They mediate a variety of physiological processes, such as cellular clearance, lipid homeostasis, energy metabolism and pathogen defence. Lysosomes can secrete their content through a process called lysosome exocytosis in which lysosomes fuse with the plasma membrane realising their content into the extracellular milieu. Lysosomal exocytosis is not only responsible for the secretion of lysosomal enzymes, but it also has a crucial role in the plasma membrane repair. Recently, it has been demonstrated that lysosome response to the physiologic signals is regulated by the transcription factor EB (TFEB). In particular, lysosomal secretion is transcriptionally regulated by TFEB which induces both the docking and fusion of lysosomes with the plasma membrane. In this work we demonstrated that TFEB nuclear translocation is accompanied by an increase of mature glycohydrolases β-hexosaminidase and β-galactosidase on cell surface. This evidence contributes to elucidate an unknown TFEB biological function leading the lysosomal glycohydrolases on plasma membrane.

  1. TFEB activation promotes the recruitment of lysosomal glycohydrolases β-hexosaminidase and β-galactosidase to the plasma membrane

    Magini, Alessandro; Polchi, Alice; Urbanelli, Lorena; Cesselli, Daniela; Beltrami, Antonio; Tancini, Brunella; Emiliani, Carla

    2013-01-01

    Highlights: •TFEB activation promotes the increase of Hex and Gal activities. •The increase of Hex and Gal activities is related to transcriptional regulation. •TFEB promotes the recruitment of mature Hex and Gal on cell surface. -- Abstract: Lysosomes are membrane-enclosed organelles containing acid hydrolases. They mediate a variety of physiological processes, such as cellular clearance, lipid homeostasis, energy metabolism and pathogen defence. Lysosomes can secrete their content through a process called lysosome exocytosis in which lysosomes fuse with the plasma membrane realising their content into the extracellular milieu. Lysosomal exocytosis is not only responsible for the secretion of lysosomal enzymes, but it also has a crucial role in the plasma membrane repair. Recently, it has been demonstrated that lysosome response to the physiologic signals is regulated by the transcription factor EB (TFEB). In particular, lysosomal secretion is transcriptionally regulated by TFEB which induces both the docking and fusion of lysosomes with the plasma membrane. In this work we demonstrated that TFEB nuclear translocation is accompanied by an increase of mature glycohydrolases β-hexosaminidase and β-galactosidase on cell surface. This evidence contributes to elucidate an unknown TFEB biological function leading the lysosomal glycohydrolases on plasma membrane

  2. Eggshell membrane hydrolyzates activate NF-κB in vitro: possible implications for in vivo efficacy

    Ruff KJ

    2015-02-01

    Full Text Available Kevin J Ruff,1 Paul L Durham,2 Austin O’Reilly,2 F Daniel Long1 1ESM Technologies, LLC, Carthage, MO, USA; 2Center for Biomedical and Life Sciences, Missouri State University, Springfield, MO, USA Purpose: Eggshell membrane (ESM has been shown to contain naturally occurring bioactive components, and biological activities such as reducing proinflammatory cytokines, liver fibrosis, and joint pain in osteoarthritis sufferers have also been reported for ESM matrix as a whole. Nuclear factor kappa-light-chain-enhancer of activated B-cells (NF-κB is a signaling protein found in the cytoplasm of nearly all human and animal cell types and is a primary regulator of immune function. The studies reported herein were designed to investigate the possible role that NF-κB activity might play in the reported biological activities of ESM. Methods: Three ESM hydrolyzates produced via fermentation, enzymatic, or chemical hydrolysis were evaluated in vitro in either human peripheral blood mononuclear cell or THP-1 (human leukemic monocyte cell cultures for NF-κB activity following 4-hour exposure. The hydrolyzates were compared with untreated control cells or cells incubated with lipopolysaccharide or ascorbic acid. The source of ESM activity was also evaluated. Results: NF-κB levels were increased above levels found in untreated cells at all three dilutions (1:100, 1:1,000, and 1:10,000 for the fermentation hydrolyzate of ESM (ESM-FH (P=0.021, P=0.020, P=0.009, respectively in peripheral blood mononuclear cells. The enzymatic hydrolyzate of ESM (ESM-EH also produced statistically significant levels of activated NF-κB at the 1:100 and 1:1,000 dilutions (P=0.004, P=0.006, respectively but fell just shy of significance at the 1:10,000 dilution (P=0.073. Similarly, ESM-FH (P=0.021, P=0.002 and ESM-EH (P=0.007, P=0.007 activated NF-κB in THP-1 cells at 1:1,000 and 1:10,000 dilutions, respectively. The chemical hydrolyzate of ESM (ESM-CH showed statistically

  3. Light- and GTP-activated hydrolysis of phosphatidylinositol bisphosphate in squid photoreceptor membranes

    Baer, K.M.; Saibil, H.R.

    1988-01-01

    Light stimulates the hydrolysis of exogenous, [ 3 H]inositol-labeled phosphatidylinositol bisphosphate (PtdInsP2) added to squid photoreceptor membranes, releasing inositol trisphosphate (InsP3). At free calcium levels of 0.05 microM or greater, hydrolysis of the labeled lipid is stimulated up to 4-fold by GTP and light together, but not separately. This activity is the biochemical counterpart of observations on intact retina showing that a rhodopsin-activated GTP-binding protein is involved in visual transduction in invertebrates, and that InsP3 release is correlated with visual excitation and adaptation. Using an in vitro assay, we investigated the calcium and GTP dependence of the phospholipase activity. At calcium concentrations between 0.1 and 0.5 microM, some hydrolysis occurs independently of GTP and light, with a light- and GTP-activated component superimposed. At 1 microM calcium there is no background activity, and hydrolysis absolutely requires both GTP and light. Ion exchange chromatography on Dowex 1 (formate form) of the water-soluble products released at 1 microM calcium reveals that the product is almost entirely InsP3. Invertebrate rhodopsin is homologous in sequence and function to vertebrate visual pigment, which modulates the concentration of cyclic GMP through the mediation of the GTP-binding protein transducin. While there is some evidence that light also modulates PtdInsP2 content in vertebrate photoreceptors, the case for its involvement in phototransduction is stronger for the invertebrate systems. The results reported here support the scheme of rhodopsin----GTP-binding protein----phospholipase C activation in invertebrate photoreceptors

  4. Correlation between protein kinase C alpha activity and membrane phase behavior.

    Micol, V; Sánchez-Piñera, P; Villalaín, J; de Godos, A; Gómez-Fernández, J C

    1999-02-01

    Lipid activation of protein kinase C alpha (PKC alpha) was studied by using a model mixture containing 1, 2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), 1, 2-dimyristoyl-sn-glycero-3-phosphoserine (DMPS), and 1, 2-dimyristoyl-sn-glycerol (1,2-DMG). This lipid mixture was physically characterized by differential scanning calorimetry (DSC), Fourier transform infrared spectroscopy (FTIR), and 31P-nuclear magnetic resonance (31P-NMR). Based on these techniques, a phase diagram was constructed by keeping a constant DMPC/DMPS molar ratio of 4:1 and changing the concentration of 1,2-DMG. This phase diagram displayed three regions and two compounds: compound 1 (C1), with 45 mol% 1,2-DMG, and compound 2 (C2), with 60 mol% 1,2-DMG. When the phase diagram was elaborated in the presence of Ca2+ and Mg2+, at concentrations similar to those used in the PKC alpha activity assay, the boundaries between the regions changed slightly and C1 had 35 mol% 1,2-DMG. The activity of PKC alpha was studied at several temperatures and at different concentrations of 1,2-DMG, with a maximum of activity reached at 30 mol% 1,2-DMG and lower values at higher concentrations. In the presence of Ca2+ and Mg2+, maximum PKC alpha activity occurred at concentrations of 1,2-DMG that were close to the boundary in the phase diagram between region 1, where compound C1 and the pure phospholipid coexisted in the gel phase, and region 2, where compounds C1 and C2 coexisted. These results suggest that the membrane structure corresponding to a mixture of 1,2-DMG/phospholipid complex and free phospholipid is better able to support the activity of PKC alpha than the 1,2-DMG/phospholipid complex alone.

  5. Preparation, characterization, biological activity, and transport study of polystyrene based calcium–barium phosphate composite membrane

    Khan, Mohammad Mujahid Ali; Rafiuddin,, E-mail: rafi_amu@rediffmail.com

    2013-10-15

    Calcium–barium phosphate (CBP) composite membrane with 25% polystyrene was prepared by co-precipitation method. Scanning electron microscopy (SEM), X-ray diffraction (XRD), Fourier transformed infrared (FTIR), and Thermogravimetric analysis (TGA) were used to characterize the membrane. The membrane was found to be crystalline in nature with consistent arrangement of particles and no indication of visible cracks. The electrical potentials measured across the composite membrane in contact with univalent electrolytes (KCl, NaCl and LiCl), have been found to increase with decrease in concentrations. Thus the membrane was found to be cation-selective. Transport properties of developed membranes may be utilized for the efficient desalination of saline water and more importantly demineralization process. The antibacterial study of this composite membrane shows good results for killing the disease causing bacteria along with waste water treatment. Highlights: • Transport properties of composite membrane are evaluated. • The composite membrane was found to be stable in all media. • TMS method is used for electrochemical characterization. • The membrane was found to be cation selective. • The order of surface charge density was found to be LiCl < NaCl < KCl.

  6. Membrane morphology is actively transformed by covalent binding of the protein Atg8 to PE-lipids.

    Roland L Knorr

    Full Text Available Autophagy is a cellular degradation pathway involving the shape transformation of lipid bilayers. During the onset of autophagy, the water-soluble protein Atg8 binds covalently to phosphatdylethanolamines (PEs in the membrane in an ubiquitin-like reaction coupled to ATP hydrolysis. We reconstituted the Atg8 conjugation system in giant and nm-sized vesicles with a minimal set of enzymes and observed that formation of Atg8-PE on giant vesicles can cause substantial tubulation of membranes even in the absence of Atg12-Atg5-Atg16. Our findings show that ubiquitin-like processes can actively change properties of lipid membranes and that membrane crowding by proteins can be dynamically regulated in cells. Furthermore we provide evidence for curvature sorting of Atg8-PE. Curvature generation and sorting are directly linked to organelle shapes and, thus, to biological function. Our results suggest that a positive feedback exists between the ubiquitin-like reaction and the membrane curvature, which is important for dynamic shape changes of cell membranes, such as those involved in the formation of autophagosomes.

  7. Phenylarsine Oxide Inhibits the Fusicoccin-Induced Activation of Plasma Membrane H+-ATPase1

    Olivari, Claudio; Albumi, Cristina; Pugliarello, Maria Chiara; De Michelis, Maria Ida

    2000-01-01

    To investigate the mechanism by which fusicoccin (FC) induces the activation of the plasma membrane (PM) H+-ATPase, we used phenylarsine oxide (PAO), a known inhibitor of protein tyrosine-phosphatases. PAO was supplied in vivo in the absence or presence of FC to radish (Raphanus sativus L.) seedlings and cultured Arabidopsis cells prior to PM extraction. Treatment with PAO alone caused a slight decrease of PM H+-ATPase activity and, in radish, a decrease of PM-associated 14-3-3 proteins. When supplied prior to FC, PAO drastically inhibited FC-induced activation of PM H+-ATPase, FC binding to the PM, and the FC-induced increase of the amount of 14-3-3 associated with the PM. On the contrary, PAO was completely ineffective on all of the above-mentioned parameters when supplied after FC. The H+-ATPase isolated from PAO-treated Arabidopsis cells maintained the ability to respond to FC if supplied with exogenous, nonphosphorylated 14-3-3 proteins. Altogether, these results are consistent with a model in which the dephosphorylated state of tyrosine residues of a protein(s), such as 14-3-3 protein, is required to permit FC-induced association between the 14-3-3 protein and the PM H+-ATPase. PMID:10677439

  8. LL-37-derived membrane-active FK-13 analogs possessing cell selectivity, anti-biofilm activity and synergy with chloramphenicol and anti-inflammatory activity.

    Rajasekaran, Ganesan; Kim, Eun Young; Shin, Song Yub

    2017-05-01

    Although the human-derived antimicrobial peptide (AMP) LL-37 has potent antimicrobial and anti-inflammatory activities, its therapeutic application is limited by its low cell selectivity and high production cost due to its large size. To overcome these problems, we tried to develop novel LL-37-derived short α-helical AMPs with improved cell selectivity and without a significant loss of anti-inflammatory activity relative to that of parental LL-37. Using amino acid substitution, we designed and synthesized a series of FK13 analogs based on the sequence of the 13-meric short FK13 peptide (residues 17-29 of LL-37) that has been identified as the region responsible for the antimicrobial activity of LL-37. Among the designed FK13 analogs, FK-13-a1 and FK-13-a7 showed high cell selectivity and retained the anti-inflammatory activity. The therapeutic index (a measure of cell selectivity) of FK-13-a1 and FK-13-a7 was 6.3- and 2.3-fold that of parental LL-37, respectively. Furthermore, FK-13-a1 and FK-13-a7 displayed more potent antimicrobial activity against antibiotic-resistant bacteria including MRSA, MDRPA, and VREF, than did LL-37. In addition, FK-13-a1 and FK-13-a7 exhibited greater synergistic effects with chloramphenicol against MRSA and MDRPA and were more effective anti-biofilm agents against MDRPA than LL-37 was. Moreover, FK-13-a1 and FK-13-a7 maintained their activities in the presence of physiological salts and human serum. SYTOX green uptake, membrane depolarization and killing kinetics revealed that FK13-a1 and FK13-a7 kills microbial cells by permeabilizing the cell membrane and damaging membrane integrity. Taken together, our results suggest that FK13-a1 and FK13-a7 can be developed as novel antimicrobial/anti-inflammatory agents. Copyright © 2017 Elsevier B.V. All rights reserved.

  9. Demonstration on endurance of ion exchange membrane immersed in high-concentration tritiated water under the Broader Approach Activities

    Iwai, Yasunori, E-mail: iwai.yasunori@jaea.go.jp; Sato, Katsumi; Kawamura, Yoshinori; Yamanishi, Toshihiko

    2013-10-15

    Highlights: • Endurance of Nafion ion exchange membrane immersed in 1.38 × 10{sup 12} Bq/kg of highly concentrated tritiated water was demonstrated. • Degradation of Nafion backbone structure by tritium beta was similar to that by gamma rays and electron beams at an equivalent dose. • Degradation directly by radiation was dominant at room temperature compared with that by reactions with radicals produced from water radiolysis. -- Abstract: The Nafion{sup ®} ion exchange membrane is a key material for electrolysis cells of the water detritiation system. Endurance of Nafion ion exchange membrane immersed in 1.38 × 10{sup 12} Bq/kg of highly concentrated tritiated water has been demonstrated at room temperature for up to 2 years under the Broader Approach Activities. The curves of percent elongation at break vs. dose and tensile strength vs. dose for the Nafion membranes immersed in tritiated water were well consistent with those for Nafion membranes irradiated to an equivalent dose with gamma rays and electron beams. This shows that the degradation of Nafion backbone structure by tritium beta is similar to that by gamma rays and electron beams. The results of ferric Fenton test indicated that the degradation directly by radiation was dominant at room temperature compared with that by reactions with radicals produced from water radiolysis. The curve of ion exchange capacity vs. dose for the Nafion membranes immersed in tritiated water was also well consistent with that for Nafion membranes irradiated to an equivalent dose with gamma rays and electron beams. These results showed irradiation tests with gamma rays and electron beams were alternative for predicting degradation of ion exchange membrane by tritium beta.

  10. Development of Novel active transport membrane devices. Phase I. Final report, 31 October 1988--31 January 1994

    Laciak, D.V.; Quinn, R.; Choe, G.S.; Cook, P.J.; Tsai, Fu-Jya

    1994-08-01

    The main objective of this program was to identify and develop a technique for fabricating Active Transport Materials (ATM) into lab-scale membrane devices. Air Products met this objective by applying thin film, multilayer fabrication techniques to support the AT material on a substrate membrane. In Phase IA, spiral-wound hollow fiber membrane modules were fabricated and evaluated. These nonoptimized devices were used to demonstrate the AT-based separation of carbon dioxide from methane, hydrogen sulfide from methane, and ammonia from hydrogen. It was determined that a need exists for a more cost efficient and less energy intensive process for upgrading subquality natural gas. Air Products estimated the effectiveness of ATM for this application and concluded that an optimized ATM system could compete effectively with both conventional acid gas scrubbing technology and current membrane technology. In addition, the optimized ATM system would have lower methane loss and consume less energy than current alternative processes. Air Products made significant progress toward the ultimate goal of commercializing an advanced membrane for upgrading subquality natural gas. The laboratory program focused on developing a high performance hollow fiber substrate and fabricating and evaluating ATM-coated lab-scale hollow fiber membrane modules. Selection criteria for hollow fiber composite membrane supports were developed and used to evaluate candidate polymer compositions. A poly(amide-imide), PAI, was identified for further study. Conditions were identified which produced microporous PAI support membrane with tunable surface porosity in the range 100-1000{Angstrom}. The support fibers exhibited good hydrocarbon resistance and acceptable tensile strength though a higher elongation may ultimately be desirable. ATM materials were coated onto commercial and PAI substrate fiber. Modules containing 1-50 fibers were evaluated for permselectivity, pressure stability, and lifetime.

  11. The cytotoxic activity of miltefosine against Leishmania and macrophages is associated with dynamic changes in plasma membrane proteins.

    Fernandes, Kelly Souza; de Souza, Paulo Eduardo Narcizo; Dorta, Miriam Leandro; Alonso, Antonio

    2017-01-01

    In this study, we combined electron paramagnetic resonance (EPR) spectroscopy with an analysis of biophysical cellular parameters to study the mechanisms underlying the in vitro anti-leishmanial activity of miltefosine (MT). A thiol-specific spin label attached to membrane-bound proteins of Leishmania amazonensis and peritoneal macrophages indicated that MT may bind to plasma membrane proteins in large quantities via a detergent-like action and cause structural changes associated with a marked increase in dynamics and exposure to an aqueous environment. EPR spectra of a spin-labeled stearic acid indicated strong interactions between the probe and membrane proteins and a marked increase in the membrane fluidity of MT-treated cells. The cytotoxicity of MT was found to depend on the cell concentration used in the assay. This dependence was described by an equation involving the 50% inhibitory concentrations of MT in the aqueous medium (c w50 ) and the cell membrane (c m50 ) and the membrane-aqueous medium partition coefficient of MT (K). With a c w50 of 8.7μM, macrophages were less sensitive to MT than amastigotes and promastigotes of Leishmania, which had c w50 values of 2.4-3.1μM. The estimated c m50 of MT for Leishmania was 1.8M, which appears sufficient to cause ruptures or formation of pores in the plasma membrane. Additionally, we demonstrated that the changes in the plasma membrane detected by EPR spectroscopy occurred at cytotoxic concentrations of MT, as assessed through in vitro assays. Copyright © 2016 Elsevier B.V. All rights reserved.

  12. Ultrasonic irradiation to modify the functionalized bionanocomposite in sulfonated polybenzimidazole membrane for fuel cells applications and antibacterial activity.

    Esmaeilzade, Banafshe; Esmaielzadeh, Sheida; Ahmadizadegan, Hashem

    2018-04-01

    In this article the new proton exchange membranes were prepared from sulfonated polybenzimidazole (s-PBI) and various amounts of sulfonated titania/cellulose nanohybrids (titania/cellulose-SO 3 H) via ultrasonic waves. The ultrasonic irradiation effectively changes the rheology and the glass transition temperature and the crystallinity of the composite polymer. Ultrasonic irradiation has a very strong mixing and dispersion effect, much stronger than conventional stirring, which can improve the dispersion of titania/cellulose-SO 3 H nanoparticles in the polymer matrix. The strong -SO 3 H/-SO 3 H interaction between s-PBI chains and titania/cellulose-SO 3 H hybrids leads to ionic cross-linking in the membrane structure, which increases both the thermal stability and methanol resistance of the membranes. After acid doping with phosphoric acid, s-PBI/titania/cellulose-SO 3 H nanocomposite membranes exhibit depressions on methanol permeability and enhancements on proton conductivity comparing to the pristine s-PBI membrane. The chemical structure of the functionlized titania was characterized with FTIR, and energy-dispersive X-ray. Imidazole and sulfonated groups on the surface of modified nanoparticles forming linkages with s-PBI chains, improved the compatibility between s-PBI and nanoparticles, and enhanced the mechanical strength of the prepared nanocomposite membranes. From SEM and TEM analysis could explain the homogeneous dispersion of titania/cellulose-SO 3 H in nanocomposite membranes. Moreover, the membranes exhibited excellent antibacterial activities against S. aureus and E. coli. A. Copyright © 2017 Elsevier B.V. All rights reserved.

  13. Role of Membrane Biophysics in Alzheimer's - related cell pathways

    Donghui eZhu

    2015-05-01

    Full Text Available Cellular membrane alterations are commonly observed in many diseases, including Alzheimer’s disease (AD. Membrane biophysical properties, such as membrane molecular order, membrane fluidity, organization of lipid rafts, and adhesion between membrane and cytoskeleton, play an important role in various cellular activities and functions. While membrane biophysics impacts a broad range of cellular pathways, this review addresses the role of membrane biophysics in amyloid-β peptide aggregation, Aβ-induced oxidative pathways, amyloid precursor protein processing, and cerebral endothelial functions in AD. Understanding the mechanism(s underlying the effects of cell membrane properties on cellular processes should shed light on the development of new preventive and therapeutic strategies for this devastating disease.

  14. Progesterone receptor membrane component 1 as the mediator of the inhibitory effect of progestins on cytokine-induced matrix metalloproteinase 9 activity in vitro.

    Allen, Terrence K; Feng, Liping; Grotegut, Chad A; Murtha, Amy P

    2014-02-01

    Progesterone (P4) and the progestin, 17α-hydroxyprogesterone caproate, are clinically used to prevent preterm births (PTBs); however, their mechanism of action remains unclear. Cytokine-induced matrix metalloproteinase 9 (MMP-9) activity plays a key role in preterm premature rupture of the membranes and PTB. We demonstrated that the primary chorion cells and the HTR8/SVneo cells (cytotrophoblast cell line) do not express the classical progesterone receptor (PGR) but instead a novel progesterone receptor, progesterone receptor membrane component 1 (PGRMC1), whose role remains unclear. Using HTR8/SVneo cells in culture, we further demonstrated that 6 hours pretreatment with medroxyprogesterone acetate (MPA) and dexamethasone (Dex) but not P4 or 17α-hydroxyprogesterone hexanoate significantly attenuated tumor necrosis factor α-induced MMP-9 activity after a 24-hour incubation period. The inhibitory effect of MPA, but not Dex, was attenuated when PGRMC1 expression was successfully reduced by PGRMC1 small interfering RNA. Our findings highlight a possible novel role of PGRMC1 in mediating the effects of MPA and in modulating cytokine-induced MMP-9 activity in cytotrophoblast cells in vitro.

  15. Scintigraphic evaluation of the osteoblastic activity of rabbit tibial defects after HYAFF11 membrane application.

    Mermerkaya, Musa Uğur; Doral, Mahmut Nedim; Karaaslan, Fatih; Huri, Gazi; Karacavuş, Seyhan; Kaymaz, Burak; Alkan, Erkan

    2016-05-03

    An unfavorable condition for bone healing is the presence of bone defects. Under such conditions, a material can play a role to cover fractured or defective bone. Technological advances now allow for the use of such material. Hyalonect(®) (Fidia Advanced Biopolymers SLR, Italy), a novel membrane comprising knitted fibers of esterified hyaluronan (HYAFF11) can be used to cover fractured or grafted bone and can also serve as a scaffold to keep osteoprogenitor cells in place. The aim of this study was to compare osteoblastic activity by the use of scintigraphic methods in defective rabbit tibias during early-phase bone healing with or without a hyaluronan-based mesh. Two groups (A and B) of New Zealand albino rabbits were used; each group included 10 animals. Operations on all rabbits were performed under general anesthesia. We also resected 10-mm bone segments from each animal's tibial diaphysis. After resection, tibias with defects were fixed using Kirschner wires. In group A, no hyaluronan-based mesh was used. In group B, tibial segmental defects were enclosed with a hyaluronan-based mesh. The rabbits were followed up for 4 weeks postoperatively, after which bone scintigraphic studies were performed on each animal to detect and compare osteoblastic activity. The mean count in the fracture side of the hyaluronan-based mesh group was significantly higher compared to that of the group A (p = 0.019). However, there was no significant difference between group B and control rabbits with respect to the mean count on the intact bone side (p = 0.437). The bone defect (fracture)/intact bone mean count ratio was significantly higher in group B compared to group A (p = 0.008). A hyaluronan-based mesh plays a role in promoting osteoblastic activity. Hyalonect(®) is suitable for restoring tissue continuity whenever the periosteal membrane is structurally impaired or inadequate. Our results demonstrated that, during early-phase bone healing, osteoblastic activity

  16. Prevention of preterm birth based on short cervix: symptomatic women with preterm labor or premature prelabor rupture of membranes.

    Ness, Amen

    2009-10-01

    The diagnosis of preterm labor (PTL) is challenging, especially in women whose cervical dilatation is 15 mm. Transvaginal ultrasound CL can also be performed in the presence of ruptured membranes and predicts latency. Although additional data are needed, the evidence so far suggests that the use of transvaginal ultrasound CL and fetal fibronectin can be used to better identify and manage women with PTL likely to have an imminent preterm delivery, and to avoid interventions in women who would not.

  17. Quantification of functional groups and modeling of their ionization behavior in the active layer of FT30 reverse osmosis membrane.

    Coronell, Orlando; Mariñas, Benito J; Zhang, Xijing; Cahill, David G

    2008-07-15

    A new experimental approach was developed to measure the concentration of charged functional groups (FGs) in the active layer of thin-film composite reverse osmosis (RO) and nanofiltration (NF) membranes as a function of solution pH. FT30 RO membrane, with a fully aromatic polyamide (PA) active layer sandwiched between a polysulfone support and a coating layer, was used. The experiments consisted of saturating charged FGs with heavy ion probes, and determining the ion probe concentration by Rutherford backscattering spectrometry (RBS). Deprotonated carboxylic groups were saturated with Ag+, and protonated amine groups with W04(2-). The ionization behavior of carboxylic and amine groups was modeled based on acid-base equilibrium theory. While the ionization behavior of amine groups was satisfactorily described by one dissociation constant (pKa = 4.74), two pKa values (5.23 and 8.97) were necessary to describe the titration curve of carboxylic groups. These results were consistent with the bimodal pore size distribution (PSD) of FT30 active layer reported in the literature. The calculated total concentrations of carboxylic and amine groups in the active layer of the FT30 RO membrane studied were 0.432 and 0.036 M, respectively, and the isoelectric point (IEP) was 4.7. The total concentration of carboxylic and amine groups revealed that the degree of cross-linking of the PA active layer of the FT30 RO membrane studied was 94%.

  18. Performance of Submerged Membrane Bioreactor Combined with Powdered Activated Carbon Addition for the Treatment of an Industrial Wastewater

    Tri Widjaja

    2010-02-01

    Full Text Available Membrane technology is one of the alternative solutions to overcome industrial wastewater treatment developed nowadays. The addition of PAC (Powdered Activated Carbon in the activated sludge using Submerged Membrane Adsorption Hybrid Bioreactor (SMAHBR is expected to increase the organic material removal. The purpose of this study was to determine the performance of submerged membrane bioreactor and activated carbon adsorption capacity of organic materials in wastewater. This study used SIER (Surabaya Industrial Estate Rungkut – Surabaya, Indonesia waste as activated sludge operated at Mixed Liquor Suspended Solid (MLSS concentrations of 8000 and 15000 mg/l, and Chemical Oxygen Demand (COD concentrations of 1500, 2500 mg/l, Sludge Retention Time (SRT of 10;20; and 30 days and activated carbon variables of 0%; 2.5%; 5%; 7.5%; 10%. The results showed that the fouling potential occurred at high MLSS where the COD removal occurred at PAC addition of 10% reaching 91.86%. High Soluble Microbial Product (SMP accumulation (± 10 mg/l occurred in short SRT and high MLSS concentration. PAC addition resulted in decreased microorganisms in the reactor and better effluent of SMAHBR, as a result, the performance of the submerged membrane bioreactor would be restored.

  19. A plasma membrane localization signal in the HIV-1 envelope cytoplasmic domain prevents localization at sites of vesicular stomatitis virus budding and incorporation into VSV virions.

    Johnson, J E; Rodgers, W; Rose, J K

    1998-11-25

    Previous studies showed that the HIV-1 envelope (Env) protein was not incorporated into vesicular stomatitis virus (VSV) virions unless its cytoplasmic tail was replaced with that of the VSV glycoprotein (G). To determine whether the G tail provided a positive incorporation signal for Env, or if sequences in the Env tail prevented incorporation, we generated mutants of Env with its 150-amino-acid tail shortened to 29, 10, or 3 amino acids (Envtr mutants). Cells infected with VSV recombinants expressing these proteins or an Env-G tail hybrid showed similar amounts of Env protein at the surface. The Env-G tail hybrid or the Envtr3 mutant were incorporated at the highest levels into budding VSV virions. In contrast, the Envtr29 or Envtr10 mutants were incorporated poorly. These results defined a signal preventing incorporation within the 10 membrane-proximal amino acids of the Env tail. Confocal microscopy revealed that this signal functioned by causing localization of human immunodeficiency virus type 1 Env to plasma membrane domains distinct from the VSV budding sites, where VSV proteins were concentrated. Copyright 1998 Academic Press.

  20. COMPARATIVE ANALYSIS OF RETINAL REATTACHMENT SURGERY WITH OR WITHOUT INTERNAL LIMITING MEMBRANE PEELING TO PREVENT POSTOPERATIVE MACULAR PUCKER.

    Forlini, Matteo; Date, Purva; Ferrari, Luisa Micelli; Lorusso, Massimo; Lecce, Gabriella; Verdina, Tommaso; Neri, Giovanni; Benatti, Caterina; Rossini, Paolo; Bratu, Adriana; DʼEliseo, Domenico; Ferrari, Tommaso Micelli; Cavallini, Gian Maria

    2017-07-18

    To determine whether internal limiting membrane (ILM) peeling during pars plana vitrectomy for rhegmatogenous retinal detachment reduces the incidence of epiretinal membrane (ERM) formation. In this retrospective study, preoperative, intraoperative, and postoperative data from all eyes undergoing pars plana vitrectomy for rhegmatogenous retinal detachment between January 2007 and December 2013 was analyzed. All cases with at least 1-year of follow-up were included. Data collection included vision, intraoperative complications, occurrence of ERM, and spectral domain optical coherence tomography characteristics. The OCTs were retrieved for all eyes and were graded by a single masked grader. Out of 159 eyes recruited, ILM peeling was done in 78 eyes (49%). Overall occurrence of ERM was 20%. Seven eyes (9%) in ILM peeling group and 25 eyes in the non-ILM peeling group (31%) showed ERM (P = 0.001). Postoperative vision was significantly better in eyes that had ILM peeling (0.48 ± 0.4 logarithm of the minimum angle of resolution [20/63] vs. 0.77 ± 0.6 logarithm of the minimum angle of resolution [20/125], P = 0.003). In multivariable models adjusting for type of tamponade, ILM peeling reduced the likelihood of ERM formation by 75% (P = 0.01). Internal limiting membrane peeling during pars plana vitrectomy for rhegmatogenous retinal detachment significantly reduces ERM formation in the postoperative period and is associated with better visual and anatomical outcomes.

  1. Local mucous membrane lesions caused by irradiation of the oropharynx can be prevented by a dental shield

    Schratter-Sehn, A.U.; Vienna Univ.; Schmidt, W.F.O.; Kaercher, K.H.; Kielhauser, R.; Langer, H.

    1992-01-01

    In patients with metallic dental fillings radiation therapy to the oral cavity can cause mucous membrane lesions, which are more severe than expected. They appear as circumscribed erosions, opposite to metallic fillings and are caused by an increase in radiation dose through secondary radiation due to the higher density and atomic number of the filling material. This dose increase can be directly measured with 0.1 mm thin sheets of graphite-loaded TLD's (LiF, Vinten). For Co-60 gamma rays a commercial amalgam filling caused a dose increase by a factor of 1.7. The half value layer for this additional radiation was measured to be approximately 0.4 mm tissue. In order to avoid painful mucous membrane ulcerations which are even more a problem if hyperfractionated treatment schedules are used, we constructed individual dental shields for each patient. As shielding material we used a dental impression material (Optosil P + , Bayer). This method was tested in 35 patients, in all of them circumscribed mucous membrane ulcerations could be avoided. The method proved to be fast and simple and was very well tolerated by all patients. (orig.) [de

  2. Electrospun polymer membrane activated with room temperature ionic liquid: Novel polymer electrolytes for lithium batteries

    Cheruvally, Gouri; Kim, Jae-Kwang; Choi, Jae-Won; Ahn, Jou-Hyeon; Shin, Yong-Jo; Manuel, James; Raghavan, Prasanth; Kim, Ki-Won; Ahn, Hyo-Jun; Choi, Doo Seong; Song, Choong Eui

    A new class of polymer electrolytes (PEs) based on an electrospun polymer membrane incorporating a room-temperature ionic liquid (RTIL) has been prepared and evaluated for suitability in lithium cells. The electrospun poly(vinylidene fluoride- co-hexafluoropropylene) P(VdF-HFP) membrane is activated with a 0.5 M solution of LiTFSI in 1-butyl-3-methylimidazolium bis(trifluoromethanesulfonyl)imide (BMITFSI) or a 0.5 M solution of LiBF 4 in 1-butyl-3-methylimidazolium tetrafluoroborate (BMIBF 4). The resulting PEs have an ionic conductivity of 2.3 × 10 -3 S cm -1 at 25 °C and anodic stability at >4.5 V versus Li +/Li, making them suitable for practical applications in lithium cells. A Li/LiFePO 4 cell with a PE based on BMITFSI delivers high discharge capacities when evaluated at 25 °C at the 0.1 C rate (149 mAh g -1) and the 0.5 C rate (132 mAh g -1). A very stable cycle performance is also exhibited at these low current densities. The properties decrease at the higher, 1 C rate, when operated at 25 °C. Nevertheless, improved properties are obtained at a moderately elevated temperature of operation, i.e. 40 °C. This is attributed to enhanced conductivity of the electrolyte and faster reaction kinetics at higher temperatures. At 40 °C, a reversible capacity of 140 mAh g -1 is obtained at the 1 C rate.

  3. Workplace suicide prevention: a systematic review of published and unpublished activities.

    Milner, Allison; Page, Kathryn; Spencer-Thomas, Sally; Lamotagne, Anthony D

    2015-03-01

    There are a number of published studies on workplace suicide prevention activities, and an even larger number of activities that are not reported on in academic literature. The aim of this review was to provide a systematic assessment of workplace suicide prevention activities, including short-term training activities, as well as suicide prevention strategies designed for occupational groups at risk of suicide. The search was based on Meta-analysis of Observational Studies in Epidemiology (MOOSE) Guidelines. The databases used for the searches were the Cochrane Trials Library and PubMed. A range of suicide prevention websites were also searched to ascertain the information on unpublished workplace suicide prevention activities. Key characteristics of retrieved studies were extracted and explained, including whether activities were short-term training programmes or developed specifically for occupations at risk of suicide. There were 13 interventions relevant for the review after exclusions. There were a few examples of prevention activities developed for at-risk occupations (e.g. police, army, air force and the construction industry) as well as a number of general awareness programmes that could be applied across different settings. Very few workplace suicide prevention initiatives had been evaluated. Results from those that had been evaluated suggest that prevention initiatives had beneficial effects. Suicide prevention has the potential to be integrated into existing workplace mental health activities. There is a need for further studies to develop, implement and evaluate workplace suicide prevention programmes. © The Author 2014. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  4. B cell activation by outer membrane vesicles--a novel virulence mechanism.

    Maria Laura A Perez Vidakovics

    2010-01-01

    Full Text Available Secretion of outer membrane vesicles (OMV is an intriguing phenomenon of Gram-negative bacteria and has been suggested to play a role as virulence factors. The respiratory pathogens Moraxella catarrhalis reside in tonsils adjacent to B cells, and we have previously shown that M. catarrhalis induce a T cell independent B cell response by the immunoglobulin (Ig D-binding superantigen MID. Here we demonstrate that Moraxella are endocytosed and killed by human tonsillar B cells, whereas OMV have the potential to interact and activate B cells leading to bacterial rescue. The B cell response induced by OMV begins with IgD B cell receptor (BCR clustering and Ca(2+ mobilization followed by BCR internalization. In addition to IgD BCR, TLR9 and TLR2 were found to colocalize in lipid raft motifs after exposure to OMV. Two components of the OMV, i.e., MID and unmethylated CpG-DNA motifs, were found to be critical for B cell activation. OMV containing MID bound to and activated tonsillar CD19(+ IgD(+ lymphocytes resulting in IL-6 and IgM production in addition to increased surface marker density (HLA-DR, CD45, CD64, and CD86, whereas MID-deficient OMV failed to induce B cell activation. DNA associated with OMV induced full B cell activation by signaling through TLR9. Importantly, this concept was verified in vivo, as OMV equipped with MID and DNA were found in a 9-year old patient suffering from Moraxella sinusitis. In conclusion, Moraxella avoid direct interaction with host B cells by redirecting the adaptive humoral immune response using its superantigen-bearing OMV as decoys.

  5. Evaluation of the damage of cell wall and cell membrane for various extracellular polymeric substance extractions of activated sludge.

    Guo, Xuesong; Liu, Junxin; Xiao, Benyi

    2014-10-20

    Extracellular polymeric substances (EPS) are susceptible to contamination by intracellular substances released during the extraction of EPS owing to the damage caused to microbial cell structures. The damage to cell walls and cell membranes in nine EPS extraction processes of activated sludge was evaluated in this study. The extraction of EPS (including proteins, carbohydrates and DNA) was the highest using the NaOH extraction method and the lowest using formaldehyde extraction. All nine EPS extraction methods in this study resulted in cell wall and membrane damage. The damage to cell walls, evaluated by 2-keto-3-deoxyoctonate (KDO) and N-acetylglucosamine content changes in extracted EPS, was the most significant in the NaOH extraction process. Formaldehyde extraction showed a similar extent of damage to cell walls to those detected in the control method (centrifugation), while those in the formaldehyde-NaOH and cation exchange resin extractions were slightly higher than those detected in the control. N-acetylglucosamine was more suitable than KDO for the evaluation of cell wall damage in the EPS extraction of activated sludge. The damage to cell membranes was characterized by two fluorochromes (propidium iodide and FITC Annexin V) with flow cytometry (FCM) measurement. The highest proportion of membrane-damaged cells was detected in NaOH extraction (26.54% of total cells) while membrane-damaged cells comprised 8.19% of total cells in the control. Copyright © 2014 Elsevier B.V. All rights reserved.

  6. Dynamic activation of basilar membrane macrophages in response to chronic sensory cell degeneration in aging mouse cochleae.

    Frye, Mitchell D; Yang, Weiping; Zhang, Celia; Xiong, Binbin; Hu, Bo Hua

    2017-02-01

    In the sensory epithelium, macrophages have been identified on the scala tympani side of the basilar membrane. These basilar membrane macrophages are the spatially closest immune cells to sensory cells and are able to directly respond to and influence sensory cell pathogenesis. While basilar membrane macrophages have been studied in acute cochlear stresses, their behavior in response to chronic sensory cell degeneration is largely unknown. Here we report a systematic observation of the variance in phenotypes, the changes in morphology and distribution of basilar membrane tissue macrophages in different age groups of C57BL/6J mice, a mouse model of age-related sensory cell degeneration. This study reveals that mature, fully differentiated tissue macrophages, not recently infiltrated monocytes, are the major macrophage population for immune responses to chronic sensory cell death. These macrophages display dynamic changes in their numbers and morphologies as age increases, and the changes are related to the phases of sensory cell degeneration. Notably, macrophage activation precedes sensory cell pathogenesis, and strong macrophage activity is maintained until sensory cell degradation is complete. Collectively, these findings suggest that mature tissue macrophages on the basilar membrane are a dynamic group of cells that are capable of vigorous adaptation to changes in the local sensory epithelium environment influenced by sensory cell status. Copyright © 2016 Elsevier B.V. All rights reserved.

  7. Activation of PVDF membranes through facile hydroxylation of the polymeric dope

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

    2017-01-01

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

  8. Activation of PVDF membranes through facile hydroxylation of the polymeric dope

    Al-Gharabli, Samer

    2017-10-30

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

  9. Active Generations: An Intergenerational Approach to Preventing Childhood Obesity

    Werner, Danilea; Teufel, James; Holtgrave, Peter L.; Brown, Stephen L.

    2012-01-01

    Background: Over the last 3 decades, US obesity rates have increased dramatically as more children and more adults become obese. This study explores an innovative program, Active Generations, an intergenerational nutrition education and activity program implemented in out-of-school environments (after school and summer camps). It utilizes older…

  10. Magnetically controlled permeability membranes

    Kosel, Jurgen

    2013-10-31

    A bioactive material delivery system can include a thermoresponsive polymer membrane and nanowires distributed within the thermoresponsive polymer membrane. Magnetic activation of a thermoresponsive polymer membrane can take place via altering the magnetization or dimensions of nanowires dispersed or ordered within the membrane matrix.

  11. Magnetically controlled permeability membranes

    Kosel, Jü rgen; Khashab, Niveen M.; Zaher, Amir

    2013-01-01

    A bioactive material delivery system can include a thermoresponsive polymer membrane and nanowires distributed within the thermoresponsive polymer membrane. Magnetic activation of a thermoresponsive polymer membrane can take place via altering the magnetization or dimensions of nanowires dispersed or ordered within the membrane matrix.

  12. Antifungal Activity of (+-2,2’-Epicytoskyrin A and Its Membrane-Disruptive Action

    Dewi Wulansari

    2016-12-01

    Full Text Available (+-2,2’-Epicytoskyrin A, a bis-anthraquinone isolated from fungal endophyte Diaporthe sp. GNBP-10 associated with Uncaria gambir Roxb., was investigated for its antifungal activity. The broth microdilution method was used to determine the minimum inhibitory concentration (MIC against 22 yeast strains and three filamentous fungi. The MICs of (+-2,2’-epicytoskyrin A ranged from 16 to 128 µg/mL, which exhibited lower activity than the antifungal nystatin. A study of the mechanism of action revealed similar effects of (+-2,2’-epicytoskyrin A and nystatin on Candida tropicalis at their MICs (16 and 8 µg/mL, respectively and 2 times of the MIC. Both compounds caused cytoplasmic material and ion leakages on fungal cell, which were characterized by an increase in absorbance at 260 nm and 280 nm as well as Ca2+ and K+ ion concentrations. The morphology of the fungal cells after (+-2,2’-epicytoskyrin A treatment was observed under a scanning electron microscope. The control cells, which were not treated with either (+-2,2’-epicytoskyrin A or nystatin, showed a smooth surface, while the cells treated with either (+-2,2’-epicytoskyrin A or nystatin shrank and displayed a donut-like shape. More shrinkage was observed in the 2 times MIC concentration and even more in the cells exposed to nystatin. The action of (+-2,2’-epicytoskyrin A was proposed through membrane disruption.

  13. Preparation of a high specific activity I-125 labeled styryl dye for leukocyte membrane labeling

    Lambert, C.; Mease, R.C.; Le, T.; Sabet, H.; Avren, L.I.; McAfee, J.G.

    1994-01-01

    The purpose of this work was to develop a high specific activity radioiodinated cell membrane probe for tracking lymphocytes in-vivo to replace the nucleus localizing, cytotoxic lipophilic chelates (In-111 oxine and Tc-99m HMPAO) currently used. Alkylation of parent dye 4-[2-[-N,N-didecylamino]phenyl]ethenyl pyridine with E-1-tributylstannyl-3-tosylpropene (prepared form E-1-tributylstannyl-1-propene-3-ol), gave a tributyltin precursor 1. Radiolabeled 3-[4-[2-[4-(N,N-didecylamino)phenyl]ethenyl]pyridino] E-[I-125]-1-iodopropene (2), was prepared from 1 using peracetic acid in acetonitrile/water. Labeling yields and specific activities achieved were 26% (∼2170 Ci/mmol), 40% (1220 Ci/mmol), and 55% (200 Ci/mmol) for nca, 0.4, and 2 nanomole carrier iodide runs respectively. Canine mixed leukocytes (0.5-1.0 x 10 8 cells) were labeled with 2 (67% and 42% yields for 200 Ci/mol and 1220 Ci/mmol preparations) and showed blood clearance similar to In 111 oxine. Radioiodinated styryl dye 2 appears to be a promising leukocyte labeling agent. Imaging studies with I-131 labeled 2 are in progress

  14. Trichomonicidal and parasite membrane damaging activity of bidesmosic saponins from Manilkara rufula.

    Patrícia de Brum Vieira

    Full Text Available The infection caused by Trichomonas vaginalis is the most common but overlooked non-viral sexually transmitted disease worldwide. Treatment relies on one class of drugs, the 5-nitroimidazoles, but resistance is widespread. New drugs are urgently needed. We reported the effect of crude and purified saponin fractions of Manilkara rufula against Trichomonas vaginalis. The compound responsible for antitrichomonal activity was isolated and identified as an uncommon bidesmosic saponin, Mi-saponin C. This saponin eliminated parasite viability without toxicity against the human vaginal epithelial line (HMVII. In addition, the isolated saponin fraction improved the metronidazole effect against a metronidazole-resistant isolate and dramatically reduced the cytoadherence of T. vaginalis to human cells. Investigation of the mechanism of death showed that the saponin fraction induced the parasite death due to profound membrane damage, inducing a disturbance of intracellular content without nuclear damage. To the best of our knowledge, this is the first report of antitrichomonal activity in the bidesmosic saponins of Manilkara rufula.

  15. Impact of coagulant and flocculant addition to an anaerobic dynamic membrane bioreactor (AnDMBR) treating waste-activated sludge

    Kooijman, G.; Lopes, Wilton; Zhou, Z.; Guo, H.; de Kreuk, M.K.; Spanjers, H.L.F.M.; van Lier, J.B.

    2017-01-01

    In this work, we investigated the effects of flocculation aid (FA) addition to an anaerobic dynamic membrane bioreactor (AnDMBR) (7 L, 35°C) treating waste-activated sludge (WAS). The experiment consisted of three distinct periods. In period 1 (day 1–86), the reactor was operated as a

  16. Na+/K(+)pump activity in photoreceptors of the blowfly Calliphora : A model analysis based on membrane potential measurements

    Gerster, U; Stavenga, DG; Backhaus, W

    Na+/K+-pump activity and intracellular Na+ and K+ concentration changes in blowfly photoreceptors are derived from intracellular potential measurements in vivo with a model based on the Goldman-Hodgkin-Katz theory for membrane currents. The relation between the intracellular Na+ concentration and

  17. Intercellular deposits of basement membrane material in active human pituitary adenomas detected by immunostaining for laminin and electron microscopy

    Holck, S; Wewer, U M; Albrechtsen, R

    1986-01-01

    and one patient with Cushing's syndrome). Concurrently, at the ultrastructural level, bunches of basement membrane-like material intermingled between the adenoma cells were demonstrated in seven of these ten active adenomas. Furthermore, secretory granules were entrapped occasionally in this intercellular...

  18. Local membrane deformations activate Ca2+-dependent K+ and anionic currents in intact human red blood cells

    Dyrda, Agnieszka; Cytlak, Urszula; Ciuraszkiewicz, Anna

    2010-01-01

    -activated transient PCa observed here under local membrane deformation is a likely contributor to the Ca(2+)-mediated effects observed during the normal aging process of red blood cells, and to the increased Ca(2+) content of red cells in certain hereditary anemias such as thalassemia and sickle cell anemia....

  19. Discursive constructions of falls prevention : Discourses of active aging versus old age as disease

    Evron, Lotte; Ulrich, Anita; Tanggaard, Lene

    2012-01-01

    information and investment in falls prevention programs, many still drop out or decline to participate in such programs. The study explores how discourses cross swords in the domain of falls prevention. We identify two main discourses in the field: Discourses of active aging opposed to discourses of old age...... as disease. In discourses of active aging falls are constructed as preventable and not necessarily related to old age; in discourses of old age as disease falls are constructed as a disease of old age. Specific agent positions are created within discourses. Discourses of active aging construct self......-responsible citizens who are physically active and motivated to participate in falls prevention programmes; discourses of old age as disease on the other hand construct “fall patients” who accept being passive in the health care system. Older citizens who are not in need of treatment or less physically active...

  20. Physical Activity Prevents Progression for Cognitive Impairment and Vascular Dementia

    Verdelho, Ana; Madureira, Sofia; Ferro, José M

    2012-01-01

    BACKGROUND AND PURPOSE: We aimed to study if physical activity could interfere with progression for cognitive impairment and dementia in older people with white matter changes living independently. METHODS: The LADIS (Leukoaraiosis and Disability) prospective multinational European study evaluates....... Physical activity was recorded during the clinical interview. MRI was performed at entry and at the end of the study. RESULTS: Six hundred thirty-nine subjects were included (74.1±5 years old, 55% women, 9.6±3.8 years of schooling, 64% physically active). At the end of follow-up, 90 patients had dementia...... (vascular dementia, 54; Alzheimer disease with vascular component, 34; frontotemporal dementia, 2), and 147 had cognitive impairment not dementia. Using Cox regression analysis, physical activity reduced the risk of cognitive impairment (dementia and not dementia: β=-0.45, P=0.002; hazard ratio, 0.64; 95...

  1. Pomegranate Extracts and Cancer Prevention: Molecular and Cellular Activities

    Syed, Deeba N.; Chamcheu, Jean-Christopher; Adhami, Vaqar M.; Mukhtar, Hasan

    2014-01-01

    There is increased appreciation by the scientific community that dietary phytochemicals can be potential weapons in the fight against cancer. Emerging data has provided new insights into the molecular and cellular framework needed to establish novel mechanism-based strategies for cancer prevention by selective bioactive food components. The unique chemical composition of the pomegranate fruit, rich in antioxidant tannins and flavonoids has drawn the attention of many investigators. Polyphenol rich fractions derived from the pomegranate fruit have been studied for their potential chemopreventive and/or cancer therapeutic effects in several animal models. Although data from in vitro and in vivo studies look convincing, well designed clinical trials in humans are needed to ascertain whether pomegranate can become part of our armamentarium against cancer. This review summarizes the available literature on the effects of pomegranate against various cancers. PMID:23094914

  2. Structural remodeling and oligomerization of human cathelicidin on membranes suggest fibril-like structures as active species

    Sancho-Vaello, Enea; François, Patrice; Bonetti, Eve-Julie

    2017-01-01

    Antimicrobial peptides as part of the mammalian innate immune system target and remove major bacterial pathogens, often through irreversible damage of their cellular membranes. To explore the mechanism by which the important cathelicidin peptide LL-37 of the human innate immune system interacts w...... that these supramolecular structures represent the LL-37-membrane active state. Collectively, our study provides new insights into the fascinating plasticity of LL-37 demonstrated at atomic resolution and opens the venue for LL-37-based molecules as novel antibiotics....

  3. Study on the antioxidant activity and membrane interaction of a multiple antioxidant

    Jodko-Jodko-Piórecka, Kasia; Klösgen, Beate; Litwinienko, Grzegorz

    /PMHC antioxidative co-operation we propose that dopamine acts mainly as a scavenger of radicals near the membrane surface while PMHC is scavenging the peroxyl radicals within the membrane. References 1) Jodko, K.; Kowalewska, E.; Sikora, B.; Litwinienko, G. Free Radic. Res. 2009, 43, 58 2) Liu, J; Mori, A. Arch...

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

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

    1995-01-01

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

  5. Prevalence and correlates of participation in fall prevention exercise/physical activity by older adults

    Merom, D.; Pye, V.; Macniven, R.; van der Ploeg, H.; Milat, A.; Sherrington, C.; Lord, S.; Bauman, A.

    2012-01-01

    Objective: To examine older people's participation in fall prevention exercise/physical activities. Methods: Participants comprised 5,681 randomly selected older people (≥ 65. years) who took part in the 2009 New South Wales (Australia) Fall Prevention telephone survey (61% response-rate). The

  6. A review of the literature on preventive occupational health and safety activities in small enterprises

    Hasle, Peter; Limborg, Hans Jørgen

    2006-01-01

    The scientific literature regarding preventive occupational health and safety activities in small enterprises has been reviewed in order to identify effective preventive approaches and to develop a future research strategy. During the last couple of years, there has been a significant increase...

  7. Effects of Iron Overload on the Activity of Na,K-ATPase and Lipid Profile of the Human Erythrocyte Membrane.

    Leilismara Sousa

    Full Text Available Iron is an essential chemical element for human life. However, in some pathological conditions, such as hereditary hemochromatosis type 1 (HH1, iron overload induces the production of reactive oxygen species that may lead to lipid peroxidation and a change in the plasma-membrane lipid profile. In this study, we investigated whether iron overload interferes with the Na,K-ATPase activity of the plasma membrane by studying erythrocytes that were obtained from the whole blood of patients suffering from iron overload. Additionally, we treated erythrocytes of normal subjects with 0.8 mM H2O2 and 1 μM FeCl3 for 24 h. We then analyzed the lipid profile, lipid peroxidation and Na,K-ATPase activity of plasma membranes derived from these cells. Iron overload was more frequent in men (87.5% than in women and was associated with an increase (446% in lipid peroxidation, as indicated by the amount of the thiobarbituric acid reactive substances (TBARS and an increase (327% in the Na,K-ATPase activity in the plasma membrane of erythrocytes. Erythrocytes treated with 1 μM FeCl3 for 24 h showed an increase (132% in the Na,K-ATPase activity but no change in the TBARS levels. Iron treatment also decreased the cholesterol and phospholipid content of the erythrocyte membranes and similar decreases were observed in iron overload patients. In contrast, erythrocytes treated with 0.8 mM H2O2 for 24 h showed no change in the measured parameters. These results indicate that erythrocytes from patients with iron overload exhibit higher Na,K-ATPase activity compared with normal subjects and that this effect is specifically associated with altered iron levels.

  8. Differences between Solution and Membrane Forms of Chitosan on the In Vitro Activity of Fibroblasts

    Bahar Uslu

    2015-03-01

    Full Text Available Background: Chitosan, a linear polysaccharide, has been recently used in biomedical applications. In vitro studies have demonstrated its effect on cellular growth and its stimulatory action on cellular layer formation. Aims: The present study aims to compare the proliferative effects of chitosan in two forms, membranous and solution forms, on Swiss 3T3 mouse embryonic fibroblasts. Study Design: In vitro study. Methods: Three experimental groups were formed: cells were cultured in a normal medium without chitosan (Control Group; cells were cultured either in a medium containing 2.0% chitosan in membranous form (Membrane Group or chitosan solution at a concentration of 2.0% (Solution Group.Two different methods were used in the experiments: cells cultured on the medium containing chitosan in solution or membranous forms (method 1; and chitosan solution or membranous forms were added into the medium containing previously cultured cells (method 2. Results: Scanning electron microscopic investigations of the experimental groups revealed cells with well-defined cellular projections, intact cellular membranes and tight intercellular junctions. They were especially prominent in the membrane group of method 1 and in the membrane and solution groups of method 2. Mouse monoclonal anti-collagen 1 primary antibody was used to indicate collagen synthesis. Prominent collagen synthesis was detected in the membrane groups on the 10th day of culture for both methods. Bromodeoxyuridine (BrdU and MTT assays were performed in order to assess cellular proliferation and viability, respectively. BrdU labelling tests indicated a higher proliferation index in the membrane group of method 1 on the 5th and 10th days. For the second method, the membranous form on the 10th day and solution form on the 5th day were the most effective groups in terms of cellular proliferation. MTT results reflected a high cellular viability in method 1 on the 5th day of treatment with the

  9. Preferential inhibition of the plasma membrane NADH oxidase (NOX) activity by diphenyleneiodonium chloride with NADPH as donor

    Morre, D. James

    2002-01-01

    The cell-surface NADH oxidase (NOX) protein of plant and animal cells will utilize both NADH and NADPH as reduced electron donors for activity. The two activities are distinguished by a differential inhibition by the redox inhibitor diphenyleneiodonium chloride (DPI). Using both plasma membranes and cells, activity with NADPH as donor was markedly inhibited by DPI at submicromolar concentrations, whereas with NADH as donor, DPI was much less effective or had no effect on the activity. The possibility of the inhibition being the result of two different enzymes was eliminated by the use of a recombinant NOX protein. The findings support the concept that NOX proteins serve as terminal oxidases for plasma membrane electron transport involving cytosolic reduced pyridine nucleotides as the natural electron donors and with molecular oxygen as the electron acceptor.

  10. Water Activated Graphene Oxide Transfer Using Wax Printed Membranes for Fast Patterning of a Touch Sensitive Device.

    Baptista-Pires, Luis; Mayorga-Martínez, Carmen C; Medina-Sánchez, Mariana; Montón, Helena; Merkoçi, Arben

    2016-01-26

    We demonstrate a graphene oxide printing technology using wax printed membranes for the fast patterning and water activation transfer using pressure based mechanisms. The wax printed membranes have 50 μm resolution, longtime stability and infinite shaping capability. The use of these membranes complemented with the vacuum filtration of graphene oxide provides the control over the thickness. Our demonstration provides a solvent free methodology for printing graphene oxide devices in all shapes and all substrates using the roll-to-roll automatized mechanism present in the wax printing machine. Graphene oxide was transferred over a wide variety of substrates as textile or PET in between others. Finally, we developed a touch switch sensing device integrated in a LED electronic circuit.

  11. Simulation of 2D Waves in Circular Membrane Using Excel Spreadsheet with Visual Basic for Teaching Activity

    Eso, R.; Safiuddin, L. O.; Agusu, L.; Arfa, L. M. R. F.

    2018-04-01

    We propose a teaching instrument demonstrating the circular membrane waves using the excel interactive spreadsheets with the Visual Basic for Application (VBA) programming. It is based on the analytic solution of circular membrane waves involving Bessel function. The vibration modes and frequencies are determined by using Bessel approximation and initial conditions. The 3D perspective based on the spreadsheets functions and facilities has been explored to show the 3D moving objects in transitional or rotational processes. This instrument is very useful both in teaching activity and learning process of wave physics. Visualizing of the vibration of waves in the circular membrane which is showing a very clear manner of m and n vibration modes of the wave in a certain frequency has been compared and matched to the experimental result using resonance method. The peak of deflection varies in time if the initial condition was working and have the same pattern with matlab simulation in zero initial velocity

  12. Osmolality, temperature, and membrane lipid composition modulate the activity of betaine transporter BetP in Corynebacterium glutamicum

    Ozcan, Nuran; Ejsing, Christer S.; Shevchenko, Andrej

    2007-01-01

    The gram-positive soil bacterium Corynebacterium glutamicum, a major amino acid-producing microorganism in biotechnology, is equipped with several osmoregulated uptake systems for compatible solutes, which is relevant for the physiological response to osmotic stress. The most significant carrier......P activity. We further correlated the change in BetP regulation properties in cells grown at different temperatures to changes in the lipid composition of the plasma membrane. For this purpose, the glycerophospholipidome of C. glutamicum grown at different temperatures was analyzed by mass spectrometry using...... quantitative multiple precursor ion scanning. The molecular composition of glycerophospholipids was strongly affected by the growth temperature. The modulating influence of membrane lipid composition on BetP function was further corroborated by studying the influence of artificial modulation of membrane...

  13. Depression of membrane-bound Na+-K+-ATPase activity induced by free radicals and by ischemia of kidney

    Kako, K.; Kato, M.; Matsuoka, T.; Mustapha, A.

    1988-01-01

    A partially purified, membrane-bound Na + -K + -ATPase fraction, prepared from the outer medulla of porcine kidney, was incubated in the presence of 0.1-100 mM H 2 O 2 for either 15 or 30 min at 37 degree C. The activity of ouabain-sensitive Na + -K + -ATPase was reduced proportionally to the concentration of H 2 O 2 and the duration of incubation. There were decreases in SH contents and turnover rates of the Na + -K + -ATPase preparation, while malondialdehyde (MDA) and conjugated dienes were generated from the membrane lipids in the course of the incubation. The concentrations of ethanolamine (E) plasmalogen and of arachidonic acid in the E glycerophospholipid molecules were reduced by the free radical reaction. Similarly, a reduction in Na + K + -ATPase activity and the formation of MDA and conjugated dienes, together with a decrease in E glycerophospholipids, were observed when the membrane fraction was exposed to ultraviolet irradiation (254 nm) for 30 min at 4 degree C. Microsomal fractions, prepared from the outer medulla of canine kidney after 1 h of unilateral ischemia and 1 h of reperfusion, showed a decreased Na + -K + -ATPase activity, a reduced amount of SH groups, and an increased MDA. These changes were normalized by the infusion of N-mercaptopropionylglycine. These results support the view (1) that free radical generation affects the enzyme protein as well as membrane lipids, and (2) that free radicals may be formed in the ischemic reperfused kidney

  14. Ubiquitination of the bacterial inositol phosphatase, SopB, regulates its biological activity at the plasma membrane.

    Knodler, Leigh A

    2009-11-01

    The Salmonella type III effector, SopB, is an inositol polyphosphate phosphatase that modulates host cell phospholipids at the plasma membrane and the nascent Salmonella-containing vacuole (SCV). Translocated SopB persists for many hours after infection and is ubiquitinated but the significance of this covalent modification has not been investigated. Here we identify by mass spectrometry six lysine residues of SopB that are mono-ubiquitinated. Substitution of these six lysine residues with arginine, SopB-K(6)R, almost completely eliminated SopB ubiquitination. We found that ubiquitination does not affect SopB stability or membrane association, or SopB-dependent events in SCV biogenesis. However, two spatially and temporally distinct events are dependent on ubiquitination, downregulation of SopB activity at the plasma membrane and prolonged retention of SopB on the SCV. Activation of the mammalian pro-survival kinase Akt\\/PKB, a downstream target of SopB, was intensified and prolonged after infection with the SopB-K(6)R mutant. At later times, fewer SCV were decorated with SopB-K(6)R compared with SopB. Instead SopB-K(6)R was present as discrete vesicles spread diffusely throughout the cell. Altogether, our data show that ubiquitination of SopB is not related to its intracellular stability but rather regulates its enzymatic activity at the plasma membrane and intracellular localization.

  15. Thrombus imaging in a primate model with antibodies specific for an external membrane protein of activated platelets

    Palabrica, T.M.; Furie, B.C.; Konstam, M.A.; Aronovitz, M.J.; Connolly, R.; Brockway, B.A.; Ramberg, K.L.; Furie, B.

    1989-01-01

    The activated platelet is a potential target for the localization of thrombi in vivo since, after stimulation and secretion of granule contents, activated platelets are concentrated at sites of blood clot formation. In this study, we used antibodies specific for a membrane protein of activated platelets to detect experimental thrombi in an animal model. PADGEM (platelet activation-dependent granule-external membrane protein), a platelet alpha-granule membrane protein, is translocated to the plasma membrane during platelet activation and granule secretion. Since PADGEM is internal in unstimulated platelets, polyclonal anti-PADGEM and monoclonal KC4 antibodies do not bind to circulating resting platelets but do interact with activated platelets. Dacron graft material incubated with radiolabeled KC4 or anti-PADGEM antibodies in the presence of thrombin-activated platelet-rich plasma bound most of the antibody. Imaging experiments with 123I-labeled anti-PADGEM in baboons with an external arterial-venous Dacron shunt revealed rapid uptake in the thrombus induced by the Dacron graft; control experiments with 123I-labeled nonimmune IgG exhibited minimal uptake. Deep venous thrombi, formed by using percutaneous balloon catheters to stop blood flow in the femoral vein of baboons, were visualized with 123I-labeled anti-PADGEM. Thrombi were discernible against blood pool background activity without subtraction techniques within 1 hr. No target enhancement was seen with 123I-labeled nonimmune IgG. 123I-labeled anti-PADGEM cleared the blood pool with an initial half-disappearance time of 6 min and did not interfere with hemostasis. These results indicate that radioimmunoscintigraphy with anti-PADGEM antibodies can visualize thrombi in baboon models and is a promising technique for clinical thrombus detection in humans

  16. Specific chlamydial inclusion membrane proteins associate with active Src family kinases in microdomains that interact with the host microtubule network.

    Mital, Jeffrey; Miller, Natalie J; Fischer, Elizabeth R; Hackstadt, Ted

    2010-09-01

    Chlamydiae are Gram-negative obligate intracellular bacteria that cause diseases with significant medical and economic impact. Chlamydia trachomatis replicates within a vacuole termed an inclusion, which is extensively modified by the insertion of a number of bacterial effector proteins known as inclusion membrane proteins (Incs). Once modified, the inclusion is trafficked in a dynein-dependent manner to the microtubule-organizing centre (MTOC), where it associates with host centrosomes. Here we describe a novel structure on the inclusion membrane comprised of both host and bacterial proteins. Members of the Src family of kinases are recruited to the chlamydial inclusion in an active form. These kinases display a distinct, localized punctate microdomain-like staining pattern on the inclusion membrane that colocalizes with four chlamydial inclusion membrane proteins (Incs) and is enriched in cholesterol. Biochemical studies show that at least two of these Incs stably interact with one another. Furthermore, host centrosomes associate with these microdomain proteins in C. trachomatis-infected cells and in uninfected cells exogenously expressing one of the chlamydial effectors. Together, the data suggest that a specific structure on the C. trachomatis inclusion membrane may be responsible for the known interactions of chlamydiae with the microtubule network and resultant effects on centrosome stability.

  17. Characteristics of Combined Submerged Membrane Bioreactor with Granular Activated Carbon (GAC) in Treating Lineal Alkylbenzene Sulphonates (LAS) Wastewater

    Guo, Jifeng; Xia, Siqing; Lu, Yanjun

    2010-11-01

    A combined MBR (cMBR) with granular activated carbon (GAC) was used as a backbone system to treat the synthetic lineal alkylbenzene sulphonates (LAS) wastewater. The GAC was added in the MBR to improve the resistance of membrane fouling. A parallel conventional MBR (pMBR) without the GAC was run to give a contrast. The results of the process demonstrate that the cMBR process was more efficient than pMBR. It was found that the TMP changes of the cMBR were slower than the pMBR. The results demonstrated that the cMBRs membrane was better than the pMBR's after a clean period run. It was the GAC scrubbing to the membrane that delayed the membrane fouling of the cMBR. Variable critical flux was found in MBR, which showed that the cMBR could make the critical flux better than pMBR in the run time, but GAC could not improve the critical flux at the end of the period for the severe membrane fouling. Based on this theory, a variable critical flux (J) of MBR was put forward, and the relationship of J with time (t) was: J = 16.081e-0.0177t.

  18. Effect of powdered activated carbon dosage on sludge properties and membrane bioreactor performance in a hybrid MBR-PAC system.

    Zhang, Shi; Zuo, Xingtao; Xiong, Juan; Ma, Cong; Hu, Bo

    2017-12-22

    An improved insight into the effect of powdered activated carbon (PAC) on membrane fouling is crucial to the MBR performance. Sludge key property, soluble microbial products (SMP) and transmembrane pressure (TMP) were monitored. The membrane fouling rate in the MBRs was also analyzed based on TMP profile and resistance-in-series model. PAC reduced the membrane filtration resistance and significantly decreased the fouling rate. The sludge filterability was improved by extending the filtration time by almost twofold. PAC affected the SMP release and protein/polysaccharide (carbohydrate) was in a lower ratio. Fourier transform infrared (FTIR) analysis indicated that PAC decreased the impact of organic carbon, and reduced the proteins' and polysaccharides' absorption and deposition on the membrane surface and in the pores. The degree of reversible and irreversible fouling was related to the PAC content added into the MBRs. At the optimum dosage of 2 g/L, the results signified the PAC potential as a mitigation strategy of membrane fouling.

  19. PREVENTION OF BLOOD ACTIVATION DURING AND COMPLICATIONS AFTER CARDIOPULMONARY BYPASS

    VANOEVEREN, W; WILDEVUUR, CRH

    1991-01-01

    The cardiopulmonary bypass (CPB) circuit for open heart surgery initiates a whole body inflammatory reaction (WBIR) resulting in impaired hemostasis and organ dysfunction. Impaired hemostasis appeared to be related to the activation of the contact system (factor XII), which can be inhibited by

  20. Effects of cytokine-suppressive anti-inflammatory drugs on inflammatory activation in ex vivo human and ovine fetal membranes.

    Stinson, Lisa F; Ireland, Demelza J; Kemp, Matthew W; Payne, Matthew S; Stock, Sarah J; Newnham, John P; Keelan, Jeffrey A

    2014-03-01

    Intrauterine infection and inflammation are responsible for the majority of early (PTBs). Anti-inflammatory agents, delivered intra-amniotically together with antibiotics, may be an effective strategy for preventing PTB. In this study, the effects of four cytokine-suppressive anti-inflammatory drugs (CSAIDs: N-acetyl cysteine (NAC), SB239063, TPCA-1 and NEMO binding domain inhibitor (NBDI)) were assessed on human and ovine gestational membrane inflammation. Full-thickness membranes were collected from healthy, term, human placentas delivered by Caesarean section (n=5). Using a Transwell model, they were stimulated ex vivo with γ-irradiation-killed Escherichia coli applied to the amniotic face. Membranes from near-term, ovine placentas were stimulated in utero with lipopolysaccharide, Ureaplasma parvum or saline control and subjected to explant culture. The effects of treatment with CSAIDs or vehicle (1% DMSO) on accumulation of PGE2 and cytokines (human interleukin 6 (IL6), IL10 and TNFα; ovine IL8 (oIL8)) were assessed in conditioned media at various time points (3-20  h). In human membranes, the IKKβ inhibitor TPCA-1 (7  μM) and p38 MAPK inhibitor SB239063 (20  μM) administered to the amniotic compartment were the most effective in inhibiting accumulation of cytokines and PGE2 in the fetal compartment. NAC (10  mM) inhibited accumulation of PGE2 and IL10 only; NBDI (10  μM) had no significant effect. In addition to the fetal compartment, SB239063 also exerted consistent and significant inhibitory effects in the maternal compartment. TPCA-1 and SB239063 suppressed oIL8 production, while all CSAIDs tested suppressed ovine PGE2 production. These results support the further investigation of intra-amniotically delivered CSAIDs for the prevention of inflammation-mediated PTB.

  1. Photodynamic Action of LED-Activated Curcumin against Staphylococcus aureus Involving Intracellular ROS Increase and Membrane Damage

    Yuan Jiang

    2014-01-01

    Full Text Available Aim. To investigate the effect of photodynamic action of LED-activated curcumin on cell viability, membrane permeability, and intracellular reactive oxygen species of Staphylococcus aureus. Methods. Staphylococcus aureus was incubated with the different concentrations of curcumin for 60 min and then irradiated by blue light with the wavelength of 470 nm and with light dose of 3 J/cm2. The colony forming unit assay was used to investigate photocytotoxicity of curcumin on Staphylococcus aureus, confocal laser scanning microscopy (CLSM and flow cytometry (FCM for assaying membrane permeability, FCM analysis with DCFH-DA staining for measuring the intracellular ROS level, and transmission electron microscopy (TEM for observing morphology and structure. Results. Blue light-activated curcumin significantly killed Staphylococcus aureus in a curcumin dose-dependent manner. TEM observed remarkable structural damages in S. aureus after light-activated curcumin. More red fluorescence of PI dye was found in S. aureus treated by blue light-activated curcumin than in those of the controlled bacterial cells. Intracellular ROS increase was observed after light-activated curcumin. Conclusion. Blue light-activated curcumin markedly damaged membrane permeability, resulting in cell death of Staphylococcus aureus and highlighted that intracellular ROS increase might be an important event in photodynamic killing of Staphylococcus aureus in the presence of curcumin.

  2. Facilitated transport of Hg(II) through novel activated composite membranes

    Paez-Hernandez, M.E. [Universidad Autonoma Metropolitana-Azcapotzalco, Departamento de Materiales, Area de Ciencia de los Materiales, Col. Reynosa-Tamaulipas (Mexico); Universidad Autonoma del Estado de Hidalgo, Centro de Investigaciones Quimicas, Pachuca, Hidalgo (Mexico); Aguilar-Arteaga, K. [Universidad Autonoma del Estado de Hidalgo, Centro de Investigaciones Quimicas, Pachuca, Hidalgo (Mexico); Valiente, M. [Universitat Autonoma de Barcelona, Departament de Quimica, Unitat Analitica, Centre GTS, Facultat de Ciencies, Bellaterra, Barcelona (Spain); Ramirez-Silva, M.T. [Universidad Autonoma Metropolitana-Iztapalapa, Departamento de Quimica, Area de Quimica Analitica, Laboratorio R-105, Col. Vicentina, Mexico D.F. (Mexico); Romero-Romo, M.; Palomar-Pardave, M. [Universidad Autonoma Metropolitana-Azcapotzalco, Departamento de Materiales, Area de Ciencia de los Materiales, Col. Reynosa-Tamaulipas (Mexico)

    2004-10-01

    The results presented in this work deal with the prime application of activated composite membranes (ACMs) for the transport of Hg(II) ions in a continuous extraction-re-extraction system using di-(2-ethylhexyl)dithiophosphoric acid (DTPA) as carrier. The effects of variables such as the pH, the nature of the acid and the concentration of the casting solutions on the transport of Hg(II) are also investigated. When the ACM was prepared with a 0.5 M DTPA solution and when the feed solution contained 2.5 x 10{sup -4} M Hg(II) in 0.1 M HCl, the amount of mercury extracted was greater than 76%. The re-extracted mercury was subsequently recovered by means of a stripping phase comprising 0.3 M thiourea solution in 2 M H{sub 2}SO{sub 4}, yielding 54% of the initial amount of mercury after transport had taken place for 180 min. (orig.)

  3. Membrane Fouling Potential of Secondary Effluent Organic Matter (EfOM) from Conventional Activated Sludge Process

    Wei, Chunhai; Amy, Gary L.

    2012-01-01

    Secondary effluent organic matter (EfOM) from a conventional activated sludge process was filtered through constant-pressure dead-end filtration tests with a sequential ultrafiltration (UF, molecular weight cut-off (MWCO) of 10k Dalton) and nanofiltration (NF, MWCO of 200 Dalton) array to investigate its membrane fouling potential. Advanced analytical methods including liquid chromatography with online carbon detection (LC-OCD) and fluorescent excitation-emission matrix (F-EEM) were employed for EfOM characterization. EfOM consisted of humic substances and building blocks, low molecular weight (LMW) neutrals, biopolymers (mainly proteins) and hydrophobic organics according to the sequence of their organic carbon fractions. The UF rejected only biopolymers and the NF rejected most humics and building blocks and a significant part of LMW neutrals. Simultaneous occurrence of cake layer and standard blocking during the filtration process of both UF and NF was identified according to constant-pressure filtration equations, which was possibly caused by the heterogeneous nature of EfOM with a wide MW distribution (several ten to several million Dalton). Thus the corresponding two fouling indices (kc for cake layer and ks for standard blocking) from UF and NF could characterize the fouling potential of macromolecular biopolymers and low to intermediate MW organics (including humics, building blocks, LMW neutrals), respectively. Compared with macromolecular biopolymers, low to intermediate MW organics exhibited a much higher fouling potential due to their lower molecular weight and higher concentration.

  4. Removal of emerging contaminants by simultaneous application of membrane ultrafiltration, activated carbon adsorption, and ultrasound irradiation

    Secondes, Mona Freda N.; Naddeo, Vincenzo; Belgiorno, Vincenzo; Ballesteros, Florencio

    2014-01-01

    Highlights: • Above 99% of the emerging contaminants were removed in the USAMe process. • Influence of PAC dose and US frequency on removal is studied. • Improved performance is due to PAC adsorption enhancement and sonolytic degradation. • US irradiation improved efficiency and delayed declines in the removal of contaminants. • Performance of the hybrid process is better under lower frequency ultrasound irradiation. -- Abstract: Advanced wastewater treatment is necessary to effectively remove emerging contaminants (ECs) with chronic toxicity, endocrine disrupting effects, and the capability to induce the proliferation of highly resistant microbial strains in the environment from before wastewater disposal or reuse. This paper investigates the efficiency of a novel hybrid process that applies membrane ultrafiltration, activated carbon adsorption, and ultrasound irradiation simultaneously to remove ECs. Diclofenac, carbamazepine, and amoxicillin are chosen for this investigation because of their assessed significant environmental risks. Removal mechanisms and enhancement effects are analysed in single and combined processes. The influence of adsorbent dose and ultrasonic frequency to EC removal are also investigated. Results suggest that adsorption is probably the main removal mechanism and is affected by the nature of ECs and the presence of other components in the mixture. Almost complete removals are achieved in the hybrid process for all ECs

  5. Removal of emerging contaminants by simultaneous application of membrane ultrafiltration, activated carbon adsorption, and ultrasound irradiation

    Secondes, Mona Freda N. [Environmental Engineering Graduate Program, Department of Chemical Engineering, University of the Philippines – Diliman, Quezon City (Philippines); Naddeo, Vincenzo, E-mail: vnaddeo@unisa.it [Sanitary and Environmental Engineering Division (SEED), Department of Civil Engineering, University of Salerno, Fisciano 84084 (Saudi Arabia) (Italy); Belgiorno, Vincenzo [Sanitary and Environmental Engineering Division (SEED), Department of Civil Engineering, University of Salerno, Fisciano 84084 (Saudi Arabia) (Italy); Ballesteros, Florencio [Environmental Engineering Graduate Program, Department of Chemical Engineering, University of the Philippines – Diliman, Quezon City (Philippines)

    2014-01-15

    Highlights: • Above 99% of the emerging contaminants were removed in the USAMe process. • Influence of PAC dose and US frequency on removal is studied. • Improved performance is due to PAC adsorption enhancement and sonolytic degradation. • US irradiation improved efficiency and delayed declines in the removal of contaminants. • Performance of the hybrid process is better under lower frequency ultrasound irradiation. -- Abstract: Advanced wastewater treatment is necessary to effectively remove emerging contaminants (ECs) with chronic toxicity, endocrine disrupting effects, and the capability to induce the proliferation of highly resistant microbial strains in the environment from before wastewater disposal or reuse. This paper investigates the efficiency of a novel hybrid process that applies membrane ultrafiltration, activated carbon adsorption, and ultrasound irradiation simultaneously to remove ECs. Diclofenac, carbamazepine, and amoxicillin are chosen for this investigation because of their assessed significant environmental risks. Removal mechanisms and enhancement effects are analysed in single and combined processes. The influence of adsorbent dose and ultrasonic frequency to EC removal are also investigated. Results suggest that adsorption is probably the main removal mechanism and is affected by the nature of ECs and the presence of other components in the mixture. Almost complete removals are achieved in the hybrid process for all ECs.

  6. Membrane-tethered peptides patterned after the TRP domain (TRPducins) selectively inhibit TRPV1 channel activity.

    Valente, Pierluigi; Fernández-Carvajal, Asia; Camprubí-Robles, María; Gomis, Ana; Quirce, Susana; Viana, Félix; Fernández-Ballester, Gregorio; González-Ros, José M; Belmonte, Carlos; Planells-Cases, Rosa; Ferrer-Montiel, Antonio

    2011-05-01

    The transient receptor potential vanilloid 1 (TRPV1) channel is a thermosensory receptor implicated in diverse physiological and pathological processes. The TRP domain, a highly conserved region in the C terminus adjacent to the internal channel gate, is critical for subunit tetramerization and channel gating. Here, we show that cell-penetrating, membrane-anchored peptides patterned after this protein domain are moderate and selective TRPV1 antagonists both in vitro and in vivo, blocking receptor activity in intact rat primary sensory neurons and their peripheral axons with mean decline time of 30 min. The most potent lipopeptide, TRP-p5, blocked all modes of TRPV1 gating with micromolar efficacy (IC(50)100 μM). TRP-p5 did not affect the capsaicin sensitivity of the vanilloid receptor. Our data suggest that TRP-p5 interferes with protein-protein interactions at the level of the TRP domain that are essential for the "conformational" change that leads to gate opening. Therefore, these palmitoylated peptides, which we termed TRPducins, are noncompetitive, voltage-independent, sequence-specific TRPV1 blockers. Our findings indicate that TRPducin-like peptides may embody a novel molecular strategy that can be exploited to generate a selective pharmacological arsenal for the TRP superfamily of ion channels.

  7. Performance enhancement with powdered activated carbon (PAC) addition in a membrane bioreactor (MBR) treating distillery effluent

    Satyawali, Yamini; Balakrishnan, Malini

    2009-01-01

    This work investigated the effect of powdered activated carbon (PAC) addition on the operation of a membrane bioreactor (MBR) treating sugarcane molasses based distillery wastewater (spentwash). The 8 L reactor was equipped with a submerged 30 μm nylon mesh filter with 0.05 m 2 filtration area. Detailed characterization of the commercial wood charcoal based PAC was performed before using it in the MBR. The MBR was operated over 200 days at organic loading rates (OLRs) varying from 4.2 to 6.9 kg m -3 d -1 . PAC addition controlled the reactor foaming during start up and enhanced the critical flux by around 23%; it also prolonged the duration between filter cleaning. Operation at higher loading rates was possible and for a given OLR, the chemical oxygen demand (COD) removal was higher with PAC addition. However, biodegradation in the reactor was limited and the high molecular weight compounds were not affected by PAC supplementation. The functional groups on PAC appear to interact with the polysaccharide portion of the sludge, which may reduce its propensity to interact with the nylon mesh.

  8. Palladium based membranes and membrane reactors for hydrogen production and purification : An overview of research activities at Tecnalia and TU/e

    Fernandez, E.; Helmi Siasi Farimani, A.; Medrano Jimenez, J.A.; Coenen, K.T.; Arratibel Plazaola, A.; Melendez Rey, J.; de Nooijer, N.C.A.; Viviente, J.L.; Zuniga, J.; van Sint Annaland, M.; Gallucci, F.; Pacheco Tanaka, D.A.

    2017-01-01

    In this paper, the main achievements of several European research projects on Pd based membranes and Pd membrane reactors for hydrogen production are reported. Pd-based membranes have received an increasing interest for separation and purification of hydrogen. In addition, the integration of such

  9. A micromachined membrane-based active probe for biomolecular mechanics measurement

    Torun, H.; Sutanto, J.; Sarangapani, K. K.; Joseph, P.; Degertekin, F. L.; Zhu, C.

    2007-04-01

    A novel micromachined, membrane-based probe has been developed and fabricated as assays to enable parallel measurements. Each probe in the array can be individually actuated, and the membrane displacement can be measured with high resolution using an integrated diffraction-based optical interferometer. To illustrate its application in single-molecule mechanics experiments, this membrane probe was used to measure unbinding forces between L-selectin reconstituted in a polymer-cushioned lipid bilayer on the probe membrane and an antibody adsorbed on an atomic force microscope cantilever. Piconewton range forces between single pairs of interacting molecules were measured from the cantilever bending while using the membrane probe as an actuator. The integrated diffraction-based optical interferometer of the probe was demonstrated to have floor for frequencies as low as 3 Hz with a differential readout scheme. With soft probe membranes, this low noise level would be suitable for direct force measurements without the need for a cantilever. Furthermore, the probe membranes were shown to have 0.5 µm actuation range with a flat response up to 100 kHz, enabling measurements at fast speeds.

  10. Prevention of childhood obesity through motivation to physical activity

    Carrillo Aguilera, Sonia

    2012-07-01

    Full Text Available This paper aims to review the current worrying situation in terms of physical activity in our country and the problem that leads us to be below the European average, with the attendant problems of obesity, particularly among children, which follow from this. We analyzed the intervention programs that are being used as PIOBIN plan (The Andalusian Plan for Childhood Obesity, effective from 2007-12, based on a national strategy called Naos Strategy and how different studies support that some intrinsic motivation toward physical activity helps to create lasting habits to the practice. We also carry out an analysis of the different Motivation theories and we base our study on the Self-determination Theory of Deci and Ryan (1985, 2000

  11. Religious Activities and Suicide Prevention: A Gender Specific Analysis

    Steven Stack

    2018-04-01

    Full Text Available The present analysis contributes to the existing literature on religion and suicide in three interrelated ways: (1 providing an analysis of suicide completions whereas most research is based on non-lethal levels of suicidality; (2 assessing the relationship with concrete individual level data on completed suicides instead of aggregated data marked by the ecological fallacy issue; and (3 providing gender specific analyses to determine if the relationship is gendered. METHODS. Data come from the U.S. Public Health Service, National Mortality Followback Survey. They refer to 16,795 deaths including 1385 suicides. Significant others of the deceased were interviewed to measure all variables. The dependent variable is a binary variable where 1 = death by suicide and 0 = all other causes. The central independent variable is an index of religious activities. Controls are included for five categories of confounders (1 psychiatric morbidity; (2 help-seeking behavior; (3 Opportunity factors such as firearms; (4 social integration; and (5 demographics. RESULTS. Multivariate logistic regression analysis determined that controlling for 16 predictors of suicide, a one unit increase in religious activities reduced the odds of a suicide death by 17% for males and by 15% for females. The difference in coefficients is not significant (Z = 0.51. Other significant predictors of suicide deaths included suicide ideation (OR = 8.87, males, OR = 11.48, females and firearm availability (OR = 4.21, males, OR = 2.83, females. DISCUSSION. Religious activities were found to lower suicide risk equally for both men and women. Further work is needed to assess pathways, including suicide ideation, between religious activities and lowered suicide risk. This is the first U.S. based study to test for a gendered association between religion and suicide at the individual level of analysis.

  12. Optimal Activation of Isopsoralen To Prevent Amplicon Carryover

    Fahle, Gary A.; Gill, Vee J.; Fischer, Steven H.

    1999-01-01

    We compared the efficiencies of activation of the photochemical isopsoralen compound 10 and its resulting amplicon neutralizations under conditions with a UV transilluminator box at room temperature (RT) and a HRI-300 UV photothermal reaction chamber at RT and at 5°C. Our data suggest that use of the HRI-300 reaction chamber at 5°C results in a statistically significantly higher degree of amplicon neutralization.

  13. Expression of a constitutively activated plasma membrane H+-ATPase in Nicotiana tabacum BY-2 cells results in cell expansion.

    Niczyj, Marta; Champagne, Antoine; Alam, Iftekhar; Nader, Joseph; Boutry, Marc

    2016-11-01

    Increased acidification of the external medium by an activated H + -ATPase results in cell expansion, in the absence of upstream activating signaling. The plasma membrane H + -ATPase couples ATP hydrolysis with proton transport outside the cell, and thus creates an electrochemical gradient, which energizes secondary transporters. According to the acid growth theory, this enzyme is also proposed to play a major role in cell expansion, by acidifying the external medium and so activating enzymes that are involved in cell wall-loosening. However, this theory is still debated. To challenge it, we made use of a plasma membrane H + -ATPase isoform from Nicotiana plumbaginifolia truncated from its C-terminal auto-inhibitory domain (ΔCPMA4), and thus constitutively activated. This protein was expressed in Nicotiana tabacum BY-2 suspension cells using a heat shock inducible promoter. The characterization of several independent transgenic lines showed that the expression of activated ΔCPMA4 resulted in a reduced external pH by 0.3-1.2 units, as well as in an increased H + -ATPase activity by 77-155 % (ATP hydrolysis), or 70-306 % (proton pumping) of isolated plasma membranes. In addition, ΔCPMA4-expressing cells were 17-57 % larger than the wild-type cells and displayed abnormal shapes. A proteomic comparison of plasma membranes isolated from ΔCPMA4-expressing and wild-type cells revealed the altered abundance of several proteins involved in cell wall synthesis, transport, and signal transduction. In conclusion, the data obtained in this work showed that H + -ATPase activation is sufficient to induce cell expansion and identified possible actors which intervene in this process.

  14. Thyroid screening in pregnancy - a compulsory preventive activity

    Scrinic Olesea

    2015-08-01

    Full Text Available Obiectives: To assess the prevalence of thyroid dysfunction in a group of pregnant women, originating from Dobrogea region of southeastern Romania, considered to be an area without iodine deficiency, including the Black Sea area. Materials and methods: We enrolled 324 pregnant women in different trimesters of pregnancy. Each case was reviewed by a detailed madical history, clinical examination and by serum dosage of thyroid hormones: TSH, FT4, and the antithyroidperoxidase. They were evaluated by comparison with trimester -specific reference range for TSH recommended by American Thyroid Association, then the results were compared with those obtained using the manufacturers reference range. Abortion rate was also analysed. Results: The prevalence of thyroid dysfunction was different in all the 3 trimesters: subclinical hypothyroidism being the most frequently approx. 24% of all cases; 7% of pregnant women had overt hypothyroidism. Incidence of thyrotoxicosis in entire study cases was approx. 5.5%. The most frecvent thyroid autoimune disorders were Hashimoto thyroiditis: 42 % - I trimester, 26,6% in II trimester and about 12,5 % in III-trimester; Graves disease have an incidence of only 0,9 % (n=3.The difference between reference methods eluded a lower number of cases using manufactures reference range for TSH (P< 0,001, but higher for recommended trimester - specific TSH value, confirming the undervalueted hypothesis. The risk of misclassifying the hypothyroidism is between 3 %-8 %. Conclusion: Necessity for thyroid hormone dosage periodic/trimesterly/ in pregnancy is a preventive measure. The reference values for hormonal dosage requires trimester-specific assessment. The possibility of hormonal disorders during pregnancy is common. The need for specific therapy at diagnosis depends on the nature of hormonal disorder. Further precautions are needed in pregnant women with known autoimmune thyroid disorder or newly diagnosed

  15. [Effect of damage integrity rat brain synaptic membranes on the functional activity GABA(A)-receptor/Cl(-)-ionophore complex in the CNC].

    Rebrov, I G; Kalinina, M V

    2013-01-01

    Functional activity of the CGABA(A)-receptor/Cl(-) ionophore complex was investigated the muscimol-stimulated entry of the radioactive isotope 36Cl(-) in synaptoneurosomes in changing the structure and permeability of neuronal membranes. Integrity of the membranes was damaged by removal of Ca(+2) and Mg(+2) from the incubation medium and by the method of freezing-thawing synaptoneurosomes. In both cases, an increase in basal 36Cl(-) entry into synaptoneurosomes, indicating increased nonspecific permeability of neuronal membranes, and decreased activity the CABA(A)-receptor/Cl(-) ionophore complex. The conclusion about the relationship of processes damage neuronal membranes and reducing the inhibitory processes in the epileptic focus.

  16. Regulation of mitogen-activated protein kinase pathways by the plasma membrane Na+/H+ exchanger, NHE1

    Pedersen, Stine Helene Falsig; Darborg, Barbara Vasek; Rentsch, Maria Louise

    2006-01-01

    activity is regulated by a three-tiered phosphorelay system, which is in turn regulated by a complex network of signaling events and scaffolding proteins. The ubiquitous plasma membrane Na(+)/H(+) exchanger NHE1 is activated by, and implicated in, the physiological/pathophysiological responses to many...... of the same stimuli that modulate MAPK activity. While under some conditions, NHE1 is regulated by MAPKs, a number of studies have, conversely, implicated NHE1 in the regulation of MAPK activity. Here, we discuss the current evidence indicating the involvement of NHE1 in MAPK regulation, the mechanisms...

  17. The effect of sodium hyaluronate-carboxymethyl cellulose membrane in the prevention of parenchymal air leaks: an experimental and manometric study in rats.

    Büyükkale, Songül; Çıtak, Necati; İşgörücü, Özgür; Sayar, Adnan

    2017-12-01

    We aimed to examine effectiveness of sodium hyaluronate-carboxymethly cellulose (NaH/CMC) for sealing pulmonary air leaks during postoperative period. The study was conducted in 16 male Sprague-Dawley rats. A linear insicion (length= 0.2 cm, depth= 0.1 cm) to the lung parenchyma on the inflated by a cutter was made. The animals were randomly divided; the control group (n= 8) and NaH/CMC-treated group (the study group, n= 8). Control group was left for physiologic healing while a NaH/CMC membrane was applied over the the incisional area in the study group. Then the pressure point where the air leakage observed was noted. No polymorphonucleer leucocytes (PMNL) infiltration was detected in control group, whereas PMNL infiltration was 0.38 ± 0.5 cell per 100 high field in study group (p= 0.234). The degree of macrophage, lymphocyte infiltration and the mean fibroblast count were found to be higher in study group compared with control group (p= 0.007, p= 0.02, p= 0.05, respectively). The mean pressure value for air leak to occur in the control group was 43.50 ± 9.55 mmHg whereas it was 73.75 ± 16.68 mmHg in the study group (p< 0.001). The data revealed that bioabsorbable NaH/CMC membrane accelerates healing with preserving the expansile character of lung parenchyma even in high ventilation pressures. However, further studies are required to assess the prevent impact of the pulmonary air-leak for NaH/CMC membrane.

  18. Persistent alterations in active and passive electrical membrane properties of regenerated nerve fibers of man and mice

    Moldovan, Mihai; Alvarez Herrero, Susana; Rosberg, Mette R.

    2016-01-01

    Excitability of regenerated fibers remains impaired due to changes in both passive cable properties and alterations in the voltage-dependent membrane function. These abnormalities were studied by mathematical modeling in human regenerated nerves and experimental studies in mice. In three adult male...... activity protocol triggered partial Wallerian degeneration in regenerated nerves but not in control nerves from age-matched mice. The current data suggest that the nodal voltage-gated ion channel machinery is restored in regenerated axons, although the electrical separation from the internodal compartment...... remains compromised. Due to the persistent increase in number of nodes, the increased activity-dependent Na+ influx could lead to hyperactivity of the Na+/K+ pump resulting in membrane hyperpolarization and neurotoxic energy insufficiency during strenuous activity....

  19. Peptides actively transported across the tympanic membrane: Functional and structural properties.

    Arwa Kurabi

    Full Text Available Otitis media (OM is the most common infectious disease of children under six, causing more antibiotic prescriptions and surgical procedures than any other pediatric condition. By screening a bacteriophage (phage library genetically engineered to express random peptides on their surfaces, we discovered unique peptides that actively transport phage particles across the intact tympanic membrane (TM and into the middle ear (ME. Herein our goals were to characterize the physiochemical peptide features that may underlie trans-TM phage transport; assess morphological and functional effects of phage peptides on the ME and inner ear (IE; and determine whether peptide-bearing phage transmigrate from the ME into the IE. Incubation of five peptide-bearing phage on the TM for over 4hrs resulted in demonstrably superior transport of one peptide, in level and in exponential increase over time. This suggests a preferred peptide motif for TM active transport. Functional and structural comparisons revealed unique features of this peptide: These include a central lysine residue, isoelectric point of 0.0 at physiological pH and a hydrophobic C-terminus. When the optimal peptide was applied to the TM independent of phage, similar transport was observed, indicating that integration into phage is not required. When 109 particles of the four different trans-TM phage were applied directly into the ME, no morphological effects were detected in the ME or IE when compared to saline or wild-type (WT phage controls. Comparable, reversible hearing loss was observed for saline controls, WT phage and trans-TM peptide phage, suggesting a mild conductive hearing loss due to ME fluid. Perilymph titers after ME incubation established that few copies of trans-TM peptide phage crossed into the IE. The results suggest that, within the parameters tested, trans-TM peptides are safe and could be used as potential agents for noninvasive delivery of drugs, particles and gene therapy

  20. Preventing anaemia in pregnancy - need for intensive IEC Activities

    A K Bhardwaj

    1995-09-01

    Full Text Available A Study to know the compliance rale of IFA tablets in respect of collection and consumption was carried out in three districts of Himachal Pradesh covering 90 clusters. Out of total women interviewed with childless than one year, only 94.8% had collected IFA tablets. 41.9% and 10% women had consumed these tablets for 60 and 100 days respectively. Majority of women did not consume these tablets with the reason that medicine is taken on ly during illness and as such they don't require these tablets. Intrusive Information, Education and Communication activities are stressed in the paper.

  1. Flunarizine Prevents Hepatitis C Virus Membrane Fusion in a Genotype-dependent Manner by Targeting the Potential Fusion Peptide within E1

    Perin, Paula M.; Haid, Sibylle; Brown, Richard J. P.; Doerrbecker, Juliane; Schulze, Kai; Zeilinger, Carsten; von Schaewen, Markus; Heller, Brigitte; Vercauteren, Koen; Luxenburger, Eva; Baktash, Yasmine M.; Vondran, Florian W. R.; Speerstra, Sietkse; Awadh, Abdullah; Mukhtarov, Furkat; Schang, Luis M; Kirschning, Andreas; Müller, Rolf; Guzman, Carlos A.; Kaderali, Lars; Randall, Glenn; Meuleman, Philip; Ploss, Alexander; Pietschmann, Thomas

    2015-01-01

    To explore mechanisms of hepatitis C virus (HCV) replication we screened a compound library including licensed drugs. Flunarizine, a diphenylmethylpiperazine used to treat migraine, inhibited HCV cell entry in vitro and in vivo in a genotype-dependent fashion. Analysis of mosaic viruses between susceptible and resistant strains revealed that E1 and E2 glycoproteins confer susceptibility to flunarizine. Time of addition experiments and single particle tracking of HCV demonstrated that flunarizine specifically prevents membrane fusion. Related phenothiazines and pimozide also inhibited HCV infection and preferentially targeted HCV genotype 2 viruses. However, phenothiazines and pimozide exhibited improved genotype coverage including the difficult to treat genotype 3. Flunarizine-resistant HCV carried mutations within the alleged fusion peptide and displayed cross-resistance to these compounds, indicating that these drugs have a common mode of action. Conclusion: These observations reveal novel details about HCV membrane fusion. Moreover, flunarizine and related compounds represent first-in-class HCV fusion inhibitors that merit consideration for repurposing as cost-effective component of HCV combination therapies. PMID:26248546

  2. Combined HLA matched limbal stem cells allograft with amniotic membrane transplantation as a prophylactic surgical procedure to prevent corneal graft rejection after penetrating keratoplasty: case report

    Paolo Capozzi

    2014-09-01

    Full Text Available Purpose. To determine if the use of combined HLA matched limbal stem cells allograft with amniotic membrane transplantation (AMT is a safe and effective prophylactic surgical procedure to prevent corneal graft after penetrating keratoplasty (PK. Methods. We report the case of a 17 years old patient with a history of congenital glaucoma, trabeculectomy and multiple corneal graft rejections, presenting total limbal cell deficiency. To reduce the possibility of graft rejection in the left eye after a new PK, a two step procedure was performed. At first the patient underwent a combined HLA matched limbal stem cells allograft (LAT and AMT and then, 10 months later, a new PK. Results. During 12 months of follow-up, the corneal graft remained stable and smooth, with no sign of graft rejection. Conclusions. In our patient, the prophylactic use of LAT from HLA-matched donors and AMT before PK, may result in a better prognosis of corneal graft survival.

  3. Complement activation and formation of the membrane attack complex on serogroup B Neisseria meningitidis in the presence or absence of serum bactericidal activity

    Drogari-Apiranthitou, M.; Kuijper, E. J.; Dekker, N. [=Nick; Dankert, J.

    2002-01-01

    Encapsulated meningococci are complement sensitive only in the presence of bactericidal antibodies by yet-unexplored mechanisms. The objective of this study was to investigate the involvement of major bacterial surface constituents on complement activation and membrane attack complex (MAC) formation

  4. Modulation of membrane potential by an acetylcholine-activated potassium current in trout atrial myocytes

    Molina, C.E.; Gesser, Hans; Llach, A.

    2007-01-01

    mV from 4.3 pA/pF to 27 pA/pF with an EC50 of 45 nM in atrial myocytes. Moreover, 3 nM ACh increased the slope conductance of Im fourfold, shifted its reversal potential from -78 ± 3 to -84 ± 3 mV, and stabilized the resting membrane potential at -92 ± 4 mV. ACh also shortened the action potential...... hypothesized that this is at least partly due to a small slope conductance of Im around the resting membrane potential in atrial myocytes. In accordance with this hypothesis, the slope conductance of Im was about sevenfold smaller in atrial than in ventricular myocytes. Interestingly, ACh increased Im at -120...... of an inwardly rectifying K+ current can modulate the membrane potential in the trout atrial myocytes and stabilize the resting membrane potential. teleost heart; IK,ACh; cholinergic modulation; action potential...

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

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

    2016-01-01

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

  6. Application of Pre-coated Microfiltration Ceramic Membrane with Powdered Activated Carbon for Natural Organic Matter Removal from Secondary Wastewater Effluent

    Kurniasari, Novita

    2012-12-01

    Ceramic membranes offer more advantageous performances than conventional polymeric membranes. However, membrane fouling caused by Natural Organic Matters (NOM) contained in the feed water is still become a major problem for operational efficiency. A new method of ceramic membrane pre-coating with Powdered Activated Carbon (PAC), which allows extremely contact time for adsorbing aquatic contaminants, has been studied as a pre-treatment prior to ceramic microfiltration membrane. This bench scale study evaluated five different types of PAC (SA Super, G 60, KCU 6, KCU 8 and KCU 12,). The results showed that KCU 6 with larger pore size was performed better compared to other PAC when pre-coated on membrane surface. PAC pre-coating on the ceramic membrane with KCU 6 was significantly enhance NOM removal, reduced membrane fouling and improved membrane performance. Increase of total membrane resistance was suppressed to 96%. The removal of NOM components up to 92%, 58% and 56% for biopolymers, humic substances and building blocks, respectively was achieved at pre-coating dose of 30 mg/l. Adsorption was found to be the major removal mechanism of NOM. Results obtained showed that biopolymers removal are potentially correlated with enhanced membrane performance.

  7. Sterol-recognition ability and membrane-disrupting activity of Ornithogalum saponin OSW-1 and usual 3-O-glycosyl saponins.

    Malabed, Raymond; Hanashima, Shinya; Murata, Michio; Sakurai, Kaori

    2017-12-01

    OSW-1 is a structurally unique steroidal saponin isolated from the bulbs of Ornithogalum saundersiae, and has exhibited highly potent and selective cytotoxicity in tumor cell lines. This study aimed to investigate the molecular mechanism for the membrane-permeabilizing activity of OSW-1 in comparison with those of other saponins by using various spectroscopic approaches. The membrane effects and hemolytic activity of OSW-1 were markedly enhanced in the presence of membrane cholesterol. Binding affinity measurements using fluorescent cholestatrienol and solid-state NMR spectroscopy of a 3-d-cholesterol probe suggested that OSW-1 interacts with membrane cholesterol without forming large aggregates while 3-O-glycosyl saponin, digitonin, forms cholesterol-containing aggregates. The results suggest that OSW-1/cholesterol interaction is likely to cause membrane permeabilization and pore formation without destroying the whole membrane integrity, which could partly be responsible for its highly potent cell toxicity. Copyright © 2017 Elsevier B.V. All rights reserved.

  8. Gene-environment interactions in considering physical activity for the prevention of dementia

    Kristyn Alissa Bates

    2015-09-01

    Full Text Available Alzheimer's disease (AD, the most common neurodegenerative disease worldwide, ranks as one of the most feared diseases in the world. Similarly, recent studies suggest that AD may be the third leading cause of death in the United States, behind heart disease and cancer. In the absence of a cure or effective treatment, strategies to prevent or delay the onset and progression of the disease are desperately needed. Decades of research have identified key risk and protective factors including genetic polymorphism in the APOE gene, age and lifestyle factors. Physical activity (PA is emerging as an attractive primary prevention strategy. This review will summarise the latest findings supporting the role of physical activity in the prevention of AD, including possible mechanisms and the influence of genetics on disease prevention. Given that AD and other dementias are recognised as a world health priority, public health strategies are needed to incorporate promoting the health benefits of physical activity across the lifespan.

  9. Prevention of corruption as administrative activity of authorities and the object of prosecutor’s observance

    Timur L. Kozlov

    2017-03-01

    Full Text Available Objective to study corruption prevention as administrative activity of state and municipal authorities. Methods dialectical systematic formallegal analysis synthesis statistical and specific sociological methods. Results basing on the study of countercorruption legislation its implementation and acts of prosecutorial response the definition of the concept quotcorruption preventionquot was formulated which refers to administrative activities of state and municipal bodies and their officials aimed at the implementation of legislation on combating corruption. The feature of the preventive component of countercorruption legislation is that it brings together the complex of binding prohibitive and restrictive rules established for the prevention of corruption. Corruption prevention is implemented through a specific type of managerial administrative activity. The legality of this activity is provided by means of prosecutorial supervision over implementation of laws on corruption prevention. Scientific novelty for the first time in the article corruption prevention is viewed not as a criminological category but as an institution of administrative law characterized by the essential features of the state municipal control. Practical significance the main provisions and conclusions of the article can be used in research and lawenforcement activities in addressing issues related to combating corruption.

  10. Telmisartan prevents weight gain and obesity through activation of peroxisome proliferator-activated receptor-delta-dependent pathways

    He, Hongbo; Yang, Dachun; Ma, Liqun

    2010-01-01

    Telmisartan shows antihypertensive and several pleiotropic effects that interact with metabolic pathways. In the present study we tested the hypothesis that telmisartan prevents adipogenesis in vitro and weight gain in vivo through activation of peroxisome proliferator-activated receptor (PPAR)-d...

  11. Evaluation of the Membrane Permeability (PAMPA and Skin) of Benzimidazoles with Potential Cannabinoid Activity and their Relation with the Biopharmaceutics Classification System (BCS)

    Alvarez-Figueroa, M. Javiera; Pessoa-Mahana, C. David; Palavecino-González, M. Elisa; Mella-Raipán, Jaime; Espinosa-Bustos, Cristián; Lagos-Muñoz, Manuel E.

    2011-01-01

    The permeability of five benzimidazole derivates with potential cannabinoid activity was determined in two models of membranes, parallel artificial membrane permeability assay (PAMPA) and skin, in order to study the relationship of the physicochemical properties of the molecules and characteristics of the membranes with the permeability defined by the Biopharmaceutics Classification System. It was established that the PAMPA intestinal absorption method is a good predictor for classifying thes...

  12. BID is cleaved by caspase-8 within a native complex on the mitochondrial membrane

    Schug, Z. T.; Gonzalvez, F.; Houtkooper, R. H.; Vaz, F. M.; Gottlieb, E.

    2011-01-01

    Caspase-8 stably inserts into the mitochondrial outer membrane during extrinsic apoptosis. Inhibition of caspase-8 enrichment on the mitochondria impairs caspase-8 activation and prevents apoptosis. However, the function of active caspase-8 on the mitochondrial membrane remains unknown. In this

  13. Effect of Light-Activated Hypocrellin B on the Growth and Membrane Permeability of Gram-Negative Escherichia coli Cells

    Yuan Jiang

    2014-01-01

    Full Text Available Aim. To investigate the effect of light-activated hypocrellin B on the growth and membrane permeability of Gram-negative bacteria. Methods. Escherichia coli (E. coli as a model bacterium of Gram-negative bacteria was incubated with various concentrations of hypocrellin B for 60 min and was subsequently irradiated by blue light with wavelength of 470 nm at the dose of 12 J/cm2. Colony forming units were counted and the growth inhibition rate of E. coli cells was calculated after light-activated hypocrellin B. Membrane permeability was measured using flow cytometry and confocal laser scanning microscopy (CLSM with propidium iodide (PI staining. Bacterial morphology was observed using transmission electron microscopy (TEM. Reactive oxygen species in bacterial cells were measured using flow cytometry with DCFH-DA staining. Results. Significant growth inhibition rate of E. coli cells was observed after photodynamic action of hypocrellin B. Remarkable damage to the ultrastructure of E. coli was also observed by TEM. Flow cytometry and CLSM observation showed that light-activated hypocrellin B markedly increased membrane permeability of E. coli. Flow cytometry showed the intracellular ROS increase in E. coli treated by photodynamic action of hypocrellin B. Conclusion. Light-activated hypocrellin B caused intracellular ROS increase and structural damages and inhibited the growth of Gram-negative E. coli cells.

  14. Cadmium induces changes in sucrose partitioning, invertase activities, and membrane functionality in roots of Rangpur lime (Citrus limonia L. Osbeck).

    Podazza, G; Rosa, M; González, J A; Hilal, M; Prado, F E

    2006-09-01

    Cadmium (Cd) uptake effects on sucrose content, invertase activities, and plasma membrane functionality were investigated in Rangpur lime roots ( CITRUS LIMONIA L. Osbeck). Cadmium accumulation was significant in roots but not in shoots and leaves. Cadmium produced significant reduction in roots DW and increment in WC. Leaves and shoots did not show significant differences on both parameters. Sucrose content was higher in control roots than in Cd-exposed ones. Apoplastic sucrose content was much higher in Cd-exposed roots than in control ones. Cd-exposed roots showed a significant decrease in both cell wall-bound and cytoplasmic (neutral) invertase activities; while the vacuolar isoform did not show any change. Alterations in lipid composition and membrane fluidity of Cd-exposed roots were also observed. In Cd-exposed roots phospholipid and glycolipid contents decreased about 50 %, while sterols content was reduced about 22 %. Proton extrusion was inhibited by Cd. Lipid peroxidation and proton extrusion inhibition were also detected by histochemical analysis. This work's findings demonstrate that Cd affects sucrose partitioning and invertase activities in apoplastic and symplastic regions in Rangpur lime roots as well as the plasma membrane functionality and H (+)-ATPase activity.

  15. [Study on membrane type leaf water evaporation inhibitors for improving effect of preventing diseases and pest controlling of Lycium barbarum].

    Wang, Dan-Dan; Lv, Zhe; Xu, Chang-Qing; Liu, Sai; Chen, Jun; Peng, Xiao; Wu, Yan

    2018-01-01

    Through indoor and field comparative experiments, the properties of membrane type leaf evaporation inhibitors and its effects on photosynthesis of Lycium barbarum and compatibility and synergistic of pesticide were studied. The evaporation inhibitors and L. barbarum were chosen to investigate the suppression of water evaporation and the compatibility with pesticides. The effect of evaporation inhibitors on photosynthesis of L. barbarum leaves was determined by the chlorophyll fluorescence imaging system. The results showed that water evaporation of L. barbarum leaves of different leaf age were evidently suppressed after treated with evaporation inhibitor. The inhibitor was well compatible with pesticide and effectively improved the pesticide efficacy,and had no significant effect on chlorophyll fluorescence parameters. It is concluded that the evaporation inhibitor has good compatibility with the pesticide, and has remarkable effect of restraining moisture evaporation, which make it can be used for reducing the dosage and improving the efficacy of the pesticide in the field of L. barbarum. Copyright© by the Chinese Pharmaceutical Association.

  16. Construction and Antiapoptosis Activities of Recombinant Adenoviral Expression Vector Carrying EBV Latent Membrane Protein 2A

    Xishuang Liu

    2011-01-01

    Full Text Available To evaluate the possible effects of LMP2A (EBV latent membrane protein 2A on human gastric cancer cell line SGC-7901, LMP2A coding gene was subcloned into shuttle plasmid pAdTrackCMV to form transfer plasmid pAdTrackCMV-2A, which was linearized with PmeI and cotransformed into E.coli BJ5183 with adenovirus genomic plasmid of pAdeasy-1. The identified recombinant adenovirus plasmid DNA was digested with PacI and transfected into 293 cells to package recombinant adenovirus particles named vAd-2A. Then the expression and antiapoptosis activities of LMP2A on SGC-7901 infected with vAd-2A were analyzed. The vAd-2A was successfully constructed and identified by PCR, restriction digestion, and sequencing. LMP2A expression in SGC was identified by strong green fluorescence expression with fluorescence microscopic photograph and Southern blotting. The growth of LMP2A expressing SGC cells was apparently improved. Both cyclin E expression and S phase ratio in LMP2A expressing SGC cells were upregulated by cell cycle analysis and confocal microscopic analysis respectively. The replication-deficient recombinant adenovirus vector can express LMP2A antigen in SGC cells and inhibit their apoptosis. The results indicate that LMP2A might play an important role in pathogenesis of EBV-associated gastric cancer (EBVaGC. This study establishes a foundation for further study on EBVaGC and its gene therapy.

  17. Reduced prostasin (CAP1/PRSS8 activity eliminates HAI-1 and HAI-2 deficiency-associated developmental defects by preventing matriptase activation.

    Roman Szabo

    Full Text Available Loss of either hepatocyte growth factor activator inhibitor (HAI-1 or -2 is associated with embryonic lethality in mice, which can be rescued by the simultaneous inactivation of the membrane-anchored serine protease, matriptase, thereby demonstrating that a matriptase-dependent proteolytic pathway is a critical developmental target for both protease inhibitors. Here, we performed a genetic epistasis analysis to identify additional components of this pathway by generating mice with combined deficiency in either HAI-1 or HAI-2, along with genes encoding developmentally co-expressed candidate matriptase targets, and screening for the rescue of embryonic development. Hypomorphic mutations in Prss8, encoding the GPI-anchored serine protease, prostasin (CAP1, PRSS8, restored placentation and normal development of HAI-1-deficient embryos and prevented early embryonic lethality, mid-gestation lethality due to placental labyrinth failure, and neural tube defects in HAI-2-deficient embryos. Inactivation of genes encoding c-Met, protease-activated receptor-2 (PAR-2, or the epithelial sodium channel (ENaC alpha subunit all failed to rescue embryonic lethality, suggesting that deregulated matriptase-prostasin activity causes developmental failure independent of aberrant c-Met and PAR-2 signaling or impaired epithelial sodium transport. Furthermore, phenotypic analysis of PAR-1 and matriptase double-deficient embryos suggests that the protease may not be critical for focal proteolytic activation of PAR-2 during neural tube closure. Paradoxically, although matriptase auto-activates and is a well-established upstream epidermal activator of prostasin, biochemical analysis of matriptase- and prostasin-deficient placental tissues revealed a requirement of prostasin for conversion of the matriptase zymogen to active matriptase, whereas prostasin zymogen activation was matriptase-independent.

  18. Effect of zinc insertion and hydrophobicity on the membrane interactions and PDT activity of porphyrin photosensitizers.

    Pavani, Christiane; Uchoa, Adjaci F; Oliveira, Carla S; Iamamoto, Yassuko; Baptista, Maurício S

    2009-02-01

    A series of photosensitizers (PS), which are meso-substituted tetra-cationic porphyrins, was synthesized in order to study the role of amphiphilicity and zinc insertion in photodynamic therapy (PDT) efficacy. Several properties of the PS were evaluated and compared within the series including photophysical properties (absorption spectra, fluorescence quantum yield Phif, and singlet oxygen quantum yield PhiDelta), uptake by vesicles, mitochondria and HeLa cells, dark and phototoxicity in HeLa cells. The photophysical properties of all compounds are quite similar (Phifmembrane potential, i.e., in uncoupled mitochondria PS binding decreases by up to 53%. The porphyrin substituted with octyl groups (TC8PyP) is the compound that is most enriched in mitochondria, and its zinc derivative (ZnTC8PyP) has the highest global uptake. The stronger membrane interaction of the zinc-substituted porphyrins is attributed to a complexing effect with phosphate groups of the phospholipids. Zinc insertion was also shown to decrease the interaction with isolated mitochondria and with the mitochondria of HeLa cells, an effect that has been explained by the particular characteristics of the mitochondrial internal membrane. Phototoxicity was shown to increase proportionally with membrane binding efficiency, which is attributed to favorable membrane interactions which allow more efficient membrane photooxidation. For this series of compounds, photodynamic efficiency is directly proportional to the membrane binding and cell uptake, but it is not totally related to mitochondrial targeting.

  19. Membrane fractions active in poliovirus RNA replication contain VPg precursor polypeptides

    Takegami, T.; Semler, B.L.; Anderson, C.W.; Wimmer, E.

    1983-01-01

    The poliovirus specific polypeptide P3-9 is of special interest for studies of viral RNA replication because it contains a hydrophobic region and, separated by only seven amino acids from that region, the amino acid sequence of the genome-linked protein VPg. Membraneous complexes of poliovirus-infected HeLa cells that contain poliovirus RNA replicating proteins have been analyzed for the presence of P3-9 by immunoprecipitation. Incubation of a membrane fraction rich in P3-9 with proteinase leaves the C-terminal 69 amino acids of P3-9 intact, an observation suggesting that this portion is protected by its association with the cellular membrane. These studies have also revealed two hitherto undescribed viral polypeptides consisting of amino acid sequences of the P2 andf P3 regions of the polyprotein. Sequence analysis by stepwise Edman degradation show that these proteins are 3b/9 (M/sub r/77,000) and X/9 (M/sub r/50,000). 3b/9 and X/9 are membrane bound and are turned over rapidly and may be direct precursors to proteins P2-X and P3-9 of the RNA replication complex. P2-X, a polypeptide void of hydrophobic amino acid sequences but also found associated with membranes, is rapidly degraded when the membraneous complex is treated with trypsin. It is speculated that P2-X is associated with membranes by its affinity to the N-terminus of P3-9

  20. Prostate cell membrane chromatography-liquid chromatography-mass spectrometry for screening of active constituents from Uncaria rhynchophylla.

    He, Jianyu; Han, Shengli; Yang, Fangfang; Zhou, Nan; Wang, Sicen

    2013-01-01

    Uncaria rhynchophylla is a traditional Chinese medicinal herb used to treat hypertension and convulsive disorders such as epilepsy. Rat prostate cell membrane chromatography combined with liquid chromatography-mass spectrometry (LC-MS) was used to identify active constituents from U. rhynchophylla extracts. Four compounds (corynoxeine, isorhynchophylline, isocorynoxeine and rhynchophylline) were discovered. Competitive binding assay results indicated that the four compounds were in direct competition at a single common binding site and interacted with α1A adrenergic receptors (α1A-AR) in a manner similar to tamsulosin. Affinity constant values of the four compounds binding with α1A-AR were also measured using rat prostate cell membrane chromatography (CMC). Finally, their pharmacodynamic effects were tested on rat caudal arteries. This CMC combined LC-MS system offers a means of drug discovery by screening natural medicinal herbs for new pharmacologically active molecules targeting specific receptors.

  1. Psychosocial Issues in Engaging Older People with Physical Activity Interventions for the Prevention of Falls

    Nyman, Samuel R.

    2011-01-01

    This article presents an overview of the psychosocial factors that influence older people's participation in physical activity interventions to prevent falls. The importance of psychosocial factors is stressed inasmuch as interventions will be rendered useless if they do not successfully gain the active participation of older people. The theory of…

  2. The membrane-topogenic vectorial behaviour of Nrf1 controls its post-translational modification and transactivation activity.

    Zhang, Yiguo; Hayes, John D

    2013-01-01

    The integral membrane-bound Nrf1 transcription factor fulfils important functions in maintaining cellular homeostasis and organ integrity, but how it is controlled vectorially is unknown. Herein, creative use of Gal4-based reporter assays with protease protection assays (GRAPPA), and double fluorescence protease protection (dFPP), reveals that the membrane-topogenic vectorial behaviour of Nrf1 dictates its post-translational modification and transactivation activity. Nrf1 is integrated within endoplasmic reticulum (ER) membranes through its NHB1-associated TM1 in cooperation with other semihydrophobic amphipathic regions. The transactivation domains (TADs) of Nrf1, including its Asn/Ser/Thr-rich (NST) glycodomain, are transiently translocated into the ER lumen, where it is glycosylated in the presence of glucose to become a 120-kDa isoform. Thereafter, the NST-adjoining TADs are partially repartitioned out of membranes into the cyto/nucleoplasmic side, where Nrf1 is subject to deglycosylation and/or proteolysis to generate 95-kDa and 85-kDa isoforms. Therefore, the vectorial process of Nrf1 controls its target gene expression.

  3. Mitigation of Membrane Biofouling in MBR Using a Cellulolytic Bacterium, Undibacterium sp. DM-1, Isolated from Activated Sludge.

    Nahm, Chang Hyun; Lee, Seonki; Lee, Sang Hyun; Lee, Kibaek; Lee, Jaewoo; Kwon, Hyeokpil; Choo, Kwang-Ho; Lee, Jung-Kee; Jang, Jae Young; Lee, Chung-Hak; Park, Pyung-Kyu

    2017-03-28

    Biofilm formation on the membrane surface results in the loss of permeability in membrane bioreactors (MBRs) for wastewater treatment. Studies have revealed that cellulose is not only produced by a number of bacterial species but also plays a key role during formation of their biofilm. Hence, in this study, cellulase was introduced to a MBR as a cellulose-induced biofilm control strategy. For practical application of cellulase to MBR, a cellulolytic ( i.e ., cellulase-producing) bacterium, Undibacterium sp. DM-1, was isolated from a lab-scale MBR for wastewater treatment. Prior to its application to MBR, it was confirmed that the cell-free supernatant of DM-1 was capable of inhibiting biofilm formation and of detaching the mature biofilm of activated sludge and cellulose-producing bacteria. This suggested that cellulase could be an effective anti-biofouling agent for MBRs used in wastewater treatment. Undibacterium sp. DM-1-entrapping beads ( i.e ., cellulolytic-beads) were applied to a continuous MBR to mitigate membrane biofouling 2.2-fold, compared with an MBR with vacant-beads as a control. Subsequent analysis of the cellulose content in the biofilm formed on the membrane surface revealed that this mitigation was associated with an approximately 30% reduction in cellulose by cellulolytic-beads in MBR.

  4. Fate of antibiotics in activated sludge followed by ultrafiltration (CAS-UF) and in a membrane bioreactor (MBR).

    Sahar, Eyal; Messalem, Rami; Cikurel, Haim; Aharoni, Avi; Brenner, Asher; Godehardt, Manuel; Jekel, Martin; Ernst, Mathias

    2011-10-15

    The fates of several macrolide, sulphonamide, and trimethoprim antibiotics contained in the raw sewage of the Tel-Aviv wastewater treatment plant (WWTP) were investigated after the sewage was treated using either a full-scale conventional activated sludge (CAS) system coupled with a subsequent ultrafiltration (UF) step or a pilot membrane bioreactor (MBR) system. Antibiotics removal in the MBR system, once it achieved stable operation, was 15-42% higher than that of the CAS system. This advantage was reduced to a maximum of 20% when a UF was added to the CAS. It was hypothesized that the contribution of membrane separation (in both systems) to antibiotics removal was due either to sorption to biomass (rather than improvement in biodegradation) or to enmeshment in the membrane biofilm (since UF membrane pores are significantly larger than the contaminant molecules). Batch experiments with MBR biomass showed a markedly high potential for sorption of the tested antibiotics onto the biomass. Moreover, methanol extraction of MBR biomass released significant amounts of sorbed antibiotics. This finding implies that more attention must be devoted to the management of excess sludge. Copyright © 2011 Elsevier Ltd. All rights reserved.

  5. Angiotensin II-induced hypertension increases plasma membrane Na pump activity by enhancing Na entry in rat thick ascending limbs.

    Gonzalez-Vicente, Agustin; Garvin, Jeffrey L

    2013-11-01

    Thick ascending limbs (TAL) reabsorb 30% of the filtered NaCl load. Na enters the cells via apical Na-K-2Cl cotransporters and Na/H exchangers and exits via basolateral Na pumps. Chronic angiotensin II (ANG II) infusion increases net TAL Na transport and Na apical entry; however, little is known about its effects on the basolateral Na pump. We hypothesized that in rat TALs Na pump activity is enhanced by ANG II-infusion, a model of ANG II-induced hypertension. Rats were infused with 200 ng·kg(-1)·min(-1) ANG II or vehicle for 7 days, and TAL suspensions were obtained. We studied plasma membrane Na pump activity by measuring changes in 1) intracellular Na (Nai) induced by ouabain; and 2) ouabain-sensitive oxygen consumption (QO2). We found that the ouabain-sensitive rise in Nai in TALs from ANG II-infused rats was 12.8 ± 0.4 arbitrary fluorescent units (AFU)·mg(-1)·min(-1) compared with only 9.9 ± 1.1 AFU·mg(-1)·min(-1) in controls (P Na pump expression, the number of Na pumps in the plasma membrane, or the affinity for Na. When furosemide (1.1 mg·kg(-1)·day(-1)) was coinfused with ANG II, no increase in plasma membrane Na pump activity was observed. We concluded that in ANG II-induced hypertension Na pump activity is increased in the plasma membrane of TALs and that this increase is caused by the chronically enhanced Na entry occurring in this model.

  6. A bioabsorbable membrane (Seprafilm®) may prevent postoperative mediastinal adhesions following mediastinoscopy: an experimental study in rats.

    Büyükkale, Songül; Çıtak, Necati; İşgörücü, Özgür; Sayar, Adnan

    2015-01-01

    The aim of this experimental study was to investigate the anti-adhesion property of a bioabsorbable membrane following mediastinoscopy in a rat model. The study was conducted in 20 male Sprague-Dawley rats. Mediastinoscopy was performed all of them. Rats were divided into two groups; control group (n=10); mediastinoscopy alone, study group (n=10); mediastinoscopy and sodiumhyaluronate-carboxymethlycellulose film (Seprafilm®; Genzyme Corporation, Cambridge, Mass. USA). It was used to the mediastinal surface at the end of the surgical procedure in study group. Re-mediastinoscopy was performed after 7 days. Adhesion and vascularity grade description scores were analyzed. The parameters evaluated were presence of polymorhphonucleer leucocyte, macrophage, lymphocyte, fibroblasts, edema, neovascularisation, collagenisation, and foreing body reaction. All the rats survived uneventfully until being sacrificed without any postoperative complications. The mean adhesion score was found to be significantly higher in control group (n=2.5±0.5) compared with study group (n=1.0±0.1) (P=0.007). Vascularity grade description score was significantly higher in control group (n=2.3±0.6) than in study group (n=1.4±0.6) (P=0.009). There were no statistical differences between the groups with regard to edema, lymphocyte and macrophage infiltration, fibroblast proliferation and foreign body reactions (P>0.05). The used of Seprafilm® during the primary procedure can reduce to the mediastinal adhesions. However, further studies are required to assess the precise impact of the anti-adhesive agents on adhesion.

  7. Calcium and Magnesium Ions Are Membrane-Active against Stationary-Phase Staphylococcus aureus with High Specificity

    Xie, Yuntao; Yang, Lihua

    2016-02-01

    Staphylococcus aureus (S. aureus) is notorious for its ability to acquire antibiotic-resistance, and antibiotic-resistant S. aureus has become a wide-spread cause of high mortality rate. Novel antimicrobials capable of eradicating S. aureus cells including antibiotic-resistant ones are thus highly desired. Membrane-active bactericides and species-specific antimicrobials are two promising sources of novel anti-infective agents for fighting against bacterial antibiotic-resistance. We herein show that Ca2+ and Mg2+, two alkaline-earth-metal ions physiologically essential for diverse living organisms, both disrupt model S. aureus membranes and kill stationary-phase S. aureus cells, indicative of membrane-activity. In contrast to S. aureus, Escherichia coli and Bacillus subtilis exhibit unaffected survival after similar treatment with these two cations, indicative of species-specific activity against S. aureus. Moreover, neither Ca2+ nor Mg2+ lyses mouse red blood cells, indicative of hemo-compatibility. This works suggests that Ca2+ and Mg2+ may have implications in targeted eradication of S. aureus pathogen including the antibiotic-resistant ones.

  8. Solubilization, partial purification, and reconstitution of glutamate- and N-methyl-D-aspartate-activated cation channels from brain synaptic membranes

    Ly, A.M.; Michaelis, E.K.

    1991-01-01

    L-Glutamate-activated cation channel proteins from rat brain synaptic membranes were solubilized, partially purified, and reconstituted into liposomes. Optimal conditions for solubilization and reconstitution included treatment of the membranes with nonionic detergents in the presence of neutral phospholipids plus glycerol. Quench-flow procedures were developed to characterize the rapid kinetics of ion flux induced by receptor agonists. [ 14 C]Methylamine, a cation that permeates through the open channel of both vertebrate and invertebrate glutamate receptors, was used to measure the activity of glutamate receptor-ion channel complexes in reconstituted liposomes. L-Glutamate caused an increase in the rate of [ 14 C]methylamine influx into liposomes reconstituted with either solubilized membrane proteins or partially purified glutamate-binding proteins. Of the major glutamate receptor agonists, only N-methyl-D-aspartate activated cation fluxes in liposomes reconstituted with glutamate-binding proteins. In liposomes reconstituted with glutamate-binding proteins, N-methyl-D-aspartate- or glutamate-induced influx of NA + led to a transient increase in the influx of the lipid-permeable anion probe S 14 CN - . These results indicate the functional reconstitution of N-methyl-D-aspartate-sensitive glutamate receptors and the role of the ∼69-kDa protein in the function of these ion channels

  9. Membrane Recruitment of the Non-receptor Protein GIV/Girdin (Gα-interacting, Vesicle-associated Protein/Girdin) Is Sufficient for Activating Heterotrimeric G Protein Signaling.

    Parag-Sharma, Kshitij; Leyme, Anthony; DiGiacomo, Vincent; Marivin, Arthur; Broselid, Stefan; Garcia-Marcos, Mikel

    2016-12-30

    GIV (aka Girdin) is a guanine nucleotide exchange factor that activates heterotrimeric G protein signaling downstream of RTKs and integrins, thereby serving as a platform for signaling cascade cross-talk. GIV is recruited to the cytoplasmic tail of receptors upon stimulation, but the mechanism of activation of its G protein regulatory function is not well understood. Here we used assays in humanized yeast models and G protein activity biosensors in mammalian cells to investigate the role of GIV subcellular compartmentalization in regulating its ability to promote G protein signaling. We found that in unstimulated cells GIV does not co-fractionate with its substrate G protein Gα i3 on cell membranes and that constitutive membrane anchoring of GIV in yeast cells or rapid membrane translocation in mammalian cells via chemically induced dimerization leads to robust G protein activation. We show that membrane recruitment of the GIV "Gα binding and activating" motif alone is sufficient for G protein activation and that it does not require phosphomodification. Furthermore, we engineered a synthetic protein to show that recruitment of the GIV "Gα binding and activating" motif to membranes via association with active RTKs, instead of via chemically induced dimerization, is also sufficient for G protein activation. These results reveal that recruitment of GIV to membranes in close proximity to its substrate G protein is a major mechanism responsible for the activation of its G protein regulatory function. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

  10. Preventive Activities of Preliminary Investigation Bodies in Respect of Crime Victims

    Svetlana A. Timko

    2017-05-01

    Full Text Available The article is devoted to the problems of the prevention of victimization by the investigation and inquiry divisions of the internal affairs bodies of the Russian Federation. It defines the main forms and methods of working with the victim during the investigation of a crime aimed at reducing the possibility of again becoming a victim of criminal assault. The organizational and legal directions of victimological prevention are analyzed, the necessity of developing effective mechanisms for assessing the activities of the units of internal affairs agencies in crime prevention is justified.

  11. Microbial activity catalyzes oxygen transfer in membrane-aerated nitritating biofilm reactors

    Pellicer i Nàcher, Carles; Domingo Felez, Carlos; Lackner, Susanne

    2013-01-01

    The remarkable oxygen transfer efficiencies attainable in membrane-aerated biofilm reactors (MABRs) are expected to favor their prompt industrial implementation. However, tests in clean water, currently used for the estimation of their oxygen transfer potential, lead to wrong estimates once biofilm...... is present, significantly complicating reactor modelling and control. This study shows for the first time the factors affecting oxygen mass transfer across membranes during clean water tests and reactor operation via undisturbed microelectrode inspection and bulk measurements. The mass transfer resistance...... of the liquid boundary layer developed at the membrane-liquid interface during clean water tests accounted for two thirds of the total mass transfer resistance, suggesting a strong underestimation of the oxygen transfer rates when it is absent (e.g. after biofilm growth). Reactor operation to attain partial...

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

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

    2018-02-01

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

  13. Double-side active TiO{sub 2}-modified nanofiltration membranes in continuous flow photocatalytic reactors for effective water purification

    Romanos, G.Em., E-mail: groman@chem.demokritos.gr [Institute of Physical Chemistry, NCSR Demokritos, 153 10 Agia Paraskevi Attikis, Athens (Greece); Athanasekou, C.P.; Katsaros, F.K.; Kanellopoulos, N.K. [Institute of Physical Chemistry, NCSR Demokritos, 153 10 Agia Paraskevi Attikis, Athens (Greece); Dionysiou, D.D. [Department of Civil and Environmental Engineering, University of Cincinnati, Cincinnati, OH 45221-0071 (United States); Likodimos, V.; Falaras, P. [Institute of Physical Chemistry, NCSR Demokritos, 153 10 Agia Paraskevi Attikis, Athens (Greece)

    2012-04-15

    Highlights: Black-Right-Pointing-Pointer A novel CVD reactor for the developments of double side active TiO{sub 2} membranes. Black-Right-Pointing-Pointer Double side active TiO{sub 2} membranes efficiently photodegrade organic pollutants. Black-Right-Pointing-Pointer A photocatalytic membrane purification device for continuous flow water treatment. - Abstract: A chemical vapour deposition (CVD) based innovative approach was applied with the purpose to develop composite TiO{sub 2} photocatalytic nanofiltration (NF) membranes. The method involved pyrolytic decomposition of titanium tetraisopropoxide (TTIP) vapor and formation of TiO{sub 2} nanoparticles through homogeneous gas phase reactions and aggregation of the produced intermediate species. The grown nanoparticles diffused and deposited on the surface of {gamma}-alumina NF membrane tubes. The CVD reactor allowed for online monitoring of the carrier gas permeability during the treatment, providing a first insight on the pore efficiency and thickness of the formed photocatalytic layers. In addition, the thin TiO{sub 2} deposits were developed on both membrane sides without sacrificing the high yield rates. Important innovation was also introduced in what concerns the photocatalytic performance evaluation. The membrane efficiency to photo degrade typical water pollutants, was evaluated in a continuous flow water purification device, applying UV irradiation on both membrane sides. The developed composite NF membranes were highly efficient in the decomposition of methyl orange exhibiting low adsorption-fouling tendency and high water permeability.

  14. Depression of membrane-bound Na sup + -K sup + -ATPase activity induced by free radicals and by ischemia of kidney

    Kako, K.; Kato, M.; Matsuoka, T.; Mustapha, A. (Univ. of Ottawa, Ontario (Canada))

    1988-02-01

    A partially purified, membrane-bound Na{sup +}-K{sup +}-ATPase fraction, prepared from the outer medulla of porcine kidney, was incubated in the presence of 0.1-100 mM H{sub 2}O{sub 2} for either 15 or 30 min at 37{degree}C. The activity of ouabain-sensitive Na{sup +}-K{sup +}-ATPase was reduced proportionally to the concentration of H{sub 2}O{sub 2} and the duration of incubation. There were decreases in SH contents and turnover rates of the Na{sup +}-K{sup +}-ATPase preparation, while malondialdehyde (MDA) and conjugated dienes were generated from the membrane lipids in the course of the incubation. The concentrations of ethanolamine (E) plasmalogen and of arachidonic acid in the E glycerophospholipid molecules were reduced by the free radical reaction. Similarly, a reduction in Na{sup +}K{sup +}-ATPase activity and the formation of MDA and conjugated dienes, together with a decrease in E glycerophospholipids, were observed when the membrane fraction was exposed to ultraviolet irradiation (254 nm) for 30 min at 4{degree}C. Microsomal fractions, prepared from the outer medulla of canine kidney after 1 h of unilateral ischemia and 1 h of reperfusion, showed a decreased Na{sup +}-K{sup +}-ATPase activity, a reduced amount of SH groups, and an increased MDA. These changes were normalized by the infusion of N-mercaptopropionylglycine. These results support the view (1) that free radical generation affects the enzyme protein as well as membrane lipids, and (2) that free radicals may be formed in the ischemic reperfused kidney.

  15. The role of physical / sports activitys as an burnout prevention for special and rehabilitation educators

    Heric, Katja

    2013-01-01

    The Diploma thesis contains an overview of literature concerning the topic of how the sport/physical activity effects the phenomenon of professional burnout with special educatiors. I would like to explore the causes for and the signs of professional burnout, does sport/physical activity have a pozitive influence on an individual and if it is possible to prevent professional burnout with the help of sport/physical activity. I have found out that numerous stress factors at work, wich every ed...

  16. Networks of enzymatically oxidized membrane lipids support calcium-dependent coagulation factor binding to maintain hemostasis

    Lauder, S.N.; Allen-Redpath, K.; Slatter, D.A.; Aldrovandi, M.; O'Connor, A.; Farewell, D.; Percy, C.L.; Molhoek, J.E.; Rannikko, S.; Tyrrell, V.J.; Ferla, S.; Milne, G.L.; Poole, A.W.; Thomas, C.P.; Obaji, S.; Taylor, P.R.; Jones, S.A.; Groot, P.G. de; Urbanus, R.T.; Horkko, S.; Uderhardt, S.; Ackermann, J.; Jenkins, P.V.; Brancale, A.; Kronke, G.; Collins, P.W.; O'Donnell, V.B.

    2017-01-01

    Blood coagulation functions as part of the innate immune system by preventing bacterial invasion, and it is critical to stopping blood loss (hemostasis). Coagulation involves the external membrane surface of activated platelets and leukocytes. Using lipidomic, genetic, biochemical, and mathematical

  17. Incorporating AIDS prevention activities into a family planning organization in Colombia.

    Vernon, R; Ojeda, G; Murad, R

    1990-01-01

    Three AIDS prevention activities were incorporated into the services offered by PROFAMILIA in two operations research projects. The activities included: (1) informative talks given both to the general public and to members of target groups by PROFAMILIA's community marketing (CM) program field workers (or instructors); (2) the establishment of condom distribution posts in meeting places of target groups; and (3) mass-media information campaigns on AIDS prevention. Community-based distributors were able to successfully provide information on AIDS to their regular audiences as well as to deliver information and condoms to special target groups without negatively affecting family planning information/education/communication activities and contraceptive sales. A radio campaign that promoted condom use for AIDS prevention did not affect public perceptions about the condom and did not jeopardize PROFAMILIA's image.

  18. Release of LHRH-activity from human fetal membranes upon exposure to PGE/sub 2/, oxytocin and isoproterenol

    Poisner, A.M.; Poisner, R.; Becca, C.R.; Conn, P.M.

    1986-03-01

    The authors have previously reported that superfused chorion laeve (fetal membranes) release LHRH-like immunoreactivity upon exposure to angiotensin II. They have now studied the effects of other agonists on the release of LHRH-activity and something of its chemical nature. Fetal membranes were obtained from placentas delivered by cesarean section, the amnion stripped from the chorion, and the chorion superfused in an Amicon thin-channel device with the maternal surface facing up. The whole device was submerged in a 37 C water bath and perfused with a modified Locke's solution at 0.4 - 1.0 ml/min. LHRH-activity was measured by radioimmunoassay using three different antisera against LHRH. The release of LHRH-activity was stimulated by 6-10 min exposure to PGE/sub 2/, oxytocin, and isoproterenol. Extracts of chorion were studied using gel filtration on Sephacryl S-200 and ultrafiltration with Amicon PM-10 filters. The bulk of the LHRH-activity appeared as a higher molecular weight form (about 70,000 daltons). Since oxytocin has been reported to release PGE/sub 2/ from chorion, it may release LHRH-activity by virtue of liberating endogenous PGE/sub 2/. The chemical nature of the LHRH-activity is presently under investigation.

  19. Estimation of active rockburst prevention effectiveness during longwall mining under disadvantageous geological and mining conditions

    Łukasz Wojtecki

    2016-01-01

    Full Text Available Underground longwall mining of coal seams in the Upper Silesian Coal Basin is currently being carried out under increasingly difficult geological and mining conditions. Mining depth, dislocations and mining remnants are the main factors responsible for the most significant rockburst hazard, which can be minimized via the use of active and passive rockburst prevention. Active rockburst prevention in longwalls is usually based on blasting, in order to either destress local stress concentrations in the rock mass or to fracture the thick layers of strong roof rocks to prevent or minimize the impact of high energy tremors on excavations. The accurate estimation of active rockburst prevention effectiveness is particularly important when mining under disadvantageous geological and mining conditions, which are associated with high levels of this hazard. The efficiency of blasting applied for this purpose is typically evaluated from the seismic effect, which is calculated based on seismic monitoring data and the weight of the charged explosive. This method, as used previously in the Czech Republic, was adopted in the present study to analyze conditions occurring in a Polish hard coal mine in the Upper Silesian Coal Basin. Parameters of long hole destress blastings in roof rocks (torpedo blastings from the face of the assigned longwall in coal seam no. 507 were correct a success according to the seismic effect method and corresponded to observations made in situ. The analytical method presented enables the rapid estimation of destress blasting effectiveness and could also be useful when determining appropriate active rockburst prevention.

  20. ATPase activity of erythrocyte membranes and their permeability for the K-ions as influenced by irradiation and serotonin

    Zhegnevskaya, V.V.; Vinogradova, M.F.; Polevoj, V.V.

    1982-01-01

    Na, K-ATPase activity of membranes of erytrocytes after 1 hour of X-ray irradiation of citrate blood of rats (25.8 Kl/kg)-increased, and after irradiation of isolated erytrocytes, placed in the isotonic solution of NaCl did not change. The exflux of K-ions out of irradiated erytrocytes increased equally in both cases. Serotonin (2x10 -4 M), added to the probes 10 minutes before irradiation, decreased the exflux of K + by irradiated erytrocytes, but Na, K-ATPase activity under the influence of amine was without changes

  1. Cathepsin activities and membrane integrity of zebrafish (Danio rerio) oocytes after freezing to -196 degrees C using controlled slow cooling.

    Zhang, T; Rawson, D M; Tosti, L; Carnevali, O

    2008-04-01

    This study investigated enzymatic activity of cathepsins and the membrane integrity of zebrafish (Danio rerio) oocytes after freezing to -196 degrees C using controlled slow cooling. Stage III oocytes (>0.5mm), obtained through dissection of anaesthetised female fish and desegregation of ovarian cumulus, were exposed to 2M methanol or 2M DMSO (both prepared in Hank's medium) for 30min at 22 degrees C before being loaded into 0.5ml plastic straws and placed into a programmable cooler. After controlled slow freezing, samples were plunged into liquid nitrogen (LN) and held for at least 10min, and thawed by immersing straws into a 27 degrees C water bath for 10s. Thawed oocytes were washed twice in Hank's medium. Cathepsin activity and membrane integrity of oocytes were assessed both after cryoprotectant treatment at 22 degrees C and after freezing in LN. Cathepsin B and L colorimetric analyses were performed using substrates Z-Arg-ArgNNap and Z-Phe-Arg-4MbetaNA-HCl, respectively, and 2-naphthylamine and 4-methoxy-2-naphthylamine were used as standards. Cathepsin D activity was performed by analysing the level of hydrolytic action on haemoglobin. Oocytes membrane integrity was assessed using 0.2% Trypan blue staining for 5min. Analysis of cathepsin activities showed that whilst the activity of cathepsin B and D was not affected by 2M DMSO treatment, their activity was lowered when treated with 2M methanol. Following freezing to -196 degrees C, the activity of all cathepsins (B, D and L) was significantly decreased in both 2M DMSO and 2M methanol. Trypan blue staining showed that 63.0+/-11.3% and 72.7+/-5.2% oocytes membrane stayed intact after DMSO and methanol treatment for 30min at 22 degrees C, respectively, whilst 14.9+/-2.6% and 1.4+/-0.8% stayed intact after freezing in DMSO and methanol to -196 degrees C. The results indicate that cryoprotectant treatment and freezing modified the activities of lysosomal enzymes involved in oocyte maturation and yolk

  2. Functional activity of Gi alpha protein in detergent resistant membrane domains from rat brain cortex

    Stöhr, Jiří; Rudajev, Vladimír; Bouřová, Lenka; Lisý, Václav; Novotný, Jiří; Svoboda, Petr

    2007-01-01

    Roč. 101, Suppl.1 (2007), s. 52-52 ISSN 0022-3042. [European Society for Neurochemistry Meeting /17./. 19.05.2007-22.05.2007, Salamanca] Institutional research plan: CEZ:AV0Z50110509 Keywords : cpo1 * GABAB receptor * Gi protein * membrane domains Subject RIV: ED - Physiology

  3. Dipolar Relaxation Dynamics at the Active Site of an ATPase Regulated by Membrane Lateral Pressure

    Fischermeier, E.; Pospíšil, Petr; Sayed, A.; Hof, Martin; Solioz, M.; Fahmy, K.

    2017-01-01

    Roč. 56, č. 5 (2017), s. 1269-1272 ISSN 1433-7851 R&D Projects: GA ČR(CZ) GBP208/12/G016 Institutional support: RVO:61388955 Keywords : fluorescence * ion pump * membrane proteins * nanodiscs * time-resolved emission Subject RIV: CF - Physical ; Theoretical Chemistry OBOR OECD: Physical chemistry Impact factor: 11.994, year: 2016

  4. In vitro and in vivo activation of mitochondrial membrane permeability transition pore using triiodothyronine

    Endlicher, R.; Drahota, Zdeněk; Červinková, Z.

    2016-01-01

    Roč. 65, č. 2 (2016), s. 321-331 ISSN 0862-8408 R&D Projects: GA ČR(CZ) GB14-36804G Institutional support: RVO:67985823 Keywords : rat liver mitochondria * membrane permeability transition pore * thyroid hormones Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 1.461, year: 2016

  5. Membrane plasmalogen composition and cellular cholesterol regulation: a structure activity study

    Su-Myat Khine K

    2010-06-01

    Full Text Available Abstract Background Disrupted cholesterol regulation leading to increased circulating and membrane cholesterol levels is implicated in many age-related chronic diseases such as cardiovascular disease (CVD, Alzheimer's disease (AD, and cancer. In vitro and ex vivo cellular plasmalogen deficiency models have been shown to exhibit impaired intra- and extra-cellular processing of cholesterol. Furthermore, depleted brain plasmalogens have been implicated in AD and serum plasmalogen deficiencies have been linked to AD, CVD, and cancer. Results Using plasmalogen deficient (NRel-4 and plasmalogen sufficient (HEK293 cells we investigated the effect of species-dependent plasmalogen restoration/augmentation on membrane cholesterol processing. The results of these studies indicate that the esterification of cholesterol is dependent upon the amount of polyunsaturated fatty acid (PUFA-containing ethanolamine plasmalogen (PlsEtn present in the membrane. We further elucidate that the concentration-dependent increase in esterified cholesterol observed with PUFA-PlsEtn was due to a concentration-dependent increase in sterol-O-acyltransferase-1 (SOAT1 levels, an observation not reproduced by 3-hydroxy-3-methyl-glutaryl-CoA (HMG-CoA reductase inhibition. Conclusion The present study describes a novel mechanism of cholesterol regulation that is consistent with clinical and epidemiological studies of cholesterol, aging and disease. Specifically, the present study describes how selective membrane PUFA-PlsEtn enhancement can be achieved using 1-alkyl-2-PUFA glycerols and through this action reduce levels of total and free cholesterol in cells.

  6. Mg(2+,ATP-dependent plasma membrane calcium pump of smooth muscle cells. ІІ. Regulation of activity

    T. О. Veklich

    2015-04-01

    Full Text Available Plasma membrane Ca2+-pump is one of key proteins, which takes part in Ca2+ exchange in smooth muscle cells. It has a lot of diverse functions from control of basal cytoplasmal Ca2+ concentration to regulation of proteins involved in Ca2+-dependent signal pathway. Ca2+ pump function is often depen­dent on the isoform or even form of alternative splicing. Allowing for a variety of Ca2+-pump functions and properties, which were reviewed in detail in the first part of our review article cycle (Ukr. Biochem. J., 2015; 87(1, the precise control of the mentioned pump activity is very important for cell functioning­. The other part of this article is dedicated to different regulation factors of smooth muscle plasma membrane Ca2+-pump activity: endogenous and exo­genous, biotic and abiotic factors. Special attention is given to literature data and own results about design and the search of selective plasma membrane Ca2+-pump inhibitor which would allow examining its functioning in smooth muscle cells more meticulously.

  7. Recovery of homogeneous polyoxometallate catalysts from aqueous and organic media by a mesoporous ceramic membrane without loss of catalytic activity.

    Roy Chowdhury, Sankhanilay; Witte, Peter T; Blank, Dave H A; Alsters, Paul L; Ten Elshof, Johan E

    2006-04-03

    The recovery of homogeneous polyoxometallate (POM) oxidation catalysts from aqueous and non-aqueous media by a nanofiltration process using mesoporous gamma-alumina membranes is reported. The recovery of Q(12)[WZn(3)(ZnW(9)O(34))(2)] (Q=[MeN(n-C(8)H(17))(3)](+)) from toluene-based media was quantitative within experimental error, while up to 97 % of Na(12)[WZn(3)(ZnW(9)O(34))(2)] could be recovered from water. The toluene-soluble POM catalyst was used repeatedly in the conversion of cyclooctene to cyclooctene oxide and separated from the product mixture after each reaction. The catalytic activity increased steadily with the number of times that the catalyst had been recycled, which was attributed to partial removal of the excess QCl that is known to have a negative influence on the catalytic activity. Differences in the permeability of the membrane for different liquid media can be attributed to viscosity differences and/or capillary condensation effects. The influence of membrane pore radius on permeability and recovery is discussed.

  8. Isolation and molecular characterization of biofouling bacteria and profiling of quorum sensing signal molecules from membrane bioreactor activated sludge.

    Lade, Harshad; Paul, Diby; Kweon, Ji Hyang

    2014-02-04

    The formation of biofilm in a membrane bioreactor depends on the production of various signaling molecules like N-acyl homoserine lactones (AHLs). In the present study, a total of 200 bacterial strains were isolated from membrane bioreactor activated sludge and screened for AHLs production using two biosensor systems, Chromobacterium violaceum CV026 and Agrobacterium tumefaciens A136. A correlation between AHLs production and biofilm formation has been made among screened AHLs producing strains. The 16S rRNA gene sequence analysis revealed the dominance of Aeromonas and Enterobacter sp. in AHLs production; however few a species of Serratia, Leclercia, Pseudomonas, Klebsiella, Raoultella and Citrobacter were also identified. The chromatographic characterization of sludge extract showed the presence of a broad range of quorum sensing signal molecules. Further identification of sludge AHLs by thin layer chromatography bioassay and high performance liquid chromatography confirms the presence of C4-HSL, C6-HSL, C8-HSL, 3-oxo-C8-HSL, C10-HSL, C12-HSL, 3-oxo-C12-HSL and C14-HSL. The occurrence of AHLs in sludge extract and dominance of Aeromonas and Enterobacter sp. in activated sludge suggests the key role of these bacterial strains in AHLs production and thereby membrane fouling.

  9. Isolation and Molecular Characterization of Biofouling Bacteria and Profiling of Quorum Sensing Signal Molecules from Membrane Bioreactor Activated Sludge

    Harshad Lade

    2014-02-01

    Full Text Available The formation of biofilm in a membrane bioreactor depends on the production of various signaling molecules like N-acyl homoserine lactones (AHLs. In the present study, a total of 200 bacterial strains were isolated from membrane bioreactor activated sludge and screened for AHLs production using two biosensor systems, Chromobacterium violaceum CV026 and Agrobacterium tumefaciens A136. A correlation between AHLs production and biofilm formation has been made among screened AHLs producing strains. The 16S rRNA gene sequence analysis revealed the dominance of Aeromonas and Enterobacter sp. in AHLs production; however few a species of Serratia, Leclercia, Pseudomonas, Klebsiella, Raoultella and Citrobacter were also identified. The chromatographic characterization of sludge extract showed the presence of a broad range of quorum sensing signal molecules. Further identification of sludge AHLs by thin layer chromatography bioassay and high performance liquid chromatography confirms the presence of C4-HSL, C6-HSL, C8-HSL, 3-oxo-C8-HSL, C10-HSL, C12-HSL, 3-oxo-C12-HSL and C14-HSL. The occurrence of AHLs in sludge extract and dominance of Aeromonas and Enterobacter sp. in activated sludge suggests the key role of these bacterial strains in AHLs production and thereby membrane fouling.

  10. Impact of ozonation and biological activated carbon filtration on ceramic membrane fouling.

    Ibn Abdul Hamid, Khaled; Sanciolo, Peter; Gray, Stephen; Duke, Mikel; Muthukumaran, Shobha

    2017-12-01

    Ozone pre-treatment (ozonation, ozonisation) and biological activated carbon (BAC) filtration pre-treatment for the ceramic microfiltration (CMF) treatment of secondary effluent (SE) were studied. Ozone pre-treatment was found to result in higher overall removal of UV absorbance (UVA 254 ) and colour, and higher permeability than BAC pre-treatment or the combined use of ozone and BAC (O3+BAC) pre-treatment. The overall removal of colour and UVA 254 by ceramic filtration of the ozone pre-treated water was 97% and 63% respectively, compared to 86% and 48% respectively for BAC pre-treatment and 29% and 6% respectively for the untreated water. Ozone pre-treatment, however, was not effective in removal of dissolved organic carbon (DOC). The permeability of the ozone pre-treated water through the ceramic membrane was found to decrease to 50% of the original value after 200 min of operation, compared to approximately 10% of the original value for the BAC pre-treated, O3+BAC pre-treated water and the untreated water. The higher permeability of the ozone pre-treated water was attributed to the excellent removal of biopolymer particles (100%) and high removal of humic substances (84%). The inclusion of a BAC stage between ozone pre-treatment and ceramic filtration was detrimental. The O3+BAC+CMF process was found to yield higher biopolymer removal (96%), lower humic substance (HS) component removal (66%) and lower normalized permeability (0.1) after 200 min of operation than the O3+CMF process (86%, 84% and 0.5 respectively). This was tentatively attributed to the chemical oxidation effect of ozone on the BAC biofilm and adsorbed components, leading to the generation of foulants that are not generated in the O3+CMF process. This study demonstrated the potential of ozone pre-treatment for reducing organic fouling and thus improving flux for the CMF of SE compared to O3+BAC pre-treatment. Copyright © 2017 Elsevier Ltd. All rights reserved.

  11. 5-Aminoimidazole-4-carboxamide ribonucleotide prevents fat gain following the cessation of voluntary physical activity.

    Ruegsegger, Gregory N; Sevage, Joseph A; Childs, Thomas E; Grigsby, Kolter B; Booth, Frank W

    2017-11-01

    What is the central question of this study? We investigated whether 5-aminoimidazole-4-carboxamide-1-beta-D-ribofuranoside (AICAR) could prevent acute increases in body fat and changes in omental and subcutaneous adipose tissue following the sudden transition from physical activity to physical inactivity. What is the main finding and its importance? AICAR prevented fat gains following the transition from physical activity to inactivity to levels comparable to rats that remained physically active. AICAR and continuous physical activity produced depot-specific changes in cyclin A1 mRNA and protein that were associated with the prevention of fat gain. These findings suggest that targeting AMP-activated protein kinase signalling could oppose rapid adipose mass growth. The transition from physical activity to inactivity is associated with drastic increases in 'catch-up' fat that in turn foster the development of many obesity-associated maladies. We tested whether 5-aminoimidazole-4-carboxamide-1-beta-D-ribofuranoside (AICAR) treatment would prevent gains in body fat following the sudden transition from a physically active state to an inactive state by locking a voluntary running wheel. Male Wistar rats were either sedentary (SED) or given wheel access for 4 weeks, at which time rats with wheels continued running (RUN), had their wheel locked (WL) or had WL with daily AICAR injection (WL + AICAR) for 1 week. RUN and WL + AICAR prevented gains in body fat compared with SED and WL (P RUN and WL + AICAR compared with SED and WL groups (P run, and that together, continuous physical activity and AICAR could, at least initially in these conditions, exert similar inhibitory effects on adipogenesis in a depot-specific manner. © 2017 The Authors. Experimental Physiology © 2017 The Physiological Society.

  12. Opportunities and supporting activities to promote preventive maintenance of NPPs in Japan

    Sanada, A.; Shinkawa, T.; Sakurada, M.

    1998-01-01

    With increase of the number of NPPs and operation history, enhancement of the safety regulation is becoming important for such long-operated NPPs. Centering on the comprehensive preventive maintenance, periodic safety reviews by utilities and its review and evaluation by MITI are in progress. The first reviews have not revealed essential and critical indications to be newly implemented. This means that the most of activities to secure reliability and safety have been done steadily. The present paper addresses the mechanism of on-going preventive maintenance and its essential elements: opportunities to identify issues and problems, and supporting activities to promote decision-makings on feedback, upgrading and modernization of NPPs. (author)

  13. Earliest Results in the Use of Activated Composite Membranes for the Transport of Silver Ions from Aqueous Solutions

    Yucundo Mendoza-Tolentino

    2014-01-01

    Full Text Available This paper presents the results concerning the first use of activated composite membranes (ACMs for the facilitated transport of silver ions containing di-(2-ethylhexyl-dithiophosphoric acid (DTPA as the carrier. DTPA was immobilized by interfacial polymerization in a dense layer that was deposited in a porous layer, which was prepared on a nonwoven fabric support by phase inversion. The influence of fundamental parameters affecting the transport of silver ion as the carrier concentration in the membrane phase and stripping agent variation of the stripping solution have been studied. In the optimal conditions, the amount of silver transported across the ACMs was greater than 50%, whereas if the content of the carrier is modified, more than the 90% of the initial silver is removed from the feed phase.

  14. Acid sphingomyelinase activity is regulated by membrane lipids and facilitates cholesterol transfer by NPC2.

    Oninla, Vincent O; Breiden, Bernadette; Babalola, Jonathan O; Sandhoff, Konrad

    2014-12-01

    During endocytosis, membrane components move to intraluminal vesicles of the endolysosomal compartment for digestion. At the late endosomes, cholesterol is sorted out mainly by two sterol-binding proteins, Niemann-Pick protein type C (NPC)1 and NPC2. To study the NPC2-mediated intervesicular cholesterol transfer, we developed a liposomal assay system. (Abdul-Hammed, M., B. Breiden, M. A. Adebayo, J. O. Babalola, G. Schwarzmann, and K. Sandhoff. 2010. Role of endosomal membrane lipids and NPC2 in cholesterol transfer and membrane fusion. J. Lipid Res. 51: 1747-1760.) Anionic lipids stimulate cholesterol transfer between liposomes while SM inhibits it, even in the presence of anionic bis(monoacylglycero)phosphate (BMP). Preincubation of vesicles containing SM with acid sphingomyelinase (ASM) (SM phosphodiesterase, EC 3.1.4.12) results in hydrolysis of SM to ceramide (Cer), which enhances cholesterol transfer. Besides SM, ASM also cleaves liposomal phosphatidylcholine. Anionic phospholipids derived from the plasma membrane (phosphatidylglycerol and phosphatidic acid) stimulate SM and phosphatidylcholine hydrolysis by ASM more effectively than BMP, which is generated during endocytosis. ASM-mediated hydrolysis of liposomal SM was also stimulated by incorporation of diacylglycerol (DAG), Cer, and free fatty acids into the liposomal membranes. Conversely, phosphatidylcholine hydrolysis was inhibited by incorporation of cholesterol, Cer, DAG, monoacylglycerol, and fatty acids. Our data suggest that SM degradation by ASM is required for physiological secretion of cholesterol from the late endosomal compartment, and is a key regulator of endolysosomal lipid digestion. Copyright © 2014 by the American Society for Biochemistry and Molecular Biology, Inc.

  15. Important factors for cell-membrane permeabilization by gold nanoparticles activated by nanosecond-laser irradiation

    Yao CP

    2017-08-01

    Full Text Available Cuiping Yao,1,2,* Florian Rudnitzki,2,* Gereon Hüttmann,2,3 Zhenxi Zhang,1 Ramtin Rahmanzadeh2 1Key Laboratory of Biomedical Information Engineering of Education Ministry, Institute of Biomedical Analytical Technology and Instrumentation, School of Life Science and Technology, Xi’an Jiaotong University, Xi’an, China; 2Institute of Biomedical Optics, University of Lübeck, Lübeck, 3Airway Research Center North (ARCN, Member of the German Center for Lung Research (DZL, Kiel, Germany *These authors contributed equally to this work Purpose: Pulsed-laser irradiation of light-absorbing gold nanoparticles (AuNPs attached to cells transiently increases cell membrane permeability for targeted molecule delivery. Here, we targeted EGFR on the ovarian carcinoma cell line OVCAR-3 with AuNPs. In order to optimize membrane permeability and to demonstrate molecule delivery into adherent OVCAR-3 cells, we systematically investigated different experimental conditions. Materials and methods: AuNPs (30 nm were functionalized by conjugation of the antibody cetuximab against EGFR. Selective binding of the particles was demonstrated by silver staining, multiphoton imaging, and fluorescence-lifetime imaging. After laser irradiation, membrane permeability of OVCAR-3 cells was studied under different conditions of AuNP concentration, cell-incubation medium, and cell–AuNP incubation time. Membrane permeability and cell viability were evaluated by flow cytometry, measuring propidium iodide and fluorescein isothiocyanate–dextran uptake. Results: Adherently growing OVCAR-3 cells can be effectively targeted with EGFR-AuNP. Laser irradiation led to successful permeabilization, and 150 kDa dextran was successfully delivered into cells with about 70% efficiency. Conclusion: Antibody-targeted and laser-irradiated AuNPs can be used to deliver molecules into adherent cells. Efficacy depends not only on laser parameters but also on AuNP:cell ratio, cell-incubation medium

  16. Behavioral Counseling to Promote a Healthful Diet and Physical Activity for CVD Prevention in Adults with Risk Factors

    Understanding Task Force Recommendations Behavioral Counseling to Promote a Healthful Diet and Physical Activity for Cardiovascular Disease Prevention in Adults with Cardiovascular Risk Factors The U.S. Preventive ...

  17. Intracellular and membrane-damaging activities of methyl gallate isolated from Terminalia chebula against multidrug-resistant Shigella spp.

    Acharyya, Saurabh; Sarkar, Prodipta; Saha, Dhira R; Patra, Amarendra; Ramamurthy, T; Bag, Prasanta K

    2015-08-01

    Shigella spp. (Shigella dysenteriae, Shigella flexneri, Shigella boydii and Shigella sonnei) cause bacillary dysentery (shigellosis), which is characterized by bloody mucous diarrhoea. Although a variety of antibiotics have been effective for treatment of shigellosis, options are becoming limited due to globally emerging drug resistance. In the present study, in vitro antibacterial activity of methyl gallate (MG) isolated from Terminalia chebula was determined by performing MIC, minimal bactericidal concentration (MBC) and time-kill kinetic studies. Bacterial membrane-damaging activity of MG was determined by membrane perturbation and transmission electron microscopy (TEM). Cellular drug accumulation, cell infection and assessment of intracellular activities of MG and reference antibiotics were performed using HeLa cell cultures. The bactericidal activity of MG against multidrug-resistant (MDR) Shigella spp. in comparison with other commonly used drugs including fluoroquinolone was demonstrated here. TEM findings in the present study revealed that MG caused the total disintegration of inner and outer membranes, and leakage of the cytoplasmic contents of S. dysenteriae. The level of accumulation of MG and tetracycline in HeLa cells incubated for 24  h was relatively higher than that of ciprofloxacin and nalidixic acid (ratio of intracellular concentration/extracellular concentration of antibiotic for MG and tetracycline>ciprofloxacin and nalidixic acid). The viable number of intracellular S. dysenteriae was decreased in a time-dependent manner in the presence of MG (4 × MBC) and reduced to zero within 20  h. The significant intracellular activities of MG suggested that it could potentially be used as an effective antibacterial agent for the treatment of severe infections caused by MDR Shigella spp.

  18. Activity ranking of synthetic analogs targeting vascular endothelial growth factor receptor 2 by an integrated cell membrane chromatography system.

    Wang, Dongyao; Lv, Diya; Chen, Xiaofei; Liu, Yue; Ding, Xuan; Jia, Dan; Chen, Langdong; Zhu, Zhenyu; Cao, Yan; Chai, Yifeng

    2015-12-01

    Evaluating the biological activities of small molecules represents an important part of the drug discovery process. Cell membrane chromatography (CMC) is a well-developed biological chromatographic technique. In this study, we have developed combined SMMC-7721/CMC and HepG2/CMC with high-performance liquid chromatography and time-of-flight mass spectrometry to establish an integrated screening platform. These systems was subsequently validated and used for evaluating the activity of quinazoline compounds, which were designed and synthesized to target vascular endothelial growth factor receptor 2. The inhibitory activities of these compounds towards this receptor were also tested using a classical caliper mobility shift assay. The results revealed a significant correlation between these two methods (R(2) = 0.9565 or 0.9420) for evaluating the activities of these compounds. Compared with traditional methods of evaluating the activities analogous compounds, this integrated cell membrane chromatography screening system took less time and was more cost effective, indicating that it could be used as a practical method in drug discovery. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  19. Healthy eating, activity and obesity prevention: a qualitative study of parent and child perceptions in Australia.

    Hesketh, K; Waters, E; Green, J; Salmon, L; Williams, J

    2005-03-01

    Preventative health strategies incorporating the views of target participants have improved the likelihood of success. This qualitative study aimed to elicit child and parent views regarding social and environmental barriers to healthy eating, physical activity and child obesity prevention programmes, acceptable foci, and appropriate modes of delivery. To obtain views across a range of social circumstances three demographically diverse primary schools in Victoria, Australia were selected. Children in Grades 2 (aged 7-8 years) and 5 (aged 10-11 years) participated in focus groups of three to six children. Groups were semi-structured using photo-based activities to initiate discussion. Focus groups with established parent groups were also conducted. Comments were recorded, collated, and themes extracted using grounded theory. 119 children and 17 parents participated. Nine themes emerged: information and awareness, contradiction between knowledge and behaviour, lifestyle balance, local environment, barriers to a healthy lifestyle, contradictory messages, myths, roles of the school and family, and timing and content of prevention strategies for childhood obesity. In conclusion, awareness of food 'healthiness' was high however perceptions of the 'healthiness' of some sedentary activities that are otherwise of benefit (e.g. reading) were uncertain. The contradictions in messages children receive were reported to be a barrier to a healthy lifestyle. Parent recommendations regarding the timing and content of childhood obesity prevention strategies were consistent with quantitative research. Contradictions in the explicit and implicit messages children receive around diet and physical activity need to be prevented. Consistent promotion of healthy food and activity choices across settings is core to population prevention programmes for childhood obesity.

  20. Bacterial membrane activity of a-peptide/b-peptoid chimeras: Influence of amino acid composition and chain length on the activity against different bacterial strains

    Hein-Kristensen, Line; Knapp, Kolja M; Franzyk, Henrik

    2011-01-01

    and subsequent killing is usually not tested. In this report, six α-peptide/β-peptoid chimeras were examined for the effect of amino acid/peptoid substitutions and chain length on the membrane perturbation and subsequent killing of food-borne and clinical bacterial isolates. RESULTS: All six AMP analogues...... acid only had a minor effect on MIC values, whereas chain length had a profound influence on activity. All chimeras were less active against Serratia marcescens (MICs above 46 μM). The chimeras were bactericidal and induced leakage of ATP from Staphylococcus aureus and S. marcescens with similar time...... of onset and reduction in the number of viable cells. EDTA pre-treatment of S. marcescens and E. coli followed by treatment with chimeras resulted in pronounced killing indicating that disintegration of the Gram-negative outer membrane eliminated innate differences in susceptibility. Chimera chain length...

  1. Potent synergistic in vitro interaction between nonantimicrobial membrane-active compounds and itraconazole against clinical isolates of Aspergillus fumigatus resistant to itraconazole.

    Afeltra, J.; Vitale, R.G.; Mouton, J.W.; Verweij, P.E.

    2004-01-01

    To develop new approaches for the treatment of invasive infections caused by Aspergillus fumigatus, the in vitro interactions between itraconazole (ITZ) and seven different nonantimicrobial membrane-active compounds--amiodarone (AMD), amiloride, lidocaine, lansoprazole (LAN), nifedipine (NIF),

  2. Novel role for proteinase-activated receptor 2 (PAR2) in membrane trafficking of proteinase-activated receptor 4 (PAR4).

    Cunningham, Margaret R; McIntosh, Kathryn A; Pediani, John D; Robben, Joris; Cooke, Alexandra E; Nilsson, Mary; Gould, Gwyn W; Mundell, Stuart; Milligan, Graeme; Plevin, Robin

    2012-05-11

    Proteinase-activated receptors 4 (PAR(4)) is a class A G protein-coupled receptor (GPCR) recognized through the ability of serine proteases such as thrombin and trypsin to mediate receptor activation. Due to the irreversible nature of activation, a fresh supply of receptor is required to be mobilized to the cell surface for responsiveness to agonist to be sustained. Unlike other PAR subtypes, the mechanisms regulating receptor trafficking of PAR(4) remain unknown. Here, we report novel features of the intracellular trafficking of PAR(4) to the plasma membrane. PAR(4) was poorly expressed at the plasma membrane and largely retained in the endoplasmic reticulum (ER) in a complex with the COPI protein subunit β-COP1. Analysis of the PAR(4) protein sequence identified an arginine-based (RXR) ER retention sequence located within intracellular loop-2 (R(183)AR → A(183)AA), mutation of which allowed efficient membrane delivery of PAR(4). Interestingly, co-expression with PAR(2) facilitated plasma membrane delivery of PAR(4), an effect produced through disruption of β-COP1 binding and facilitation of interaction with the chaperone protein 14-3-3ζ. Intermolecular FRET studies confirmed heterodimerization between PAR(2) and PAR(4). PAR(2) also enhanced glycosylation of PAR(4) and activation of PAR(4) signaling. Our results identify a novel regulatory role for PAR(2) in the anterograde traffic of PAR(4). PAR(2) was shown to both facilitate and abrogate protein interactions with PAR(4), impacting upon receptor localization and cell signal transduction. This work is likely to impact markedly upon the understanding of the receptor pharmacology of PAR(4) in normal physiology and disease.

  3. Activation of AMPK in human fetal membranes alleviates infection-induced expression of pro-inflammatory and pro-labour mediators.

    Lim, R; Barker, G; Lappas, M

    2015-04-01

    In non-gestational tissues, the activation of adenosine monophosphate (AMP)-activated kinase (AMPK) is associated with potent anti-inflammatory actions. Infection and/or inflammation, by stimulating pro-inflammatory cytokines and matrix metalloproteinase (MMP)-9, play a central role in the rupture of fetal membranes. However, no studies have examined the role of AMPK in human labour. Fetal membranes, from term and preterm, were obtained from non-labouring and labouring women, and after preterm pre-labour rupture of membranes (PPROM). AMPK activity was assessed by Western blotting of phosphorylated AMPK expression. To determine the effect of AMPK activators on pro-inflammatory cytokines, fetal membranes were pre-treated with AMPK activators then stimulated with bacterial products LPS and flagellin or viral dsDNA analogue poly(I:C). Primary amnion cells were used to determine the effect of AMPK activators on IL-1β-stimulated MMP-9 expression. AMPK activity was decreased with term labour. There was no effect of preterm labour. AMPK activity was also decreased in preterm fetal membranes, in the absence of labour, with PROM compared to intact membranes. AMPK activators AICAR, phenformin and A769662 significantly decreased IL-6 and IL-8 stimulated by LPS, flagellin and poly(I:C). Primary amnion cells treated with AMPK activators significantly decreased IL-1β-induced MMP-9 expression. The decrease in AMPK activity in fetal membranes after spontaneous term labour and PPROM indicates an anti-inflammatory role for AMPK in human labour and delivery. The use of AMPK activators as possible therapeutics for threatened preterm labour would be an exciting future avenue of research. Copyright © 2015 Elsevier Ltd. All rights reserved.

  4. Concurrent Transient Activation of Wnt/β-Catenin Pathway Prevents Radiation Damage to Salivary Glands

    Hai Bo; Yang Zhenhua; Shangguan Lei; Zhao Yanqiu; Boyer, Arthur; Liu, Fei

    2012-01-01

    Purpose: Many head and neck cancer survivors treated with radiotherapy suffer from permanent impairment of their salivary gland function, for which few effective prevention or treatment options are available. This study explored the potential of transient activation of Wnt/β-catenin signaling in preventing radiation damage to salivary glands in a preclinical model. Methods and Materials: Wnt reporter transgenic mice were exposed to 15 Gy single-dose radiation in the head and neck area to evaluate the effects of radiation on Wnt activity in salivary glands. Transient Wnt1 overexpression in basal epithelia was induced in inducible Wnt1 transgenic mice before together with, after, or without local radiation, and then saliva flow rate, histology, apoptosis, proliferation, stem cell activity, and mRNA expression were evaluated. Results: Radiation damage did not significantly affect activity of Wnt/β-catenin pathway as physical damage did. Transient expression of Wnt1 in basal epithelia significantly activated the Wnt/β-catenin pathway in submandibular glands of male mice but not in those of females. Concurrent transient activation of the Wnt pathway prevented chronic salivary gland dysfunction following radiation by suppressing apoptosis and preserving functional salivary stem/progenitor cells. In contrast, Wnt activation 3 days before or after irradiation did not show significant beneficial effects, mainly due to failure to inhibit acute apoptosis after radiation. Excessive Wnt activation before radiation failed to inhibit apoptosis, likely due to extensive induction of mitosis and up-regulation of proapoptosis gene PUMA while that after radiation might miss the critical treatment window. Conclusion: These results suggest that concurrent transient activation of the Wnt/β-catenin pathway could prevent radiation-induced salivary gland dysfunction.

  5. Concurrent Transient Activation of Wnt/{beta}-Catenin Pathway Prevents Radiation Damage to Salivary Glands

    Hai Bo; Yang Zhenhua; Shangguan Lei; Zhao Yanqiu [Institute for Regenerative Medicine, Scott and White Hospital, Molecular and Cellular Medicine Department, Texas A and M Health Science Center, Temple, Texas (United States); Boyer, Arthur [Department of Radiology, Scott and White Hospital, Temple, Texas (United States); Liu, Fei, E-mail: fliu@medicine.tamhsc.edu [Institute for Regenerative Medicine, Scott and White Hospital, Molecular and Cellular Medicine Department, Texas A and M Health Science Center, Temple, Texas (United States)

    2012-05-01

    Purpose: Many head and neck cancer survivors treated with radiotherapy suffer from permanent impairment of their salivary gland function, for which few effective prevention or treatment options are available. This study explored the potential of transient activation of Wnt/{beta}-catenin signaling in preventing radiation damage to salivary glands in a preclinical model. Methods and Materials: Wnt reporter transgenic mice were exposed to 15 Gy single-dose radiation in the head and neck area to evaluate the effects of radiation on Wnt activity in salivary glands. Transient Wnt1 overexpression in basal epithelia was induced in inducible Wnt1 transgenic mice before together with, after, or without local radiation, and then saliva flow rate, histology, apoptosis, proliferation, stem cell activity, and mRNA expression were evaluated. Results: Radiation damage did not significantly affect activity of Wnt/{beta}-catenin pathway as physical damage did. Transient expression of Wnt1 in basal epithelia significantly activated the Wnt/{beta}-catenin pathway in submandibular glands of male mice but not in those of females. Concurrent transient activation of the Wnt pathway prevented chronic salivary gland dysfunction following radiation by suppressing apoptosis and preserving functional salivary stem/progenitor cells. In contrast, Wnt activation 3 days before or after irradiation did not show significant beneficial effects, mainly due to failure to inhibit acute apoptosis after radiation. Excessive Wnt activation before radiation failed to inhibit apoptosis, likely due to extensive induction of mitosis and up-regulation of proapoptosis gene PUMA while that after radiation might miss the critical treatment window. Conclusion: These results suggest that concurrent transient activation of the Wnt/{beta}-catenin pathway could prevent radiation-induced salivary gland dysfunction.

  6. Liaison of 3H 5-HT and adenyl cyclasic activation induced by the 5-HT in preparations of brain glial membranes

    Fillion, Gilles; Beaudoin, Dominique; Rousselle, J.-C.; Jacob, Joseph

    1980-01-01

    Purified glial membrane preparations have been isolated from horse brain striatum. Tritiated 5-HT bound to these membranes with a high affinity (K(D)=10 nM); the corresponding bindings is reversible and appears specific of the serotoninergic structure. In parallel, 5-HT activates an adenylate cyclase with a low affinity (K(D)=1 μM). The sites involved in this binding and in this adenylate cyclase activation appear different from the serotoninergic sites reported in the neuronal membrane preparations [fr

  7. [Prevention of spontaneous preterm birth (excluding preterm premature rupture of membranes): Guidelines for clinical practice - Text of the Guidelines (short text)].

    Sentilhes, L; Sénat, M-V; Ancel, P-Y; Azria, E; Benoist, G; Blanc, J; Brabant, G; Bretelle, F; Brun, S; Doret, M; Ducroux-Schouwey, C; Evrard, A; Kayem, G; Maisonneuve, E; Marcellin, L; Marret, S; Mottet, N; Paysant, S; Riethmuller, D; Rozenberg, P; Schmitz, T; Torchin, H; Langer, B

    2016-12-01

    To determine the measures to prevent spontaneous preterm birth (excluding preterm premature rupture of membranes)and its consequences. The PubMed database, the Cochrane Library and the recommendations from the French and foreign obstetrical societies or colleges have been consulted. In France, premature birth concerns 60,000 neonates every year (7.4 %), half of them are delivered after spontaneous onset of labor. Among preventable risk factors of spontaneous prematurity, only cessation of smoking is associated to a decrease of prematurity (level of evidence [LE] 1). This is therefore recommended (grade A). Routine screening and treatment of vaginal bacteriosis in general population is not recommended (grade A). Asymptomatic women with single pregnancy without history of preterm delivery and a short cervix between 16 and 24 weeks is the only population in which vaginal progesterone is recommended (grade B). A history-indicated cerclage is not recommended in case of only past history of conisation (grade C), uterine malformation (Professional consensus), isolated history of pretem delivery (grade B) or twin pregnancies in primary (grade B) or secondary (grade C) prevention of preterm birth. A history-indicated cerclage is recommended for single pregnancy with a history of at least 3 late miscarriages or preterm deliveries (grade A).). In case of past history of a single pregnancy delivery before 34 weeks gestation (WG), ultrasound cervical length screening is recommended between 16 and 22 WG in order to propose a cerclage in case of lengthpremature rupture of membranes. Maintenance tocolysis is not recomended (grade B). Antenatal corticosteroid administration is recommended to every woman at risk of preterm delivery before 34 weeks of gestation (grade A). After 34 weeks, evidences are not consistent enough to recommend systematic antenatal corticosteroid treatment (grade B), however, a course might be indicated in the clinical situations associated with the

  8. Preparation of robust braid-reinforced poly(vinyl chloride) ultrafiltration hollow fiber membrane with antifouling surface and application to filtration of activated sludge solution.

    Zhou, Zhuang; Rajabzadeh, Saeid; Fang, Lifeng; Miyoshi, Taro; Kakihana, Yuriko; Matsuyama, Hideto

    2017-08-01

    Braid-reinforced hollow fiber membranes with high mechanical properties and considerable antifouling surface were prepared by blending poly(vinyl chloride) (PVC) with poly(vinyl chloride-co-poly(ethylene glycol) methyl ether methacrylate) (poly(VC-co-PEGMA)) copolymer via non-solvent induced phase separation (NIPS). The tensile strength of the braid-reinforced PVC hollow fiber membranes were significantly larger than those of previously reported various types of PVC hollow fiber membranes. The high interfacial bonding strength indicated the good compatibility between the coating materials and the surface of polyethylene terephthalate (PET)-braid. Owing to the surface segregation phenomena, the membrane surface PEGMA coverage increased upon increasing the poly(VC-co-PEGMA)/PVC blending ratio, resulting in higher hydrophilicities and bovine serum albumin (BSA) repulsion. To compare the fouling properties, membranes with similar PWPs were prepared by adjusting the dope solution composition to eliminate the effect of hydrodynamic conditions on the membrane fouling performance. The blend membranes surface exhibited considerable fouling resistance to the molecular adsorption from both BSA solution and activated sludge solution. In both cases, the flux recovered to almost 80% of the initial flux using only water backflush. Considering their great mechanical properties and antifouling resistance to activated sludge solution, these novel membranes show good potential for application in wastewater treatment. Copyright © 2017 Elsevier B.V. All rights reserved.

  9. Athermal alterations in the structure in the canalicular membrane and ATPase activity induced by thermal levels of microwave radiation

    Phelan, A.M.; Neubauer, C.F.; Timm, R.; Neirenberg, J.; Lange, D.G.

    1994-01-01

    Sprague-Dawley rats (200-250 g) were exposed 30 min/day for 4 days to thermogenic levels (rectal temperature increase of 2.2 degrees C) of microwave radiation [2.45 GHz, 80 mW/cm 2 , continuous-wave mode (CW)] or to a radiant heat source resulting in an equivalent increase in body temperature of 2.2 degrees C. On the fifth day the animals were sacrificed and their livers removed. The canalicular membranes were isolated and evaluated for adenosinetriphosphatase (ATPase) activity, total fatty acid composition and membrane fluidity characteristics. Mg ++ -ATPase activity (V max ) decreased by 48.5% in the group exposed to microwave radiation, with no significant change in the group exposed to radiant heat. The decrease in Mg ++ -ATPase was partially compensated by a concomitant increase in Na + /K + -ATPase activity (170% increase in V max over control) in animals exposed to microwave radiation, while no change occurred in the group exposed to radiant heat. This alteration in ATPase activity in the group exposed to microwave radiation is associated with a large decrease in the ratio of saturated to unsaturated fatty acids. Conversely, the group exposed to radiant heat had an increase in the ratio of saturated to unsaturated fatty acids. The most dramatic changes were found in the levels of arachidonic acid. Finally, the electron paramagnetic resonance (EPR) spin label technique used to measure the fluidity of the canalicular membranes of the animals in the three groups (sham, microwave radiation and radiant heat) indicated that the results were different in the three groups, reflecting the changes found in their fatty acid composition. The physiological response to open-quotes equivalentclose quotes thermal loads in rats is expressed differently for different types of energy sources. Possible mechanisms producing these divergent thermogenic responses are discussed. 34 refs., 3 figs., 2 tabs

  10. Phosphotyrosine-mediated LAT assembly on membranes drives kinetic bifurcation in recruitment dynamics of the Ras activator SOS.

    Huang, William Y C; Yan, Qingrong; Lin, Wan-Chen; Chung, Jean K; Hansen, Scott D; Christensen, Sune M; Tu, Hsiung-Lin; Kuriyan, John; Groves, Jay T

    2016-07-19

    The assembly of cell surface receptors with downstream signaling molecules is a commonly occurring theme in multiple signaling systems. However, little is known about how these assemblies modulate reaction kinetics and the ultimate propagation of signals. Here, we reconstitute phosphotyrosine-mediated assembly of extended linker for the activation of T cells (LAT):growth factor receptor-bound protein 2 (Grb2):Son of Sevenless (SOS) networks, derived from the T-cell receptor signaling system, on supported membranes. Single-molecule dwell time distributions reveal two, well-differentiated kinetic species for both Grb2 and SOS on the LAT assemblies. The majority fraction of membrane-recruited Grb2 and SOS both exhibit fast kinetics and single exponential dwell time distributions, with average dwell times of hundreds of milliseconds. The minor fraction exhibits much slower kinetics, extending the dwell times to tens of seconds. Considering this result in the context of the multistep process by which the Ras GEF (guanine nucleotide exchange factor) activity of SOS is activated indicates that kinetic stabilization from the LAT assembly may be important. This kinetic proofreading effect would additionally serve as a stochastic noise filter by reducing the relative probability of spontaneous SOS activation in the absence of receptor triggering. The generality of receptor-mediated assembly suggests that such effects may play a role in multiple receptor proximal signaling processes.

  11. Differential effects of near-UV and visible light on active transport and other membrane processes in Escherichia coli

    Sprott, G.D.; Martin, W.G.; Schneider, N.

    1976-01-01

    The effects of monochromatic near-UV and visible light on active transport and several other membrane processes in Escherichia coli were investigated. Using mercury lines at 366, 405, 435, 546 and 578 nm, large differential effects were observed. Transport systems with photosensitive initial rates of uptake were classified into three groups on the basis of wavelength dependence. Three, and possibly four photosensitizers may be involved; three active under aerobic conditions and the fourth in the absence of oxygen. Respiration rate exhibited the same sensitivity as one of the groups, suggesting that the active uptake of member amino acids (e.g. glycine) is largely dependent on oxidation energy. The photosensitivity of glycine transport at 405 nm was not the result of inhibition of the membrane-bound Ca-Mg adenosine triphosphates as shown using an isogenic mutant strain. Cell viability was not affected at the highly active wavelength, 405 nm. Photoeffects on transport of α-methylglucoside were minimal at 366 and 405 nm, contrasting to most of the amino acids investigated. The relative photosensitivity of respiration and several amino acid transport systems depended on carbon source. (author)

  12. Membrane proteins involved in potassium shifts during muscle activity and fatigue

    Kristensen, Michael; Hansen, T.; Juel, C.

    2006-01-01

    while trying to manipulate the opening probability or transport capacity of these proteins during electrical stimulation of isolated soleus muscles. All experiments were made with excised muscle from male Wistar rats. Kir2.1 channels were almost undetectable in the sarcolemmal membrane but present...... muscle contractions, whereas Kir2.1 and NKCC1 may have a role in K+ reuptake. channels and cotransporters; T tubule...

  13. Membrane dynamics

    Bendix, Pól Martin

    2015-01-01

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

  14. Effectiveness of a school-based physical activity injury prevention program

    Collard, D.C.M.; Verhagen, E.A.L.M.; Chin A Paw, M.J.M.; Knol, D.L.; van Mechelen, W.

    2010-01-01

    Objective: To study the effects of a school-based injury prevention program on physical activity injury incidence and severity. Design: Cluster randomized controlled trial performed from January 1, 2006, through July 31, 2007. Setting: Forty Dutch primary schools. Participants: Atotal of 2210

  15. Assessment of the Hindlimb Membrane Musculature of Bats: Implications for Active Control of the Calcar.

    Stanchak, Kathryn E; Santana, Sharlene E

    2018-03-01

    The striking postcranial anatomy of bats reflects their specialized ecology; they are the only mammals capable of powered flight. Bat postcranial adaptations include a series of membranes that connect highly-modified, or even novel, skeletal elements. While most studies of bat postcranial anatomy have focused on their wings, bat hindlimbs also contain many derived and functionally important, yet less studied, features. In this study, we investigate variation in the membrane and limb musculature associated with the calcar, a neomorphic skeletal structure found in the hindlimbs of most bats. We use diffusible iodine-based contrast-enhanced computed tomography and standard histological techniques to examine the calcars and hindlimb membranes of three bat species that vary ecologically (Myotis californicus, a slow-flying insectivore; Molossus molossus, a fast-flying insectivore; and Artibeus jamaicensis, a slow-flying frugivore). We also assess the level of mineralization of the calcar at muscle attachment sites to better understand how muscle contraction may enable calcar function. We found that the arrangement of the calcar musculature varies among the three bat species, as does the pattern of mineral content within the calcar. M. molossus and M. californicus exhibit more complex calcar and calcar musculature morphologies than A. jamaicensis, and the degree of calcar mineralization decreases toward the tip of the calcar in all species. These results are consistent with the idea that the calcar may have a functional role in flight maneuverability. Anat Rec, 301:441-448, 2018. © 2018 Wiley Periodicals, Inc. © 2018 Wiley Periodicals, Inc.

  16. Therapeutic drug monitoring of flucytosine in serum using a SERS-active membrane system

    Berger, Adam G.; White, Ian M.

    2017-02-01

    A need exists for near real-time therapeutic drug monitoring (TDM), in particular for antibiotics and antifungals in patient samples at the point-of-care. To truly fit the point-of-care need, techniques must be rapid and easy to use. Here we report a membrane system utilizing inkjet-fabricated surface enhanced Raman spectroscopy (SERS) sensors that allows sensitive and specific analysis despite the elimination of sophisticated chromatography equipment, expensive analytical instruments, and other systems relegated to the central lab. We utilize inkjet-fabricated paper SERS sensors as substrates for 5FC detection; the use of paper-based SERS substrates leverages the natural wicking ability and filtering properties of microporous membranes. We investigate the use of microporous membranes in the vertical flow assay to allow separation of the flucytosine from whole blood. The passive vertical flow assay serves as a valuable method for physical separation of target analytes from complex biological matrices. This work further establishes a platform for easy, sensitive, and specific TDM of 5FC from whole blood.

  17. Impact of Coagulant and Flocculant Addition to an Anaerobic Dynamic Membrane Bioreactor (AnDMBR) Treating Waste-Activated Sludge.

    Kooijman, Guido; Lopes, Wilton; Zhou, Zhongbo; Guo, Hongxiao; de Kreuk, Merle; Spanjers, Henri; van Lier, Jules

    2017-03-23

    In this work, we investigated the effects of flocculation aid (FA) addition to an anaerobic dynamic membrane bioreactor (AnDMBR) (7 L, 35 °C) treating waste-activated sludge (WAS). The experiment consisted of three distinct periods. In period 1 (day 1-86), the reactor was operated as a conventional anaerobic digester with a solids retention time (SRT) and hydraulic retention time (HRT) of 24 days. In period 2 (day 86-303), the HRT was lowered to 18 days with the application of a dynamic membrane while the SRT was kept the same. In period 3 (day 303-386), a cationic FA in combination with FeCl₃ was added. The additions led to a lower viscosity, which was expected to lead to an increased digestion performance. However, the FAs caused irreversible binding of the substrate, lowering the volatile solids destruction from 32% in period 2 to 24% in period 3. An accumulation of small particulates was observed in the sludge, lowering the average particle size by 50%. These particulates likely caused pore blocking in the cake layer, doubling the trans-membrane pressure. The methanogenic consortia were unaffected. Dosing coagulants and flocculants into an AnDMBR treating sludge leads to a decreased cake layer permeability and decreased sludge degradation.

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

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

    2016-01-15

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

  19. Leisure activities in the home for the elderly to prevent immobilization syndrome

    MARKVARTOVÁ, Kateřina

    2012-01-01

    This bachelor thesis with the topic: "Leisure time activities in the Home for seniors as prevention of the immobilization syndrome" is divided into two sections - theoretical and practical. In the theoretical section the terms relating to old age and types of leisure time activities offered by the Home for seniors are described. In the thesis conclusion I describe the immobilization syndrome also occurring in the elderly. Recently, there has been a large increase in senior population, and man...

  20. Prevention of ligneous conjunctivitis by topical and subconjunctival fresh frozen plasma.

    Tabbara, Khalid F

    2004-08-01

    To present a case of ligneous conjunctivitis where the recurrence of membranous conjunctivitis was prevented by subconjunctival and topical instillation of fresh frozen plasma. Interventional case report. A case of ligneous conjunctivitis with multiple recurrences since the age of 3 years developed recurrent membranous conjunctivitis after transconjunctival levator recession. Blood plasminogen activity was determined. The membrane was excised, and the membrane reappeared 4 days later. The patient was treated with excision of the membrane and subconjunctival injection of fresh frozen plasma and topical fresh frozen plasma. Plasminogen activity of the fresh frozen plasma was normal. Plasminogen blood functional activity was 52% (normal is 80%-120%). The patient had complete remission with no recurrences of membranous conjunctivitis after topical and subconjunctival fresh frozen plasma. Prophylactic use of topical and subconjunctival fresh frozen plasma may help in the prevention of membranes in susceptible patients with plasminogen deficiency.

  1. Measuring trauma center injury prevention activity: an assessment and reporting tool.

    Sise, Michael J; Sise, Carol Beth

    2006-02-01

    To develop an assessment and reporting tool for a trauma center's community partnership strategy to deliver injury prevention programs in a large metropolitan area. The tool was designed to track prevention activity and serve as a reporting format for the parent health system, county designating agency, and the American College of Surgeons' Trauma Center Verification Process. The tool collected data including trauma center paid and volunteer personnel time, equipment, resource, and financial costs, community group and public agency contributions, number of community members receiving prevention material or presentations, impact on public policy, and print and broadcast media coverage. These measurements were incorporated in a reporting grid format. Six youth injury prevention programs were evaluated over a recent 2-year interval to demonstrate the tool's usefulness. Of six programs, three focused on motor vehicle injuries, one on teen suicide, one on firearm injuries, and one on drug and alcohol abuse. Trauma Center personnel asset allocation included 3% full-time equivalent by the Trauma Medical Director, 30% by the Injury Prevention and Community Outreach Coordinator, and 473 person hours (both work and volunteer) by physicians, nurses, and other personnel. Trauma Center equipment and fixed asset expenses totaled $3,950 and monetary contribution totaled $4,430. Community groups and public agencies contributed 20,400 person hours with estimated in-kind costs exceeding $750,000. Five of the six programs continued during the 2-year period. A gun-lock giveaway program was suspended because of a product recall. A total of over 29,000 youth received prevention material and presentations. Four public policy initiatives and 18 Trauma Center media stories with over 50 mentions and 37 new community partnerships resulted. The reports generated were easily incorporated in the trauma center's reports to local and national organizations and agencies. This assessment tool

  2. Role of Religion in Preventing Youth Sexual Activity in Malaysia: A Mixed Methods Study.

    Muhammad, Noor Azimah; Shamsuddin, Khadijah; Sulaiman, Zaharah; Amin, Rahmah Mohd; Omar, Khairani

    2017-12-01

    One of the popular approaches of preventing youth sexual activity in Malaysia is using religion to promote premarital sexual abstinence. Despite this intervention, youth continue to practise premarital sex. Thus, the purpose of this exploratory mixed methods study was to understand the role of religion on sexual activity among college students in Klang Valley, Malaysia. A self-administered questionnaire survey to determine the relationship between religiosity and youth sexual activity was carried out on 1026 students recruited from 12 randomly selected colleges. Concurrently, face-to-face interviews were conducted on 15 students to explore how religiosity had influenced their decision on sexual activity. The survey data were analysed using logistic regression, while the qualitative data from the interviews were examined using thematic analysis with separate analysis for each gender. Both quantitative and qualitative results were then compared and integrated. Religious activity significantly reduced the risk of continuing sexual activity among female students (AOR = 0.67, CI = 0.47, 0.95, p = 0.02) but not male students. There was no significant relationship of religious affiliation and intrinsic religiosity (inner faith) to sexual activity by gender. Having faith in religion and strong sexual desire were the main themes that explained participants' sexual behaviour. Engaging in religious activity might be effective at preventing female students from being sexually active. However, when sexual urges and desires are beyond control, religiosity might not be effective.

  3. Sealing of Corneal Lacerations Using Photo Activated Rose Bengal Dye and Amniotic Membrane

    2017-04-22

    detected nor was there any appearance o f fluorescein leakage from 234 the retinal vessels (Figure 4 B). 235 Irreversible damage to the photoreceptors...unaffected and no evidence of 35 breakdown of the blood retina barrier was detected . Retina from grccnlight laser treated eyes 36 showed normal RPE...formation of a 55 water -tight seal is critical to preventing infection and preventing surface epithelium from gaining 56 entry into the eye. This stabi

  4. Continuous Hydrolysis and Liquid–Liquid Phase Separation of an Active Pharmaceutical Ingredient Intermediate Using a Miniscale Hydrophobic Membrane Separator

    Cervera Padrell, Albert Emili; Morthensen, Sofie Thage; Lewandowski, Daniel Jacob

    2012-01-01

    Continuous hydrolysis of an active pharmaceutical ingredient intermediate, and subsequent liquid–liquid (L-L) separation of the resulting organic and aqueous phases, have been achieved using a simple PTFE tube reactor connected to a miniscale hydrophobic membrane separator. An alkoxide product......, obtained in continuous mode by a Grignard reaction in THF, reacted with acidic water to produce partially miscible organic and aqueous phases containing Mg salts. Despite the partial THF–water miscibility, the two phases could be separated at total flow rates up to 40 mL/min at different flow ratios, using...

  5. Gas separation membranes

    Schell, William J.

    1979-01-01

    A dry, fabric supported, polymeric gas separation membrane, such as cellulose acetate, is prepared by casting a solution of the polymer onto a shrinkable fabric preferably formed of synthetic polymers such as polyester or polyamide filaments before washing, stretching or calendering (so called griege goods). The supported membrane is then subjected to gelling, annealing, and drying by solvent exchange. During the processing steps, both the fabric support and the membrane shrink a preselected, controlled amount which prevents curling, wrinkling or cracking of the membrane in flat form or when spirally wound into a gas separation element.

  6. Constraints imposed by transmembrane domains affect enzymatic activity of membrane-associated human CD39/NTPDase1 mutants.

    Musi, Elgilda; Islam, Naziba; Drosopoulos, Joan H F

    2007-05-01

    Human CD39/NTPDase1 is an endothelial cell membrane-associated nucleotidase. Its large extracellular domain rapidly metabolizes nucleotides, especially ADP released from activated platelets, inhibiting further platelet activation/recruitment. Previous studies using our recombinant soluble CD39 demonstrated the importance of residues S57, D54, and D213 for enzymatic/biological activity. We now report effects of S57A, D54A, and D213A mutations on full-length (FL)CD39 function. Enzymatic activity of alanine modified FLCD39s was less than wild-type, contrasting the enhanced activity of their soluble counterparts. Furthermore, conservative substitutions D54E and D213E led to enzymes with activities greater than the alanine modified FLCD39s, but less than wild-type. Reductions in mutant activities were primarily associated with reduced catalytic rates. Differences in enzymatic activity were not attributable to gross changes in the nucleotide binding pocket or the enzyme's ability to multimerize. Thus, composition of the active site of wild-type CD39 appears optimized for ADPase function in the context of the transmembrane domains.

  7. No consensus on restrictions on physical activity to prevent incisional hernias after surgery

    Pommergaard, H-C; Burcharth, J; Danielsen, Anne Kjaergaard

    2014-01-01

    of restrictions on physical activity recommended for patients operated for colorectal cancer and to evaluate the agreement among surgical specialists. METHODS: A questionnaire was sent to 60 general surgeons (specialists) in Denmark and Sweden working in academic departments of surgery with a high volume......PURPOSE: In the postoperative phase after colorectal surgery, restrictions on physical activity are often recommended for patients to prevent incisional hernias. However, evidence does not support that restrictions may prevent such hernias. The purpose of this study was to evaluate the extent...... of colorectal cancer resections. The questionnaire was case based and contained questions regarding possible restrictions on physical activity recommended for patients 0-2, 2-6 and >6 weeks after resection for colorectal cancer. Agreement among the surgeon on whether restrictions should be recommended...

  8. Top 10 Research Questions Related to Preventing Sudden Death in Sport and Physical Activity.

    Katch, Rachel K; Scarneo, Samantha E; Adams, William M; Armstrong, Lawrence E; Belval, Luke N; Stamm, Julie M; Casa, Douglas J

    2017-09-01

    Participation in organized sport and recreational activities presents an innate risk for serious morbidity and mortality. Although death during sport or physical activity has many causes, advancements in sports medicine and evidence-based standards of care have allowed clinicians to prevent, recognize, and treat potentially fatal injuries more effectively. With the continual progress of research and technology, current standards of care are evolving to enhance patient outcomes. In this article, we provided 10 key questions related to the leading causes and treatment of sudden death in sport and physical activity, where future research will support safer participation for athletes and recreational enthusiasts. The current evidence indicates that most deaths can be avoided when proper strategies are in place to prevent occurrence or provide optimal care.

  9. Prevention of LDL-suppression of HMG-CoA reductase (HMGR) activity by progesterone (PG): evidence for cytochrome P-450 involvement

    Sexton, R.C.; Gupta, A.; Panini, S.R.; Rudney, H.

    1987-01-01

    Incubation of rat intestinal epithelial cells (IEC-6) with PG has been reported by us to prevent the suppression of HMGR activity by LDL. In the present study, addition of LDL and PG to IEC-6 cells resulted in a 2 fold increase in cellular free cholesterol (CH) in 24 h, while HMGR activity remained elevated. PG did not affect the internalization and degradation of [ 125 I] LDL nor the accumulation of free [ 3 H] CH in cells incubated with [ 3 H-cholesteryl linoleate]-LDL. Also, PG did not affect the intracellular transport of LDL-derived [ 3 H] CH to the plasma membrane nor the efflux of the [ 3 H] CH into medium containing human high density lipoprotein. Addition of LDL to cells, in which the cellular CH was radiolabeled from [ 3 H] acetate, resulted in an increased formation of radiolabeled oxysterols, detected by HPLC, and a corresponding decrease in HMGR activity. PG attenuated both the LDL-induced formation of oxysterols and suppression of HMGR activity. PG inhibited cytochrome P-450 dependent oxidation of benzphetamine, aminopyrine and aniline by liver microsomes from phenobarbitol treated rats. These results suggest PG may prevent LDL suppression of HMGR activity in IEC-6 cells by inhibiting cytochrome P-450 dependent formation of regulatory oxysterols

  10. Ca2+ Channel Re-localization to Plasma-Membrane Microdomains Strengthens Activation of Ca2+-Dependent Nuclear Gene Expression

    Krishna Samanta

    2015-07-01

    Full Text Available In polarized cells or cells with complex geometry, clustering of plasma-membrane (PM ion channels is an effective mechanism for eliciting spatially restricted signals. However, channel clustering is also seen in cells with relatively simple topology, suggesting it fulfills a more fundamental role in cell biology than simply orchestrating compartmentalized responses. Here, we have compared the ability of store-operated Ca2+ release-activated Ca2+ (CRAC channels confined to PM microdomains with a similar number of dispersed CRAC channels to activate transcription factors, which subsequently increase nuclear gene expression. For similar levels of channel activity, we find that channel confinement is considerably more effective in stimulating gene expression. Our results identify a long-range signaling advantage to the tight evolutionary conservation of channel clustering and reveal that CRAC channel aggregation increases the strength, fidelity, and reliability of the general process of excitation-transcription coupling.

  11. Electrochemical activation and inhibition of neuromuscular systems through modulation of ion concentrations with ion-selective membranes

    Song, Yong-Ak; Melik, Rohat; Rabie, Amr N.; Ibrahim, Ahmed M. S.; Moses, David; Tan, Ara; Han, Jongyoon; Lin, Samuel J.

    2011-12-01

    Conventional functional electrical stimulation aims to restore functional motor activity of patients with disabilities resulting from spinal cord injury or neurological disorders. However, intervention with functional electrical stimulation in neurological diseases lacks an effective implantable method that suppresses unwanted nerve signals. We have developed an electrochemical method to activate and inhibit a nerve by electrically modulating ion concentrations in situ along the nerve. Using ion-selective membranes to achieve different excitability states of the nerve, we observe either a reduction of the electrical threshold for stimulation by up to approximately 40%, or voluntary, reversible inhibition of nerve signal propagation. This low-threshold electrochemical stimulation method is applicable in current implantable neuroprosthetic devices, whereas the on-demand nerve-blocking mechanism could offer effective clinical intervention in disease states caused by uncontrolled nerve activation, such as epilepsy and chronic pain syndromes.

  12. Preparation of wheat root plasma membrane vesicles and effect of water stress on 45Ca2+ transport activity

    Lu Jinyin; Gao Junfeng; Cao Cuiling

    1998-01-01

    The wheat roots plasma membrane (PM) vesicles were obtained by sucrose gradient centrifugation. The experiment results shows that the wheat roots of Zhengyin No.1 PM H + -ATPase latent activity was 24%, and PM inside-out vesicle (IOV) accounts for 76%. With -1.0 MPa stress of 24h, PM Ca 2+ -ATPase activity of both orientation wheat roots were increased. Under normal water condition and PEG stress, 62% and 53% of the enzyme activity was inhibited respectively by EGTA, radioactive calcium-45 transport amount was 22.09 nmol/mg pro and 4.17 nmol/mg pro. respectively with PM-IOV.PEG stress results in a decrease of 45 Ca 2+ transport amount of wheat roots PM-IOV by 81%

  13. Physical activity in children: prevention of obesity and type 2 diabetes.

    Rush, Elaine; Simmons, David

    2014-01-01

    There is strong evidence that increased physical activity is beneficial for blood glucose homeostasis and the prevention of obesity and type 2 diabetes mellitus. This chapter takes a life course approach with an emphasis on the intrauterine and childhood stages of life. Firstly, growth and development at critical periods with a focus on skeletal muscle and adipose tissue; then, obesity and type 2 diabetes mellitus are considered in relation to physical activity and sedentary behaviour. The importance of the development of fundamental movement skills in early childhood for both physical fitness and also growth and development is emphasised. Physical activity guidelines in westernised countries are examined for commonalities. Finally, the effective translation of the evidence base for the benefits of physical activity into randomised controlled trials and then into real-world public health services that are sustainable is addressed with a case study from New Zealand of Project Energize--a through-school physical activity and nutrition intervention. Physical activity, alongside a 'healthy diet' is arguably the best preventive measure and treatment for both obesity and type 2 diabetes. It is an essential and normal activity of daily life, and all aspects of the life course and the environment should support physical activity.

  14. Fragile X Mental Retardation Protein Regulates Activity-Dependent Membrane Trafficking and Trans-Synaptic Signaling Mediating Synaptic Remodeling

    Sears, James C.; Broadie, Kendal

    2018-01-01

    Fragile X syndrome (FXS) is the leading monogenic cause of autism and intellectual disability. The disease arises through loss of fragile X mental retardation protein (FMRP), which normally exhibits peak expression levels in early-use critical periods, and is required for activity-dependent synaptic remodeling during this transient developmental window. FMRP canonically binds mRNA to repress protein translation, with targets that regulate cytoskeleton dynamics, membrane trafficking, and trans-synaptic signaling. We focus here on recent advances emerging in these three areas from the Drosophila disease model. In the well-characterized central brain mushroom body (MB) olfactory learning/memory circuit, FMRP is required for activity-dependent synaptic remodeling of projection neurons innervating the MB calyx, with function tightly restricted to an early-use critical period. FMRP loss is phenocopied by conditional removal of FMRP only during this critical period, and rescued by FMRP conditional expression only during this critical period. Consistent with FXS hyperexcitation, FMRP loss defects are phenocopied by heightened sensory experience and targeted optogenetic hyperexcitation during this critical period. FMRP binds mRNA encoding Drosophila ESCRTIII core component Shrub (human CHMP4 homolog) to restrict Shrub translation in an activity-dependent mechanism only during this same critical period. Shrub mediates endosomal membrane trafficking, and perturbing Shrub expression is known to interfere with neuronal process pruning. Consistently, FMRP loss and